Title of Invention

A PROCESS OF PREPARING 4-[PHENYL-(PIPERIDIN-4-YL)-AMINO]-BENZAMIDE DERIVATIVES

Abstract

Compounds of general formula I [Chemical formula should be inserted here. Please see paper copy] R₁ is selected from any one of phenyl, pyridinyl, pyrrolyl, thienyl, furanyl, imidazolyl, triazolyl, and pyridine N-oxide; where each R₁ phenyl ring and R₁ heteroaromatic ring may optionally and independently be further substituted by 1,2 or 3 substituents selected from straight and branched C₁-C6alkyl, NO₂, CF₃, C₁-C6alkoxy, chloro, fluoro, bromo, and iodo. The substitutions on the phenyl ring and on the heteroaromatic ring may take place in any position on said ring systems; are disclosed and claimed in the present application, as well as their pharmaceutically acceptable salts and pharmaceutical compositions comprising the novel compounds and their use in therapy, in particular in the management of pain, anxiety and functional gastrointestinal disorders.

Full Text

FORM 2 THE PATENTS ACT 1970 [39 OF 1970] The Patents Rule, 2003 COMPLETE SPECIFICATION [See Section 10 and Rule 13] "A PROCESS OF PREPARING 4-[PHENYL-(PIPERIDIN-4-YL) AMINOJ-BENZAMIDE DERIVATIVES ASTRAZENECA AB, a Swedish company of S-151 85 Sodertalje, Sweden, The following specification particularly describes the nature of the invention and the manner in which it is to be performed:- WO 02/094782 PCT/SE02/00944 Field of the Invention 5 The present invention is directed to novel compounds, to a process for their preparation, their . use and pharmaceutical compositions comprising the novel compounds. The novel compounds are useful in therapy, and in particular for the treatment of pain, anxiety and functional gastrointestinal disorders. 10 Background of the Invention The S receptor has been identified as having a role in many bodily functions such as circulatory and pain systems. Ligands for the 5 receptor may therefore find potential use as analgesics, and/or as antihypertensive agents. Ligands for the 5 receptor have also been shown 15 to possess immunomodulatory activities. The identification of at least three different populations of opioid receptors (fx, 5 and K) is now well established and all three are apparent in both central and peripheral nervous systems of many species including man. Analgesia has been observed in various animal models when one 20 or more of these receptors has been activated. With few exceptions, currently available selective opioid 5 ligands are peptidic in nature and are unsuitable for administration by systemic routes. One example of a non-peptidic 5-agonist is SNC80 (Bilsky E.J. et al, Journal of Pharmacology and Experimental 25 Therapeutics, 273(1), pp. 359-366 (1995)). There is however still a need for selective 5-agonists having not only improved selectivity, but also an improved side-effect profile. Thus, the problem underlying the present invention was to find new analgesics having improved analgesic effects, but also with an improved side-effect profile over current \i 30 agonists, as well as having improved systemic efficacy. WO 02/094 782 PCT/SE02/0Q944 Analgesics that have been identified and are existing in the prior art have many disadvantages in that they suffer from poor pharmacokinetics and are not analgesic when administered by systemic routes. Also, it has been documented that preferred 5 agonist compounds, described within the prior art, show significant convulsive effects when administered systemically. 5 We have now found certain compounds that exhibit surprisingly improved properties, i.a. improved 5-agonist potency, in vivo potency, pharmacokinetic, bioavailability, in vitro stability and/or lower toxicity. o Outline of the Invention The novel compounds according to the present invention are defined by the formula I 0 wherein R is selected from any one of (i) phenyl; WO 02/094782 PCT/SE02/00944 where each R phenyl ring and R heteroaromatic ring may optionally and independently be 5 further substituted by 1, 2 or 3 substituents independently selected from straight and branched C1-C6 alkyl, N02, CF3, Ci-Cg alkoxy, chloro, fluoro, bromo, and iodo. The substitutions on the phenyl ring and on the heteroaromatic ring may take place in any position on said ring systems; 10 When the R phenyl ring and the R heteroaromatic ring(s) are substituted, the preferred substituents are selected from anyone of CF3, methyl, iodo3 bromo, fluoro and chloro. R is as defined above and each R phenyl ring and R heteroaromatic ring may A further embodiment of the present invention is a compound according to figure I wherein R is as defined above and each R phenyl ring and R 15 independently be further substituted by a methyl group A further embodiment of the present invention is a compound according to figure I wherein R is phenyl, pyrrolyl, pyridinyl, thienyl or furanyl, optionally with 1 or 2 of the preferred substituents on the R phenyl or R heteroaromatic ring. 20 Another embodiment of the present invention"is a compound according to figure I wherein R is phenyl, pyrrolyl or pyridinyl, optionally with 1 or 2 of the preferred substituents on the R phenyl or R heteroaromatic ring. 1 25 Another embodiment of the present invention is a compound"according to figure I wherein RJ is thienyl or furanylO ptionaly 1or2 of the preferred substitutes on the R1 heteroaromatic ring. r- WO 02/094782 PCT/SE02/00944 > Within the scope of the invention are also salts and enantioiners of the compounds of the formula I, including salts of enantioraers. ) b in Scheme 1, vide infra, is performed by reacting an intermediate compound 1 formula H 10 wherein PG is a urethane protecting group, such as Boc and CBZ,benzyl or substituted benzyl protecting group, such as 2t4~dimethoxybenzyl;with 3-bromobenzonitrile,using a palladium catalyst, e.g. tris(dibenzylideneacetone) dipalladium(0) (Pd2(dba)3), in the presence of abase, e.g. tertBuON phosphine ligand such as bis-diphenylphosphanyl-dimethyl- 9H-xanthene (xantphos), to give the compounds of general formula HI, 0 "^ m which is thereafter deprotected, under standard conditions hydrolysed under basic conditions and alkylated using either: 15 i) a compound of the general foimulaR1 CH2rein R1 is as defined above and X is a halogen preferlbly chlorine and a smtable base or ii) a compound of the general formula R1 C HOwherein K.1 is as defined above, and a suitable reducing agent, to sive compounds of the general formula I. WO 02/094782 PCT/SE02/00944 Suitable bases to be used in the standard alkylation step 1) above include, but are not limited to, triethylamine and potassium carbonate. Suitable reducing agents to be used in the standard reduction step ii) include, but are not 5 limited to, sodium cyanoborohydride and sodium triacetoxyborohydride. Reaction step b is alternatively performed as in Scheme 2, vide infra, by reacting an intermediate compound of the general formula IV IV 10 wherein PG is a urethane protecting group, such as Boc and CBZ, or benzyl or substituted benzyl protecting group, such as 2,4-dimethoxybenzyl; with N,N-diethyl-4-bromobei(2amidey using a palladium catalyst, e.g. Pd2(dba)3., in the presence of a base, e.g. tert-BuONa and a phosphine liganoVsuch as xantphos, to give the compounds of general formula IE, above. 15 The novel compounds of the present invention are useful in therapy, especially for the treatment of various pain conditions such as chronic pain, neuropathic pain, acute pain, cancer pain, pain caused by rheumatoid arthritis, migraine, visceral pain etc. This list should however not be interpreted as exhaustive. 20 Compounds of the invention are useful as immunomodulators, especially for autoimmune diseases, such as arthritis, for skin grafts, organ transplants and similar-surgical needs, for collagen diseases, various allergies, for use as anti-tumour agents and anti viral agents. Compounds of the invention are useful in disease states where degeneration or dysfunction of 25 opioid receptors is present or implicated in that paradigm. This may involve the use of WO 02/094782 PCT/SE02/00944 WO 02/094782 PCT/SE02/00944 PG PG PG wherein PG is"a urethane protecting group, such as Boc and CBZ or benzyl or substituted benzyl protecting group, such as 2,4-dimethoxybenzyl. 5 A further aspect of the present invention is intermediates of Formula X; O wherein R is as described above in relation to Formula I 10 . In a alternative synthetic route, depicted in Scheme 4 infra, the steps a and b of Scheme 1 infra, or steps a and b of Scheme 2 infra, are accomplished in a "one-pot" protocol whereby intermediates of general formulae H or IV are not isolated. -Utilizing this protocol, the initial palladium catalyzed coupling is performed in the same manner as for reactions according to is step a of Schemes 1 and 2, However, when the reaction is complete, rather than isolating an intermedate of formulla II or IV ,the secound aryl bromide and additional base (sodium tert butoxide) are added, and the temperature conditions of step b (Schemes 1 & 2) are employed, yielding products of general formula HI above. WO 02/094782 PCT/SE02/00944 In another alternative synthetic route, Reaction step b in Scheme 5, vide infra, is performed by reacting an intermediate compound of the general formula V. wherein PG is a urethane or benzyl-like protecting group, such as Boc,N,N diethy-4- bromobenzamide, using a palladium catalyst, e.g. tris(dibenzylideneacetone) dipalladium(0 [ Pd2 (dba)3esence of a base, e.g. tert BuONa and hosphine ligand such as bis-diphenyIphosphanyl-dimethyI-9H-xanthene (xantphos), to give the compounds of general 10 formula VI, which is thereafter deprotected and alkylated by means described above either reductively with a compound of the general formula R -CHO, or directly, using a compound of general formula R"CH2 -X,onversion of the ketal functionality to a primary amide 15 under standard conditions via i) hydrolysis of the ketal to the aldehyde (formula VII), followed by iii) amidation with ammonium chloride to the primary amide to give compounds of the general formula I WO 02/094782 PCT/SE02/00944 10 Suitable hydrolysis conditions to be used in the standard hydrolysis step (i) include, but are not limited to aqueous hydrochloric acid in tetrahydrofuran. Suitable conditions for the oxidation step (ii) include, but are not limited to stirring at 0°C in aqueous sodium dihydrogen phosphate and sodium chlorite in the presence of excess 2- methyl-2-butene. Suitable conditions for the amidation step (iii) include but are not limited to treatment with excess ammonium chloride in the presence of a coupling agent such as benzotriazole-1 -yloxy- trisphosphonium hexafluorophosphate (hereinafter Py-BOP). Methods of preparation EXAMPLES The invention will now be described in more" detail by the following Schemes and Examples, 15 which are not to be construed as limiting the invention. Scheme 1: Synthesis Of Intermediate 1 (Method 1) WO 02/094782 PCT/SE02/00944 Method la (compound 3): 4-(4-DietHaYlcarbamovl-Phenvlamino")-piperidine-l-carboxvlic acid tert-butvl ester. 5 To a dry flask containingN,N-diethyl-4-bromobenzaniide (5.33g; 1 .Ocq) and 2 (5.0g; 1.2eq) in dry toluene (80mL) was added BINAP (390mg; 0,03eq), palladium acetate (94mg; 0.02eq) and sodium tert-butoxide (2.80g; 1.4eq). The reaction was heated to 80 C under nitrogen. After 2 hours the solution was cooled, diluted with ethyl acetate and washed with 10 one portion water. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with 65% to 70% ethyl acetate in hexanes. The residue obtained was slurried in ethyl acetate/hexanes and colleGted^by-fil-tration. A-yellow-solid was obtained(5J^69g; 66% yieid^. i5 Method lb (compound4): 4-rC3-Cvano-phenvlVr4-diethvlcarbamovTphenvl)-amino]-piperidine-1 -carboxvlic acid tert-butvl ester. WO 02/094782 PCT/SE02/00944 To a dry flask containing amine 3 (5.1 Og) in dry toluene (SOmL) was added 3-bromobenzonitrile (3.47g; 1.4eq), xantphos (472mg; 0.06sq), Pd2(dba)3 (374mg; 0.03eq) and sodium tert-butoxide (1.83g; 1.4eq). The reaction was heated to 110 C under nitrogen. After .5 21 hours the solution was cooled, diluted with ethyl acetate and washed with one portion water. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with 50% ethyl acetate in hexanes. Apale yellow foam was obtained (5.60g; 86% yield). io Method 1c ("compound 1): rr3-Cvano-phenvl)-piperidin-4-vI-aniinoy]N,N-diethyl-benzamide. To a solution of 4 (5.43g) in dry dichloromethane (70mL) was added trifluoroacetic acid (8.8mL; lO.Oeq). The reaction was stirred overnight under nitrogen. The solution was washed with 2N sodium hydroxide and the aqueous was extracted with two portions 15 dichloromethane. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified through a small plug of silica eluting with 50% methanol in dichloromethane. A colorless foam was obtained (3.159g; 74% yield). WO 02/094782 PCT/SE02/00944 Scheme 2: Synthesis Of Intermediate i (Method 2) Method 2a (compound 5): 4-(3-Cvaao-phenvlaminoVpiperidine-l-carboxylic acid tert-butyl ester. 5 To a dry flask containing amine 2 (5.0g; 1.2eq) and 3-bromobenzonitrile (3.79g; " 1 .Oeq) in dry toluene (80mL) was added BINAP (390mg; 0.03eq), palladium acetate (94mg; 0.02eq) and sodium tert-butoxide (2.8g; 1.4eq). The reaction was heated to 80°C and was stirred for 24 hours under nitrogen. The solution was cooled, diluted with ethyl acetate and washed with two portions water. The organics were dried over anhydrous magnesium sulfate, 10 filtered and concentrated. The residue was purified by flash chromatography eluting with 100% hexanes to 30% ethyl acetate in hexanes. A colorless oil was obtained (5.12g; 82% yield). Method 2b: (Compound 4) 44[(3-Cyano-phenvl-(4-diethylcarbamoyl-phenyl)-amino")-15 piperidine-1-cafboxvlic acid tert-butvl ester. To a flask containing amine 5 (5.12g) in dry toluene (80mL) was N,N-diethyl-4-bromobenzamide (6.10g; 1.4eq), xantphos (590mg; 0.06eq), Pd2(dba)3 (467mg; 0.03eq) and 14 WO 02/094782 PCT/SE02/00944 sodium tert-butoxide (2.29g; 1.4eq). The reaction was heated to 110 C and stirred overnight under nitrogen. After 20 hours the reaction was diluted with ethyl acetate/water and filtered through diatomaceous earth. The organics were separated then dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was loaded on to a column in 5 dichloromethane-and eluted with 100% hexanes to 60% ethyl acetate in hexanes. An orange foam was obtained (6.67g; 82% yield). Method 2c ("compound 1); [(3-Cyano-phenyl)"piperidin-4-yl-amino]-N.N-diethyl-benzamide. To a solution of 4 (5.43g) in dry dichloromethane (70mL) was added trifluoroacetic acid ID (S.8mL; lO.Oeq). The reaction was stirred overnight under nitrogen. The solution was washed with 2N sodium hydroxide and the aqueous was extracted with two portions dichloromethane. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified through a small plug of silica eluting with 50% methanol in dichloromethane. A colorless foam was obtained (3.159g; 74% yield). 15 15 Scheme 3: Synthesis Of Example 1 WO 02/094782 PCT/SE02/00944 Intermediate 7: 4--(l-Benzyl-(piperdin-4-vlamino)-N,N-diethyl-benzimide To a dry flask containing l-benzyl-piperidin-4-ylarnine 6 (5.0mL; 1.2eq) in dry toluene (75mL) was added N,N-diethyl-4-bromobenzamide (5.24g; 1.0eq),BINAP (383mg; 0.03eq), palladium acetate (92mg; 0.02eq) and sodium rerr-butoxide (2.75g; 1.4eq). The reaction was 5 heated to 80°C under nitrogen. After 2 hours the reaction was cooled, diluted with ethyl acetate and washed with two portions water. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with 5% to 10% methanol in dichloromethane. The residue obtained was boiled in ethyl acetate, cooled and collected by filtration. A pale yellow solid was to obtained (5.724g; 77% yield). Intermediate 8: [(Benzyl -piperidine-4-yl)-(3-cyano-phenyl)-amino]-N,N-diethyl-benzamide To a dry flask containing amine 7 (1.767g; 1 .Oeq) in dry toluene (30mL) was added 3-brornobenzonitrile (1.234g; 1.4eq), xantphos (224mg; O.OSeq), Pd2(dba)3 (177mg; 0.04eq) and sodium rerr-butoxide (650mg; 1.4eq). The reaction was heated to110°C overnight under 15 nitrogen. After 16 hours the solution was cooled, diluted with ethyl acetate and washed with one portion water. The organics were dried over anhydrous" magnesium sulfate, filtered and concentrated. The residue was initially purified by flash chromatography eluting with 100% ethyl acetate. It was purified a second time eluting with 2.5% methanol in dichloromethane. A yellow foam was obtained (1.665g; 73%). 20 Example 1: 3-[(1-Benzyl-diethylcarbamoyl-phenyl)-amino1-benzamide. To a solution of 8 (1.58g) in tert-butanol (50mL) was added potassium hydroxide (475mg; 2.5eq). The reaction was heated to reflux for 2 hrs then was cooled. The solution was diluted with dichloromethane and washed with one portion water. The aqueous was then neutralized and extracted with two portions dichloromethane. The combined organics were 25 " dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with 5% to 10% methanol in dichloromethane. A pale yellow foam was obtained (1.556g; 95% yield; >99% purity). The residue was suspended in ether (15mL) and enough dichloromethane was added to give a homogeneous solution. IN HCl/ether (4.9mL; = 1.5eq) was added and after 30 minutes the suspension was concentrated 30 and the solid dried under high vacuum overnight. 16 WO 02/094782 PCT/SE02/00944 Alternative Synthesis of Intermediate 8: N,N-diethyl-4-[(3-cvanophenyl)[l-(phenylmethy)-4-piperidinyl]amino]benzamide Scheme 4: Performance of two consecutive Palladium catalyzed couplings in "One-pot" 5 To a solution of 1.07g of 3-bromobenzonitrile (5.88 mmol) in 15 ml dry toluene was added 1.2 mL of 4-amino-jV-benzyl piperidrne, 6 (5.89 mmol), 293 mg racemic BINAP (0.47 mmol), 10 215 mgtris(dibenzylideneacetone)dipalladium(0) (0.23 mmol) and 790 mg sodium tert butoxide (8.23 mmol).- -The reaction was heated at 80C under a nitrogen atmosphere for 4 hours. The reaction was cooled to room temperature and 2.26 g of N, 7V-diethyl-4-brornobenzamide (8.83 mmol) and 790 mg of sodium tert butoxide (8.23 mmol) were added and the reaction heated to reflux. After 20 hours the solution was cooled to room temperature 15 and the reaction diluted with ethyl acetate (50 ml) and water (30 ml) was added. The mixture was filtered through diatomaceous earth and then the organic layer was separated, dried (MgS04), filtered and concentrated. The residue was purified by flash chromatography, eluring 2% dichloromethane in methanol to yield an orange oil (2.54g, 5.45 mmol; 93%). (400MHz, CDC13) 5H 1.36 (br s, 6H, CH3) ; 1.44-1.54 (m, 2H, CH2); 1.91 (d, >12.5Hz, 20 2H, CH2); 2.12 (t, J=12Hz, 2H, NCH2) ; 2.97 (d, J=12H2, 2H, NCH2); 3.26-3.60 (m, 4H, NCH); 3.51 (s, 2H, NCH2Ar); 3.79-3.86 (m, 1H, NCH); 6.86-6.89 (m, 1H, Ar-H); 6.92- 6.97 (m, 2H, Ar-H); 7.12-7.15 (m, IB, Ar-H); 7.23-7.32 (m, 6H, Ar^Hjy7.37-7:4r(m, 3HT Ar-H). 17 WO 02/094782 PCT/SE02/00944 Intermediate 9; fl-Benzyl-piperidin-4-yl)-(3-[l,3]dioxolans2-yl-phenyl)-amine. To a dry flask containing 2-(3-bromophenyl)-l,3-dioxaolane (l.OmL; l.Oeq) and amine 6, 5 (1.6mL; 1.2eq) in dry toluene (20mL) was added BINAP (l24mg; 0.03eq), palladium acetate (30mg; 0.02eq) and sodium tert-butoxide (0.89g; 1.4eq). The reaction was heated to 80°C under nitrogen. After 2 hours the solution was cooled, diluted with ethyl acetate and washed" with one portion water. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography, eluting with-5% io methanol in dichloromethane. An orange oil was obtained (2.035s; 91% yield). Intermediate 10:;[(benzyl-piperidin-4-yl)-(3-[1,3]dioxolan-2-yl)-amino]-n,n-diethyl-benzamide. WO 02/094782 PCT/SE02/00944 To a dry flask containing amine 9 (1.61g)m dry toluene (14OmL) was added ary] bromide (8.07g; i.4eq), xantphos (780mg; 0.06eq), Pd2(dba)3 (6l9mg; 0.03eq) and sodium tert-butoxide (3.03g; 1 -4eq). The reacrion was heated to 110°C under nitrogen. Aiter 23 hours"the solution was cooled, diluted with ethyl acetate and washed with one portion water. The 5 organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by -flash chromatography, eluting with 3 % methanol in dichloromethane. An orange oil was obtained (10.16g; 88% yield). Intermediate 11: r(l-Benzvl-piperidin-4-ylV(3-formyl-phenyl)-arnino]-N.N-diethyl- 10 benzamide To a solution of acetal 10 (10.16g) in tetrahydroluran (270mL) was added 2N HC1 solution (110mL). After 16 hours at room temperature, dichloromethane (200mL) was added and the aqueous layer was neutralized with aqueous saturated sodium bicarbonate solution. The organic layer was removed and the aqueous layer extracted with dichloromethane (2 x 15 200mL). The combined organic extracts were dried (MgSO4), filtered and concentrated and the residue was purified by flash chromatography, eluting with ethyl acetate. An orange oil . was obtained (8.3 lg; 89% yield). Intermediate 12: 3-[J-Benzyl-pjperidin-4-yl)-(4-diethvicarbamoyl-phenyl)-aminol-bernzoic 20 acid. To a solution of aldehyde l l (490mg; l.Qeq) in tert-butauol (25mL) was added 2-methyl-2-butene (l.lmL; lO.Oeq) and the solution was cooled to 0°C. A solution of sodium dihydrogen phosphate (1.30g; 9eq) and sodium chlorite (1.06g; 9eq) in water (8mL) was added and the reaction was stirred for 30 minutes at 0°C. The lert-butano\ was removed and the aqueous 25 was extracted with dichloromethane (5 x 15mL). The combined organic extracts were dried (MgSCO)4 filtered and concentrated and the residue was purified by flash chromatography, " eluting 15% methanol to 30% methanol in dichloromethane. A pink foam (340.5mg; 67% yield was obtained). 30 Example 1: 3-[(l-Benzyl-piperidin-4-yl)-(4-diethvlcarbamovl-phenvl)-ainmo1-benzamide (altemative synthesis). WO 02/094782 PCT/SE02/00944 To a solution of acid 12 (420mg; 1 .Oeq) in DMF (8mL) was added pyBOP (675mg; 1.5eq); HOBt (175mg; 1.5eq), diisppropylethylamine (0.6mL; 4.0eq) and ammonium chloride (93mg; 2eq). After 20 hours at room temperature the reaction was concentrated. The residue was dissolved in. ethyl acetate (30mL) and was washed with water (2 x 20mL) and saturated 5. sodium bicarbonate solution (20mL). The organic layer was dried (MgSO4)5 filtered and concentrated and the residue was purified by flash chromatography, eluting 5% methanol in dichloromethane. A pale pink oil was obtained (271.8mg; 65% yield). Additional examples were synthesized via the general procedures described below. 10 To a solution of the amine, 1, in dry tetrahydrofuran (THF) or 1,2-dichloroethane is added the aldehyde (1-1.5 eg) followed by sodium triacetoxy borohydride (1-1.6 eg.). The reaction is 15 stirred at room temperature under a nitrogen atmosphere for an extended period of time (6-48 hours) to ensure complete reaction. The reaction mixture is then subjected to a standard work-up procedure and a standard purification. The amount of THF or 1,2-dichloroethane is not crucial. An amount corresponding to about lmL/30 mg is preferred. 20 Procedure 2A in the synthesis of Example 2 below is typical. B. Hydrolysis ot the intermediate cyano compound: 20 WO 02/094782 PCT/SE02/00944 R1 R1 To a solution of the cyano intermediate in teit-butanol, is added ground potassium hydroxide (KOH) (2.5 eq.) and the resulting mixture is heated to reflux for about two hours. The mixture was then cooled to room temperature and subjected to a standard work-up procedure and a standard purification. The amount oftert-butanol is not crucial. An amount corresponding to about lmL/30 mg is preferred. Procedure 2B in the synthesis of Example 2 below is typical. Example 2; 3-r[l-(2.4-Dichloro-benzyl)-piperidin-(4-diethylcarbamoy-phenyl)-amino]-10 benzamide. I 13 2A:{( 3-Cyano-phenyl)-[l--(2.4-dichloro-benzyl)-piperidn-4-yl]-amino}-.N,N-diethyl-benzamide. 15 To a solution of amine l (556mg) in dry tetrahydromran (15mL) was added 2,4- diclorobenzaldebyde(285ing;" 1.1 eq). After 15 rninutes at room temperature, sodium triacetoxyborohydride (407.5mg; 1,3eq) was added and stirring was continued at room temperature under nitrogen. After 23 hours the reaction was quenched with saturated aqueous 21 WO 02/094782 PCT/SE02/00944 sodium bicarbonate and extracted with three portions dichlorornethane. The organics were dried over anhydrous magnesium sulfate, filtered and concentrated (LC/MS showed 8% starting material left). The residue was purified by flash chromatography during with 2% to 4% methanol in dichloromethane. A colorless foam was obtained (644mg; 81% yield). 13 Example 2 2B: 3-[l-(2.4-Dichloro-benzyl)-piperidin-4-y]-(4-diethylcarbamoyl-phenyl)-aminol-benzamide. 10 To a solution of 13 (644mg) in tert-butanol (20mL) was added ground potassium hydroxide " (169mg; 2.5eq) and the reaction was heated to reflux for 90 minutes, then cooled and diluted with dichloromethane. The solution was washed with one portion water and the organic was separated. The aqueous layer was neutralized with 2N HC1 and extracted with two portions 15 dichloromethane. The combined organics were dried over anhydrous magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with 5% methanol in dichloromethane; A colorless foam was obtained (610mg; 92% yield; >99% purity). The material was suspended in ether (12mL) and enough dichloromethane was added to obtain a homogeneous solution. IN HCl/ether (1.7mL; = 1 ."5eq) was added and after 2 20 hours the suspension was concentrated and the solid dried under high vacuum. Additional Exampleare prepared analogously. Analytical data for synthetic Examples is shown in Table 1 on the following pages. 22 WO 02/094782 PCT/SE02/00944 WO 02/094782 PCT/SE02/00944 WO 02/094782 PCT/SE02/00944 Table 1 (continued): Analytical data for synthetic Examples. &" WO 02/094782 PCT/SE02/00944 26 WO 02/094782 PCT/SE02/00944 Pharmaceutical compositions The novel compounds according to the present invention may be administered orally, sublingually, intramuscularly, subcutaneously topically, intranasally,intraperitoneally, 5 intrathoracially, intravenously, epidurally, intrathecally,intracerebroventricularly and by injection into the joints. A preferred route of administration is orally, intravenously or intramuscularly. The dosage.will depend on. the route of administration, the severity of the disease, age and weight of the patient and other factors normally considered by the attending physician, when determining the individual regimen and dosage level as the most appropriate for a particular patient. 15 For preparing pharmaceutical compositions from the compounds of this invention, inert, pharmaceutically acceptable carriers can be either solid or liquid. Solid form preparations include powders, tablets, dispersible granules, capsules, cachets, and suppositories. A solid carrier can be one or more substances which may also act as diluents, flavoring agents, 20 solubilizers, lubricants, suspending agents, binders, or tablet disintegrating agents; it can also be an encapsulating material. In powders, the carrier is a finely divided solid which is in a mixture with the finely divided active, component In tablets, the active component is mixed with the carrier having the 25 necessary binding properties in suitable proportions and compacted in the shape and size desired. For preparing suppository compositions, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for 30 example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify. WO 02/094782 PCT/SE02/00944 Suitable carriers are magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methyl cellulose, sodium carboxymethyl cellulose, a low-melting wax, cocoa butter, and the like. 5 Salts include, but are not limited to, pharrnaceutically acceptable salts. Examples of pharmaceutically acceptable salts within the scope of the present invention include: acetate, benzenesulfonate, benzoate, bicarbonate bitartrate, bromide, calcium acetate, camsylate, carbonate, chloride, citrate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, glucaptate, gluconate, glutamate, glycollylarsanilate, hexyiresorcinate, hydrabamine, 10 hydrobromide, hydrochloride, hydroxynaphthoate, isethionate, lactate, lactobionate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, pamoate (embonate), pantothenate, phosphate/diphosphate, polygalacturonate, sahcylate, stearate, subacetate, succinate, sulfate, tannate, tartrate, teoclate, triethiodide, and benzathine. 15 Examples of pharmaceutically unacceptable salts within the scope of the present invention include:"hydroiodide, perchlorate, tetrafluoroborate. Pharmaceutically unacceptable salts could be of use because of their advantageous physical and/or chemical properties, such as crystallinity. 20 Preferred pharmaceutically acceptable salts are hydrochlorides, sulfates and bitartrates. The hydrochloride and sulfate salts are particularly preferred The term composition is intended to include the formulation of the active component with 25 encapsulating material as a carrier providing a capsule in which the active component (with or without other carriers) is surrounded by a carrier which is thus in association with it. Similarly, cachets are included. Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral 30 administration. 29 WO 02/094782 PCT7SE02/00944 Liquid from compositions include solutions, suspensions, and emulsions. Sterile water or " water-propylene glycol solutions of the active compounds may be mentioned as an example of liquid preparations suitable for parenteral administration. Liquid compositions" can also be formulated in solution in aqueous polyethylene glycol solution. 5 Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use can be made by dispersing the finely divided active component in water together with a viscous material such as natural synthetic gums, resins, methyl cellulose, sodium carboxymethyl cellulose, and other suspending agents known to the 10 pharmaceutical formulation art. Preferably the pharmaceutical compositions is in unit dosage form. In such form, the composition is divided into unit doses containing appropriate quantities of the active component. The unit dosage form can be a packaged preparation, the package containing 15 discrete quantities of the preparations, for example, packeted tablets, capsules, and powders in vials or ampoules. The unit dosage form can also be a capsule, cachet, or tablet itself, or it can be the appropriate number of any of these packaged forms. BIOLOGICAL EVALUATION 20 In vitro model Cell culture 25 A. Human 293S cells expressing cloned human u., 5, and K receptors and neomycin resistance were grown in suspension at 3 7°C and 5% C02 in shaker flasks containing calcium-free DMEM10% FBS, 5% BCS, 0.1% Pluronic F-68, and 600 jig/ml geneticin. B. Mouse and rat brains were weighed and rinsed in ice-cold PBS (containing 2.5mM EDTA, pH 7.4). The brains were homogenized with a polytron for 15 sec (mouse) or 30 30 sec (rat) in ice-cold lysis buffer (50mM Tris, pH 7.0, 2.5mM EDTA, with WO 02/094782 PCT/SE02/00944 phenylmethylsulfonyl fluoride added just prior use to 0.5MmM from a 0.5M stock in DMSO:ethanol). Membrane preparation 5 Cells were pelleted and resuspended in lysis buffer (50 mM Tris, pH 7.0, 2.5 mM EDTA, with PMSF added just prior to use to 0.1 mM from 0.1 M stock in ethanol), incubated on ice for 15 min, then homogenized with a polytron for 30 sec. The suspension was spun at lOOOg (max) for 10 min at 4°C. The supernatant was saved on ice and the pellets resuspended and spun as before. The supernatants from both spins were combined and spun at 46,000 g(max) 10 for 30 min. The pellets were resuspended in cold Tris buffer (50 .mM Tris/Cl, pH 7.0) and spun again. The final pellets were resuspended in membrane buffer ( 50 mM Tris, 0.32 M sucrose, pH 7.0), Aliquots (1 ml) in polypropylene tubes were frozen in dry ice/ethanol and stored at -70°C until use. The protein concentrations were determined by a modified Lowry assay with SDS. 15 Binding assays Membranes were thawed at 37°C, cooled on ice, passed 3 times through a 25-gauge needle, and diluted into binding buffer (50 mM Tris, 3 mM MgCl2, 1 mg/ml BSA (Sigma A-7888), 20 pH 7.4, which was stored at 4°C after filtration through a 0.22 m filter, and to which had been freshly added 5 ug/rni aprotinin, 10uM bestatin, ] 10uM diprotin A, no DTT). Aliquots of 100 ul were added to iced 12x75 mm polypropylene tubes containing 100 ul of the appropriate radioligand and 100 ul of test compound at various concentrations. Total (TB) and nonspecific (NS) binding were determined in the absence and presence of 10uM 25 naloxone respectively. The tubes were vortexed and incubated at 25°C for 60-75 rnin, after which time the contents are rapidly vacuum-filtered and. washed with about 12 ml/tube iced wash buffer (50 mM Tris, pH 7.0, 3 mM MgCL2 ) through GF/B filters (Whatman) presoaked for at least 2h in 0.1 % polyethyleneixnine. The radioactivity (dpm) retained on the filters was measured with a beta counter after soaking the filters for at least 12h in minivials containing 30 6-7 ml scintillation fluid. If the assay is set up in 96-place deep well plates, the filtration is over 96-place PEI-soaked unifilters, which were washed with 3 x 1 ml wash buffer, and dried WO 02/094782 PCT/SE02/00944 in an oven at 55°C for 2h. The filter plates were counted in a TopCount (Packard) after adding 50 ul MS-20 scintillation fluid/well. Functional Assays 5 The agonist activity of the compounds is measured by deterrriining the degree to which the compounds receptor complex activates the" binding of GTP to G-proteins to which the receptors are coupled. In the GTP binding assay, GTP[Y]35S is combined with test compounds and membranes from HEK-293S cells expressing the cloned human opioid receptors or from 10 homogenised rat and mouse brain. Agonists stimulate GTP[y] S binding in these membranes. The EC50 and Emax values of compounds are determined from dose-response curves. Right shifts of the dose response curve by the delta antagonist naltrindole are performed to verify that agonist activity is mediated through delta receptors. 15 The plates are vortexed and incubated for 60 minutes at room temperature, filtered on Unifilters GF/B (presoaked in water) with the Tomtec or Packard harvester using 4 ml of wash buffer (50 mM Tris, 5 mM MgCl2; 50 mM NaCl, pH 7.0). The filters are dried for 1 hour at 55°C. The radioactivity (cpm) is counted in a TopCount (Packard) after adding 65 ul/well of MS-20 scintillation liquid. The stimulation (SB) in the presence of ligands is 20 expressed as percentage of Emax of the reference agonist. Values of EC50 for ligands in stimulating radioligand binding are calculated in Activity Base. Mean ± S.E.M. values of EC50 and % Emax are reported for ligands tested in at least three dose-response curves. Procedure for rat brain GTP 25 Rat brain membranes are thawed at 37°C, passed 3 times through a 25-gauge blunt-end needle and diluted in the GTPyS binding (50 mM Hepes, 20 mM NaOH, 100 mM NaCl, 1 mM EDTA, 5 mM MgCl2, pH 7.4, Add fresh: 1 mM DTT, 0.1% BSA). 120uM GDP final is added membranes dilutions. The EC50 and Emax of compounds are evaluated from 10-point dose-response curves done in 300ul with the appropriate amount of membrane protein 30 (20ug/well) and 100000-130000 dpm of GTPy35S per well (0.11 -0.14nM). The basal and maximal stimulated binding are determined in absence and presence of 3uM SNC-80 32 WO 02/094782 PCT/SE02/00944 Data analysis . The specific binding (SB) was calculated as TB-NS, and the SB in the presence of various 5 test compounds was expressed as percentage of control SB. Values of IC50 and Bill coefficient (nH) for ligands in displacing specifically bound radioligand were calculated from logit plots or curve fitting programs such as Ligand, GraphPad Prism, SigmaPlot, or ReceptorFit. Values of Ki were calculated from the Cheng-Prussoff equation. Mean ± S.E.M. values of IC50, Ki and nH were reported for ligands tested in at least three displacement 10 curves. Biological activity of the compounds of the present invention is indicated in Table 2. Table 2: Biological data. 15 Receptor saturation experiments Radioligand K5 values were determined by performing the binding assays on cell membranes with the appropriate radioligands at concentrations ranging from 0.2 to 5 times the estimated K5(up to 10 times if amounts of radioligand required are feasible). The specific radioligand 20 " binding was expressed as pmole/mg membrane protein. Values of K5 and Bmax from individual experiments were obtained from nonlinear fits of specifically bound (B) vs. nM free (F) radioligand from individual according to a one-site model. DETERMINATION OF MECHANO-ALLODYNIA USING VON FREY TESTING 25 Testing was performed between 08:00 and 16:00h using the method described by Chaplan et al. (1994). Rats were placed in Plexiglas cages on top of a wire mesh bottom which allowed access to the paw, and were left to habituate for 10-15 min. The area tested was the mid- 33 WO 02/094782 PCT/SE02/00944 plantar left hind paw, avoiding the less sensitive foot pads. The paw was touched with a series of 8 Von Frey hairs with logarithmically incremental stiffness (0.41, 0.69, 1.20, 2.04, 3.63, 5.50, 8.51, and 15.14 grams; Stoelting, 111, USA). The von Frey hair was applied from underneath the rnesh floor perpendicular to the plantar surface with sufficient force to cause a 5 slight buckling against the paw, and held for approximately 6-8 seconds. A positive response was noted if the paw was sharply withdrawn. Flinching immediately upon removal of the hair was also considered a positive response. Ambulation was considered an ambiguous response, and in such cases the stimulus was repeated. 10 Testing Protocol The animals were tested on postoperative day 1 for the FCA-treated group. "The 50% withdrawal threshold was determined using the up-down method of Dixon (1980). Testing was started with the 2.04 g hair, in the middle of the series. Stimuli were always presented in a consecutive way, whether ascending or descending. In the absence of a paw withdrawal is response to the initially selected hair, a stronger stimulus was presented; in the event of paw withdrawal, the next weaker stimulus was chosen. Optima] threshold calculation by this method requires 6 responses in the immediate vicinity of the 50% threshold, and counting of these 6 responses began when the first change in response occurred, e.g. the threshold was first crossed. In cases where thresholds fell outside the range of stimuli, values of 15.14 20 (normal sensitivity) or 0.41 (maximally allodynic) were respectively assigned. The resulting pattern of positive and negative responses was tabulated using the convention, X =" no withdrawal; O = withdrawal, and the 50% withdrawal threshold was interpolated using the formula: 25 50% g threshold = 10(Xf+k8) / 10,000 where Xf= value of the last von Frey hair used (log units); k - tabular value (from Chaplan et al. (1994)) for the pattern of positive / negative responses; and 5 = mean difference between stimuli (log units). Here 5 = 0.224. 30 34- WO 02/094782 PCT/SE02/00944 Von Frey thresholds were converted to percent of maximum possible effect (% MFE), according to Chaplan et al. 1994. The following equation was used to compute % MPE: % MPE = Drug treated threshold (g) - allodynia threshold (g) X 100 5 Control threshold (g) - allodynia threshold (g) ADMINISTRATION OF TEST SUBSTANCE Rats were injected (subcutaneously, intraperitoneally, intravenously or orally) with a test substance prior to von Frey testing, the time between administration of test compound and the 10 von Frey test varied depending upon the nature of the test compound. WRITHING TEST Acetic acid will bring abdominal contractions when administered intraperitoneally in mice. These will then extend their body in a typical pattern. When analgesic drugs are administered, 15 this described movement is less frequently observed and the drug selected as a potential good candidate. A complete and typical Writhing reflex is considered only when the following elements are present: the animal is not in movement; the lower back is slightly depressed; the plantar aspect of both paws is observable. In this assay, compounds of the present invention demonstrate 20 significant inhibition of writhing responses after oral dosing of l-100gm01/kg. (i) Solutions preparation Acetic acid (AcOHY 120 uL of Acetic Acid is added to 19.88 ml of distilled water in order to obtain a final volume of 20 ml with a final concentration of 0.6% AcOH. The solution is then is mixed (vortex) and ready for injection. Compound (drug): Each compound is prepared and dissolved in the most suitable vehicle according to standard procedures. 30 (ii) Solutions administration 35 The compound (drug) is administered orally, intraperitoneally (i.p.) , subcutaneously (s.c.) or • intravenously (i.v.)) at 3 0 rnl/kg (considering the average mice body weight) 20, 3 0 or 40 minutes (according to the class of compound and its characteristics) prior to testing. When the compound is delivered centrally: Intraventricularly (i.c.v.) or intrathecally (i.t.) a volume of 5 uL is administered. The AcOH is administered intraperitoneally (i.p.) in two sites at 10 ml/kg (considering the average mice body weight) immediately prior to testing. (in) Testing The animal (mouse)" is observed for a period of 20 minutes and the number of occasions (Writhing reflex) noted and compiled at the end of the experiment. Mice are kept in individual - "shoe box" cages with contact bedding. A total of 4 mice are usually observed at the same time: one control and three doses of drug. For the anxiety and anxiety-like indications, efficacy has been established in the geller-seifter . conflict test in the rat. For the functional gastrointestina disorder indication, efficacy can be established in the assay described by Coutinho SV et al, in American Journal of Physiology - Gastrointestinal & Liver -Physiology. 282(2):G307-16, 2002 Feb, in the rat Co-pending application numbers: 1011/MUMNP/2003, 1018/MUMNP/2003, 1012/MUMNP/2003, 1014/MUMNP/2003, 1015/MUMNP/2003, 1016/MUMNP/2003 and 1017/MUMNP/2003. WE CLAIM:- 1. A process for preparing a compound 4-[phenyl-(piperidin-4-yl)-arnino-benzamide derivatives of formula I, comprising reacting a compound of the general formula II O wherein PG is a urethane protecting group, such as Boc and CBZ or benzyl or substituted benzyl protecting group, such as 2,4-dimethoxybenzyl, with 3-bromobenzonitrile, using a palladium catalyst, Pd2(dba)3, in the presence of a base, preferably sodium tert-butoxide, and a phosphine ligand such as heTein described to give the compounds of general formula III, which is thereafter deprotected, under standard conditions and alkylated under reductive conditions with a compound of the general formula R1 CHO to give the compounds of the general formula I, 38 wherein R1 is selected from any one of phenyl, pyridinyl, thienyl, furyl, imidazolyl, triazolyl, pyrrolyl, thiazolyl and pyridyl-N-oxide; wherein each R1 phenyl ring and R1 heteroaromatic ring may independently be further substituted by 1, 2 or 3 substituents independently selected from straight and branched C1-C6 alkyl, N02, CF3, C1-C6 alkoxy, chloro, fluoro, bromo and iodo, as well as salts thereof.

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01013-mumnp-2003-abstract(11-01-2005).doc

01013-mumnp-2003-abstract(11-01-2005).pdf

01013-mumnp-2003-cancelled pages(11-01-2005).pdf

01013-mumnp-2003-claims(granted)-(11-01-2005).doc

01013-mumnp-2003-claims(granted)-(11-01-2005).pdf

01013-mumnp-2003-correspondence(11-12-2005).pdf

01013-mumnp-2003-correspondence(ipo)-(07-01-2005).pdf

01013-mumnp-2003-form 19(03-11-2003).pdf

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01013-mumnp-2003-form 2 (granted)-(11-01-2005).doc

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01013-mumnp-2003-form-form-pct-ipea-409(11-01-2005).pdf

01013-mumnp-2003-form-form-pct-ipea-416(11-01-2005).pdf

01013-mumnp-2003-form-pct-isa-210(11-01-2005).pdf

01013-mumnp-2003-petition under rule 137(01-11-2004).pdf

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01013-mumnp-2003-petition under rule 138(01-11-2004).pdf

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1013-mumnp-2003-abstract(24-12-2007).pdf

1013-mumnp-2003-claims(amanded)-(1-11-2004).pdf

1013-MUMNP-2003-CLAIMS(COMPLETE)-(3-11-2003).pdf

1013-mumnp-2003-claims(granted)-(24-12-2007).pdf

1013-mumnp-2003-correspondence(ipo)-(21-1-2008).pdf

1013-MUMNP-2003-DESCRIPTION(COMPLETE)-(3-11-2003).pdf

1013-mumnp-2003-description(granted)-(24-12-2007).pdf

1013-mumnp-2003-form 2(complete)-(3-11-2003).pdf

1013-mumnp-2003-form 2(granted)-(24-12-2007).pdf

1013-MUMNP-2003-FORM 2(TITLE PAGE)-(COMPLETE)-(3-11-2003).pdf

1013-mumnp-2003-form 2(title page)-(granted)-(24-12-2007).pdf

1013-mumnp-2003-wo international publication report(2-11-2003).pdf