PROCESS FOR SYNTHESIS OF ELIGLUSTAT AND INTERMEDIATE COMPOUNDS THEREOF

Abstract

The present invention relates to a process for synthesis of Eliglustat and intermediate compounds thereof. In particular, the present invention belongs to the field of synthesis of organic compounds, relates to a process for synthesis of Eliglustat and pharmaceutically acceptable salts thereof, and relates to the intermediate compounds used in the process and a process for preparation of the intermediate compounds. Compared with the existing synthetic processes, the process for preparing Eliglustat according to the present invention utilizes novel intermediates and steps for synthesis, has advantages of convenient operations, high yield, good purity of intermediates and desired product, and it is easy to apply the process in the industrial production.

Claims

1-50. (canceled)

51. A process for preparation of Eliglustat and pharmaceutically acceptable salts thereof, ##STR00099## the process comprising the following steps: (a-1) sulfonylation reaction of Compound V to give Compound VI, ##STR00100## wherein R.sub.5 is hydrogen or hydroxy-protecting group, said hydroxy-protecting group is selected from alkyl, haloalkyl, aralkyl, alkoxyalkyl, allyl, acyl or silyl protective group, such as t-BuMe.sub.2Si, t-BuPh.sub.2Si, (i-Pr).sub.3Si, Et.sub.3Si, Me.sub.3Si, allyl, 2-tetrahydropyranyl, methoxymethyl, formyl, acetyl, benzyl or CH.sub.2Ar, wherein Ar is unsubstituted or substituted aryl, such as p-methoxyphenyl or halogen-substituted phenyl, M is alkyl, aryl, substituted aryl or substituted alkyl, (a-2) reacting Compound VI with pyrrolidine to give Compound VII, ##STR00101## (a-3) reducing Compound VII by catalytic hydrogenation with metal catalyst or reducing Compound VII with organophosphorus reagent, to give Compound VIII, ##STR00102## (a-4) amidation reaction of Compound VIII with Compound IX to give Compound X, ##STR00103## wherein R.sub.6 is selected from hydroxy, halogen or succimidyloxy, when R.sub.5 is hydrogen, Compound X is Eliglustat, or when R.sub.5 is hydroxy-protecting group, the following step is further carried out: (a-5) deprotecting Compound X to remove hydroxy-protecting group, to give Eliglustat ##STR00104##

52. The process according to claim 51, wherein R.sub.5 is hydrogen or silyl protective group.

53. The process according to claim 51, wherein R.sub.6 is chloro or succimidyloxy.

54. The process according to claim 51, wherein in the step (a-1), the sulfonylation reaction is carried out with sulfonyl halide, such as alkylsulfonyl chloride, arylsulfonyl chloride, substituted arylsulfonyl chloride or substituted alkylsulfonyl chloride, the reaction can be carried out without catalyst or with appropriate amount of acylation catalyst, and the catalyst used may be DMAP; the base is an organic base.

54. The process according to claim 51, wherein in the step (a-2), pyrrolidine is reacted with the sulfonate of formula VI, and the reaction solvent is an organic aprotic solvent or solvents.

56. The process according to claim 51, wherein in the step (a-3), the metal catalyst used in the hydrogenation is Pd catalyst or Ni catalyst and the organophosphorus reagent is triphenylphosphine.

57. The process according to claim 51, wherein in the step (a-4): when R.sub.6 is hydroxy, the reaction of Compound VIII with Compound IX is carried out under catalysis of coupling agent to give Compound X; and the coupling agent is a coupling agent or agents for amidation; or when R.sub.6 is chloro or succimidyloxy, amidation reaction of Compound VIII with Compound IX yields Compound X; the reaction is carried out without catalyst or with an appropriate amount of deacid reagent.

58. The process according to claim 51, wherein in the step (a-5), the reaction conditions are: when R.sub.5 is silyl protective group, the reaction of step a-5 is carried out in the presence of base, acid or a fluorine-containing salt, the base is selected from alkali metal or alkaline earth metal hydroxide or carbonate, and the fluorine-containing salt is tetrabutylammonium fluoride (TBAF); when R.sub.5 is alkyl, haloalkyl, alkoxyalkyl or allyl protective group, the reaction of step a-5 is carried out in the presence of an acid; when R.sub.5 is aralkyl protective group, the reaction of step a-5 is carried out under catalytic hydrogenation with a Pd catalyst or Ni catalyst; and the organophosphorus reagent is triphenylphosphine; when R.sub.5 is p-methoxybenzyl, the reaction of step a-5 is carried out in the presence of an oxidizing agent; or when R.sub.5 is acyl protective group, the reaction of step a-5 is carried out under the condition of conventional deprotection to remove the acyl.

59. The process according to claim 51, wherein R.sub.5 is hydrogen, and the process only includes the above reaction steps (a-1), (a-2), (a-3) and (a-4).

60. The process according to claim 51, wherein the process comprises the following steps before step (a-3): (c-1) coupling reaction of Compound I with Compound II in the presence of Lewis acid, deacid reagent and coordination agent, to give Compound III, ##STR00105## wherein each of R.sub.1, R.sub.2 and R.sub.4, independent of each other, is selected from hydrogen, alkyl, aryl or aralkyl, such as phenyl, isopropyl or benzyl, (c-2) optionally, reacting Compound III with hydroxy-protecting reagent in the presence of base to give Compound IV, ##STR00106## wherein R.sub.5 is hydroxy-protecting group, said hydroxy-protecting group is selected from alkyl, haloalkyl, aralkyl, alkoxyalkyl, allyl, acyl or silyl protective group, such as t-BuMe.sub.2Si, t-BuPh.sub.2Si, (i-Pr).sub.3Si, Et.sub.3Si, Me.sub.3Si, allyl, 2-tetrahydropyranyl, methoxymethyl, formyl, acetyl, benzyl or CH.sub.2Ar, wherein Ar is unsubstituted or substituted aryl, such as p-methoxyphenyl or halogen-substituted phenyl; and (c-3) preparing the compound of formula V by the following steps: (c-3.1) reducing Compound III to obtain the Compound V wherein R.sub.5 is hydrogen ##STR00107## or reducing Compound IV to obtain the Compound V wherein R.sub.5 is hydroxy-protecting group as defined above ##STR00108## or (c-3.2) oxidizing Compound III to give Compound XI-1, and reducing Compound XI-1 to obtain the Compound V wherein R.sub.5 is hydrogen (i.e, Compound XI) ##STR00109## or oxidizing Compound IV to give Compound V-1, and reducing Compound V-1 to obtain the Compound V wherein R.sub.5 is hydroxy-protecting group as defined above ##STR00110##

61. The process according to claim 60, wherein R.sub.4 is isopropyl and R.sub.1 and R.sub.2 are phenyl.

62. The process according to claim 60, wherein R.sub.6 is chloro or succimidyloxy.

63. The process according to claim 60, wherein R.sub.5 is hydrogen or silyl protective group.

64. The process according to claim 60, wherein: In step c-1, the Lewis acid is titanium tetrachloride or tin dichloride, the deacid reagent is organic base, and the coordination agent is N-methyl pyrrolidone; In the optional step c-2, the reaction solvent is an organic aprotic solvent or solvents; In step c-3.1, the reducing agent is selected from sodium borohydride, potassium borohydride, boron trifluoride etherate, boranes, or a mixture of any two or more thereof; In step c-3.2, the oxidizing agent is peroxide or a manganese-containing salt or salts, the reaction is carried out with or without catalysis of a base selected from alkali metal or alkaline earth metal hydroxide or carbonate; In step c-3.2, the reducing agent is selected from sodium borohydride, potassium borohydride, boron trifluoride etherate, boranes, or a mixture of any two or more thereof.

65. A process for preparation of the compound of formula IV, ##STR00111## wherein R.sub.5 is hydroxy-protecting group, said hydroxy-protecting group is selected from alkyl, haloalkyl, aralkyl, alkoxyalkyl, allyl, acyl or silyl protective group, wherein Ar is unsubstituted or substituted aryl, the process comprising the following steps: (c-1) coupling reaction of Compound I with Compound II in the presence of Lewis acid, deacid reagent and coordination agent, to give Compound III, ##STR00112## and (c-2) reacting compound III with hydroxy-protecting reagent in the presence of base to give Compound IV ##STR00113## wherein each of R.sub.1, R.sub.2 and R.sub.4, independent of each other, is selected from hydrogen, alkyl, aryl or aralkyl.

66. The process according to claim 65, wherein R.sub.1 and R.sub.2 are hydrogen and R.sub.4 is phenyl or benzyl.

67. The process according to claim 65, wherein R.sub.4 is isopropyl and R.sub.1 and R.sub.2 are phenyl.

68. The process according to claim 65, wherein the reaction conditions are: In step c-1, the Lewis acid is titanium tetrachloride or tin dichloride, the deacid reagent is organic base, In step c-2, the reaction solvent is an organic aprotic solvent or solvents.

68. Crystal form A of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol monohydrate, wherein the crystal form A has a X-ray powder diffraction pattern having characteristic diffraction peaks at 2-theta angle (): 17.5, 18.5, and 21.3.

70. The crystal form A according to claim 65, wherein the crystal form A has a X-ray powder diffraction pattern having the characteristic diffraction peaks at 2-theta angle (): 17.5, 18.5, 19.9, 21.3, and 25.7.

Description

DESCRIPTION OF FIGURES

[0207] FIGS. 1 and 2 show the results of crystal form A of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol monohydrate in X-ray powder diffraction experiment.

[0208] FIG. 3 shows the result of crystal form A of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol monohydrate in thermogravimetric analysis (TGA) experiment.

[0209] FIG. 4 shows the result of crystal form A of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol monohydrate in differential scanning calorimetry (DSC) experiment.

EXAMPLES

[0210] The embodiments of the present invention will be further illustrated by the following examples. It should be understood that the following examples are provided to help further understand the present invention, not intended to limit the scope of the present invention in any manner.

Example 1

Preparation of 3-((2S,3R)-2-azido-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxypropanoyl)-4-phenyloxazolidin-2-one (Compound III-1)

[0211] ##STR00077##

[0212] 10 mmol Compound I (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl) was dissolved in 100 ml dichloromethane, to which 12 mmol titanium tetrachloride was added dropwise at 78 C. After the mixture was stirred for 20 minutes, 15 mmol DIPEA was added. After the mixture was further stirred for 3 hours, 20 mmol N-methyl pyrrolidone was added. After the addition was completed, the reaction mixture was stirred for 30 minutes, and then 15 mmol

[0213] 2,3-dihydrobenzo[b][1,4]dioxin-6-formaldehyde was added. After the addition was completed, the reaction mixture was maintained for 30 minutes. The reaction mixture was warmed to 20 to 30 C. and maintained for 2 hours at the temperature. Subsequently, saturated ammonium chloride aqueous solution was added to quench the reaction. The reaction mixture was extracted with dichloromethane twice. The organic phases were combined, washed with water, dried and concentrated to give 2.5 g of the product Compound III-1.

[0214] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.40-7.26 (m, 5H), 6.99 (d, 1H), 6.91 (dd, 1H), 6.84 (d, 1H), 5.33 (dd, 1H), 5.18 (d, 1H), 5.15-5.13 (m, 1H), 4.59 (t, 1H), 4.25 (dd, 1H), 4.22 (s, 4H); .sup.13C NMR (101 MHz, CDCl.sub.3) 168.06, 153.28, 143.56, 143.49, 138.15, 132.56, 129.31, 129.20, 128.96, 126.05, 125.81, 119.24, 117.29, 115.32, 73.70, 70.65, 65.69, 64.35, 64.29, 57.90.; HR-MS (ESI) calcd for C.sub.20H.sub.19O.sub.6N.sub.4 (M+H).sup.+: 411.1305, found 411.1312.

Example 2

Preparation (S)-3-((2S,3R)-2-azido-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-((trimethylsilyl)oxy)propanoyl)-4-phenyloxazolidin-2-one (Compound IV, wherein R.SUB.1 .and R.SUB.2 .are hydrogen, R.SUB.4 .is phenyl, R.SUB.5 .is trimethylsilyl)

[0215] ##STR00078##

[0216] 10 mmol Compound III-1 (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is hydrogen) was dissolved in 200 ml dichloromethane, 10 which 12 mmol triethylamine was added al 0 C., and then 11 mmol trimethylsilyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 4.3 g of Compound IV (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is trimethylsilyl).

[0217] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.40-7.26 (m, 5H), 6.99 (d, 1H), 6.91 (dd, 1H), 6.83 (d, 1H), 5.24 (dd, 1H), 5.12 (d, 1H), 5.05 (d, 1H), 4.51 (t, 1H), 4.29-4.26 (m, 1H), 4.24 (s, 4H) , 0.07 (s, 9H); .sup.13C NMR (101 MHz, CDCl.sub.3) 167.86, 153.04, 143.34, 143.28, 138.11, 133.37, 129.30, 128.96, 125.78, 119.41, 116.98, 115.48, 75.30, 70.46, 66.08, 64.33, 64.30, 57.90, 0.29.; HR-MS (ESI) calcd for C.sub.23H.sub.27O.sub.6N.sub.4 (M+H).sup.+: 483.1700, found 483.1717.

Example 3

Preparation of (S)-3-((2S,3R)-2-azido-3-(tert-butyldimethylsilyl)oxy)-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propanoyl)-4-phenyloxazolidin-2-one (Compound IV, wherein R.SUB.1 .and R.SUB.2 .are hydrogen, R.SUB.4 .is phenyl, R.SUB.5 .is tert-butyldimethylsilyl)

[0218] ##STR00079##

[0219] 10 mmol Compound III-1 (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is hydrogen) was dissolved in 200 ml dichloromethane, to which 12 mmol triethylamine was added at 0 C. and then 11 mmol tert-butyldimethylsilyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 4.8 g of Compound IV (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is tert-butyldimethylsilyl).

[0220] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.33-7.22 (m, 3H), 7.20-7.13 (m, 3H), 6.97 (d, 1H), 6.90 (d, 1H), 5.67 (ddt, 1H), 5.44 (dt, 1H), 4.47-4.40 (m, 2H), 4.28-4.17 (m, 4H), 4.10 (dd, 1H), 0.85 (s, 7H), 0.06 (s,4H); .sup.13C NMR (101 MHz, CDCl.sub.3) 171.90, 154.26. 143.28, 143.18, 137.83, 133.03, 128.25, 128.02, 127.45, 120.01, 114.41, 112.33, 76.73, 69.67, 63.72, 63.26, 63.20, 57.36, 25.61, 17.98, 4.77.; HR-MS (ESI) calcd for C.sub.26H.sub.32O.sub.6N.sub.4Si (M+H).sup.+: 524.2091, found 524.2084.

Example 4

Preparation of (1R,2R)-2-azido-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propane-1,3-diol (Compound XI)

[0221] ##STR00080##

[0222] 7 mmol Compound III-1 was dissolved in a mixed solvent of 60 ml tetrahydrofuran and 10 ml water, to which 35 mmol sodium borohydride was added at 0 C. After the addition was completed, the reaction mixture was warmed to 25 C. and maintained for 3 hours at the temperature. After the reaction was completed, saturated mixture was extracted with ethyl acetate twice. The combined organic phases were washed with sodium bicarbonate solution, dried and concentrated to give 1.7 g of Compound XI.

[0223] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.80-6.89 (m, 3H), 4.66 (d, J=6.8 Hz, 1H), 4.25 (m, 4H), 3.59-3.66 (m, 2H), 3.50-3.57 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) 143.4, 133.5, 118.9, 117.7, 114.8, 74.1. 68.8, 64.0, 62.9; HR-MS (ESI)calcd for C.sub.11H.sub.14O.sub.4N.sub.3 (M+H).sup.+: 252.0984, found 252.0988.

Example 5

Preparation of (2R,3R)-2-azido-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-((trimethylsilyl)oxy)propane-1-ol (Compound V, wherein R.SUB.5 .is trimethylsilyl)

[0224] ##STR00081##

[0225] 7 mmol Compound IV was dissolved in a mixed solvent of 60 ml tetrahydrofuran and 10 ml water, to which 35 mmol sodium borohydride was added at 0 C. After the addition was completed, the reaction mixture was warmed to 25 C. and maintained for 3 hours at the temperature. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with sodium bicarbonate solution, dried and concentrated to give 2.5 g of Compound V. 1H NMR (400 MHz, CDCl.sub.3) 7.09-7.00 (m, 2H), 6.88 (d, 1H), 4.73-4.68 (m, 1H), 4.28-4.17 (m, 4H), 3.62-3.53 (m, 1H), 3.42-3.31 (m, 2H), 2.68 (t, 1H), 0.16 (s, 7H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.14, 143.12, 133.45, 119.83, 114.35, 112.44, 76.61, 68.15, 63.63, 63.60, 62.78, 0.26., HR-MS (ESI) calcd for C.sub.14H.sub.21O.sub.4N.sub.3Si (M+H).sup.+: 323.1301, found 323.1321.

Example 6

Preparation of (2R,3R)-2-azido-3-((tert-butyldimethylsilyl)oxy)-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propane-1-ol (Compound V, wherein R.SUB.5 .is tert-butyldimethylsilyl)

[0226] ##STR00082##

[0227] 7 mmol Compound IV was dissolved m a mixed solvent of 60 ml tetrahydrofuran and 10 ml water, to which 35 mmol sodium borohydride was added at 0 C. After the addition was completed, the reaction mixture was warmed to 25 C. and maintained for 3 hours at the temperature. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with sodium bicarbonate solution, dried and concentrated to give 2.8 g of Compound V.

[0228] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.09-7.02 (m, 2H), 6.88 (d, 1H), 4.85-4.80 (m, 1H), 4.28-4.17 (m, 4H), 3.69 (ddd, 1H), 3.60 (td, 1H), 3.50 (ddd, 1H), 2.68 (t, 1H), 0.84 (s, 6H), 0.02 (s, 4H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.14, 143.14, 133.42, 119.89, 114.34, 112.39, 76.07, 68.60, 63.47, 63.44, 62.45, 25.59, 18.00, 4.76.; HR-MS (ESI) calcd for C.sub.17H.sub.27O.sub.4N.sub.3Si (M+H).sup.+: 365.1771, found 365.1782.

Example 7

Preparation of (2R,2S)-2-azido-1(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-oxo-3-((S)-2-oxo-4-phenyloxazolidin-3-yl)propyl benzoate (Compound IV, wherein R.SUB.1 .and R.SUB.2 .are hydrogen, R.SUB.4 .is phenyl, R.SUB.5 .is benzoyl)

[0229] ##STR00083##

[0230] 10 mmol Compound III-1 (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is hydrogen) was dissolved in 200 ml dichloromethane, to which 12 mmol triethylamine was added at 0 C., and then 11 mmol benzoyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 4.9 g of Compound VI (wherein R.sub.1 and R.sub.2 are hydrogen, R.sub.4 is phenyl, R.sub.5 is benzoyl).

[0231] .sup.1H NMR (400 MHz, CDCl.sub.3) 8.17-8.07 (m, 2H), 7.62-7.56(m. 1H), 7.49-7.45(m, 2H), 7.40-7.33 (m, 3H), 7.30-7.27 (m, 2H), 7.13 (s, 1H), 7.06 (dd, 1H), 6.89 (d, 1H), 6.62 (d, 1H), 5.30-5.27 (m, 2H), 4.80 (t, 1H), 4.35 (dd, 1H), 4.25 (s, 4H); .sup.13C NMR (101 MHz, CDCl.sub.3) 167.55, 165.53, 153.74, 143.86, 143.58, 138.17, 134.56, 133.66, 130.56, 130.03, 129.28, 129.19, 128.96, 128.94, 128.88, 128.61, 125.91, 119.49, 117.54, 115.48, 74.88, 70.88, 68.31, 64.42, 64.34, 64.30, 58.11.; HR-MS (ESI) calcd for C.sub.27H.sub.23O.sub.7N.sub.4 (M+H).sup.+: 515.1567, found 515.1558.

Example 8

Preparation of (1R,2R)-2-azido-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propane-1,3-diol (Compound XI)

[0232] ##STR00084##

[0233] 7 mmol Compound XI-1 was dissolved in 60 ml tetrahydrofuran, to which 14 mmol sodium borohydride and 8 mmol boron trifluoride etherate were added at 0 C. After the addition was completed, the reaction mixture was stirred for 3 hours while the temperature was maintained. After the reaction was completed, dilute hydrochloric acid was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with sodium bicarbonate solution, dried and concentrated to give 1.4 g of Compound XI. .sup.1H NMR (400 MHz, CDCl.sub.3) 6.80-6.89 (m, 3H), 4.66 (d, J=6.8 Hz, 1H), 4.25 (m, 4H), 3.59-3.66 (m, 2H), 3.50-3.57 (m, 1H); .sup.13C NMR (101 MHz, CDCl.sub.3) 143.4, 133.5, 118.9, 117.7, 114.8, 74.1, 68.8, 64.0, 62.9; HR-MS (ESI) calcd for C.sub.11H.sub.14O.sub.4N.sub.3 (M+H).sup.+: 252.0984, found 252.0988.

Example 9

Preparation of (2R,3R)-2-azido-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-((trimethylsilyl)oxy)propyl 4-methylbenzenesulfonate (Compound VI, wherein R.SUB.5 .is trimethylsilyl)

[0234] ##STR00085##

[0235] 9 mmol Compound V (wherein R5 is trimethylsilyl) was dissolved in 200 ml dichloromethane, to which 12 mmol triethylamine and 10 mg DMAP were added at 0 C. 10 mmol p-toluenesulfonyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 3.7 g of Compound VI (wherein R.sub.5 is trimethylsilyl).

[0236] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.75-7.73 (m, 2H), 7.34-7.32 (m, 2H), 6.78-6.76 (m, 2H), 6.68(dd, 1H), 4.58 (d, 1H), 4.24 (s, 4H), 3.93 (dd, 1H), 3.71 (dd, 1H), 3.55-3.51 (m, 1H), 2.44 (s, 3H), 0.00 (s, 9H); .sup.13C NMR (101 MHz, cdcl.sub.3) 145.07, 143.56, 143.49, 133.13, 132.44, 129.88, 127.97, 119.43, 117.30, 115.32, 74.63, 68.37, 65.95, 64.29, 64.27, 26.89, 0.12.; HR-MS (ESI) calcd for C.sub.21H.sub.28O.sub.6N.sub.3SSi (M+H).sup.+: 478.1468, found 478.1463.

Example 10

Preparation of (2R,3R)-2-azido-3-((tert-butyldimethylsilyl)oxy)-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)propyl 4-methylbenzenesulfonate (Compound VI, wherein R.SUB.5 .is tert-butyldimethylsilyl)

[0237] ##STR00086##

[0238] 9 mmol Compound V (wherein R.sub.5 is teri-butyldimethylsilyl) was dissolved in 200 ml dichloromethane, to which 12 mmol triethylamine and 10 mg DMAP were added at 0 C. 10 mmol p-toluenesulfonyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 4.2 g of Compound VI (wherein R.sub.5 is tert-butyldimethylsilyl).

[0239] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.75-7.69 (m, 2H), 7.48 (dq, 2H), 7.12 (dd, 1H), 6.95 (d, 1H), 6.90 (d, 1H), 4.74 (d, 1H), 4.28-4.17 (m, 5H), 3.96 (dd, 1H), 3.61 (q, 1H), 2.37 (d, 3H), 0.85 (s, 7H), 0.06 (s, 4H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.59, 143 21, 143.16, 134.03, 133.24, 129.69, 128.09, 120.03, 114.38, 112.44, 76.17, 68.10, 66.52, 63.59, 63.56, 25.61, 21.53, 17.98, 4.80.; HR-MS (ESI) calcd for C.sub.24H.sub.33O.sub.6N.sub.3SSi (M+H).sup.+: 519.1859, found 519.1866.

Example 11

Preparation of (2R,3R)-2-azido-3-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-hydroxypropyl 4-methylbenzenesulfonate (Compound VI, wherein R.SUB.5 .is hydrogen)

[0240] ##STR00087##

[0241] 9 mmol Compound XI (wherein R.sub.5 is hydrogen) was dissolved in 200 ml dichloromethane, to which 12 mmol triethylamine and 10 mg DMAP were added at 0 C. 10 mmol p-toluenesulfonyl chloride was added slowly. After the addition was completed, the reaction was maintained for 3 hours. The reaction mixture was quenched by adding water. Organic phase was dried and concentrated to give 3.5 g of Compound VI (wherein R.sub.5 is hydrogen).

[0242] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.81 (d, J=8.4 Hz, 2H), 7.37 (d, J=8.4 Hz, 2H), 6.75-6.87 (m, 3H), 4.60 (d, J=6.4 Hz, 1H), 4.28 (s, 4H), 4.11 (dd, J=3.6 Hz, 10.4 Hz, 1H), 3.90 (dd, J=7.2 Hz, 10.4 Hz, 1H), 3.67-3.71 (m, 1H), 2.45 (s, 3H); .sup.13C NMR (101 MHz, CDCl.sub.3) 145.5, 144.2, 143.9, 132.1, 129.7, 128.3, 119.0, 117.9, 114.8, 73.1, 69.1, 65.7, 64.1; HR-MS (ESI) calcd for C.sub.18H.sub.20O.sub.6N.sub.3S (M+H).sup.+: 406.1037, found 406.1035.

Example 12

Preparation of 1-((2R,3R)-2-azido-3(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3((trimethylsilyl)oxy)propyl)pyrrolidine (Compound VII, wherein R.SUB.5 .is trimethylsilyl)

[0243] ##STR00088##

[0244] 5 mmol Compound VI (wherein R.sub.5 is trimethylsilyl) was dissolved in 50 ml DMF, to which 15 mmol pyrrolidine was added. The reaction was stirred at 60 C. for 10 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 1.0 g of Compound VII (wherein R.sub.5 is trimethylsilyl).

[0245] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.82-5.74 (m, 3H), 4.60 (d, 1H), 4.23 (s, 4H), 3.44-3.39 (m, 1H), 2.52-2.41 (m, 5H), 2.33 (dd, 1H), 1.75-1.72 (m, 4H), 0.03 (s, 9H); .sup.13C NMR (101 MHz, CDCl.sub.3) 160.05, 139.96, 128.98, 128.47, 125.97, 72.29, 64.26, 64.19, 56.28, 54.07, 23.45, 21.74, 0.09.; HR-MS (ESI) calcd for C.sub.18H.sub.31O.sub.3N.sub.2Si (M+H).sup.+: 376.1931, found 376.1940.

Example 13

Preparation of (1R,2R)-2-azido-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol (Compound XII)

[0246] ##STR00089##

[0247] 5 mmol Compound VI (wherein R.sub.5 is hydrogen) was dissolved in 50 ml DMF, to which 15 mmol pyrrolidine was added. The reaction was stirred at 60 C. for 10 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 1.1 g of Compound XII.

[0248] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.82-6.92 (m, 3H), 4.93 (m, 1H), 4.24 (s, 4H), 4.10(m, 1H), 3.15-3.19 (m, 1H), 3.07 (m, 4H), 2.81-2.87 (m, 1H), 1.96 (m, 4H); .sup.13C NMR (101 MHz, CDCl.sub.3) 143.7, 134.1, 119.8, 117.5, 115.9, 75.5, 64.1, 63.9, 56.7, 54.4, 23.8; HR-MS (ESI) calcd for C.sub.15H.sub.21O.sub.3N.sub.4 (M+H).sup.+: 305.1608, found 305.1611.

Example 14

Preparation of 1-((2R,3R)-2-azido-3-((tert-butyldimethylsilyl)oxy)-3-(2-dihydrobenzo[b][1,4]dioxin-6-yl)propyl)pyrrolidine (Compound XII)

[0249] ##STR00090##

[0250] 5 mmol Compound VI (wherein R.sub.5 is tert-butyldimethylsilyl) was dissolved in 50 ml DMF, to which 15 mmol pyrrolidine was added. The reaction was stirred al 60 C. for 10 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 2.1 g of Compound XII.

[0251] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.09-7.02 (m, 2H), 6.88 (d, 1H), 4.75-4.70 (m, 1H), 4.28-4.17 (m, 4H), 3.47 (q, 1H), 2.93-2.79 (m, 4H), 2.53 (dd, 1H), 2.02 (dd, 1H), 1.84 (hept, 4H), 0.84 (s, 7H), 0.02 (s, 4H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.18, 143.15, 132.60, 120.03, 114.34, 112.38, 78.23, 63.80, 63.78, 63.69, 59.99, 54.51, 25.61, 23.73, 18.03, 4.78.; HR-MS (ESI) calcd for C.sub.21H.sub.34O.sub.3Si (M+H).sup.+: 418.2400, found 418.2413.

Example 15

Preparation of (1R,2R)-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)-1-((trimethylsilyl)oxy)propan-2-amine (Compound VIII, wherein R.SUB.5 .is trimethylsilyl)

[0252] ##STR00091##

[0253] 4 mmol Compound VII (wherein R.sub.5 is trimethylsilyl) was dissolved in 200 ml methanol, to which 20 mg of 5% Pd/C was added. The reaction was carried out under 0.1 MPa of hydrogen pressure at 25 C. for 10 hours. After the reaction was completed, the reaction mixture was filtrated and concentrated to give 1.3 g of Compound VIII (wherein R.sub.5 is trimethylsilyl).

[0254] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.15-7.10 (m, 1H), 7.07-7.00 (m, 1H), 6.88 (d, 1H), 4.46 (d, 1H), 4.28-4.17 (m, 4H), 3.59 (d, 2H), 3.25 (qt, 1H), 2.91-2.77 (m, 4H), 2.49 (dd, 1H), 2.40 (dd, 1H), 1.89- 1.80 (m, 4H), 0.16 (s, 7H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.16, 142.99, 134.17, 120.00, 114.33, 112.41, 78.06, 63.52, 63.50, 61.58, 54.57, 53.62, 23.64, 0.26.; HR-MS (ESI) calcd for C.sub.18H.sub.30O.sub.3N.sub.2SI (M+H).sup.+: 350.2026. found 350.2032.

Example 16

Preparation of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol (Compound VIII, wherein R.SUB.5 .is hydrogen)

[0255] ##STR00092##

[0256] 2 mmol Compound XII (wherein R.sub.5 is hydrogen) was dissolved in 100 ml methanol, to which 10 mg of 5% Pd/C was added. The reaction was carried out under 0.1 MPa of hydrogen pressure at 25 C. for 10 hours. After the reaction was completed, the reaction mixture was filtrated and concentrated to give 0.6 g of Compound VIII (wherein R.sub.5 is hydrogen).

[0257] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.77-6.85 (m, 3H), 5.85 (d, J=7.2 Hz, 1H), 4.91 (d, J=3.6 Hz, 1H), 4.23 (s, 4H), 4.15-4.21 (m, 1H), 2.74-2.83 (m, 2H), 2.61-2.67 (m, 4H), 2.07-2.11 (m, 2H), 1.79-1.81 (m, 4H), 1.51-1.56 (m, 2H), 1.22-1.33 (m, 10H). 0.89 (t, J=6.8 Hz, 3H); .sup.13C NMR (101 MHz, CDCl.sub.3) 174.1, 143.2, 142.7, 134.6, 119.2, 117.3, 115.1, 75.6, 64.5, 57.9, 55.1, 52.0, 36.5, 31.7, 29.5, 29.1, 25.8, 23.4, 22.7, 13.9. HR-MS (ESI) calcd for C.sub.23H.sub.37O.sub.4N.sub.2 (M+H).sup.+: 405.2748, found 405.2741.

Example 16a

Preparation of Crystal Form A of (1R,2R)-2-amino-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propane-1-ol:

[0258] ##STR00093##

[0259] Preparation of 5% NaOH solution: add 10 g sodium hydroxide to 190 g water, dissolve, and store the solution until use.

[0260] In a clean and dry 1000 ml reaction bottle, 100 g ELG-4-PTA and 200 g drinking water were added, and the 5% NaOH solution prepared in advance was added under stirring. The resulting solution was stirred adequately for 0.5 h and different layers were formed. The organic layer was washed with 200 g drinking water, and the layers were separated to obtain the organic layer, to which 10 g triethylamine and 6 g Pd/C were added. The reaction was performed at controlled temperature of 20 to 30 C. and at pressure of 0.3 to 0.5 Mpa for 12-18 hours. After the reaction was completed, the reaction atmosphere was replaced with nitrogen gas, then the mixture was warmed to 35-40 C., dissolved, filtered to yield filtrate. The filtrate was concentrated to about 240 g of the solution of ELG-5 in isopropyl acetate under reduced pressure while controlling the external temperature as 45 C. To the solution 17 g drinking water was added. The resulting solution was stirred to precipitate crystal, the temperature was reduced to 5 to 5 C. and maintained for 2 hours to crystallize. The resulting mixture was filtered, rinsed with hexane, and dried under reduced pressure at controlled external temperature of 40 C. to give 55 g to 65 g white solid (as monohydrate).

[0261] 1. X-ray Diffraction Measurement of Crystal Form A:

TABLE-US-00001 TABLE 1 Instruments of X-ray diffraction and the corresponding parameters Instrument name X-ray Diffractometer (XRD) Instrument type Germany, Bruker D8 advance Instrument Catalog No. LY-01-005 Light source Cu Method and parameter Voltage and current 40 kV, 40 mA Starting and ending positions 3-40 2 stepping 0.02 2 Time for each stepping 0.5 s

[0262] The result showed that the crystal form A has a X-ray powder diffraction pattern having the characteristic diffraction peaks at 2-theta angle (): 17.5, 18.5, and 21.3.

[0263] In particular, the crystal form A has a X-ray powder diffraction pattern having the characteristic diffraction peaks at 2-theta angle (): 17.5, 18.5, 19.9, 21.3, and 25.7.

[0264] More particularly, the crystal form A has a X-ray powder diffraction pattern having the characteristic diffraction peaks at 2-theta angle (): 6.4, 11.4, 12.3, 17.5, 18.5, 19.9, 21.3, 22.9, 24.1, and 25.7.

[0265] The results of X-ray diffraction experiment of the crystal form A were shown in FIGS. 1 and 2.

[0266] 2. Thermogravimetric Analysis (TGA) of the Crystal Form A

TABLE-US-00002 TABLE 2 Instruments of TGA and the corresponding parameters Instrument name Thermogravimetric analyser (TGA) Instrument type TA Q500 Instrument Catalog No. LY-01-003 Method and parameter 1. Raising the temperature to 300.0 C. in a rate of 10.0 C./min 2. N.sub.2 flowing rate of 40 mL/min

[0267] TGA showed that the crystal lost about 6.0% of total weight progressively before 100 C., the temperature at which the solvent was removed was about 50 C., and the sample was decomposed after 150 C.

[0268] The result of TGA experiment of the crystal form A was shown in FIG. 3.

[0269] 3. Differential Scanning Calorimetry (DSC) Experiment of the Crystal Form A

TABLE-US-00003 TABLE 3 Instruments of DSC and the corresponding parameters Instrument name Differential scanning calorimeter (DSC) Instrument type TA Q200 Instrument Catalog No. LY-01-002 Method and parameter 1. Equilibrating at 20.0 C. 2. Raising the temperature to 200.0 C. in a rate of 10.0 C./min 3. N.sub.2 flowing rate of 40 mL/min 4. Aluminum pan with lid.

[0270] DSC showed that the sample has an endothermal peak between 68 C. and 91 C., the onset value of the endothermal peak is 82.9 C. and the peak value is 85.9 C. The result of DSC experiment of the crystal form A was shown in FIG. 4.

Example 17

Preparation of (1R,2R)-1-((tert-butyldimethylsilyl)oxy)-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-2-amine (Compound VIII, wherein R.SUB.5 .is tert-butyldimethylsilyl)

[0271] ##STR00094##

[0272] 2 mmol Compound XII (wherein R.sub.5 is tert-butyldimethylsilyl) was dissolved in 100 ml methanol, to which 10 mg of 5% Pd/C was added. The reaction was carried out under 0.1 MPa of hydrogen pressure at 25 C. for 10 hours. After the reaction was completed, the reaction mixture was filtrated and concentrated to give 1.1 g of Compound VIII (wherein R.sub.5 is tert-butyldimethylsilyl).

[0273] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.14-7.09 (m, 1H), 7.03 (dd, 1H), 6.88 (d, 1H), 4.67 (dt, 1H), 4.28-4.17 (m, 4H), 3.59 (d, 2H), 3.24 (qt, 1H), 2.92-2.77 (m, 4H), 2.74 (dd, 1H) 2.40 (dd, 1H), 1.89-1.80 (m, 4H), 0.84 (s, 7H), 0.01 (s, 4H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 143.17, 143.04, 134.13, 120.03, 114.33, 112.46, 76.86, 63.55, 63.52, 61.53, 54.58, 53.69, 25.61, 23.74, 17.89, 4.79.; HR-MS (ESI) calcd for C.sub.21H.sub.36O.sub.3N.sub.2Si (M+H).sup.+: 392.2495, found 392.2483.

Example 18

Preparation of N-((1R,2R)-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-1-((trimethylsilyl)oxy)-3-(pyrrolidin-1-yl)propan-2-yl)octanamide (Compound X, wherein R.SUB.5 .is trimethylsilyl)

[0274] ##STR00095##

[0275] 1 mmol Compound VIII (wherein R.sub.5 is trimethylsilyl) was dissolved in 30 ml DMF, to which 1.5 mmol DIPEA and 1.2 mmol Compound IX (wherein R.sub.6 is succimidyloxy) were added. After the addition was completed, the reaction was maintained at 25 C. for 24 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 0.5 g of Compound X.

[0276] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.76-6.59 (m, 3H), 5.77 (d, 1H), 5.01 (d, 1H), 4.16 (s, 3H), 4.06 (q, 1H), 3.99 (dd, 1H), 2.63 (dd, 1H), 2.58-2.38 (m, 4H), 2.24 (dd, 1H). 2.03 (td, 2H), 1.98 (S, 1H), 1.76-1.64 (m, 4H), 1.45 (p, 2H), 1.27-1.09 (m, 8H), 0.82 (t, 3H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 172.55, 171.02, 142.91, 142.34, 135.90, 118.63, 116.57, 114.67, 71.33, 64.26, 64.18, 60.31, 56.47, 55.06, 54.26, 53.94, 36.76, 31.61, 29.03, 29.01, 25.64, 23.59, 22.59, 20.97, 14.14, 14.05, 1.36.; HR-MS (ESI) calcd for C.sub.18H.sub.31O.sub.3N.sub.2Si (M+H).sup.+: 477.3149. found 477.3155.

Example 19

Preparation of N-((1R,2R)-1-((tert-butyldimethylsilyl)oxy-1-(2,3-dihydrobenzo[b][1,4]dioxin-6-yl)-3-(pyrrolidin-1-yl)propan-2-yl)octanamide (Compound X, wherein R.SUB.5 .is tert-butyldimethylsilyl)

[0277] ##STR00096##

[0278] 1 mmol Compound VIII (wherein R.sub.5 is tert-butyldimethylsilyl) was dissolved in 30 ml DMF, to which 1.5 mmol DIPEA and 1.2 mmol Compound IX (wherein R.sub.6 is succimidyloxy) were added. After the addition was completed, the reaction was maintained at 25 C. for 24 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 0.9 g of Compound X.

[0279] .sup.1H NMR (400 MHz, CDCl.sub.3) 7.04 (ddd, 1H), 6.91-6.81 (m, 2H), 6.18 (d, 1H). 5.19 (d, 1H), 4.28-4.17 (m, 4H), 4.11 (dq. 1H), 2.91-2.83 (m, 2H), 2.86-2.76 (m, 2H), 2.59 (dd, 1H), 2.51 (dd, 1H), 2.26 (dt, 1H), 2.19 (dt, 1H), 1.83 (p, 4H), 1.63 (dt, 1H), 1.47 (dt, 1H), 1.39-1.14 (m, 7H), 0.94-0.84 (m, 3H), 0.85 (s, 6H), 0.06 (s, 4H).; .sup.13C NMR (101 MHz, CDCl.sub.3) 174.41, 143.16, 143.13, 134.01, 120.36, 114.37, 112.72, 76.42, 63.59, 63.56, 60.11, 54.61, 54.20, 35.54, 31.26, 28.68, 28.40, 25.62, 25.07, 23.79, 22.79, 17.98, 14.06, 4.76.; HR-MS (ESI) calcd for C.sub.29H.sub.50O.sub.4N.sub.2Si (M+H).sup.+: 518.3540, found 518.3549.

Example 20

Preparation of Eliglustat

[0280] ##STR00097##

[0281] 1 mmol Compound VIII (wherein R.sub.5 is hydrogen) was dissolved in 30 ml DMF, to which 1.5 mmol DIPEA and 1.2 mmol Compound IX (wherein R.sub.6 is succimidyloxy) were added. After the addition was completed, the reaction was maintained at 25 C. for 24 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. The combined organic phases were washed with water, dried and concentrated to give 0.3 g Eliglustat.

[0282] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.72-6.90 (m, 3H), 5.83 (d, 1H), 4.89 (d, 1H), 4.25 (m, 4H), 4.15-4.19 (m, 1H), 2.71-2.77 (m, 2H), 2.56-2.70 (m, 4H) , 2.10 (t, 2H), 1.73-1.82 (m, 4H), 1.46-1.57 (m, 2H), 1.15-1.30 (m, 8H), 0.88 (t, 3H); .sup.13C NMR (101 MHz, CDCl.sub.3) 174.47, 143.25, 143.17, 135.21, 120.41, 114.34, 112.85, 74.96, 63.81, 63.60, 59.47, 54.48, 53.67, 35.86, 31.39, 28.44, 28.27, 25.39, 23.75, 22.71, 14.08. HR-MS (ESI) calcd for C.sub.23H.sub.37O.sub.2N.sub.4 (M+H).sup.+: 405.2753. found 405.2760.

Example 21

Preparation of Eliglustat

[0283] ##STR00098##

[0284] 1 mmol Compound X (wherein R.sub.5 is trimethylsilyl) was dissolved in 30 ml THF, to which 20 ml of 2N HCl was added. After the addition was completed, the reaction was maintained at 25 C. for 2 hours. After the reaction was completed, water was added to quench the reaction, and the reaction mixture was extracted with ethyl acetate twice. Organic phases were combined, and washed with saturated sodium bicarbonate solution once. The resulting organic phase was dried and concentrated to give 0.4 g Eliglustat.

[0285] .sup.1H NMR (400 MHz, CDCl.sub.3) 6.72-6.90 (m, 3H), 5.83 (d, 1H), 4.89 (d, 1H), 4.25 (m, 4H), 4.15-4.19 (m, 1H), 2.71-2.77 (m, 2H), 2.56-2.70 (m, 4H), 2.10 (t, 2H), 1.73-1.82 (m, 4H), 1.46-1.57 (m, 2H), 1.15-1.30 (m, 8H), 0.88 (t, 3H); .sup.13C NMR (101 MHz, CDCl.sub.3) 174.47, 143.25, 143.17, 135.21, 120.41, 114.34, 112.85, 74.96, 63.81, 63.60, 50.47, 54.48, 53.67, 35.86, 31.39, 28.44, 28.27, 25.39, 23.75, 22.71, 14.08. HR-MS (ESI) calcd for C.sub.23H.sub.37O.sub.2N.sub.4 (M+H).sup.+: 405.2753, found 405.2760.