Title of Invention	INDOLYLMALEIMIDE DERIVATIVES AND PROCESS FOR THEIR PRODUCTION
Abstract	ABSTRACT "INPOLYLMALEIMIDE DERIVATIVES AND PROCESS FOR THEIR PRODUCTION" The invention related to compounds of formula I where R, Ra and Rb are as defined in the claims, which have valuable pharmacological properties e.g. inhibiting Protein Kinase C, inhibiting T-cell activation and proliferation, etc.

Full Text

Indolvlmaleimide Derivatives The present invention relates to indolylmaleimide derivatives, process for their production and pharmaceutical compositions containing them. More particularly the present invention provides a compound of formula I R1 is piperazin-1 -yl optionally substituted by CH3 in position 3 or 4; or 4,7-diaza-spiro [2.5] oct-7-yl; R2 is CI; Br; CF3; or CH3; and Rs is H; CH3; or CF3; R2 being other than CH3 or CI when R3 is H, Ra is H or CH3, Rb is R4 is 1 -piperazinyl substituted in positions 3 and/or 4 by CH3; or 4,7-diaza-spiro [2.5] oc±-7-yl; R2 being other than H or CH3 when R4 is 4-methyl-1-piperazinyl; or 3 by methyl or ethyl and optionally In position 4 by methyl. The compounds of formula I may exist in free form or in salt form, e.g. addition salts with e.g. organic or Inorganic acids, for example, hydrochloric acid, acetic add, trifluoroacetic acid. It will be appreciated that the compounds of formula I may exist in the form of optical isomers, racemates or diastereoisomers. For example, a ring carbon atom bearing a substltuent in the position 3 of the piperazinyl residue Is asymmetric and may have the R- or S- configuration. It is to be understood that the present Invention embraces all enantiomers and their mixtures. Enantiomers are preferred over racemates. Similar considerations apply in relation to starting materials exhibiting asymmetric carbon atoms as mentioned. Alkyl or alkoxy may be straight or branched. Phenyl-C1-4alkyl is preferably benzyl or phenethyl. In C1-4alkoxy-C1-4alkyl the alkoxy moiety is preferably methoxy or ethoxy and the alkyl moiety preferably methyl or ethyl; a suitable example is e.g. 2-methoxyethyl. Halogen may be F, CI, Br or I, preferably F, CI or Br. Halogeno-C1-4alkyl is alkyl wherein one or more H are replaced by halogen, e.g. CI or F, e.g. CH2CI, CH2F or CF3 R is preferably a radical of formula (a), (c) or (e), preferably (e). In the radical of formula (a) or (c), R2 or R15 is preferably in para to R1 or R14, respectively. R3 is preferably in meta to Ri. In the radical or formula (e), R9 is preferably 4,7-diaza-spiro [2.5] and, where required, converting the resulting compound of formula I obtained in free form to a salt form or vice versa, as appropriate. The process may conveniently be effected in the presence of a strong base, e.g. t-BuOK, e.g. as disclosed in WO02/38561, the contents of which being incorporated herein by reference, and as illustrated in the Examples. Compounds of formula II and III may be prepared in accordance with known methods, e.g. as disclosed in WO02/38561, the contents of which being incorporated herein by reference, and as illustrated in the Examples. Insofar as the production of the starting materials Is not particularly described, the compounds are known or may be prepared analogously to methods known in the art or as described hereafter. 2-[2-Chloro-3-methyl-5-(4-methyl-piperazin-1-yl)-phenyl]-acetamide (211 mg, 0.75 mmol) and 3-indoleglyoxylate (270 mg, 1.35 mmol) are dissolved in THF (5 mL). A solution of 1.0 M tert-BuOK in THF (2.98 mL, 4 eq) is added and the mixture is stirred at 35°C overnight. The reaction is diluted with AcOEt (20 mL) and washed with H2O (20 mL) and brine (10 mL). The organic phase is dried over Na2S04 and the solvent is evaporated. The residue is purified by FCC (AcOEt/AcOH/H20 7:1:1) to afford the title compound in the form of its acetate salt. 1H NMR (DMSO, 400 MHz) 5 2.16 (s, 3H), 2.31 (s, 3H), 2.32-2.38 (m, 4H), 2.97-3.10 (m, 4H), 6.61 (d, J = 8.0 Hz, 1H), 6.71 -6.77 (m, 2H), 7.04 (d, J = 2.8 Hz, 1H), 7.08 (dd, J = 7.4, 7.2 Hz, 1H), 7.41 (d, J = 8.2 Hz, 1H), 7.98 (d, J = 1.5 Hz, 1H), 11.05 (br s, 1H), 11.90 (br s, 1H); ES-MS: 435 [M+Hf. used as starting material may be prepared as follows: (5-Bromo-2-chloro-3-methyl-phenyl)-acetic acid methyl ester (2.0 g, 7.2 mmol), N-methylpiperazine (960 nL, 8.6 mmol) and CS2CO3 (3.3 g, 10.1 mmol) are suspended in toluene (80 mL). Pd(0Ac)2 (81 mg, 0.36 mmol) and BINAP (224 mg, 0.36 mmol) are added and the reaction is stirred at 100°C overnight. The mixture is filtered through Celite and the solvent Is evaporated. The residue is purified by FCC (AcOEt/AcOH/HaO 60:15:15) to afford 2-[2-chloro-3-methyl-5-(4-methyl-piperazin-1-yl)-phenyl]-acetic acid methyl ester. The ester is suspended in NH4OH 25% (60 mL). The mixture is stirred overnight at RT and the precipitate is filtered off to yield the amide. 1H NMR (DMSO, 400 MHz) 6 2.20 (s, 3H), 2.27 (s, 3H), 2.40-2.45 (m, 4H), 3.07-3.13 (m, 4H), 3.48 (s, 2H), 6.78 (s, 1H), 6.82 (s, 1H), 6.91 (brs, 1H), 7.34(brs, 1H). CuCIa (8.0 g, 0.06 mol) and tert-butylnitrile (8.9 mL, 0.074 mol) are suspended in acetonitrile (60 mL). 1,1-dlchloroethylene is added dropwise at 20°C. 5-Bromo-2-chloro-3-methyl-phenylamine (11.0 g, 0.05 mol), dissolved in acetonitrile (60 mL), is added and the reaction is stirred at RT for 3 h. The mixture is poured into aqueous 20% HCI (150 mL) and the aqueous phase is extracted with CH2CI2 (2 x 150 mL). The organic phase is dried over Na2S04 and the solvent is evaporated. The residue is purified by FCC (hexane/CH2Cl2 9:1) to afford 5-bromo-2-chloro-1-methyl-3-(2,2,2-trichloro-ethyl)-benzene. The intermediate is redissolved in MeOH (80 mL) and heated to 70°C. A solution of 5.4 M NaOMe in MeOH (28.4 mL) is added dropwise and the reaction is stirred at 70°C for 3 h. The reaction is cooled to RT and concentrated H2SO4 (10 mL) is added. The reaction is stirred at reflux for 1 h. The mixture is diluted with H2O (200 mL) and extracted with CH2CI2 (2 x 200 mL). The combined organic phases are dried over Na2S04 and the solvent is evaporated. The residue is purified by FCC (hexane/CH2Cl2 9:1 to 1:1) to afford (5-bromo-2-chloro-3-methyl-phenyl)-acetic acid methyl ester. 1H NMR (CDCI3, 400 MHz) 5 2.26 (s, 3H), 3.61 (s, 3H), 3.63 (s, 2H), 7.15 (s, 1H), 7.21 (s, 1H). By following the procedure of Example 1 but using the appropriate starting materials, the compounds of formula I wherein R is a residue of formula (a), as indicated in Table 1 below may be obtained. 2-[3-(4,7-Diaza-spiro[2.5]oct-7-yl)-naphthaIen-1-yl]-acetannide (100 mg, 0.30 mmol) and (1-methyl-1H-indol-3-yl)-oxo-acetic acid methyl ester (97 mg, 0.44 mmol) are azeotroped three times with dry THF and then dissolved in dry THF (3 ml). A solution of 1.0 M KOtBu in THF (1.2 ml) is added dropwise over 20 minutes at RT. After 5 minutes, TLC analysis indicates complete conversion of starting materials. The reaction is quenched by the additon of water (5 ml). The mixture is diluted with EtOAc and washed twice with saturated aq. NH4CI. The aqueous layers are back extracted twice with EtOAc. The combined organic layers are dried over Na2S04 and the solvent is evaporated. The residue is purified by FCC (EtOAc/AcOH/HaO 800:55:45) to afford the title compound as its acetate salt. The compound is dissolved in MeOH/TFA and the solvent is removed to yield the title compound as its trifluoroacetate salt. 1H NMR (DMSO, 400 MHz) 6 0.95 (M. 4H), 3.39 (br, 4H), 3.49 (br, 2H), 3.86 (s, 3H), 6.16 (d, J = 7.9 Hz, 1H), 6.46 (dd, J = 6.6 / 7.9 Hz, 1H), 7.00 (dd, J = 6.6 / 7.9 Hz, 1H). 7.13 (dd, J = 7.6/7.6 Hz, 1H), 7.40 (m, 4H), 7.55 (d, J = 8.5 Hz, 1H), 7.78 (d, J = 7.6 Hz, 1H), 8.14 (s, 1H), 9.10 (br, 2H), 11.17 (s, 1H); ES-MS: 463 [M+H]2-[3-(4,7-Diaza-spiro[2.5]oct-7-yl)-naphthalen-1 -yl]-acetamide, used as starting material may a) 2-[3-(4-Benzyl-4,7-dlaza-spiro[2.5]oct-7-yl)-naphthalen-1-yl]-acetamide (280 mg, 0.73 mmol) is azeotroped twice with a 1.25 M solution of HCI in MeOH. The residue is dissolved in Eton (10 ml). Palladium on charcoal (10%, 77 mg) is added, and the mixture is stirred under an atmosphere of hydrogen (1 atm) at RT for 14 h and at 50 °C for 2 h. The mixture is filtered, and the filtrate is concentrated. The residue is purified by FCC (EtOAc/AcOH/HaO 750:83.68 to 600:150:150) to yield the title compound containing 0.7 eq of AcOH. 1H NMR (DMSO, 400 MHz) 5 0.53 (m, 4H), 1.78 (s, AcOH), 2.93 (ddd, 2H), 3.04 (s, 2H), 3.16 (ddd, 2H), 3.76 (s, 2H), 6.94 (br, 1H), 6.99 (s, 1H), 7.25 (dd, IN), 7.26 (s, 1H), 7.35 (dd, J = 6.7/ 7.8, 1H), 7.49 (br, 1H), 7.69 (d, J = 7.8, 1H), 7.88 (d, J = 10.0, 1H); ES-MS: 296 [M+H]^ b)2-[3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-naphthalen-1-yl]-acetamide [3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-naphthalen-1-yl]-acetic acid ethyl ester (347 mg, 0.84 mmol) and formamide (126 mg, 2.80 mmol) are dissolved under an atmosphere of argon in DMF (1 ml). The solution is heated to 105 °C, and NaOMe (155 ^L of a 5.4 M solution in MeOH, 45 mg, 0.84 mmol) is added dropwise during 15 minutes. After 30 minutes at 105 °C, TLC analysis indicates complete consumption of starting material. The reaction mixture is cooled to RT, diluted with water, and extracted with EtOAc. The EtOAc layers are washed twice with water. Removal of solvent and purification by FCC (EtOAc/MeOH 98:2 to 96:4 to 90:10) yielded the title compound. 1H NMR (CDCI3, 400 MHz) 5 0.71 (ddd, 2H), 0.89 (ddd, 2H), 3.10 (ddd, 2H), 3.16 (s, 2H), 3.31 (ddd, 2H), 3.93 (s, 2H), 3.99 (s, 2H), 5.33 (br, 1H), 5.42 (br, 1H), 7.11 (s, 1H), 7.23 (d, J = 2.0, 1H), 7.32 (m, 5H), 7.37 (dd, J = 6.7 Hz, 1H), 7.45 (dd. J = 6.7 Hz, 1H), 7.73 (d, J = 8.9 Hz, 1H), 7.85 (d, J = 8.9 Hz, 1H); ES-MS: 386 [M+H]^ c) [3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-naphthalen-1 -yl]-acetic acid ethyl ester (3-Trifluoromethanesulfonyloxy-naphthalen-1 -yl)-acetic acid ethyl ester (500 mg, 1.38 mmol) is dissolved under an atmosphere of argon in dry THF (10 ml). 4-Benzyl-4,7-diaza-spiro[2.5]octane (325 mg, 1.61 mmol) is added, followed by the addition of K3PO4 (410 mg, 1.93 mmol), Pd2(dba)3 (62 mg, 0.069 mmol) and biphenyl-2-yl-di-tert-butyl-phosphane (21 mg, 0.069 mmol). The reaction mixture is heated to 80 °C. After 4 h, TLC analysis indicates complete consumption of starting materials. The reaction mixture is cooled to RT, filtered and concentrated. The residue is purified by FCC (hexanes/EtOAc 100:0 to 90:10 to 80:20 to 70:30 to 0:100) to afford the title compound. 1H NMR (CDCI3, 400 MHz) 5 0.69 (ddd, 2H), 0.86 (ddd, 2H). 1.24 (t, J = 7.0 Hz, 3H), 3.09 (ddd, 2H), 3.19 (s, 2H), 3.31 (ddd, 2H), 3.93 (s. 2H), 4.02 (s, 2H), 4.17 (q, J = 7.0 Hz, 2H), 7.08 (s, 1H), 7.21 (s, 1H), 7.33 (m, 6H), 7.42 (dd, J = 8.8 Hz, 1H), 7.71 (d, J = 8.8 Hz, 1H), 7.88 (d, J = 8.8 Hz, 1H); ES-MS: 415 [M+Hr. (3-Hydroxy-naphthaIen-1-yl)-acetic acid ethyl ester (1.67 g, 7.25 mmol) is dissolved in CH2CI2 (20 ml) under an atmosphere of argon. Pyridine (1.17 ml, 1.15 g, 14.50 mmol) is added, and the reaction mixture is cooled to 0 °C, whereupon trifluoromethanesulfonic anhydride (1.79 ml, 3.07 g, 10.88 mmol) is added dropwise. The reaction mixture is warmed to RT, and after 1 h a RT, TLC analysis indicates complete consumption of starting material. The reaction mixture is diluted with EtOAc and washed twice with H2O. The combined organic layers are dried over Na2S04, the solvent is removed, and the residue is purified by FCC (hexanes/EtOAc 100:0 to 97:3 to 95:5 to 93:7 to 90:10) to yield the title compound. 1H NMR (DMSO, 400 MHz) 5 1.19 (t, J = 7.2 Hz, 3H), 4.11 (q, J = 7.2 Hz, 2H), 4.38 (s, 2H), 7.60 (d, J = 3.0 Hz, 1H), 7.70 (m, 2H), 8.03 (m, 1H), 8.12 (m, 2H); ES-MS: 362 [M+H]*. e) (3-Hydroxy-naphthalen-1-yl)-acetic acid ethyl ester (3-Benzyloxy-naphthalen-1-yl)-acetic acid ethyl ester (2.43 g, 7.58 mmol) is dissolved in MeOH (50 ml). Palladium on charcoal (807 mg) is added, and the reaction mixture is stirred at RT under an atmosphere of hydrogen (1 atm) for 14 h. The reaction mixture is filtered. Concentration yields the pure title compound . 1H NMR (DMSO, 400 MHz) 5 1.18 (t, J = 7.2 Hz, 3H), 4.05 (s, 2H), 4.10 (q, J = 7.2 Hz, 2H), 7.04 (d, J = 1.8 Hz, 1H), 7.06 (d, J = 1.8, 1H), 7.30 (t, J = 7.8, 1H), 7.40 (t, J = 7.8, 1H), 7.70 (d, J = 7.8, 1H), 7.80 (d, J = 7.8 Hz, 1H); ES-MS: 230 [M+H]^ f) (3-Benzyloxy-naphthalen-1 -yl)-acetic acid ethyl ester 3-Benzyloxy-1-bromo-naphthalene (5.64 g, 18.01 mmol) is dissolved under an atmosphere of argon in dry DMF (100 ml). Tributylstannanyl-acetic acid ethyl ester (7.47 g, 19.81 mmol) is added, as well as [bis(tri-ortho-tolyl-phosphine)]palladium (II) dichloride (2.83 g, 3.60 mmol) and zinc(ll) bromide (5.27 g, 23.41 mmol). The reaction mixture is heated to 80 °C for 3 h. The reaction mixture is diluted with EtOAc and washed twice with diluted brine (back extracted). The combined organic layers are dried over Na2S04, the solvent is removed, and the residue is purified by FCC (hexane/EtOAc 100:0 to 97.5:2.5 to 95:5 to 90:10). The resulting oil, still containing tin residues, is stirred in a 1:1 mixture of EtOAc/IN NaOH (200 ml) for 1 h. The mixture is extracted twice with EtOAc. The combined organic layers are washed twice with H2O (back extracted), dried over Na2S04 and concentrated. The residue is purified by FCC (hexane/EtOAc 100:0 to 97:3 to 95:5 to 93:7 to 92:8 to 90:10) to yield the title compound. 1H NMR (DMSO, 400 MHz) 6 1.18 (t, J = 7.2 Hz, 3H), 4.10 (q, J = 7.2 Hz, 2H), 4.11 (s, 2H), 5.25 (s, 2H), 7.21 (d, J = 3.0 Hz, 1H), 7.41 (m, 8H), 7.53 (d, J = 6.6 Hz, 1H), 7.84 (dd, J = 7.2 Hz, 1H); ES-MS: 320 [M+H]^ g) 3-Benzyloxy-1 -bromo-naphthalene 4-Bromo-naphthaIen-2-ol (5.0 g, 22.41 mmol) is dissolved in dry DMF (50 ml) under an atmosphere of argon. Sodium hydride (986 mg of a 60% suspension in mineral oil, 592 mg, 24.65 mmol) is added and the mixture is stirred at 50 °C for 1 h. After re-cooling to RT, benzyl bromide (3.46 ml, 4.98 g, 29.14 mmol) and tetrabutyl ammoniumiodide (828 mg, 2.24 mmol) are added. After 16 h at RT, the reaction mixture is diluted with EtOAc. The solution is washed twice with semi-concentrated brine (back extracted). The combined organic layers are dried over Na2S04, the solvent is removed, and the residue is purified by FCC (hexane/EtOAc 100:0 to 95:5 to 90:10) to yield the title compound. 1H NMR (CDCI3, 400 MHz) 5 5.22 (s, 2H), 7.25 (d, J = 1.5 Hz, 1H), 7.45 (m, 7H), 7.64 (d, J = 2.4, 1H), 7.75 (d, J = 7.8, 1H), 8.17 (d, J = 7.8, 1H); ES-MS: 312 [M+H]*. By following procedure of example 20, the compounds of formula I wherein R is a residue of formula (b), as indicated in Table 2 may be obtained. TABLE 2 By following the procedure as disclosed above or in Example 56 in WO02/38561, but using the appropriate starting materials the compounds of formula I wherein R is a residue of formula (c), as indicated in Table 3 below may be obtained. By following the procedure as disclosed above or in Example 163 in WO 02/38561, but using the appropriate starting material the compounds of formula I wherein R is a residue of formula (d), as indicated in Table 4 below, may be obtained. 2-[3-(4,7-Diaza-spiro[2.5]oct-7-yl)-isoquinolin-1-yl]-acetamide (4.95 g, 16.70 mmol) and (7-Methyl-1H-indol-3-yl)-oxo-acetic acid methyl ester (5.44 g, 25.05 mmol) are azeotroped twice with dry THF. Dry THF (100 ml) is then added, and under an atmosphere of argon KOtBu (1.0 M in THF, 50 ml, 50 mmol) is added dropwise during 20 min. After an additional 90 min, TLC analysis indicates complete conversion of starting materials. The reaction mixture is diluted with H2O and extracted twice with EtOAc. The combined organic layers are washed twice with saturated aq NH4CI solution (back extracted), dried over Na2S04 and concentrated. Purification by FCC (CHgClz/MeOH 100:0 to 98:2 to 96:4 to 94:6 to 92:8 to 90:10) yields the title compound, which is converted to its acetate salt by concentration of a EtOH/AcOH solution. 1H NMR (DMSO, 400 MHz) 5 0.26 - 053 (br, 4 H), 1.89 (s, 3H, CH3COOH), 2.36 (s, 3H), 2.80 (br m, 2 H), 3.15 - 3.48 (br m, 2H), 6.14 (d, J = 8.2 HZ, 1H), 6.44 (dd, J = 8.2 / 7.4 Hz, 1H), 6.75 (d, J = 7.4 Hz, 1H), 7.00 (s, 1H), 7.02 (dd, J = 8.2 / 8.2 Hz, 1H), 7.40 (dd, J = 8.2 / 8.2 Hz, 1H), 7.59 (d, J = 8.2 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.63 (d, J = 8.2 Hz, 1H), 7.97 (d, J = 2.9 Hz, 1H), 11.04 - 11.21 (br, 1H), 11.86 (d, J = 2.9 a) 2-[3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-isoquinolin-1-yl]-acetamide (490 mg, 1.27 mmol) is dissolved in absolute MeOH (5 ml). Pd on charcoal (140 mg) is added, as well as ammonium formate (200 mg, 3.17 mmol). After refluxing for 1 h (T = 75 °C), an additional charge of ammonium formate (200 mg, 3.17 mmol) is added. 1 h later, TLC analysis indicates complete conversion of the starting material. After filtration and concentration, the residue is taken up in CH2CI2 and washed with water (pH 10 by addition of 2 N NaOH). The organic layer is dried over Na2S04 and the solvent is removed. Purification by FCC (EtOAc/AcOH/HgO 750:83:68 to 700:110:90 to 650:130:120 to 600:150:150) affords the title compound as its bis-acetate salt. 1H NMR (DMSO, 400 MHz) 10.46 - 0.52 (m, 4H), 2.88 (t, J = 5.5, 2H), 3.35 (s, 2H), 3.49 (t, J = 5.5, 2H), 3.94 (s, 2H), 6.77 (s, 1H), 7.00 (br s, 1H), 7.18-7.25 (m, 1H), 7.45 - 7.56 (m, 2H), 7.60 - 7.65 (m, 1H), 7.95 (d, J = 9.9,1H). ES-MS: 297 [M+nr. b)2-[3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-isoquinolin-1-yl]-acetamide [3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yl)-isoquinolin-1-yl]-acetic acid ethyl ester (700 mg, 1.68 mmol) is dissolved in dry DMF under an atnnosphere of argon. Formamide (224 ^1, 254 mg, 5.64 mmol) is added, and the reaction mixture is heated to 105 °C. At this temperature, NaOMe (312 \i\ of a 5.4 M solution in MeOH, 91 mg, 1.68 mmol) is added dropwise during 20 minutes. After 30 minutes at 105 °C, TLC analysis indicates complete conversion of starting material. Cooling the reaction mixture to RT is followed by addition of water and extraction with EtOAc. The organic layers are washed with H2O (twice), dried over Na2S04 and concentrated. The residue is purified by FCC (hexanes/EtOAc 1:1 to 1:3 to 0:100 to EtOAc/MeOH 98:2) to afford the title compound. 'H NMR (DMSO, 400 MHz) 5 0.68 - 0.70 (m, 2H), 0.82 - 0.88 (m, 2H), 3.08 (t, J = 4.4, 2H), 3.45 (s, 2H), 3.58 (t, J = 4.4, 2H), 3.96 (s, 2H), 4.27 (s, 2H). 5.3 - 5.5 (br, 1H), 6.55 - 6.7 (br, 1H), 6.72 (s, 1H), 7.26 - 7.36 (m, 6H), 7.51 (t, J = 8.8, 1H), 7.60 d, J = 9.9, 1H), 8.02 (d, J = 9.9, 1H); ES-MS: 387 [M+Hf. c) [3-(4-Benzyl-4,7-diaza-spiro[2.5]oct-7-yI)-isoquinolin-1-yl]-acetic acid ethyl ester (3-Chloro-isoquinolin-1-yl)-acetic acid ethyl ester (2.50 g, 10.01 mmol), 4-benzyl-4,7-diaza-spiro[2.5]octane (2.23 g, 11.01 mmol), NaOtBu (1.06 g, 11.01 mmol), BINAP (249 mg, 0.40 mmol) and Palladium(ll)-acetate (180 mg, 0.80 mmol) are mixed under argon. After addition of degassed, dry dioxane (36 ml), the suspension is heated to 85 °C. After 25 minutes at 85 °C, HPLC analysis indicates a conversion of 71%. The mixture is cooled to RT, diluted with EtOAc, and washed with H2O and sat. aq. NH4CI (back extracted). The combined organic layers are dried over Na2S04, the solvent is removed and the residue purified by FCC (hexane/EtOAc 100:0 to 96:4 to 93:7 to 90:10 to 85:15) to afford the title compound. 1H NMR (DMSO, 400 MHz) 6 0.58 - 0.61 (m, 2H), 0.70 - 0.73 (m, 2H), 1.18 (t, J = 8.8, 3H), 2.98 (t, J = 5.5, 2H), 3.39 (s, 2H), 3.49 (t, J = 5.5, 2H), 3.86 (s, 2H), 4.12 (s, 2H), 4.12 (q, J = 8.8, 2H), 5.59 (s, 1H), 7.14 - 7.19 (m, 6H), 7.39 (t, J = 8.8, 1H), 7.51 (d, J = 9.9, 1H), 7.78 (d, J = 9.9, 1H);ES-MS:417[M+Hr. d) (3-Chloro-isoquinolin-1-yl)-acetic acid ethyl ester 1,1,1,3,3,3-Hexamethyl-disilazane (27.4 ml, 20.37 g, 126.2 mmol) are dissolved in dry toluene (150 ml). After cooling to -78 °C, n-BuLi (79 ml of a 1.6 M solution in hexanes, 126.2 mmol) is slowly added during 20 minutes. The white suspension is stirred at -78 °C for 15 minutes and at RT for 15 minutes, after which time a clear bright yellow solution is obtained. This solution is canulated into a second two-necl After stirring at RT for 10 minutes, the clear darl The compounds of formula I in free form or in pliarmaceuticaliy acceptable salt form exhibit valuable pharmacological properties, e.g. inhibiting Protein Kinase C (PKC), e.g. PKC isoforms like a, P, 5, e, TI or 9 activity, inhibiting T-ce!l activation and proliferation, e.g. by inhibiting production by T-cells or cytokines, e,g, IL-2, by inhibiting the proliferative response of T-cells to cytokines, e.g. IL-2, e.g. as indicated in in vitro and in vivo tests and are therefore indicated for therapy. A. in vitro 1. Protein Kinase C assay The compounds of formula I are tested for their activity on different PKC isoforms according to a published method (D. Geiges et al. Biochem. Pharmacol. 1997;53:865-875) The assay is performed in a 96-well polypropylene microtiterplate (Costar 3794) that has been previously siliconized with Sigmacote (Sigma SL-2). The reaction mixture (50 ixl) contains 10 nl of the relevant PKC isozyme together with 25 ^1 of the test compound and 15 nl of a mix solution that contains 200 ^g/mj protamine sulfate, 10 mM Mg(N03)2,10 ^M ATP (Boehringer 519987) and 3750 Bq of ^P-ATP (Hartmann Analytic SFC301,110TBq/mmol) in 20 mM Tris-buffer pH 7.4 + 0.1% BSA. Incubation is performed for 15 min at 32°C in a microtiterplate shaking incubator (Biolabo Scientific Instruments). Reaction Is stopped by adding 10 |il of 0.5 M NaaEDTA, pH 7.4. 50 \i\ of mixture are pipetted onto a pre-wetted phosphocellulose paper (Whatmann 3698-915) under gentle pressure. Non-incorporated ATP is washed away with 100 nl bi-dist H2O. The paper is washed twice in 0.5% H3PO4 for 15 min followed by 5 min in EtOH. Thereafter the paper is dryed and placed in an omnifliter (Packard 6005219), and overlayed with 10 ^I/well of l\/licroscint-0 (Packard 6013611) before counting in a Topcount radioactivity counter (Pacl 2. Protein Kinase C 9 Assav Human recombinant PKCe is used under the assay conditions as described above. In this assay, compounds of formula I inhibit PKC 6 with an IC50 3. Protein Kinase Ca Assav Human recombinant PKCa was obtained from Oxford Biomedical Research and is used under the assay conditions as described under Section A.I above. In this assay, compounds of formula I inhibit PKC 9 with an IC50 ^ 1 ^M. Compound of Example 29 inhibits PKCa in this assay with an IC50 of 4.3 nM. 4. Protein Kinase CB1 Assav Human recombinant PKCpi was obtained from Oxford Biomedical Research and is used under the assay conditions as described under Section A.I above. In this assay, compounds of formula I inhibit PKC 9 with an IC50 5. Protein Kinase 05 Assav Human recombinant PKC5 was obtained from Oxford Biomedical Research and is used under the assay conditions as described under Section A.1 above. In this assay, compounds of formula I inhibit PKC 9 with an IC50 6. Protein Kinase Ce Assav Human recombinant PKCe was obtained from Oxford Biomedical Research and is used under the assay conditions as described under Section A.I above. In this assay, compounds of formula I inhibit PKC 9 with an IC50 7. Protein Kinase On Assav Human recombinant PKCTI was obtained from PanVera and is used under the assay conditions as described under Section A.I above. In this assay, compounds of formula I inhibit PKC 9 with an IC50 8. CD28 costimulation assay 22 The assay is performed with Jurl CD28- or PMA/ionomycin-stimulated cells without any test sample. Low controls are subtracted from all values. The Inhibition obtained in the presence of a test compound is calculated as percent inhibition of the high control. The concentration of test compounds resulting in 50% inhibition (IC50) is determined from the dose-response curves. In this assay, compounds of formula I inhibit anti-T cell receptor/anti-CD28 and PMA/ionomycin stimulated Jurkat cells with an IC50 9. Allogeneic Mixed Lymphocyte Reaction (MLR) The two-way MLR is performed according to standard procedures (J. Immunol. Methods, 1973, 2. 279 and Meo T. et al., Immunological Methods, New York, Academic Press, 1979, 227-39). Briefly, spleen cells from CBA and BALB/c mice (1.6x10^ cells from each strain per well in flat bottom tissue culture microtiter plates, 3.2 x 10^ in total) are incubated in RPMI medium containing 10% PCS, 100 U/ml penicillin, 100//g/ml streptomycin (Gibco BRL, Basel, Switzerland), 50/vM 2-mercaptoethanol (Fluka, Buchs, Switzerland) and serially diluted compounds. Seven three-fold dilution steps in duplicates per test compound are performed. After four days of incubation 1 pCi 1H-thymidine is added. Cells are harvested after an additional five-hour incubation period, and incorporated 1H-thymidine is determined according to standard procedures. Background values (low control) of the MLR are the proliferation of BALB/c cells alone. Low controls are subtracted from all values. High controls without any sample are taken as 100% proliferation. Percent inhibition by the samples is calculated, and the concentrations required for 50% inhibition (IC50 values) are determined. In this assay. Compound of Example 29 has an IC50 of 28 nM. Compounds of formula I also exhibit IC50 values in the nM range when tested in the human MLR. 10. Inhibition of GSK-3B The GSK-3P binding assay is performed in 50^1 reactions in 96 well polypropylene plate, each reaction containing 20mM magnesium chloride, 40 nM ATP, 2mM DTT, 88.5jiM biotinylated and phosphorylated CREB-peptide substrate (biotin-KRREILSRRPS(P04)YR-OH ; Q. M. Wang et al., J. Biol. Chem. 269, 14566-14574,1994), [y-^PJATP (1^Ci) and 2ii\ of the compound to be tested in DMSO(various concentrations). 15nl of GSK-3P (various concentrations) is added and the mixture is incubated at 30°C for 1 hour. The reaction is stopped by transferring 25 nl of the mixture to a phosphocellulose plate containing 130^1 of 1.85% phosphoric acid. The free radionucleotides in the membrane are washed off under vacuum with 1.85% phosphoric acid (5 times). After the last wash, the plate is transferred to an adaptor plate and 50^1 of scintillation cocktail (Microscint-20, Packard, cat. #20-133) is added to each well and the amount of radioactivity is counted in a top counter. Compounds of formula I are active in this assay. Compounds of formula I may also be tested in other standard GSK-3P binding assays using other substrates, e.g. as commercially available, B. In vivo Rat Heart transplantation The strain combination used: Male Lewis (RT^ haplotype) and BN (RT^ haplotype). The animals are anaesthetised using inhalational isofluorane. Following heparinisation of the donor rat through the abdominal inferior vena cava with simultaneous exsanguination via the aorta, the chest is opened and the heart rapidly cooled. The aorta is ligated and divided distal to the first branch and the brachiocephalic trunk is divided at the first bifurcation. The left pulmonary artery is ligated and divided and the right side divided but left open. All other vessels are dissected free, ligated and divided and the donor heart is removed into iced saline. The recipient is prepared by dissection and cross-clamping of the infra-renal abdominal aorta and vena cava. The graft is implanted with end-to-side anastomoses, using 10/0 monofilament suture, between the donor brachiocephalic trunk and the recipient aorta and the donor right pulmonary artery to the recipient vena cava. The clamps are removed, the graft tethered retroabdominally, the abdominal contents washed with warm saline and the animal is closed and allowed to recover under a heating lamp. Graft survival is monitored by daily palpation of the beating donor heart through the abdominal wall. Rejection is considered to be complete when heart beat stops. Increases of graft survival are obtained in animals treated with a compound of formula I administered orally at a daily dose of 1 to 30 mg/kg bid. Graft V. Host Model Spleen cells (2x10^) from Wistar/F rats are injected subcutaneously into the right hind footpad of (Wistar/F x Fischer 344)Fi hybrid rats. The left footpad is left untreated. The animals are treated with the test compounds on 4 consecutive days (0-3). The popliteal lymph nodes are removed on day 7, and the weight differences between two corresponding lymph nodes are determined. The results are expressed as the inhibition of lymph node enlargement (given in percent) comparing the lymph node weight differences in the experimental groups to the weight difference between the corresponding lymph nodes from a group of animals left untreated with a test compound. In this assay, an inhibition of 100% is obtained with compound of Example 29 when administered at a dose of 30 mg/kg/day bid. The compounds of formula I are, therefore, useful in the treatment and/or prevention of diseases or disorders mediated by T lymphocytes and/or PKC, e.g. acute or chronic rejection of organ or tissue alio- or xenografts, graft versus host diseases, atherosclerosis, vascular occlusion due to vascular injury such as angioplasty, restenosis, obesity, syndrome X, impaired glucose tolerance, polycystic ovary syndrome, hypertension, heart failure, chronic obstructive pulmonary disease, CNS diseases such as Alzheimer disease or amyotrophic lateral sclerosis, cancer, infectious diseases such as AIDS, septic shock or adult respiratory distress syndrome, ischemia/reperfusion injury e.g. myocardial infarction, stroke, gut ischemia, renal failure or hemorrhage shock, or traumatic shock, e.g. traumatic brain injury. The compounds of formula I are also useful in the treatment and/or prevention of T-cell mediated acute or chronic inflammatory diseases or disorders or autoimmune diseases e.g. rheumatoid arthritis, osteoarthritis, systemic lupus erythematosus, Hashimoto's thyroidis, multiple sclerosis, myasthenia gravis, diabetes type I or II and the disorders associated therewith, e.g. angiopathy, diabetic proliferative retinopathy, diabetic macular edema, nephropathy, neuropathy and dawn phenomenon, respiratory diseases such as asthma or inflammatory lung injury, inflammatory liver injury, inflammatory glomerular injury, cutaneous manifestations of immunologically-mediated disorders or illnesses, inflammatory and hyperproliferative skin diseases (such as psoriasis, atopic dermatitis, allergic contact dermatitis, irritant contact dermatitis and further eczematous dermatitises, seborrhoeic dermatitis), inflammatory eye diseases, e.g. Sjoegren's syndrome, keratoconjunctivitis or uveitis, inflammatory bowel disease, Crohn's disease or ulcerative colitis. For the above uses the required dosage will of course vary depending on the mode of administration, the particular condition to be treated and the effect desired. In general, satisfactory results are indicated to be obtained systemically at daily dosages of from about 0.1 to about 100 mg/kg body weight. An indicated daily dosage in the larger mammal, e.g. humans, is in the range from about 0.5 mg to about 2000 mg, conveniently administered, for example, in divided doses up to four times a day or in retard form. The compounds of formula I may be administered by any conventional route, in particular enterally, e.g. orally, e.g. in the form of tablets or capsules, or parenterally, e.g. in the form of injectable solutions or suspensions, topically, e.g. in the form of lotions, gels, ointments or creams, or in a nasal or a suppository form. Pharmaceutical compositions comprising a compound of formula I in free form or in pharmaceutically acceptable salt form in association with at least one pharmaceutical acceptable carrier or diluent may be manufactured In conventional manner by mixing with a pharmaceutically acceptable carrier or diluent. Unit dosage forms for oral administration contain, for example, from about 0.1 mg to about 500 mg of active substance. Topical administration is e.g. to the skin. A further form of topical administration is to the eye. The compounds of formula I may be administered in free form or in pharmaceutically acceptable salt form e.g. as indicated above. Such salts may be prepared in conventional manner and exhibit the same order of activity as the free compounds. In accordance with the foregoing the present invention further provides: 1.1 A method for preventing or treating disorders or diseases mediated by T lymphocytes and/or PKC or GSK-3P, e.g. such as indicated above, in a subject in need of such treatment, which method comprises administering to said subject an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof; 1.2 A method for preventing or treating acute or chronic transplant rejection or T-cell mediated inflammatory or autoimmune diseases, e.g. as indicated above, in a subject in need of such treatment, which method comprises administering to said subject an effective amount of a compound of formula I or a pharmaceutically acceptable salt thereof; 2. A compound of formula I, in free form or in a pharmaceutically acceptable salt form for use as a pharmaceutical, e.g. in any of the methods as indicated under 1.1 and 1.2 above. 3. A pharmaceutical composition, e.g. for use in any of the methods as in 1.1 and 1.2 above comprising a compound of formula I in free form or pharmaceutically acceptable salt form in association with a pharmaceutically acceptable diluent or carrier therefor. 4. A compound of formula I or a pharmaceutically acceptable salt thereof for use in the preparation of a pharmaceutical composition for use in any of the method as in 1.1 and 1.2 above. Compounds of formula I may be administered as the sole active ingredient or together with other drugs in immunomodulating regimens or other anti-Inflammatory agents e.g. for the treatment or prevention of alio- or xenograft acute or chronic rejection or inflammatory or autoimmune disorders. For example, they may be used in combination with oyolosporines, or ascomycines or their immunosuppressive analogs or derivatives, e.g. cyclosporin A, ISA Tx247, FK-506, ABT-281, ASM 981; an mTOR inhibitor, e.g. rapamycin, 40-O-(2-hydroxy)ethyl-rapamycin, CCI779, ABT578 or a rapalog, e.g. AP23573 etc.; corticosteroids; cyclophosphamide; azathioprene; methotrexate; an EDG receptor agonist having accelerating lymphocyte homing properties, e.g. FTY 720 or an analogue thereof; leflunomide or analogs thereof; mizoribine; mycophenolic acid; mycophenolate mofetil; 15-deoxyspergualine or analogs thereof; immunosuppressive monoclonal antibodies, e.g., monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3, CD4, CD 11a/CD18, CD7, CD25, CD 27, B7, CD40, CD45, CD58, CD 137, ICOS, CD150 (SLAM), OX40, 4-1 SB or their ligands, e.g. GDI 54; or other immunomodulatory compounds, e.g. a recombinant binding molecule having at least a portion of the extracellular domain of CTLA4 or a mutant thereof, e.g. an at least extracellular portion of CTLA4 or a mutant thereof joined to a non-CTLA4 protein sequence, e.g. CTLA4lg (for ex. designated ATCC 68629) or a mutant thereof, e.g. LEA29Y, or other adhesion molecule inhibitors, e.g. mAbs or low molecular weight inhibitors including LFA-1 antagonists, Selectin antagonists and VLA-4 antagonists. Compounds of formula I may also be administered together with an antiproliferative drug, e.g. a chemotherapeutic drug, e.g. as used in cancer treatment, including but not limited to aromatase inhibitors, antiestrogens, topoisomerase I inhibitors, topoisomerase II inhibitors, microtubule active agents, alkylating agents, histone deacetylase inhibitors, farnesyl transferase inhibitors, COX-2 inhibitors, MMP inhibitors, mTOR inhibitors, antineoplastic antimetabolites, platin compounds, compounds decreasing the protein kinase activity and further anti-angiogenic compounds, gonadorelin agonists, anti-androgens, bengamides, bisphosphonates, antiproliferative antibodies and temozolomide, or with an anti-diabetic drug, an insulin secretagogue or insulin secretion enhancer, e.g. a sulphonyl urea, e.g. tolbutamide, chlorpropamide, tolazamide, acetohexamide, 4-chloro-N-[(1-pyroIidinylamino)carbonyl]-benzensulfonamide (glycopyramide), glibenclamide (glyburide), gliclazide, 1-butyl-3-metanilylurea, carbutamide, glibonuride, glipizide, gliquidone, glisoxepid, glybuthiazole, glibuzole, glyhexamide, glymidine, glypinamide, phenbutamide or tolylcyclamide, an oral insulinotropic agent derivative, e.g. a short acting insulin enhancer, e.g. meglitinide, repaglinide, a phenyl acetic acid derivative, e.g.nateglinide, a DPP IV inhibitor, e.g. 1 -{2-[(5-cyanopyridin-2-yl)amino]ethylamino}acetyl-(2S)-cyano-pyrrolidine dihydrochloride, LAF237, GLP-1 or a GLP-1 agonist analog, or an insulin sensitizer e.g. a peroxisome proliferator activated receptor y agonist (PPARy), e.g. a glitazone, a non- glitazone type such as a N-(2-benzoylphenyl)-L-tyrosine analogue, e.g. GI-262570, or an oxolidinedione, e.g. JTT501, a dual PPARy/PPARa agonist, e.g. DRF-554158, NC-2100 or NN-622, a retinoid X receptor agonist or a rexinoid, e.g. 2-[1 -(3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyl)-cyclopropyl]-pyridine-5-carboxylic acid, 4-[(3,5,5,8,8-pentamethyl-5,6,7,8-tetrahydro-2-naphthyl)-2-carbonyl]-benzoic acid, 9-cis retlnolc acid or an analog, derivative or a pharmaceutically acceptable salt thereof, in diabetes therapy, In accordance with the foregoing the present invention provides in a yet further aspect: 5. A method as defined above comprising co-administration, e.g. concomitantly or in sequence, of a therapeutically effective amount of an inhibitor of GSK-sp, PKC or of T-cell activation and proliferation, e.g. a compound of formula I in free form or in pharmaceutically acceptable salt form, and a second drug substance, said second drug substance being an immunosuppressant, immunomodulatory, anti-inflammatory, antiproliferative or anti-diabetic drug, e.g. as indicated above. 6. A therapeutic combination, e.g. a kit, comprising a) an inhibitor of GSK-SP, PKC or of T-cell activation and proliferation, e.g. a compound of formula I in free form or in pharmaceutically acceptable salt form, and b) at least one second agent selected from an immunosuppressant, immunomodulatory, anti-inflammatory, antiproliferative and anti-diabetic drug. Component a) and component b) may be used concomitantly or in sequence. The kit may comprise instructions for its administration. Where an inhibitor of GSK-3P, PKC or of T-cell activation and proliferation, e.g. a compound of formula I, is administered in conjunction with other immunosuppressive/immunomo¬dulatory, anti-inflammatory, antiproliferative or anti-diabetic therapy, e.g. for preventing or treating acute or chronic graft rejection or inflammatory or autoimmune disorders as hereinabove specified, dosages of the co-administered immunosuppressant, immunomodulatory, anti-inflammatory, antiproliferative or anti-diabetic compound will of course vary depending on the type of co-drug employed, e.g. whether it is a steroid or a cyclosporine, on the specific drug employed, on the condition being treated and so forth. Compounds of formula I have an interesting pharmacokinetic profile and interesting in vitro and in vivo activities. We claim: 1. A compound of formula I Rb is H; halogen; C1-4alkoxy; or C1-6alkyl, and either R1 Is piperazin-1-yl optionally substituted by CH3 in position 3 or 4; or 4,7-dlaza-spiro [2.5] oct-7-yl; R2 Is CI; Br; CF3; or CH3; and Ra Is H; CHa; or CF3; R2 being other than CH3 or CI when R3 Is H, R4 is H or CH3, Rb is H and Ri is 4-methyl-1-plperazinyl; R4 is plperazin-1-yl substituted In positions 3 and/or 4 by CH3; or 4,7-diaza-spiro [2.5] oct-7-yl; R2 being other than H or CH3 when R4 Is 4-methyi-1-piperazlnyi; or spiro [2.5] oct-7-yl; R15 is halogen; CF3; or CH3; R15 being other than CH3 when R18 is CH3 or H, R2 is H or CH3, Rb is H and R14 is 4-methyl-1-piperazinyl; and R16 is H; CH3; CH2-CH3; or CF3; R16 being other than H when Rie is Ci, Ra is H or CH3, Rb is H and R14 is 4-methyl-1-piperazinyl; or Rg is 4,7-diaza-splro [2.5] oct-7-yl; or 1-piperazinyl substituted in position 3 by ethyl and optionally In position 4 by methyl; or 1-piperazlnyl substituted in positions 3 and 4 by methyl; or a salt thereof. 2. The compound as claimed in claim 1 which is 3-[3-(4,7-dlaza-spiro[2.5]oct-7-yl)-lsoquinolin-1 -yl]-4-(7-methyl-1 H-indol-3-yl)-pyrrole-2,5-dlone or a salt thereof. 3. The compound as claimed in claim 2 which Is the acetate salt. 4. The compound as claimed in claim 1 which is selected from 3-[4-chloro-5-methyi-2-(4- methyl-piperazin-1-yl)-pyrimldln-4-yl]-4-(1H-lndol-3-yl)-pyrrole-2,5-dlone and 3-(1H-indol-3- yl)-4-[2-(4-methyl-piperazin-1 -yl)-4-trifluoromethyl-pyrimidin-4-yl]pyrrole-2,5-dione or a salt thereof. b) converting the resulting compound of formula I obtained in free form to a salt form or vice versa, as appropriate. 6. A pharmaceutical composition comprising a compound of formula I as claimed in any one of claims 1 to 4 in free form or a pharmaceutically acceptable salt thereof in association with a pharmaceutically acceptable diluent or carrier therefor. 7. The composition as claimed in claim 6 wherein the compound of formula I is 3-[3- (4,7-diaza-spiro[2.5]oct-7-yl)-isoquinolin-1 -yl]-4-(7-methyl-1 H-indol-3-yl)-pyrrole-2,5-dione or a salt thereof. 8. The composition as claimed in claim 6 or 7 for topical administration route to the skin or to the eye. 9. A pharmaceutical combination comprising a) a compound of formula I as claimed in any one of claims 1 to 4 in free form or in pharmaceutically acceptable salt form, and b) at least one second agent selected from an immunosuppressant .immunomodulatory, anti¬inflammatory, antiproliferative, chemotherapeutic and anti- diabetic drug. 10. The combination as claimed in claim 9 wherein the second agent is selected from cyclosporine or immunosuppressive analog or derivative thereof, ascomycine or immunosuppressive analog or derivative thereof, mTOR inhibitor, corticosteroid, cyclophosphamide , azathioprene, methotrexate, EDG receptor agonist having accelerating lymphocyte homing properties, leflunomide or analog thereof, mizoribine, mycophenolic acid, mycophenolate mofetil, 15-deoxysperguaiine or analog thereof, immunosuppressive monoclonal antibody, immunomodulatory compound, and adhesion molecule inhibitor. 11. The combination as claimed in claim 9 wherein the second agent is selected from cyclosporine A, ISATx247, FK-506, ABT-281, ASM 981, rapamycin, 40- 0-(2-hydroxy)ethyl-rapamycin, FTY 720, CTLA4lg, LFA-1 antagonist, selectin antagonist, VLA-4 antagonist, monoclonal antibody to MHC, CD2, CD3, CD4, CD 11a/CD18, CD7, CD26, CD 27, B7, CD40, CD45, CD58, CD 137, ICOS, CD150, OX40, 4-1BB and ligand thereof.

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2172-chenp-2004 abstract duplicate.pdf

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2172-chenp-2004 abstract.pdf

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2172-chenp-2004 correspondence-others.pdf

2172-chenp-2004 description (complete) duplicate.pdf

2172-chenp-2004 description (complete).pdf

2172-chenp-2004 form-1.pdf

2172-chenp-2004 form-3.pdf

2172-chenp-2004 form-5.pdf

2172-chenp-2004 pct search report.pdf

2172-chenp-2004-mas-97 form-19.pdf

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2172-chenp-2004-mas-97 others.pdf

2172-chenp-2004-mas-97 pct.pdf

2172-chenp.rtf