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 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 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 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. but not by Ouabain, or 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 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 () 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. Sensitive effects on the functions of various tissues, includ FIGS. 3A-3D Capsaicin (1 uM) caused a positive ino ing ileum Smooth muscles, cardia muscles, and airways are tropic and chronotropic response on Spontaneously beating known to be caused by the activation of sensory C-fibres (C. guinea-pig right atrium and electrically-driven guinea-pig A. Maggi et al., Gen. Pharmacol. 19, 1-43, 1988). The left atrium. CAPBZ and NVABI pretreatment for 30 min positive inotropic action of capsaicin is believed to be 35 decreased the effect of capsaicin. Each data was the associated with the release of CGRP (calcitonin gene-related meani-S.E.M. of six experiments (*P-0.05. **P-0.01, Stu peptide) from intracardiac neurons, and has shown that capsaicin and CGRP prolong action potential duration in dent's t-test) guinea pig atria (Franco-Cereceda et al., Acta Physiol. FIG. 4 The effects of CAPBZ on response of guinea-pig Scand. 132, 181–90, 1988). 40 ileum to capsaicin (10 uM), substance P (SP, 0.1 uM), and The effects of capsaicin on rat urethra, urinary bladder, carbachol (0.5uM). Note concentration-dependently reduc gastrointestinal, renal pelvis, genitourinary tract, and ureters tion of response to SP, and carbachol. Thirty minutes fol have been reviewed by C. A. Maggi, et al.(Gen. Pharmacol. lowing washout, responses to SP, and carbachol were recov 19, 1-43, 1988). The activation of this proton-gated cation ered. conductance allows Sodium, calcium, and potassium ion to 45 FIGS. 5A-5B Effect of CAPBZ on the positive chrono flow down its concentration gradient into the dorsal root tropic effects of increasing CaCl concentrations in the ganglion cells causing depolarization and action potential isolated guinea-pig right atria and electrically-driven generation (C. A. Forbes et al., Soc. Neurosci. Abstract. 14, guinea-pig left atria. Each point represents the mean of Six 642. 1988). The inhalation of capsaicin can be used to experiments; vertical bars represent the S.E.M. increase airway resistance and leads to bronchoconstriction 50 FIGS. 6A-6B Effect of CAPCNC6 (10 uM) on the (J. G. Collier et al., Br. J. Pharmacol. 81, 113–7, 1984). positive chronotropic (spontaneously beating right atrium) The synthetic capsaicin derivatives with cardioinhibitory and inotropic (electrically-driven left atria) effect of increas and antibronchoconstrictory properties have rarely been ing Ca" concentrations in guinea pigs. Beating rate and described. Cap S a Zep in e 24-chlor phenyl) tension were expressed as % of control values. Data repre ehtylaminothiocarbonyl-7, 8-dihydroxy-2, 3, 4, 55 5-tetrahydro-1-H-2-benzazepine), first synthesized by sents the meant S.E.M. (n=7). Whalloe et al. (Br. J. Pharmacol. 107,544–552, 1992) as an FIG. 7 Cumulative concentration-response curves for the antagonist of capsaicin (S. Bevan et al., Br. J. Pharmacol. chronotropic effects of hCGRP in the absence and presence 107,544–52, 1992), was found to be effective in inhibiting of CAPBZ (50 uM) and benzocaine (10 uM) on spontaneous the contractile response evoked by capsaicin or by electrical 60 beatings of guinea-pig right atria. Each value is the field stimulation in guinea-pig bronchi (M. G. Belvisi et al., meantS.E.M. of six experiments. Eur. J. Pharmacol. 215, 341-344, 1992). Capsazepine, FIGS. 8A-8C Percentage of inhibition of capsaicin (10 chemically with a bicyclic benzazepine moiety similar to the Aug/kg, i.v.) induced triphasic response during a 15 min beZodiazepine moiety of diazepam which is effective in infusion of capsazepine or NVABI or NVADA (100 tug/kg/ inhibiting GABA receptor hypothesized locating on preter 65 min) and recovery from A, B and C phase of blood pressure minal region of capsaicin-sensitive Sensory nerves (D. A. after termination of each infusion. Each data was the Brown et al., Brain Rev. 156, 187–91, 1978), has been meantS.E.M. of six experiments. 5,840,720 3 4 DETAILED DESCRIPTION Adult guinea pigs (Hartley), weighing 350–450 g, were The present invention describes a series of 4-O & sacrificed by a blow on the head followed by cervical 5-aminomethylated capsaicin derivatives which have the dislocation. The bronchi and trachea were removed from the formula A: lungs, cleaned of all parenchyma, and immediately placed in Krebs solution. As the method reported by M. G. Belvisi et 1n R Formula A 1 al. (Eur. J. Pharmacol. 215,341-4, 1992). O N OH Cumulative addition of 0.01-30 uM capsaicin to the organ bath caused a concentration-dependent increase in contrac CHO H3CO CHN-R tion of isolated guinea-pig bronchi and trachea. These effects were shifted rightward in bronchi in the presence of com 1O pound 1 (CAPBZ, 1.0-100, uM)), compound 3 (CAPBI, O O 1.0-100 uM) and capsazepine (1-10 uM). This inhibitory CHNHC-R1. CHNH-C-R1. effect of compound 1 and 3 were more effective in bronchi Wherein R is than in trachea (FIGS. 2A-C). 15 B. Antagonist activity of compound on capsaicin-induced contraction in the isolated guinea-pig atrium Also as shown in FIGS. 3A-3D, 1.0 uM capsaicin R1, induced a positive inotropic and chronotropic effect in the -()- COOR, -K)- COOR2, isolated right and left guinea-pig atria, respectively. These effects were significantly inhibited in the presence of 0.1-10 R1OOC uM compound 1 (CAPBZ), compound 6 (CAPCNC6), com pound 11 (NVABI), compound 12 (NVADA), concentra tiondependently. The ICso value against capsaicin-induced COOH, contractility for these compounds can be estimated from the 25 data presented in FIGS. 3A-3D. C. Antagonist activity of compound on capsaicin and Substance Pinduced contractilities of isolated guinea -COOR, -COOR2, -R1COOR, pig ileum -()- R1COOR2, The abdominal incisions for the guinea pig were made and ilea were isolated and placed in cold (4 C.) Tyrode's R1OC Solution. 2.5 cm Segments of ileum were then trimmed and Suspended in an organ bath containing 20 ml low calcium Kreb's solution, aerated with 95% O and 5% CO at 37° C. - ) s -( ) COR1, -K)- R1COOR1, As the method reported by M. Takaki et al. (Eur. J. Phar 35 macol. 174, 5762, 1989). R1 represents C112 alkyl, C, 12 alkenyl; AS shown in FIG.4, carbachol was used as a control agent to induce maximum contraction of ileum Smooth muscle; 10 R represents H, C-NRR. I. Methods of Preparation uM capsaicin and 0.1 uM substance P (SP) induced weaker The methods of preparation for this invention (formula A) contractions. This effect of capsaicin or Substance P was was shown in the section of Synthesis and Its Scheme (IV) 40 inhibited in the presence of 1.0-100 uM of compound 1 in Schemes 1-4. The methods might be performed using (CAPBZ), concentration-dependently. amino benzyl, amino alkyl, amino phenylacetic acid, D. Effects of compound on the calcium channel in the 4-aminopropiophenone and anthranilic acid compound and isolated guinea-pig atrium their esters to react with N-nonanoylvanillylamide (NVA) or Cumulative addition of CaCl concentrations in the other Substituted guaiacol compounds via the Mannich reac 45 Kreb's Solution could concentration-dependently produce a tion to give 4-O- and 5-amino-methylated derivatives of positive chronotropic effect in the isolated Spontaneously capsaicin or other guaiacol compounds (Formula A and FIG. beating right atrium, and a positive inotropic effect in the 1). isolated electrically driven left guinea-pig atrium. Pretreat The other methods of preparation comprise reacting NVA ment with compound 1 and 6 could inhibit these calcium (N-nonanoylvanillylamide) or other guaiacol derivatives 50 effects (FIGS. 5A-5B and FIGS. 6A-6B). with dialkylamine to give 4-guaiacolic and E. Ineffective effect on CGRP releasing in the isolated 5-aminomethylated compounds (e.g. Compound 4, Scheme atrium 4) via Mannich reaction as described shown in Formula A Cumulative addition of hCGRP to the organ bath concen and Scheme of Synthesis Section (IV), Schemes 1-4. trationdependently increased the beating in the right atrium In the invention of novel capsaicin derivative compounds most of the reacting materials were first amine, Second 55 and the tension in the electrically driven left atrium of the amine, and/or Substituted with Straight or branched alkyl, guinea pig. These effects were not inhibited by compound 1 Straight or branched alkenyl. The guaiacoxyl group com (CAPBZ). After the pretreatment with CGRP for 30 minutes, pounds including 4-O- and 5-aminoethylated capsaicin compound 1 also could not change CGRP-induced contrac (CAP) nonvamide and other guaiacol derivatives were sub tilities (FIG. 7). Stituted with Straight or branched alkyl group, alkenyl group. 60 F. Effects of compounds on capsaicin-induced blood These structures of compounds of formula A described preSSure and heart rate changes above were assigned according to the H-NMR, IR, MS, After 15 minutes, intravenous perfusion of NVABI, elemental analytical data, C-NMR. NVABZ, NVADA and capsazepine (100 tug/kg/min), bolus II. Pharmacological Activity capsaicin (10 ug/kg, i.v.) was given at 5th, 10th, 15th, 30th, A. Antagonist activity of compounds on capsaicin 65 45th and 60th minutes in the pentobarbital-anesthesized induced contractility of isolated guinea-pig bronchi and Wistar rats. This experiment was carried out to study the trachea antagonistic effects of these compounds on capsaicin elicit 5,840,720 S 6 triphasic response of blood pressure. The results indicated EXAMPLE 2 the antagonist activities of NVABI, NVABZ, and cap Sazepine were recovered to basal level by Saline infusion for NIEthylamino-(4-oxymethylene 5-methylene)-3- 30, 45, and 60 minutes gradually. Estimated recovery half methyoxybenzyl nonanamide (compound 2) life of capsaicin-induced blood pressure changes (A, B, and 5 10 g nonivamide was dissolved in 1000 ml ethanol, added C effect) for NVABI was 10.67, 4.30, and 21.75 min, for to 3.5 mole times ethylamine, 4 mole times , NVABZ. 55.13 min, for NVADA 4.30, 9.01, and 4.49, and and drops of acetic acid, then refluxed under room tempera for capsazepine 2.73, 2.89, and 3.78 min (Table 1 & 2). The ture for 24 hrs. After cooling, the Solvent was evaporated. duration order of their antagonist activities was The residue was recrystallized from n-hexane to give com NVABZ>NVABI>capsazepine. The efficacy order of their pound 2. antagonist activities was capsazepined-NVABZ>NVABI in the A effect, NVABI>capsazepines-NVABZ in the B effect, EXAMPLE 3 and capsazepines-NVABI>NVABZ in the C effect. These compounds could not inhibit calcitonin gene-related peptide, N-hexylamino-(4-oxymethene, 5-methylene)-3- Substance P, and bradykinininduced hypotensive effects. 15 methoxybenzyl nonanamide (compound 6) Intrathecal perfusion of these compounds also reversed 10 g nonivamide and 5 ml hexylamine was used as the epigastric intraarterial capsaicin (10 ug/kg)-induced Starting material and treated according to the procedure hypotensive reflex in rats (Table 3). It is concluded that described in example 1 to give compound 6. NVABI, NVABZ, NVADA, and capsazepine all modulate the presynaptic capsaicin-Sensitive Sensory neurons and thus EXAMPLES 4-12 may inhibit capsaicin-induced release of neuropeptides, but are different from each other in their pharmacokinetic prop The compounds 3-5, 7-12 can be prepared in a manner erties. analogous to those described in example 1-3. The physical III. Pharmaceutical Compositions constants and Spectral data are shown in Section IV. The novel compounds of this invention together with a 25 conventional adjuvant, carrier, or diluent, and if desired in Scheme 1: the form of pharmaceutically acceptable Salts, may be pre OH pared in the form of pharmaceutical compositions and unit dosages. In Such forms, they may be employed as Solids, or H3CO liquids, for oral use; in the form of Suppositories for rectal -- administration; in the form of Sterile injectable Solutions for O parental (including Subcutaneous) use. The Solid pharmaceutical dosages may comprise disinte CHNHC- (CH2) 7CH3 NVA grating age in tS Such as Starch, Sodium 35 carboxymethylcellulose, and/or binderS Such as ethyl O alcohol, glycerin, and/or carrierS Such as magnesium HCHO, CH3COOH Stearate, lactose, which are prepared by conventional phar HN C-O-CH-CH, Cotton to c. Gd maceutical methods. The Sterile injectable Solutions, dosages, or other liquid preparations can be adjusted with Benzocaine buffers, Such as phosphate Solutions, if desired, with auxil 40 iary agents, emulsifiers, which particularly comprise acque O ous Solutions or Salt Solutions of the novel compounds. The 1n C-O-CH2CH3 novel pharmaceutical compositions and unit dosages thereof O N allow the formation of a pharmaceutically acceptable Salt, are extremely useful in Selectively antagonizing capsaicin 45 H3CO Sensitive Sensory neurons, inhibiting innervation of the atrium, airway and ileum Smooth muscles in Vitro, and as well as producing direct cardioinhibitory effects, tachyphy O CHNHC-(CH2)-CH3 laxis. The novel compounds of the invention may accord 50 ingly be administered to a Subject, e.g. a living animal body, CAPBZ (Compound 1) including a human, and should be adjusted according to the complexity of the Symptoms. IV. Synthesis and its Scheme Scheme 2: 55 EXAMPLE 1. OH H3CO NP-(carboxylic acid ethyl ester) phenylamine-(4- oxymethylene, 5-methylene)-3-methoxybenzyl O nonanamide, capsazocaine, (compound 1) 60 10 g nonivamide was dissolved in a mixture of absolute CHNH-C-(CH2)-CH3 20 ml, mixed with 5.7 g benzocaine in 10 ml NVA absolute methanol Solution, then poured into a three-neck HCHO, CH3COOH flask, adding 10 ml 34-37% formaldehyde solution, 5 ml NH2-CH2-CH3 CH3CHOH, 70° C. Gd acetic acid, then refluxing under 70-75 C. for 24 hrs. After 65 Ethylamine cooling, the Solvent was evaporated. The residue was recryS tallized from ethylacetate to give compound 1, Yield 77%. 5,840,720 7 8 -continued V. Pharmaceutical Formulation A typical tablet which may be prepared by conventional Scheme 2: tabletting techniques contains

active compound 40 mg lactose 30 mg starch 8 mg mag.stearate 10 mg corn starch 12 mg O CHNH-C-(CH2)-CH3 VI. Physical Constants and Spectral Data of Synthesized NVABI (Compound 2) Examples The physical constants and Spectral data are shown as follows: 15 Compound 1 Scheme 3: NP-(carboxylic acid ethyl ester)-phenylamine-(4- OH oxymethylene, 5-methylene)-3-methoxybenzyl nonanamide, capsazocaine, (compound 1) mp: 145°-147.5°C.,

O 1H-NMR(CDC1): 8 0.87 (t, 3H, CH.), 1.32-2.20 (m, -(CH)x7), NVA 25 3.84 (s, 3H, OCH), O 4.32 (q, 2H, -COO-CHCH), HCHO, CH3COOH HN C-O-CH2-CH(CH3)2 CH3CHOH, 70° C. Gd 4.32 (d. 2H, ArCH-NHCO-), 4.65 (S, 2H, ArCH-N=), p-aminobenzoic acid isobutyl ester 5.45 (S, 2H, ArOCH=N-), 5.71 (m, 1H, NH), O 6.57 (d. 1H, Ar), 1n C-O-CH2-CH(CH3)2 O N 6.66 (d. 1H, Ar), 7.08 (m, 2H, Ar-COO-), 7.94 (m, 2H, Ar-N); IR(KBr) u (cm) O 1650 cm (C=O), CHNH-C-(CH2)-CH3 1720 cm (-COOR) 40 NVABI (Compound 10) 3300 cm (NH-CO-); MS(FAB+):MS m/z 483(M+H)+ (FIGS. 5A-5B); Anal. Calcd for CHNOs, Scheme 4: C, 69.57%, H, 7.88%, N, 5.80%, O, 16.75%, 45 Found: OH C, 69.53%, H, 7.90%, N, 5.75%, O, 6.82%, UV(EtOH) max nm (log e): (FIGS. 6A-6B) 211 (2.45), 289 (2.36) Compound 2 O NIEthylamino-(4-oxymethylene, 5-methylene)-3- methyoxybenzyl nonanamide (compound 2) NVA mp: 94°-97° C. HCHO, CH3COOH H-NMR; (CDC1,) (FIGS. 3A-3D): CH3CHOH, 70° C. G 8 0.87 (t, 3H, CH.), 55 1.3–1.61 (m, CH), diethylamine 2.21 (t, 2H, J=7.58, NH-CO-CH-), 2.83 (tert, 2H, J=10.71, N-CH), CHN1 3.85 (s, 3H, OCH), YOHs 60 3.98 (s, 2H, Ar-CH), 4.32 (d. 2H, J=2.84, Ar-CH-N), O 4.95 (s, 2H.O-CH-N), CHNH-C-(CH2)-CH3 5.63 (s, 1H, NH), NVADA (Compound 4) 65 6.65-6.66 (d. 2H, Ar-H); IR(KBr) u (cm) 1650 cm (C=O),

5,840,720 11 12 5.48 (s, 2H, ArOCHN=) IR(KBr) (cm) 6.58–6.70 (q, 2H, ArH) 1650 cm (CO) 7.16–7.20 (d. 2H, ArCOO) 2950 cm (CH) 7.78–7.82 (d. 2H, ArCOO) MS (FAB+):MS m/z. 378 (M+H)+ 8.14–8.19 (t, 1H, CH-NHCO) Anal. Calcd for CHNO 12.44 (s, 1H, ArCOOH). Compound 12 Compound 9 NIp-ethylketone -phenylamino-(4-oxymethylene, No-(carboxylic acid methylester)-phenylamino-(4- 5-methylene)-3-methoxybenzyl) nonamide oxymethylene, 5-methylene)-3-methoxybenzyl) nona- 10 Melting point: 124°–126° C. mide NMR 1H-NMR (CDC1): 8 0.87 (3H, t, CH-(CH) ) 0.87 (t, 3H, CH) 1.256 (3H, t, CH-CHCO-) 1.1.0-2.20 (m, 14H, CHx7) 15 1.25-1.33 (12H, m, 6x(CH)) 3.73 (s, 3H, COOCH) 2.20 (2H, t, CO-CH-) 2.92 (2H, q, CHCHCO) 3.84-3.87 (d, 3H, OCH) 3.85 (3H, s, OCH) 4.30 (d. 2H, ArCH-NHCO) 4.68, 5.47 (each 2H, s, -CH-N-CH-O-) 4.42 (s, 2H, ArCHN=) 5.65 (1H, t, —CH-NHCO-) 5.70 (b 1H, NH) 2O 6.58 (1H, s, 3–H) 6.74 (m, 2H, 2xArH) 6.67 (1H, s, 5-H) 7.08-7.10 (d. 1H, ArH) 7.10 (2H, d. 2' and 6'-H) 7.13–7.14 (d. 12H, ArH) 25 7.91 (2H.d, 3'and 5'-H) 7.74–7.75 (d. 2H, ArH) Compound 10 TABLE 1. Np-carboxylic acid isobutylester-phenylamino-(4- Recovery half-life of capsaicin-induced blood oxymethylene, 5-methylene)-3-methoxybenzyl nona pressure (A, B and C phase) changes during a 15 min mide 3O infusion of NVABI, NVADA or capsazepine mp: 110-110.5 C. 1H-NMR (CDC1): 8 0.87 (t, 3H, CH) Compd. A phase B phase C phase NVABI 10.67 15.75 21.75 & 1.27 (m, 12H, (CH)6 CH) 35 NVADA 4.30 9.OO 4.49 8 2.18 (d. 2H, -CO-CH-, J-8.2 Hz) Capsazepine 2.73 2.89 3.78 & 3.84 (s, 3H, OCH) & 4.05 (d. 2H, COOCHCH, J=6.6 Hz) *t was calculated by one compartment model. & 4.34 (d. 2H, -ArCH-NH-, J-5.7 Hz) & 4.67 (s, 1H, N-H) 40 TABLE 2 & 6.62 (d. 1H, Ar-H, J=3.4 Hz) Recovery half-life of capsaicin-induced heart rate changes (fast and slow phase) during a 15 min infusion of NVABI, & 7.09 (d. 2H, Ar-H-COO-, J=9.1 Hz) NVADA or capsazepine. & 7.96 (d. 2H, ArH, J–9.0 Hz) IR (KBr) (cm) 45 L (min)" 1650 (CO) Compd. Fast phase Slow phase 1720 (COOR) NVABI 11.95 15.75 3300 (NH) NVADA 5.33 6.OO MS(FAB+): MS m/z 510 (M+H) 50 Capsazepine 1.36 3/52 Anal. Calcd. for CoHNOs *t was calculated by one compartment model. Found: C, 70.18%, H, 8.25%, N, 5.65% Compound 11 TABLE 3 55 N-(4-hydroxy, 5- diethylamino methyl)-3- Effects of intrathecal infusion (i.t.) of 1 nmole of NVABI, methyoxybenzyl nonamide NVADA or capsazepine on the depressor reflex responses to intraarterial injection of capsaicin (10 ug/kg) induced blood mp: 218–218.5 C. pressure changes. 1H-NMR (CDC1) & 0.87 (t, 3H, CH, J=6.2 Hz) 60 Pretreatment (i.t. 1 nmole) Capsaicin (i.a., 10 ug/kg) Ö 1.24 (m, 12H, (CH),CH) None -19.6 1.9 NVABI 19.2 2.5* & 2.12 (t, 2H, -NHCOCH-, J-7.3 Hz) NVADA -10.6 - 2.5 & 2.83 (m, 2H, NCH-) Capsazepine 25.83.O* & 3.72 (s, 3H, OCH) 65 & 4.14 (d. 2H, Ar-CH-, J-5.8 Hz) All values after pretreatment differ significantly (* P-0.05) & 6.54 (d. 1H, Ar-H, J=1.7 Hz) from control. Each datum represents the meani-S.E.M. (n=6) 5,840,720 13 What we claim is: 1. A capsaicin derivative having the formula

CHO O CHO O CH-NHCR 1O wherein R is Selected from the group consisting of -R wherein R is a member Selected from the group consisting of -R 15 -()- COOR -()- COOR2 -()- COOR -()- COOR2 R1OOC R1OOC () -()- COOH () -()- COOH HOOC 25 HOOC -COOR R1COOR R1COOR2 -COOR R1COOR -COOR R1COOR2 -COOR2 R1OC

R1OC

-()- R1COOR2 () 35 -()- R1COOR -()- COR -()- R1COOR -()- COR 40 COORSO3H

COORSO3H

45 wherein R is Selected from the group consisting of C wherein R is a member Selected from the group consist alkyl, C112 alkenyl, C112 alkylene, and C112 ing of C2 alkyl, C. 12 alkenyl, C. 12 alkylene, and C - alkenylene, and alkenylene, and wherein R is a member Selected from the group consist wherein R is selected from the group consisting of H, ing of H, C alkylene-NRR, and C alkenylene 50 Calkylene-NRR and Coalkenylene-NRR, and NRR. wherein Said composition contains an amount of Said 2. The capsaicin derivative as defined in claim 1 compound Sufficient to Selectively antagonize wherein R is Selected from the group consisting of Cs capsaicin-Sensitive Sensory neurons of a living animal alkyl, Cs alkenyl, Cs alkylene, and Cs body. alkenylene, and 55 wherein R is Selected from the group consisting of H, 7. A pharmaceutical composition comprising a compound C alkylene-NRR, and C alkenylene-NRR. having the following formula 3. The capsaicin derivative as defined in claim 1 wherein R is selected from the group consisting of C. alkyl, Co alkenyl, Co alkylene, and Co alk 60 enylene. 4. The compound as defined in claim 1 wherein R and R2 are Straight-chained or branched. CHO 5. A pharmaceutical Salt prepared from the compound as O defined in claim 1. 65 6. A pharmaceutical composition comprising a compound CH-NHCR having the following formula 5,840,720 15 wherein R is Selected from the group consisting of -R -continued HOOC

-COOR R1COOR R1COOR2 -()- COOR -()- COOR2 -COOR R1OOC R1OC () -()- COOH -()- R1COOR2 () HOOC

-COOR R1COOR 15 R1COOR2 -()- R1COOR -()- COR -COOR

COORSO3H

-()- R1COOR2 () wherein R is Selected from the group consisting of C alkyl, C112 alkenyl, C112 alkylene, and C112 25 alkenylene, and wherein R is Selected from the group consisting of H, -()- R1COOR -()- COR C alkylene-NRR, and C alkenylene-NRR, and wherein Said composition contains an amount of Said COORSO3H compound sufficient to inhibit innervation of the ileum Smooth muscles of a living animal body. 9. A pharmaceutical composition comprising a compound having the following formula wherein R is Selected from the group consisting of C2 alkyl, C112 alkenyl, C112 alkylene, and C112 35 alkenylene, and wherein R is Selected from the group consisting of H, C alkylene-NR, and Coalkenylene-NRR, and CHO wherein Said composition contains an amount of Said 40 compound Sufficient to inhibit innervation of an airway O in a living animal body. 8. A pharmaceutical composition comprising a compound CH-NHCR having the following formula 45 wherein R is selected from the group consisting of -R -()- COOR -()- COOR2 CHO 50 O CH-NHCR wherein R is Selected from the group consisting of -R 55 HOOC

COOR COOR2 -COOR R1COOR R1COOR2 60 -COOR2

R1OC

COOH 65 -()- R1COOR2 5,840,720 17 -continued -continued R1OOC

R1COOR COR COOH

HOOC COORSO3H

1O -COOR R1COOR R1COOR2 -COOR2 wherein R is Selected from the group consisting of C2 alkyl, C112 alkenyl, C112 alkylene, and C112 R1OC alkenylene, and 15 wherein R is selected from the group consisting of H, C alkylene-NRR, and C alkenylene-NRR, and R1COOR2 () wherein Said composition contains an amount of Said compound Sufficient to produce direct negative chro notropic and/or negative inotropic cardioinhibitory effects. R1COOR -()- COR 10. A pharmaceutical composition comprising a com pound having the following formula 25 COORSO3H

CHO O wherein R is Selected from the group consisting of C alkyl, C112 alkenyl, C112 alkylene, and C112 CH-NHCR alkenylene, and wherein R is Selected from the group consisting of H, wherein R is selected from the group consisting of -R 35 C alkylene-NRR, and C alkenylene-NRR, and COOR COOR2 wherein Said composition contains an amount of Said compound Sufficient to produce tachyphylaxis. 40