Method for the preparation of (4S)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1-6-naphthyridine-3-carboxamide and the purification thereof for use as an active pharmaceutical ingredient

Abstract

The present invention related to a novel and improved process for preparing (4S)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide of the formula (I) ##STR00001##

Claims

.[.1. A process for preparing a compound of the formula (I) ##STR00069## comprising separating a racemic compound of the formula (XIII) into its enantiomers, wherein the compound of the formula (XIII) ##STR00070## is prepared by reacting a compound of the formula (XVIII) ##STR00071## with an orthoester (XX) ##STR00072## where R may be H or methyl, wherein the compound of the formula (XVIII) ##STR00073## is prepared by reacting compounds of the formula (XVI a,b) ##STR00074## with a compound of the formula (IX) ##STR00075## and wherein the compound of the formula (XVI a,b) is prepared by reacting a compound of the formula (VI) ##STR00076## with a compound of the formula (XVII) ##STR00077## .].

.[.2. A process for preparing a compound of the formula (I) ##STR00078## comprising separating a racemic compound of the formula (XIII) into its enantiomers, where the compound of the formula (XIII) ##STR00079## is prepared by reacting a compound of the formula (XVIII) ##STR00080## with an orthoester (XX) ##STR00081## where R may be H or methyl, and wherein the compound of the formula (XVIII) ##STR00082## is prepared by reacting compounds of the formula (XVI a,b) ##STR00083## with a compound of the formula (IX) ##STR00084## .].

.[.3. A process for preparing a compound of the formula (I) ##STR00085## comprising separating a racemic compound of the formula (XIII) into its enantiomers, where the compound of the formula (XIII) ##STR00086## is prepared by reacting a compound of the formula (XVIII) ##STR00087## with an orthoester (XX) ##STR00088## where R may be H or methyl..].

.[.4. A process for preparing a compound of the formula (I) ##STR00089## comprising using the compound of the formula (XVIII) ##STR00090## .].

.[.5. A process for preparing a compound of the formula (I) ##STR00091## comprising using the compounds of the formula (XVI a,b) ##STR00092## .].

.[.6. A process for preparing a compound of the formula (I) ##STR00093## comprising using a compound of the formula (XVIII) ##STR00094## and compounds of the formula (XVI a,b) ##STR00095## .].

.[.7. A compound of the formula (XVIII) ##STR00096## and the salts, solvates and solvates of the salts thereof..].

.[.8. A compound of the formula (XVI a,b) as an E/Z mixture ##STR00097## and the salts, solvates and solvates of the salts thereof..].

.[.9. A process for preparing a compound of the formula (XVIII) ##STR00098## comprising reacting compounds of the formula (XVI a,b) ##STR00099## with a compound of the formula (IX) ##STR00100## .].

.[.10. A process for preparing compounds of the formula (XVI a,b) ##STR00101## comprising reacting a compound of the formula (VI) ##STR00102## with a compound of the formula (XVII) ##STR00103## .].

.[.11. A process for preparing a compound of the formula (XIII) ##STR00104## comprising reacting a compound of the formula (XVIII) ##STR00105## with an orthoester (XX) ##STR00106## where R may be H or methyl..].

.[.12. A process for preparing a compound of the formula (XIII) ##STR00107## comprising reacting a compound of the formula (XVIII) ##STR00108## with an orthoester (XX) ##STR00109## where R may be H or methyl, and wherein the compound of the formula (XVIII) is prepared by reacting compounds of the formula (XVI a,b) ##STR00110## with a compound of the formula (IX) ##STR00111## .].

.[.13. A process for preparing a compound of the formula (I) in crystalline polymorph I ##STR00112## comprising preparing a compound of the formula (I) ##STR00113## according to claim 1 and then stirring the compound of the formula (I), present in one or more polymorphs or as a solvate in an inert solvent, at a temperature of 20 C.-120 C. and isolating the compound of the formula (I) in crystalline polymorph I..].

.Iadd.14. A compound of the formula (I) in crystalline form of polymorph I ##STR00114## wherein the x-ray diffractogram of the compound exhibits peak maxima of the 2 theta angle at 8.5, 14.1, and 19.0. .Iaddend.

.Iadd.15. The compound of formula (I) in crystalline form of polymorph I according to claim 14, wherein the x-ray diffractogram of the compound further exhibits peak maxima of the 2 theta angle at 17.2, 20.5, 25.6, and 26.5. .Iaddend.

.Iadd.16. The compound of the formula (I) in crystalline form of polymorph I according to claim 14, wherein the IR spectrum of the compound exhibits band maxima at 3475, 2230, 1681, 1658, 1606, 1572, 1485, 1255, 1136 and 1031 cm.sup.1. .Iaddend.

.Iadd.17. The compound of the formula (I) in crystalline form of polymorph I according to claim 14, wherein the Raman spectrum of the compound exhibits band maxima at 3074, 2920, 2231, 1601, 1577, 1443, 1327, 1267, 827 and 155 cm.sup.1. .Iaddend.

.Iadd.18. The compound of the formula (I) in crystalline form of polymorph I according to claim 14, wherein the compound has a melting point of 252 C. .Iaddend.

.Iadd.19. A compound of the formula (I) in crystalline form of polymorph I ##STR00115## wherein the IR spectrum of the compound exhibits band maxima at 3475, 2230, 1681, 1658, 1606, 1572, 1485, 1255, 1136 and 1031 cm.sup.1. .Iaddend.

.Iadd.20. The compound of the formula (I) in crystalline form of polymorph I according to claim 19, wherein the x-ray diffractogram of the compound exhibits peak maxima of the 2 theta angle at 8.5, 14.1, 17.2, 19.0, 20.5, 25.6, and 26.5. .Iaddend.

.Iadd.21. The compound of the formula (I) in crystalline form of polymorph I according to claim 19, wherein the Raman spectrum of the compound exhibits band maxima at 3074, 2920, 2231, 1601, 1577, 1443, 1327, 1267, 827 and 155 cm.sup.1. .Iaddend.

.Iadd.22. The compound of the formula (I) in crystalline form of polymorph I according to claim 19, wherein the compound has a melting point of 252 C. .Iaddend.

.Iadd.23. A compound of the formula (I) in crystalline form of polymorph I ##STR00116## wherein the Raman spectrum of the compound exhibits band maxima at 3074, 2920, 2231, 1601, 1577, 1443, 1327, 1267, 827 and 155 cm.sup.1. .Iaddend.

.Iadd.24. The compound of the formula (I) in crystalline form of polymorph I according to claim 23, wherein the x-ray diffractogram of the compound exhibits peak maxima of the 2 theta angle at 8.5, 14.1, 17.2, 19.0, 20.5, 25.6, and 26.5. .Iaddend.

.Iadd.25. The compound of the formula (I) in crystalline form of polymorph I according to claim 23, wherein the IR spectrum of the compound exhibits band maxima at 3475, 2230, 1681, 1658, 1606, 1572, 1485, 1255, 1136 and 1031 cm.sup.1. .Iaddend.

.Iadd.26. The compound of the formula (I) in crystalline form of polymorph I according to claim 23, wherein the compound has a melting point of 252 C. .Iaddend.

.Iadd.27. A compound of the formula (I) in crystalline form of polymorph I ##STR00117## prepared by a process comprising obtaining the compound of formula XIII ##STR00118## separating the compound of formula XIII by chiral separation to obtain the compound of formula I, dissolving the compound of formula I in an inert solvent, stirring at a temperature of 20 C.-120 C. and isolating the compound of the formula (I) in crystalline form of polymorph I. .Iaddend.

.Iadd.28. The compound of the formula (I) in crystalline form of polymorph I according to claim 27, where the compound of the formula (XIII) ##STR00119## is prepared by reacting a compound of the formula (XVIII) ##STR00120## with an orthoester (XX) ##STR00121## where R may be H or methyl. .Iaddend.

.Iadd.29. A compound of the formula (I) in crystalline form of polymorph I ##STR00122## wherein the compound is isolated in crystalline form by a process comprising: filtering a solution of the compound of the formula (XIII) ##STR00123## in a mixture of acetonitrile and methanol or a mixture of acetonitrile and ethanol; performing a solvent exchange with ethanol; concentrating the solution until the compound crystallizes; cooling the solution to 0 C.; and isolating the compound in crystalline form. .Iaddend.

.Iadd.30. A compound of the formula (I) in crystalline form of polymorph I ##STR00124## wherein the compound is crystallized from a solvent selected from the group consisting of methanol, ethanol, tetrahydrofuran, acetonitrile, and mixtures thereof with water. .Iaddend.

Description

EXPERIMENTAL

Abbreviations and Acronyms

(1) MS: mass from mass spectrometry

(2) HPLC: high-performance liquid chromatography

(3) DMF: dimethylformamide

(4) Red-Al solution in toluene: sodium bis(2-methoxyethoxy) aluminium dihydride in toluene

(5) THF: tetrahydrofuran

(6) Aqu. HCl: aqueous hydrochloric acid

(7) DMAP: 4-(dimethylamino)pyridine

EXAMPLES

Example 1

Methyl 4-bromo-2-methoxybenzoate (XV)

(8) ##STR00061##

(9) 3.06 kg (22.12 mol) of potassium carbonate were initially charged in 3.6 l of acetone and heated to reflux. To this suspension were added 1.2 kg of 4-bromo-2-hydroxybenzoic acid (5.53 mol), suspended in 7.8 l of acetone, and were rinsed in with 0.6 l of acetone. The suspension was heated under reflux for 1 hour (vigorous evolution of gas!). 2.65 kg (21.01 mol) of dimethyl sulphate were then added over 4 hours while boiling. The mixture was subsequently stirred under reflux for 2.5 hours. The solvent was largely distilled off (to the point of stirrability) and 12 l of toluene were added and the residual acetone was then distilled off at 110 C. About 3 l of distillate were distilled off, this being supplemented by addition of a further 3 l of toluene to the mixture. The mixture was allowed to cool to 20 C. and 10.8 l of water were added and vigorously stirred in. The organic phase was separated off and the aqueous phase was extracted once more with 6.1 l of toluene. The combined organic phases were washed with 3 l of saturated sodium chloride solution and the toluene phase is concentrated to about 4 l. Determination of the content by evaporation of a portion resulted in a converted yield of 1.306 kg (96.4% of theory). The solution was used directly in the subsequent stage.

(10) HPLC method A: RT about 11.9 min.

(11) MS (Elpos): m/z=245 [M+H].sup.+

(12) .sup.1H NMR (400 MHz, CD.sub.2Cl.sub.2): =3.84 (s, 3H), 3.90 (s, 3H), 7.12-7.20 (m, 2H), 7.62 (d, 1H).

Example 2

4-Bromo-2-methoxybenzaldehyde (XIX)

(13) ##STR00062##

(14) 1.936 kg (6.22 mol) of a 65% Red-Al solution in toluene were charged with 1.25 l of toluene at 5 C. To this solution was added 0.66 kg (6.59 mol) of 1-methylpiperazine, which was rinsed in with 150 ml of toluene, keeping the temperature between 7 and 5 C. The mixture was then left to stir at 0 C. for 30 minutes. This solution was then added to a solution of 1.261 kg (5.147 mol) of methyl 4-bromo-2-methoxybenzoate (XV), dissolved in 4 l of toluene, keeping the temperature at 8 to 0 C. After rinsing in twice with 0.7 l of toluene, the mixture was then stirred at 0 C. for 1.5 hours. For the work-up, the solution was added to cold aqueous sulphuric acid at 0 C. (12.5 l of water+1.4 kg of conc. sulphuric acid). The temperature should increase at maximum to 10 C. (slow addition). The pH was adjusted to pH 1, if necessary, by addition of further sulphuric acid. The organic phase was separated off and the aqueous phase was extracted with 7.6 l of toluene. The combined organic phases were washed with 5.1 l of water and then substantially concentrated and the residue taken up in 10 l of DMF. The solution was again concentrated to a volume of about 5 l. Determination of the content by evaporation of a portion resulted in a converted yield of 1.041 kg (94.1% of theory). The solution was used directly in the subsequent stage.

(15) HPLC method A: RT about 12.1 min.

(16) MS (EIpos): m/z=162 [M+H].sup.+

(17) .sup.1H NMR (CDCl.sub.3, 400 MHz): =3.93 (3H, s), 7.17 (2H, m), 7.68 (1H, d), 10.40 (1H, s)

Example 3

4-Formyl-3-methoxybenzonitrile (VI)

(18) ##STR00063##

(19) 719 g (3.34 mol) of 4-bromo-2-methoxybenzaldehyde (XVI) as a solution in 4.5 l of DMF were charged with 313 g (0.74 mol) of potassium hexacyanoferrate (K.sub.4[Fe(CN).sub.6]) and 354 g (3.34 mol) of sodium carbonate and a further 1.2 l of DMF and 3.8 g (0.017 mol) of palladium acetate were added. The mixture was stirred at 120 C. for 3 hours. The mixture was left to cool to 20 C. and 5.7 l of water were added to the mixture. The mixture was extracted with 17 l of ethyl acetate and the aqueous phase was washed once more with 17 l of ethyl acetate. The organic phases were combined and substantially concentrated, taken up in 5 l of isopropanol and concentrated to about 2 l. The mixture was heated to boiling and 2 l of water were added dropwise. The mixture was allowed to cool to 50 C. and another 2 l of water were added. The mixture was cooled to 3 C. and stirred at this temperature for one hour. The product was filtered off and washed with water (21.2 l). The product was dried at 40 C. under vacuum.

(20) Yield: 469 g (87% of theory) of a beige solid.

(21) HPLC method A: RT about 8.3 min.

(22) MS (EIpos): m/z=162 [M+H]+

(23) 1H NMR (300 MHz, DMSO-d6): =3.98 (s, 3H), 7.53 (d, 1H), 7.80 (s, 1H), 7.81 (d, 1H), 10.37 (s, 1H).

Example 4

(2E/2Z)-2-(4-Cyano-2-methoxybenzylidene)-3-oxobutanamide (XVI a,b)

(24) ##STR00064##

(25) 1000 g (6204.95 mmol) of 4-formyl-3-methoxybenzonitrile (VI), 721.5 g (7135.7 mmol) of 3-oxobutanamide (XVII), 53 g (620 mmol) of piperidine and 37.3 g (620 mmol) of glacial acetic acid were heated under reflux in 15 l of dichloromethane for 4 hours on a water separator. Subsequently, about 10 l of dichloromethane were distilled off and the mixture was left to cool to room temperature. The mixture was cooled to 0 C. and left to stir for 4 hours, and the product was filtered off and washed twice with 1000 ml each time of cold dichloromethane. The product was dried at 40 C. under vacuum under entraining gas.

(26) Yield: 1439.8 g (95.0% of theory) of a yellow solid.

(27) HPLC method A: RT about 3.55 min.

(28) MS (EIpos): m/z=245 [M+H].sup.+

(29) .sup.1H NMR (500 MHz, DMSO-d.sub.6): =2.35 (s, 3H), 3.30 (s, 2H), 3.90 (s, 3H), 7.45 (d, 1H), 7.7 (m, 3H), 7.75 (d, 1H), 8.85 (d, 1H)

Example 5

4-(4-Cyano-2-methoxyphenyl)-2,8-dimethyl-5-oxo-1,4,5,6-tetrahydro-1,6-naphthyridine-3-carboxamide (XVIII)

(30) ##STR00065##

(31) 2.128 kg (8.712 mol) of (2E/2Z)-2-(4-cyano-2-methoxybenzylidene)-3-oxobutanamide (XVI a,b) were taken up with 29 l of 2-butanol, 1.277 kg (7.92 mol) of 4-amino-5-methylpyridone were added, and then the mixture was heated in a closed stirred tank under elevated pressure at internal temperature 120 C. for 12 h. The mixture was then cooled to 0 C. by means of a gradient over a period of 5 h and then stirred at 0 C. for 3 hours. The product was then filtered off and washed with 2.1 l of cold isopropanol. The product was dried at 60 C. under vacuum.

(32) Yield: 2.081 kg (75% of theory based on 4-amino-5-methylpyridone, since this component is used substoichiometrically) of a pale yellow solid.

(33) HPLC method A: RT about 3.64 min.

(34) MS (EIpos): m/z=351 [M+H].sup.+

(35) .sup.1H H NMR (500 MHz, DMSO-d.sub.6): =2.00 (s, 3H), 2.10 (s, 3H), 3.78 (s, 3H), 5.22 (s, 1H), 6.65 (s(broad), 1H), 6.85 (s(broad), 1H), 6.91 (s, 1H), 7.11 (d, 1H), 7.28 (d, 1H), 7.35 (s, 1H), 7.52 (s, 1H), 10.61 (s, 1H)

Example 5

4-(4-Cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (XIII)

(36) ##STR00066##

(37) 1.857 kg (5.3 mol) of 4-(4-cyano-2-methoxyphenyl)-2,8-dimethyl-5-oxo-1,4,5,6-tetrahydro-1,6-naphthyridine-3-carboxamide (XVIII) and 4.70 kg (29 mol) of triethyl orthoacetate were dissolved in 12.15 l of dimethylacetamide, and 157.5 g of concentrated sulphuric acid were added. The mixture was heated at 115 C. for 1.5 hours and then cooled to 50 C. At 50 C., 12.15 l of water were added dropwise over 30 minutes. After completion of the addition, the mixture was seeded with 10 g of the title compound (XI) and a further 12.15 l of water were added dropwise over 30 minutes at 50 C. The mixture was cooled to 0 C. (gradient, 2 hours) and then stirred at 0 C. for two hours. The product was filtered off, washed twice with 7.7 l each time of water and dried at 50 C. under vacuum.

(38) Yield: 1.845 kg (92.0% of theory) of a pale yellow solid.

(39) HPLC method B: RT about 10.2 min.

(40) MS (EIpos): m/z=433 [M+H].sup.+

(41) .sup.1H NMR (300 MHz, DMSO-d.sub.6): =1.11 (t, 3H), 2.16 (s, 3H), 2.42 (s, 3H), 2.78 (m, 2H), 3.77 (s, 3H), 4.01-4.13 (m, 4H), 5.37 (s, 1H), 7.25 (d, 1H), 7.28-7.33 (m, 2H), 7.60 (s, 1H), 8.35 (s, 1H).

Example 6

(4S)-4-(4-Cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (I) as a Solution in 40:60 acetonitrile/methanol

(42) ##STR00067##

(43) Enantiomer Separation on an SMB System

(44) The feed solution was a solution corresponding to a concentration consisting of 50 g of racemic 4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (XIII), dissolved in 1 liter of a mixture of 60:40 methanol/acetonitrile.

(45) The solution was chromatographed by means of an SMB system on a stationary phase: Chiralpak AS-V, 20 m. The pressure was 30 bar and a mixture of methanol/acetonitrile 60:40 was used as eluent.

(46) 9.00 kg of 4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (XIII) were dissolved in 180 l of a mixture consisting of methanol/acetonitrile 60:40 and chromatographed by means of SMB. After concentrating the product-containing fractions, 69.68 liters of a 6.2% solution (corresponding to 4.32 kg of (4S)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (I)) were obtained as a solution in acetonitrile/methanol 40:60.

(47) Yield: 4.32 kg (48% of theory), as a colourless fraction dissolved in 69.68 liters of acetonitrile/methanol 40:60.

(48) Enantiomeric purity: >98.5% e.e. (HPLC, Method D)

(49) A sample is concentrated under vacuum and gives: MS (EIpos): m/z=379 [M+H].sup.+

(50) .sup.1H NMR (300 MHz, DMSO-d.sub.6): =1.05 (t, 3H), 2.12 (s, 3H), 2.18 (s, 3H), 3.82 (s, 3H), 3.99-4.07 (m, 2H), 5.37 (s, 1H), 6.60-6.84 (m, 2H), 7.14 (d, 1H), 7.28 (dd, 1H), 7.37 (d, 1H), 7.55 (s, 1H), 7.69 (s, 1H).

Example 7

(4S)-4-(4-Cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (I)

(51) ##STR00068##

(52) Crystallization and Polymorph Adjustment

(53) 64.52 liters of a 6.2% solution from Example 6 in a mixture of acetonitrile/methanol 40:60 (corresponding to 4.00 kg of compound 1) were filtered through a filter cartridge (1.2 um) and subsequently sufficiently concentrated at 250 mbar such that the solution was still stirrable. 48 l of ethanol, denatured with toluene, was added and distilled again at 250 mbar up to the limit of stirrability (redistillation in ethanol). A further 48 l of ethanol, denatured with toluene, were added and then distilled off at atmospheric pressure down to a total volume of about 14 l (jacket temperature 98 C.). The mixture was cooled via a gradient (4 hours) to 0 C., stirred at 0 C. for 2 hours and the product filtered off. The product was washed twice with 4 l of cold ethanol each time and then dried at 50 C. under vacuum.

(54) Yield: 3.64 kg (91% of theory) of a colourless crystalline powder.

(55) Enantiomeric purity: >>99% e.e. (HPLC Method D); retention times/RRT: (4S)-4-(4-cyano-2-me thoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (I) about 11 min. RRT: 1.00; (4R)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (I) about 9 min. RRT: 0.82

(56) Purity: >99.8% (HPLC Method B), RT: about 6.7 min.

(57) Content: 99.9% (relative to external standard)

(58) specific rotation (chloroform, 589 nm, 19.7 C., c=0.38600 g/100 ml): 148.8.

(59) MS (EIpos): m/z=379 [M+H].sup.+

(60) .sup.1H NMR (300 MHz, DMSO-d.sub.6): =1.05 (t, 3H), 2.12 (s, 3H), 2.18 (s, 3H), 3.82 (s, 3H), 3.99-4.07 (m, 2H), 5.37 (s, 1H), 6.60-6.84 (m, 2H), 7.14 (d, 1H), 7.28 (dd, 1H), 7.37 (d, 1H), 7.55 (s, 1H), 7.69 (s, 1H).

(61) Melting point: 252 C. (compound of the formula (I) in crystalline form of polymorph I)

(62) Physicochemical Characterization of Compound of the Formula (I) in Crystalline Form of Polymorph I

(63) Compound of the formula (I) in crystalline form of polymorph I melts at 252 C., H=95-113 Jg.sup.1 (heating rate 20 Kmin.sup.1).

(64) A depression of the melting point was observed depending on the heating rate.

(65) The melting point decreases at a lower heating rate (e.g. 2 Kmin.sup.1) since decomposition occurs.

(66) No other phase transitions were observed. A loss of mass of about 0.1% was observed up to a temperature of 175 C.

(67) Stability and Moisture Absorption

(68) Samples of compound of the formula (I) in crystalline form of polymorph I were stored at 85% and 97% rel. humidity (25 C.). The samples were evaluated after 12 months by DSC, TGA and XRPD. After 12 months, a mass change of <0.1% is observed in both cases. This means that compound of the formula (I) in crystalline form of polymorph I shows no significant absorption of water under these storage conditions. According to DSC, TGA and XRPD, no difference exists in compound of the formula (I) in crystalline form of polymorph I.

(69) HPLC Conditions/Methods

(70) Method A

(71) YMC Hydrosphere C18

(72) 150*4.6 mm, 3.0 m

(73) 25 C., 1 ml/min, 270 nm, 4 nm

(74) 0: 70% TFA 0.1%*; 30% acetonitrile

(75) 17: 20% TFA 0.1%*; 80% acetonitrile

(76) 18: 70% TFA 0.1%*; 30% acetonitrile

(77) *: TFA in water

(78) Method B

(79) YMC Hydrosphere C18

(80) 150*4.6 mm, 3.0 m

(81) 25 C., 1 ml/min, 255 nm, 6 nm

(82) 0: 90% TFA 0.1%*; 10% acetonitrile

(83) 20: 10% TFA 0.1%*; 90% acetonitrile

(84) 18: 10% TFA 0.1%*; 90% acetonitrile

(85) Method C

(86) Nucleodur Gravity C18

(87) 150*2 mm, 3.0 m

(88) 35 C., 0.22 ml/min, 255 nm, 6 nm

(89) Solution A: 0.58 g of ammonium hydrogen phosphate and 0.66 g of ammonium dihydrogen phosphate in 1 l of water (ammonium phosphate buffer pH 7.2)

(90) Solution B: acetonitrile

(91) 0: 30% B; 70% A

(92) 15: 80% B; 20% A

(93) 25: 80% B; 20% A

(94) Method D

(95) Column length: 25 cm

(96) Internal diameter: 4.6 mm

(97) Packing: Chiralpak IA, 5 m

(98) Reagents: 1. Acetonitrile HPLC grade 2. Methyl tert-butyl ether (MTBE), p.a.

(99) Test solution The sample is dissolved at a concentration of 1.0 mg/ml in acetonitrile.

(100) (e.g. about 25 mg of sample, weighed accurately, dissolved in acetonitrile to 25.0 ml).

(101) Eluent A. acetonitrile

(102) B. Methyl tert-butyl ether (MTBE), p.a.

(103) Flow rate 0.8 ml/min

(104) Column oven temperature 25 C.

(105) Detection measurement wavelength: 255 nm

(106) Bandwidth: 6 nm

(107) Injection volumes 5 l

(108) Mix composition of eluents A and B in ratio by volume of 90:10

(109) Chromatogram run time 30 min

(110) Retention times/RRT:

(111) (4S)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (1) about 11 min. RRT: 1.00

(112) (4R)-4-(4-cyano-2-methoxyphenyl)-5-ethoxy-2,8-dimethyl-1,4-dihydro-1,6-naphthyridine-3-carboxamide (1) about 9 min. RRT: 0.82

(113) Measuring Parameters of the X-Ray Diffractometry for the Analysis of the Compound of the Formula (I) in Crystalline Form of Polymorph I

(114) TABLE-US-00001 Dataset name 2429-08a r2 Scan axis 2Theta-Omega Start position [2th.] 2.0000 End position [2th.] 37.9900 Type of divergence slit fixed Size of divergence slit [] 1.0000 Measurement temperature [ C.] 25 Anode material Cu K-alpha1 [] 1.54060 Generator setting 35 mA, 45 kV Diffractometer type transmission diffractometer Goniometer radius [mm] 240.00 Focus-div. slit gap [mm] 91.00 Primary beam monochromator yes Sample rotation yes Peak maximum [2 theta] Polymorph I 8.5 11.4 11.9 13.4 14.1 14.8 15.0 15.4 16.0 17.2 18.5 19.0 19.8 20.5 20.8 22.1 22.7 23.0 23.1 23.6 23.9 24.6 24.9 25.2 25.6 26.0 26.5 27.1 27.3 28.3 28.5 28.8 29.6 30.1 30.6 31.5 31.9 32.4 32.9 33.1 33.4 33.7 34.5 34.7 35.0 35.8 36.2 36.5 37.2 37.4

(115) Measuring Conditions for the IR and Raman Spectroscopy for the Measurement of the Compound of the Formula (I) in Crystalline Form of Polymorph I:

(116) TABLE-US-00002 IR: Instrument Perkin Elmer Spectrum One Number of scans 32 Resolution 4 cm.sup.1 Technique Diamond ATR unit Raman: Instrument Bruker Raman RFS 100/S Number of scans 64 Resolution 2-4 cm.sup.1 Laser power 350 mW Laser wavelength 1064 nm Band maximum [cm.sup.1] IR-ATR Polymorph I Raman Polymorph 1 3475 3074 3416 2997 3366 2970 3074 2941 2992 2920 2952 2836 2835 2231 2230 1659 1681 1641 1658 1623 1606 1601 1572 1577 1485 1487 1464 1443 1454 1383 1431 1362 1420 1327 1407 1303 1381 1267 1355 1230 1341 1191 1325 1161 1303 1123 1285 1093 1267 1032 1255 991 1229 883 1222 827 1161 810 1136 759 1097 734 1031 708 991 671 976 613 967 528 924 505 909 471 875 442 847 346 827 320 810 297 776 186 758 155 746 114 733 723 706 697 670