USOO584O72OA United States Patent (19) 11 Patent Number: 5,840,720 Chen (45) Date of Patent: Nov. 24, 1998 54) 4-O AND 5-AMINOMETHYLATION OF wherein R is a member Selected from the group consisting SYNTHETIC CAPSAICIN DERIVATIVES, A of NEW DISCOVERY OF CAPSAICN ANTAGONIST 75 Inventor: Ing-Jun Chen, Kaohsiung, Taiwan -()- COOR -()- COOR2 73 Assignee: Tong-Ho Lin, Taipei, Taiwan 21 Appl. No.: 547,040 22 Filed: Oct. 23, 1995 (51) Int. Cl. .............................................. C07D 273/01 52 U.S. Cl. .................. ... 544/230.5; 54.4/90 58 Field of Search ............................. 544/90; 514/230.5 HOOC 56) References Cited -COOR R1COOR U.S. PATENT DOCUMENTS R1COOR2 -COOR2 3,082,112 3/1963 Hemwall ................................... 544/90 4,035,363 7/1977 Oka et al. ................................. 544/90 OTHER PUBLICATIONS Ruthenium red antagonism of the effect of capsaicin on the motility of the isolated guinea-pig ileum, Takaki, Jin, and Nakayama, European Journal of Pharmacology, 174 (1989). -()- R1COOR2 () (List continued on next page.) Primary Examiner Donald G. Daus Attorney, Agent, or Firm-Beveridge, DeGrandi, Weilacher & Young LLP -()- R1COOR -()- COR 57 ABSTRACT A Series of 4-O & 5-aminomethylation of Synthetic capsaicin derivatives Selectively reveal antagonist activity on COORSO3H capsaicin-Sensitive Sensory neurons, and inhibit its inner Vating atrium, airway, and ileum Smooth muscles in vitro. The compound of this invention has the following formula wherein R is a member Selected from the group consisting 1n R of C112 alkyl, C, 12 alkenyl, C, 12 alkylene, and C. 12 alkenylene, and CHO wherein R is a member Selected from the group consisting O of H, C alkylene-NRR, and Coalkenylene-NRR. 10 Claims, 19 Drawing Sheets or )-chnic, )- CH-CHCH CH 3 Capsaicin (Cap. ; A) 8-methyl-N-Vanilly-nonenamide to D-chi-cho-chO nonanoylvanillylamideNonivamide (NVA; B) 5,840,720 Page 2 OTHER PUBLICATIONS guinea-pig heart: correlation with effects on cardiac con CapSazepine inhibits low pH- and lactic acid-evoked tractility, Cereceda, Lou, and Lundberg, Br. J. Pharmacol. release of calcitonin gene-related peptide from Sensory (1991). nerves in guinea-pig heart, Cereceda and Lundberg, Euro A comparison of capsaZepine and ruthenium red as capsaicin pean Journal of Pharmacology, 221 (1992). antagonists in the rat isolated urinary bladder and Vas Ruthenium-red inhibits CGRP release by capsaicin and deferens, Maggi, Bevan, Walpole, Rang, and Giuliani, Br. J. resiniferatoxin but not by Ouabain, bradykinin or nicotine in Pharmacol. (1993). U.S. Patent Nov. 24, 1998 Sheet 1 of 19 5,840,720 H,CO O HO CH, NHC-(CH,)— CH-CHCHK CH, 3 Capsaicin (Cap. ; A) 8-methyl-N-Vanilly-nonenamide H,CO O to O-ch, NHC-(CH,)-CH, nonanoylvanillylamideNonivamide (NVA; B) F.G. 1 U.S. Patent Nov. 24, 1998 Sheet 2 of 19 5,840,720 Bronchi 2 O pA: 6.01 + 0.02 100 8 O 6 O 4 O O Control O CAPBZ 1.0 g M 20 V CAPBZ 10 (1 M V CAPBZ 100 u M -Log ( Capsaicin M FIG.2A U.S. Patent Nov. 24, 1998 Sheet 3 of 19 5,840,720 Trachea > 80 S. C !-- 60 - kill 1 - S. / 1 5 40 / 5 / s // O Control s 2 O CAPBZ 10 (1 M & -Log ( Capsaicin M FIG.2B U.S. Patent Nov. 24, 1998 Sheet 4 of 19 5,840,720 Bronchi 100 4.68OOO 2 O -Log ( Capsaicin M FIG.2C U.S. Patent Nov. 24, 1998 Sheet 5 of 19 5,840,720 Right Atria O Capsaicin l (l. M Za Capsaicin 1 u M + NVABI 0. 1 (1 M 100 Capsaicin 1 u M + NVABI 1 (1 M El Capsaicin 1 u M + NVABI 10 (1 M 8 O 6 O 4 O 2 O U.S. Patent Nov. 24, 1998 Sheet 6 of 19 5,840,720 Right Atria O CAP 1.0 (1 M CAP 1.0 u M + CAPBZ 1.0 t, M CAP 1.0 u M + CAPBZ 10 i? M to ill CAP 1.0 u M + CAPBZ 100 M U.S. Patent Nov. 24, 1998 Sheet 7 of 19 5,840,720 Left Atria O Capsaicin 1 (1 M 4. Capsaicin 1 u M + NVABI 0.1 g M (2 Capsaicin 1 (1 M + NVABI 1 (1 M 8E Capsaicin 1 u M+NVABI 10 a M 1.6 IC50=23.98 (1 M - 1.4 59 3 1.2 1.0 0.8 3 2 0.6 O 0. O U.S. Patent Nov. 24, 1998 Sheet 8 of 19 5,840,720 Light Atria L CAP 1.0 u M Z2 CAP 1.0 (1 M + CAPBZ 1.0 (1 M CAP 1.0 (1 M + CAPBZ 10 (1 M Hill CAP 1.0 u M + CAPBZ 100 t, M IC50-35. 14 Al M g Ke Y w w FIG.3D U.S. Patent Nov. 24, 1998 Sheet 12 of 19 5,840,720 Right Atria O Control O CAPBZ 1.0 (1 M V CAPBZ 10 (1 M S.C. ap E 30 CVS CD S. CD 20 CM CVS 9 OS 10 O U.S. Patent Nov. 24, 1998 Sheet 13 of 19 5,840,720 Left Atria 100. 2;ult 80 /. / 60 O Control O CAPBZ 1.0 (1 M V CAPBZ 10 (1 M V CAPBZ 100 (1 M 5 6 7 8 CaCl2(mM) FIGSB U.S. Patent Nov. 24, 1998 Sheet 14 of 19 5,840,720 O Control O CAPCNC (10 u M) Pretreatment Right Atria 120 u 100 -11 8 O 6 O 4 O U.S. Patent Nov. 24, 1998 Sheet 15 of 19 5,840,720 1 2 O O Left Atria 800 400 0.9 1.8 3.6 5.4 7.2 Ca Concentration(mM) FIG.6B U.S. Patent Nov. 24, 1998 Sheet 16 of 19 5,840,720 Rightt Atria O Control O CAPBZ 50 (1 M 50 V Benzocaine 10 (1 M 23OO 1 O -Log hCGRP M FIG.7 U.S. Patent Nov. 24, 1998 Sheet 17 of 19 5,840,720 O CapSZepine (100 (1 g/kg/min) O NVABI (100 u g/kg/min) 90 - V NVADA (100 (1 g/kg/min) 70 -- 60 5 O O 0 5 10 15 20 25 30 35 40 45 50 55 60 Time(min) FIG.8A U.S. Patent Nov. 24, 1998 Sheet 18 of 19 5,840,720 O CapSzepine (100 u g/kg/min) 10 O NVABI (100 u g/kg/min) 100 -- V NVADA (100 u g/kg/min) 0 5 10 15 20 25 30 35 40 45 50 55 60 Time(min) FIG.8B U.S. Patent Nov. 24, 1998 Sheet 19 of 19 5,840,720 90 O CapSzepine (100 g g/kg/min) 80 O NVABI (100 u g/kg/min) VNVADA (100 g g/kg/min) 70 2 O 0 5 10 15 20 25 30 35 40 45 50 55 60 Time(min) FIG.8C 5,840,720 1 2 4-O AND 5-AMINOMETHYLATION OF evaluated as a competitive capsaicin antagonist (S. Bevan et SYNTHETIC CAPSAICIN DERIVATIVES, A al., 1992). Our previous product, glyceryl nonivamide, was NEW DISCOVERY OF CAPSAICN also proven to be a selective capsaicin agonist (I. J. Chen et ANTAGONIST al., Eur. J. Med. Chem. 27, 187-92, 1992). The positive results of S. Bevan et al. (1992) have encouraged us to TECHNICAL FIELD OF THE INVENTION Search for other new capsaicin antagonist. The present invention relates to new and useful capsaicin Previous studies on the structure-activity relationship of antagonist derivative S, especially to 4-O & nonivamide (N-nonanoyl Vanillylamide), the Synthetic 5-aminomethylation of Synthetic capsaicin derivatives, and capsaicin, indicated that Substitution for the OH group of other 3-methoxy, 4-hydroxy compounds. capsaicin or nonivamide may lead to less pungency in the capsaicin derivatives (I. J. Chen et al. (1992)), including a BACKGROUND OF THE INVENTION non-pungent beta adrenergic blocker derivative with cardio Capsaicin is the principle of Capsium annuum Linne, a tonic and CGRP releasing properties (I. J. Chen et al., J. medicinal plant of Solanaceae. Capsaicin-Sensitive func Med. Chem. 37,938-43, 1994). In a similar way, a series of tional change has been found in cardia, aorta, trachea, and 15 4-O & 5-aminomethylation of capsaicin derivatives (FIG. 1) animal tissues (Maggi & Meli, 1988). Capsaicin has been another bicyclic capsaicin derivative like capsaZepine, was shown to have potent positive chronotropic and inotropic first Synthesized by a chemical reaction both masking the effects when applied to isolated guinea-pig atria (N. Fukada phenolic OH of and bicyclic nonivamide with a series of & M. Fumjiwara et al., J. Pharmacol. 21, 622-24, 1969; J. amino benzyl and amino alkyl compounds. Molnar et al., Acta Physiolo. Acad. Sci Hung. 35, 369-74. 1969). BRIEF DESCRIPTION OF THE DRAWINGS The contraction effect of capsaicin on the isolated guinea pig ileum was Suggested to be caused by parasympathetic FIG. 1 Chemical Structures of capsaicin and nonivamide transmission due to SP (substance P) release (L. A. Chahl, 25 FIGS. 2A-2C Cumulative concentration-response curves Naunyn-Schmiedebergs Arch. Pharmacol. 310, 212-15, to capsaicin (10-10M) in the absence and presence of 1982). Capsaicin has been reported to inhibit neuronal CAPBZ and NVABI in the isolated guinea-pig trachea, sodium currents (K. Yamanaka et al., Brain Res. 300, 113-9, expressed as a % of the maximum contraction to carbachol 1984) also inhibit neuronal calcium currents (M. Petersenet (1 uM). Each point represents meantS.E.M of six experi al., Pflvgers Arch. 409, 403-10, 1987). The capsaicin mentS.