LIPID COMPOUNDS AND LIPID NANOPARTICLE COMPOSITIONS

Abstract

Provided herein are lipid compounds that can be used in combination with other lipid components, such as neutral lipids, cholesterol and polymer conjugated lipids, to form lipid nanoparticles for delivery of therapeutic agents (e.g., nucleic acid molecules) for therapeutic or prophylactic purposes, including vaccination. Also provided herein are lipid nanoparticle compositions comprising said lipid compounds.

Claims

1-67. (canceled)

68. A compound of Formula (I): ##STR00381## or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein: G.sup.1 and G.sup.2 are each independently a bond, C.sub.2-C.sub.12 alkylene, or C.sub.2-C.sub.12 alkenylene; L.sup.1 is —OC(═O)R.sup.1, —C(═O)OR.sup.1, —OC(═O)OR.sup.1, —C(═O)R.sup.1, —OR.sup.1, —S(O).sub.xR.sup.1, —S—SR.sup.1, —C(═O)SR.sup.1, —SC(═O)R.sup.1, —NR.sup.aC(═O)R.sup.1, —C(═O)NR.sup.bR.sup.c, —NR.sup.aC(═O)NR.sup.bR.sup.c, —OC(═O)NR.sup.bR.sup.c, —NR.sup.aC(═O)OR.sup.1, —SC(═S)R.sup.1, —C(═S)SR.sup.1, —C(═S)R.sup.1, —CH(OH)R.sup.1, —P(═O)(OR.sup.b)(OR.sup.c), —(C.sub.6-C.sub.10 arylene)-R.sup.1, -(6- to 10-membered heteroarylene)-R.sup.1, or R.sup.1; L.sup.2 is —OC(═O)R.sup.2, —C(═O)OR.sup.2, —OC(═O)OR.sup.2, —C(═O)R.sup.2, —OR.sup.2, —S(O).sub.xR.sup.2, —S—SR.sup.2, —C(═O)SR.sup.2, —SC(═O)R.sup.2, —NR.sup.dC(═O)R.sup.2, —C(═O)NR.sup.eR.sup.f, —NR.sup.dC(═O)NR.sup.eR.sup.f, —OC(═O)NR.sup.eR.sup.f, —NR.sup.dC(═O)OR.sup.2, —SC(═S)R.sup.2, —C(═S)SR.sup.2, —C(═S)R.sup.2, —CH(OH)R.sup.2, —P(═O)(OR.sup.e)(OR.sup.f), —(C.sub.6-C.sub.10 arylene)-R.sup.2, -(6- to 10-membered heteroarylene)-R.sup.2, or R.sup.2; R.sup.1 and R.sup.2 are each independently C.sub.5-C.sub.32 alkyl or C.sub.5-C.sub.32 alkenyl; R.sup.a, R.sup.b, R.sup.d, and R.sup.e are each independently H, C.sub.1-C.sub.24 alkyl, or C.sub.2-C.sub.24 alkenyl; R.sup.c and R.sup.f are each independently C.sub.1-C.sub.32 alkyl or C.sub.2-C.sub.32 alkenyl; R.sup.0 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; G.sup.3 is C.sub.2-C.sub.12 alkylene or C.sub.2-C.sub.12 alkenylene; R.sup.4 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; R.sup.5 is C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; x is 0, 1, or 2; s is 0 or 1, provided that when s is 0, then R.sup.4 is not C.sub.1-C.sub.12 alkyl; and wherein each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, arylene, and heteroarylene, is independently optionally substituted.

69. The compound of claim 68, wherein G.sup.3 is C.sub.2-C.sub.4 alkylene.

70. The compound of claim 68, which is a compound of Formula (V): ##STR00382## wherein t is an integer from 2 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

71. The compound of claim 68, wherein R.sup.0 is C.sub.1-C.sub.6 alkyl.

72. The compound of claim 68, which is a compound of Formula (IV): ##STR00383## or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

73. The compound of claim 68, wherein R.sup.4 is C.sub.3-C.sub.8 cycloalkyl.

74. A compound of Formula (II): ##STR00384## or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein: G.sup.1 and G.sup.2 are each independently a bond, C.sub.2-C.sub.12 alkylene, or C.sub.2-C.sub.12 alkenylene; L.sup.1 is —OC(═O)R.sup.1, —C(═O)OR.sup.1, —OC(═O)OR.sup.1, —C(═O)R.sup.1, —OR.sup.1, —S(O)XR.sup.1, —S—SR.sup.1, —C(═O)SR.sup.1, —SC(═O)R.sup.1, —NR.sup.aC(═O)R.sup.1, —C(═O)NR.sup.bR.sup.c, —NR.sup.aC(═O)NR.sup.bR.sup.c, —OC(═O)NR.sup.bR.sup.c, —NR.sup.aC(═O)OR.sup.1, —SC(═S)R.sup.1, —C(═S)SR.sup.1, —C(═S)R.sup.1, —CH(OH)R.sup.1, —P(═O)(OR.sup.b)(OR.sup.c), —(C.sub.6-C.sub.10 arylene)-R.sup.1, -(6- to 10-membered heteroarylene)-R.sup.1, or R.sup.1; L.sup.2 is —OC(═O)R.sup.2, —C(═O)OR.sup.2, —OC(═O)OR.sup.2, —C(═O)R.sup.2, —OR.sup.2, —S(O).sub.xR.sup.2, —S—SR.sup.2, —C(═O)SR.sup.2, —SC(═O)R.sup.2, —NR.sup.dC(═O)R.sup.2, —C(═O)NR.sup.eR.sup.f, —NR.sup.dC(═O)NR.sup.eR.sup.f, —OC(═O)NR.sup.eR.sup.f, —NR.sup.dC(═O)OR.sup.2, —SC(═S)R.sup.2, —C(═S)SR.sup.2, —C(═S)R.sup.2, —CH(OH)R.sup.2, —P(═O)(OR.sup.e)(OR.sup.f), —(C.sub.6-C.sub.10 arylene)-R.sup.2, -(6- to 10-membered heteroarylene)-R.sup.2, or R.sup.2; R.sup.1 and R.sup.2 are each independently C.sub.5-C.sub.32 alkyl or C.sub.5-C.sub.32 alkenyl; Y is H, C.sub.1-C.sub.14 alkyl, or —C(═O)(C.sub.1-C.sub.14 alkyl); R.sup.a, R.sup.b, R.sup.d, and R.sup.e are each independently H, C.sub.1-C.sub.24 alkyl, or C.sub.2-C.sub.24 alkenyl; R.sup.c and R.sup.f are each independently C.sub.1-C.sub.32 alkyl or C.sub.2-C.sub.32 alkenyl; G.sup.3 is C.sub.2-C.sub.12 alkylene or C.sub.2-C.sub.12 alkenylene; R.sup.3 is —N(R.sup.4)R.sup.5 or —OR.sup.6; R.sup.4 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; R.sup.5 is C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; or R.sup.4, R.sup.5, together with the nitrogen to which they are attached form a cyclic moiety; R.sup.6 is hydrogen, C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, or C.sub.6-C.sub.10 aryl; x is 0, 1, or 2; and wherein each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, arylene, heteroarylene, and cyclic moiety is independently optionally substituted.

75. The compound of claim 68, wherein L.sup.1 is —OC(═O)R.sup.1, —C(═O)OR.sup.1, —C(═O)R.sup.1, —C(═O)NR.sup.bR.sup.c, or R.sup.1; and L.sup.2 is —OC(═O)R.sup.2, —C(═O)OR.sup.2, —C(═O)R.sup.2, —C(═O)NR.sup.eR.sup.f, or R.sup.2.

76. The compound of claim 68, which is a compound of Formula (IV-A), (IV-B), (IV-C), (IV-D), (IV-E), (IV-F), (IV-G), or (IV-H): ##STR00385## wherein y and z are each independently an integer from 2 to 12, and t is an integer from 2 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

77. The compound of claim 68, which is a compound of Formula (V-A), (V-B), (V C), (V-D), (V-E), (V-F), (V-G), or (V-H): ##STR00386## wherein y and z are each independently an integer from 2 to 12, and t is an integer from 2 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

78. The compound of claim 74, which is a compound of Formula (VI-A), (VI-B), (VI-C), or (VI-D): ##STR00387## wherein z is an integer from 2 to 12, t is an integer from 2 to 12, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

79. A compound of Formula (III): ##STR00388## or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof, wherein: R.sup.1 and R.sup.2 are each independently C.sub.5-C.sub.32 alkyl or C.sub.5-C.sub.32 alkenyl; R.sup.0 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; G.sup.3 is C.sub.2-C.sub.12 alkylene or C.sub.2-C.sub.12 alkenylene; G.sup.4 is C.sub.2-C.sub.12 alkylene or C.sub.2-C.sub.12 alkenylene; R.sup.3 is —N(R.sup.4)R.sup.5 or —OR.sup.6; R.sup.4 is C.sub.1-C.sub.12 alkyl, C.sub.2-C.sub.12 alkenyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; R.sup.5 is C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, C.sub.6-C.sub.10 aryl, or 4- to 8-membered heterocycloalkyl; or R.sup.4, R.sup.5, together with the nitrogen to which they are attached form a cyclic moiety; R.sup.6 is hydrogen, C.sub.1-C.sub.12 alkyl, C.sub.3-C.sub.8 cycloalkyl, C.sub.3-C.sub.8 cycloalkenyl, or C.sub.6-C.sub.10 aryl; and wherein each alkyl, alkenyl, cycloalkyl, cycloalkenyl, heterocycloalkyl, aryl, alkylene, alkenylene, and cyclic moiety is independently optionally substituted.

80. The compound of claim 68, wherein R.sup.1 and R.sup.2 are each independently branched C.sub.6-C.sub.24 alkyl or branched C.sub.6-C.sub.24 alkenyl.

81. A compound in Table 1, Table 2, or Table 3, or a pharmaceutically acceptable salt, prodrug or stereoisomer thereof.

82. A composition comprising the compound of claim 68, and a therapeutic or prophylactic agent.

83. A lipid nanoparticle comprising the compound of claim 68.

84. A pharmaceutical composition comprising the compound of claim 68, and a pharmaceutically acceptable excipient or diluent.

85. A composition comprising the compound of claim 74, and a therapeutic or prophylactic agent.

86. A lipid nanoparticle comprising the compound of claim 74.

87. A pharmaceutical composition comprising the compound of claim 74, and a pharmaceutically acceptable excipient or diluent.

88. A composition comprising the compound of claim 79, and a therapeutic or prophylactic agent.

89. A lipid nanoparticle comprising the compound of claim 79.

90. A pharmaceutical composition comprising the compound of claim 79, and a pharmaceutically acceptable excipient or diluent.

Description

7. EXAMPLES

[0436] The examples in this section are offered by way of illustration, and not by way of limitation.

General Methods.

[0437] General preparative HPLC method: HPLC purification is carried out on an Waters 2767 equipped with a diode array detector (DAD) on an Inertsil Pre-C8 OBD column, generally with water containing 0.1% TFA as solvent A and acetonitrile as solvent B.

[0438] General LCMS method: LCMS analysis is conducted on a Shimadzu (LC-MS2020) System. Chromatography is performed on a SunFire C18, generally with water containing 0.1% formic acid as solvent A and acetonitrile containing 0.1% formic acid as solvent B.

7.1 Example 1: Preparation of Starting Materials and Intermediates

Preparation of Compound A

[0439] ##STR00200##

Preparation of Compound B

[0440] ##STR00201##

Preparation of Compound C

[0441] ##STR00202##

Preparation of Compound D

[0442] ##STR00203##

Preparation of Compound E

[0443] ##STR00204##

Preparation of Compound F

[0444] ##STR00205##

[0445] To a solution of compound B (446.0 mg, 1.0 mmol, 1.0 eq) and ethanolamine (180.0 mg, 3.0 mmol, 3.0 eq) dissolved in ACN (10.0 mL) was added Cs.sub.2CO.sub.3 (97.5 mg, 0.3 mmol, 0.3 eq), K.sub.2CO.sub.3 (414.0 mg, 3.0 mmol, 3.0 eq) and NaI (14.6 mg, 0.1 mmol, 0.1 eq) at RT. The mixture was stirred for 16 hours at 85° C. LCMS showed the reaction was completed. The mixture was evaporated under reduced pressure and purified by FCC (DCM/MeOH=I/O-20/1) to provide compound F (0.35 g, 82% yield) as yellow oil.

Preparation of Compound G

[0446] ##STR00206##

Step 1: Preparation of Compound G-1

[0447] To a solution of compound A (1.87 g, 12.38 mmol) in EtOH (50 mL) was added 2-(benzyloxy)ethan-1-amine (2.0 g, 8.25 mmol). The reaction was stirred at RT for 12 hours. The reaction mixture was concentrated in vacuo and purified by flash column chromatography (CH.sub.2Cl.sub.2: MeOH=10:1) to give the target product as yellow oil (1.4 g, yield: 43%). LCMS: Rt: 0.824 min; MS m/z (ESI): 394.3 [M+H].sup.+.

Step 2: Preparation of Compound G

[0448] To a solution of compound G-1 (1.4 g, 3.56 mmol) in CH.sub.3CN (50 mL) was added K.sub.2CO.sub.3 (1.47 g, 10.67 mmol), Cs.sub.2CO.sub.3 (350 mg, 1.07 mmol), NaI (160 mg, 1.07 mmol), and compound B (2.24 g, 5.34 mmol). The reaction was stirred at 80° C. for 10 hours. The reaction mixture was concentrated in vacuo. The crude product was purified by flash column chromatography (CH.sub.2Cl.sub.2: MeOH=20:1) to give the target product as yellow oil (2.0 g, yield: 76%). LCMS: Rt: 1.750 min; MS m/z (ESI): 732.6 [M+H].sup.+.

Preparation of Compound H

[0449] ##STR00207##

Preparation of Compound I

[0450] ##STR00208##

Preparation of Compound J

[0451] ##STR00209##

[0452] To a solution of Compound H-1 (2.0 g, 4.0 mmol, 1.0 eq) in DMF (40 mL) was added NaH (320 mg, 8.0 mmol, 2.0 eq) at room temperature. The mixture was stirred at room temperature for 1 hour. Then 1-iodohexane (1.7 g, 8.0 mmol, 2.0 eq) and NaI (120 mg, 0.8 mmol, 0.2 eq) were added. The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=40/1) to give the title compound (1.2 g, 52% yield) as yellow oil. LCMS: Rt: 1.070 min; MS m/z (ESI): 584.4 [M+H].sup.+.

Preparation of Compound K

[0453] ##STR00210##

Preparation of Compound L

[0454] ##STR00211##

Preparation of Compound M

[0455] ##STR00212##

Step 1: Preparation of Compound M-2

[0456] To a solution of compound M-1 (30.0 g, 98.25 mmol) in DMF (800 mL) was added NaCN (9.63 g, 196.5 mmol). The reaction was stirred at 60° C. for 10 hours. The reaction mixture was poured into water (500 ml) and extracted with EtOAc (3×500 mL). The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The crude product was purified by flash column chromatography (EtOAc: PE=1:20) to give the target product as yellow oil (18.3 g, yield: 74%).

Step 2: Preparation of Compound M-3

[0457] To a solution of Compound M-2 (17.0 g, 67.61 mmol) in EtOH (200 mL) was added H.sub.2SO.sub.4 (40 mL). The reaction was stirred at 90° C. for 48 hours. The reaction mixture was poured into water (500 ml) and extracted with EtOAc (3×500 mL). The combined organic layer was washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to give the target product as yellow oil (15 g, yield: 75%).

Step 3: Preparation of Compound M-4

[0458] To a solution of Compound M-3 (14 g, 46.90 mmol) in MeOH (240 mL) and H.sub.2O (60 mL) was added LiOH.Math.H2O (9.84 g, 234.5 mmol). The reaction was stirred at 50° C. for 10 hours. The reaction mixture concentrated in vacuo to give the target product. The crude product was dissolved in water. The residue was adjusted to PH=2 with 6M HCl and extracted with EtOAc (3×500 mL). The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo to give the target product as yellow oil (15 g, yield: 75%).

Step 4: Preparation of Compound M

[0459] To a solution of Compound M-4 (1.0 g, 4.0 mmol) in DCM (10 mL) was added SOCl.sub.2 (0.9 g, 8.0 mmol). The reaction was stirred at 35° C. for 2 hours. Removal of solvent to get the compound M (1.2 g, crude) as brown oil.

Preparation of Compound N

[0460] ##STR00213##

Preparation of compound O

##STR00214##

Preparation of compound P

##STR00215##

Preparation of compound Q

##STR00216##

Step 1: Preparation of Compound Q-1

[0461] To a mixture of NaH (3 g, 74.07 mmol, 2.5 eq) in DMF (30 mL) was added dimethyl malonate (4 g, 30 mmol, 1.0 eq) at 0° C. under N.sub.2. The reaction mixture was stirred at 0° C. for 0.5 hour. The 1-iodohexane (16 g, 75 mmol, 2.5 eq) in DMF (30 ml) was added to it. The reaction mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was poured in water and washed with EA. The combined organic layers were separated and dried over Na.sub.2SO.sub.4. Removal of solvent, FCC to get the compound Q-1 (5.4 g, 59.92%) as colorless oil.

[0462] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 3.70 (s, 6H), 1.88-1.84 (m, 4H), 1.63 (s, 1H), 1.27 (s, 10H), 1.13 (s, 5H), 0.88-0.86 (m, 6H).

Step 2: Preparation of Compound Q-2

[0463] To a solution of compound Q-1 (5.4 g, 17.97 mmol, 1.0 eq) in DMF (100 mL) was added LiCl (7.6 g, 179.7 mmol, 10.0 eq). The reaction mixture was stirred at 120° C. for 12 hours. TLC showed the reaction was complete. The mixture was poured in water and washed with EA. The combined organic layers were separated and dried over Na.sub.2SO.sub.4. Removal of solvent, FCC to get the compound Q-2 (3.1 g, 71.17%) as colorless oil.

[0464] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 3.67 (s, 3H), 2.35-2.31 (m, 1H), 1.61-1.54 (m, 2H), 1.47-1.40 (m, 2H), 1.26 (s, 16H), 0.89-0.86 (m, 6H).

Step 3: Preparation of Compound Q-3

[0465] To a solution of compound Q-2 (3.1 g, 12.79 mmol, 1.0 eq) in THE (100 mL) was added slowly LiAlH.sub.4 (972 mg, 25.58 mmol, 2.0 eq) in 0° C. The reaction mixture was stirred at reflux for 1 hour. TLC showed the reaction was complete. After being cooled to 0° C., the mixture was quenched with successive addition of water (1.3 ml), 15% aq NaOH (1.3 ml) and water (3.9 ml). The resulting mixture was diluted with EA and the precipitate was removed by titration. The filtrate was evaporated under reduced pressure and FCC to get the compound Q-3 (2.7 g, 98.46%) as yellow oil.

[0466] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 3.54 (d, J=5.2 Hz, 2H), 1.47-1.43 (m, 2H), 1.28 (s, 20H), 0.90-0.87 (m, 6H).

Step 4: Preparation of Compound Q

[0467] To a solution of compound Q-3 (1 g, 4.664 mmol, 1.0 eq) in DCM (15 mL) was added 6-bromohexanoic acid (1.1 g, 5.597 mmol, 1.2 eq), EDCI (1.34 g, 6.996 mmol, 1.5 eq), DMAP (114 mg, 0.9328 mmol, 0.2 eq), DIEA (1.2 g, 9.328 mmol, 2.0 eq). The reaction mixture was stirred at 50° C. for 16 hours. TLC showed the reaction was complete. Removal of solvent, FCC to get the compound Q (650 mg, 35.61%) as yellow oil.

Preparation of Compound SM2

[0468] ##STR00217##

Preparation of Compound R

[0469] ##STR00218##

Preparation of Compound S

[0470] ##STR00219##

Preparation of Compound SM5

[0471] ##STR00220##

Step 1: Preparation of Compound SM5-3

[0472] To a solution of compound SM5-1 (5 g, 47.56 mmol, 1.0 eq.) in MeOH (200 mL) was added compound SM5-2 (9.33 g, 95.11 mmol, 2.0 eq.) and Pd/C (1.0 g). The mixture was stirred at RT for 10 hours under H.sub.2. The mixture was filtered, concentrated and purified by column chromatography on silica gel (DCM/MeOH=10/1) to give the title compound (5.0 g, 56% yield) as colorless oil.

Step 2: Preparation of Compound SM5

[0473] To a solution of compound SM5-3 (500 mg, 2.67 mmol, 1.0 eq.) and DIPEA (690 mg, 5.34 mmol, 2.0 eq.) in DCM (20 mL) was added MsCl (305 mg, 2.67 mmol, 1.0 eq.). The mixture was stirred at room temperature for 2 hours. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (450 mg, 63% yield) as yellow oil. It was used in next step without further purification.

Preparation of Compound SM6

[0474] ##STR00221##

7.2 Example 2: Preparation of Compound 1

[0475] ##STR00222##

Step 1: Preparation of Compound 1-1

[0476] A mixture of cyclobutanone (463 mg, 6.6 mmol, 1.0 eq), 2-(benzyloxy)ethan-1-amine (1.0 g, 6.6 mmol, 1.0 eq) and titanium tetraisopropanolate (2.3 g, 7.9 mol, 1.2 eq) in methanol (50 mL) was stirred under argon at room temperature for 16 h. Sodium borohydride (250 mg, 6.6 mmol, 1.0 eq) was then added and the resulting mixture was stirred for an additional 2 h. LCMS showed the reaction was complete. The reaction was quenched with water and then filtered through a pad of celite and washed with MeOH. The filtration was concentrated and extracted with EA. The combined organic layers were dried over Na.sub.2SO.sub.4 and evaporated under reduced pressured and purified by column chromatography on silica gel (DCM/MeOH=50/1) to give the title compound (930 mg, 69% yield) as yellow oil. LCMS: Rt: 0.750 min; MS m/z (ESI): 206.2 [M+H].sup.+.

Step 2: Preparation of Compound 1-2

[0477] To a solution of compound A (915 mg, 3.77 mmol, 1.0 eq) in EtOH (40 mL) was added compound 1-1 (930 mg, 4.53 mmol, 1.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was evaporated under reduced pressured and purified by column chromatography on silica (DCM/MeOH=100/1) to give the title compound (920 mg, 54%) as yellow oil. LCMS: Rt: 0.870 min; MS m/z (ESI): 448.3 [M+H].sup.+.

Step 3: Preparation of Compound 1-3

[0478] To a stirred solution of compound 1-2 (500 mg, 1.12 mmol, 1.0 eq) in DMF (10 mL) was added NaH (58 mg, 1.46 mmol, 1.3 eq) in 0° C. The mixture was stirred at RT for 2 hours. Then 1-bromododecane 416 mg, 1.68 mmol, 1.5 e.q) was added to this mixture. The mixture was stirred at 120° C. for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=2/1) to give the title compound (300 mg, 43% yield) as yellow oil. LCMS: Rt: 1.360 min; MS m/z (ESI): 616.5 [M+H].sup.+.

Step 4: Preparation of Compound 1

[0479] To a solution of 1-3 (300 mg, 0.48 mmol, 1.0 eq) in MeOH (10 mL) were added Pd/C (64 mg) and concentrated HCl (4 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by prep-HPLC to give the title compound (16 mg, 6.5% yield) as yellow oil.

[0480] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.79-0.83 (m, 6H), 1.14-1.26 (m, 38H), 1.47-1.61 (m, 6H), 1.86-1.96 (m, 4H), 2.51-2.58 (m, 4H), 3.17 (s, 1H), 3.32-3.44 (m, 5H), 3.51-3.66 (m, 3H). LCMS: Rt: 0.94 min; MS m/z (ESI): 526.5 [M+H].sup.+.

7.3 Example 3: Preparation of Compound 2

[0481] ##STR00223##

Step 1: Preparation of Compound 2-1

[0482] To a stirred solution of 1-2 (300 mg, 0.67 mmol, 1.0 eq), DIPEA (260 mg, 2.01 mmol, 3.0 eq) in DCM (10 mL) were added dodecanoyl chloride (293 mg, 1.34 mmol, 2.0 eq) and DMAP (16 mg, 0.134 mmol, 0.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=150/1) to give the title compound (320 mg, 76% yield) as yellow oil. LCMS: Rt: 1.340 min; MS m/z (ESI): 630.5 [M+H].sup.+.

Step 2: Preparation of Compound 2

[0483] To a solution of Compound 2-1 (320 mg, 0.51 mmol, 1.0 eq) in MeOH (10 mL) were added Pd/C (64 mg) and concentrated HCl (4 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by prep-HPLC to give the title compound (75 mg, 27% yield) as yellow oil.

[0484] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 6H), 1.25-1.33 (m, 35H), 1.50-1.69 (m, 7H), 1.87-1.99 (m, 1H), 2.00-2.08 (m, 2H), 2.33 (t, J=7.6 Hz, 2H), 2.56-2.81 (m, 4H), 3.17-3.27 (m, 1H), 3.38-3.48 (m, 3H), 3.50-3.65 (m, 3H), 5.08-5.14 (m, 1H). LCMS: Rt: 1.180 min; MS m/z (ESI): 540.4 [M+H].sup.+.

7.4 Example 4: Preparation of Compound 3

[0485] ##STR00224##

Compound 3-1

[0486] LCMS: Rt: 1.140 min; MS m/z (ESI): 546.4 [M+H].sup.+.

Compound 3

[0487] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.93 (m, 6H), 1.25-1.33 (m, 23H), 1.49-1.69 (m, 7H), 1.90-1.99 (m, 1H), 2.00-2.12 (m, 2H), 2.34 (t, J=7.6 Hz, 2H), 2.56-2.79 (m, 4H), 3.20-3.29 (m, 1H), 3.38-3.48 (m, 3H), 3.50-3.67 (m, 3H), 5.09-5.17 (m, 1H). LCMS: Rt: 0.890 min; MS m/z (ESI): 456.4 [M+H].sup.+.

7.5 Example 5: Preparation of Compound 4

[0488] ##STR00225##

Compound 4-1

[0489] LCMS: Rt: 1.380 min; MS m/z (ESI): 786.6 [M+H].sup.+.

Compound 4

[0490] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.80-0.83 (m, 13H), 1.20-1.26 (m, 58H), 1.48-1.51 (m, 2H), 1.52-1.55 (m, 7H), 2.24 (m, 1H), 3.34 (m, 1H), 3.46-3.48 (m, 2H), 3.89-3.90 (m, 1H). LCMS: Rt: 1.050 min; MS m/z (ESI): 696.6 [M+H].sup.+.

7.6 Example 6: Preparation of Compound 5

[0491] ##STR00226##

Step 1: Preparation of Compound 5-1

[0492] A mixture of compound A (1.0 g, 4.1 mmol, 1 equiv) and N-methyl-1-phenyl-methanamine (1 g, 8.2 mmol, 2 equiv) in EtOH (15 mL) was stirred overnight at room temperature. The mixture was concentrated under vacuum and purified by silica gel column chromatography (PE:EA=1:0 to 5:3) to give the desired product (1.3 g, 86.7% yield) as yellow oil. LCMS: Rt: 0.880 min; MS m/z (ESI): 364.3 [M+H].sup.+.

Step 2: Preparation of Compound 5-2

[0493] To a stirred solution of compound 5-1 (1 g, 2.8 mmol, 1 equiv) in DMF (10 ML) was added sodium hydride (164 mg, 4.1 mmol, 1.5 equiv) at room temperature. After 0.5 h, 1-iodohexane (875 mg, 4.1 mmol, 1.5 equiv) was added to the mixture. The mixture was stirred overnight at 110° C. The mixture was quenched with water (10ML), extracted over EA (3×10 ML). The combined organic layers were washed with brine (2×10ML), dried over anhydrous sodium sulfate, and then concentrated under vacuum. The residual was purified by silica gel column chromatography (PE:EA=1:0 to 10:1) to give the desired product 9001-119-3 (340 mg, 27.6% yield) as yellow oil. LCMS: Rt: 1.080 min; MS m/z (ESI): 448.4 [M+H].sup.+.

Step 3: Preparation of Compound 5-3

[0494] A mixture of compound 5-2 (340 mg, 0.76 mmol, 1 equiv) and Pd/C (90 mg, 10%) in MeOH (10 ML) was stirred overnight. The mixture was filtered, the filtrate was concentrated under vacuum to give the desired product (250 mg, crude) as colorless oil. LCMS: Rt: 0.997 min; MS m/z (ESI): 358.4 [M+H].sup.+.

Step 4: Preparation of Compound 5-4

[0495] A mixture of compound 5-3 (250 mg, 0.7 mmol, 1.0 equiv), 2-bromoethanol (175 mg, 1.4 mmol, 2 equiv) and DIEA (181 mg, 1.4 mmol, 2 equiv) in EtOH (16 ML) was stirred overnight at 70° C. The mixture was concentrated under vacuum to give the desired product (444 mg, crude) as colorless semi-solid. LCMS: Rt: 1.020 min; MS m/z (ESI): 402.4 [M+H].sup.+.

Step 5: Preparation of Compound 5-5

[0496] A mixture of compound 5-4 (330 mg, 0.82 mmol, 1 equiv) and thionyl chloride (294 mg, 02.46 mmol, 3 equiv) in DCM (6 ML) was stirred overnight at room temperature. The mixture was concentrated under vacuum to give the desired product (331 mg, crude) as brown oil. LCMS: Rt: 1.127 min; MS m/z (ESI): 420.3 [M+H].sup.+.

Step 6: Preparation of Compound 5

[0497] A mixture of 5-5 (220 mg, 0.52 mmol, 1 equiv), compound D (203 mg, 1.56 mmol, 3 equiv), DIEA (338 mg, 2.60 mmol, 5 equiv) and catalyst of NaI in THE (6 ML) was stirred overnight at 70° C. The mixture was concentrated under vacuum, the residual was purified by prep-HPLC to give the desired product (23 mg, 12.8% yield) as yellow oil.

[0498] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.84-0.93 (m, 6H), 1.13-1.40 (m, 27H), 1.48-1.61 (m, 7H), 1.66-1.77 (m, 2H), 1.80-1.94 (m, 2H), 2.48 (s, 3H), 2.67-3.05 (m, 7H), 3.35-3.51 (m, 5H), 3.54-3.81 (m, 4H). LCMS: Rt: 0.960 min; MS m/z (ESI): 513.5 [M+H].sup.+.

7.7 Example 7: Preparation of Compound 6

[0499] ##STR00227##

Step 1: Preparation of Compound 6-1

[0500] To a stirred solution of 5-1 (1.4 g, 3.9 mmol, 1 equiv) in DMF (14 mL) was added sodium hydride (231 mg, 5.8 mmol, 1.5 equiv) at room temperature. After 0.5 h, 1-bromododecane (1.45 g, 5.8 mmol, 1.5 equiv) and catalyst of NaI were added to the mixture. The mixture was stirred overnight at 110° C. The mixture was quenched with water (15 mL), extracted over EA (3×15 mL). The combined organic layers were washed with brine (2×15 mL), dried over anhydrous sodium sulfate, and then concentrated under vacuum. The residual was purified by silica gel column chromatography (PE:EA=1:0 to 1:1) to give the desired product (244 mg, 11.9% yield) as brown oil. LCMS: Rt: 1.680 min; MS m/z (ESI): 532.5 [M+H].sup.+.

Step 2: Preparation of Compound 6-2

[0501] A mixture of 6-1 (244 mg, 0.46 mmol, 1 equiv) and Pd/C (100 mg, 10%) in MeOH (5 mL) was stirred overnight under hydrogen atmosphere. The mixture was filtered, the filtrate was concentrated under vacuum to give the desired product (146 mg, crude) as light yellow oil. LCMS: Rt: 1.250 min; MS m/z (ESI): 442.4 [M+H].sup.+.

Step 3: Preparation of Compound 6-3

[0502] A mixture of 6-2 (146 mg, 0.33 mmol, 1.0 equiv), 2-bromoethanol (83 mg, 0.66 mmol, 2 equiv) and DIEA (86 mg, 0.66 mmol, 2 equiv) in EtOH (5 mL) was stirred overnight at 70° C. The mixture was concentrated under vacuum to give the desired product (438 mg, crude) as yellow oil. LCMS: Rt: 1.270 min; MS m/z (ESI): 486.5 [M+H].sup.+.

Step 4: Preparation of Compound 6-4

[0503] A mixture of 6-3 (219 mg, 0.45 mmol, 1 equiv) and thionyl chloride (161 mg, 1.35 mmol, 3 equiv) in DCM (6 mL) was stirred overnight at 35° C. The mixture was concentrated under vacuum to give the desired product (133 mg, crude) as yellow oil. LCMS: Rt:1.860 min; MS m/z (ESI): 504.4 [M+H].sup.+.

Step 5: Preparation of Compound 6

[0504] A mixture of compound 6-4 (133 mg, 0.26 mmol, 1 equiv), compound E (114 mg, 0.79 mmol, 3 equiv), DIEA (170 mg, 1.32 mmol, 5 equiv) and catalyst of NaI in THE (3 ML) was stirred overnight at 70° C. The mixture was concentrated under vacuum, the residual was purified by prep-HPLC to give the desired product compound 6 (19 mg, 11.8% yield) as yellow oil.

[0505] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.81-0.93 (m, 6H), 0.99-1.14 (m, 2H), 1.26 (s, 39H), 1.49-1.59 (m, 4H), 1.61-1.70 (m, 1H), 1.76-1.87 (m, 2H), 1.90-2.19 (m, 9H), 2.53-2.95 (m, 6H), 3.35-3.53 (m, 5H), 3.54-3.75 (m, 3H). LCMS: Rt: 1.500 min; MS m/z (ESI): 611.6 [M+H].sup.+.

7.8 Example 8: Preparation of Compound 7

[0506] ##STR00228##

Step 1: Preparation of Compound 7-1

[0507] To a solution of N1,N1-dimethylethane-1,2-diamine (1.2 g, 13.6 mmol, 2.0 eq) and compound B (2.8 g, 6.8 mmol, 1.0 eq) in ACN (100 mL) were added K.sub.2CO.sub.3 (2.8 g, 20.4 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (665 mg, 2.04 mmol, 0.3 eq) and NaI (306 mg, 2.04 mmol, 0.3 eq). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=50/1-3011) to give the title compound (900 mg, 31% yield) as yellow oil. LCMS: Rt: 0.780 min; MS m/z (ESI): 427.4 [M+H].sup.+.

Step 2: Preparation of Compound 7

[0508] To a solution of compound A (190 mg, 0.78 mol, 1.0 eq) in EtOH (20 mL) was added 7-1 (400 mg, 0.94 mmol, 1.2 eq). The mixture was stirred at room temperature for 48 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure and purified by prep-HPLC to give the title compound (25 mg, 5% yield) as white solid.

[0509] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.35 (m, 45H), 1.41-1.67 (m, 7H), 2.28-2.32 (m, 3H), 2.36-2.70 (m, 11H), 2.79-2.83 (m, 2H), 3.35-3.46 (m, 4H), 3.77-3.85 (m, 1H), 3.96-3.97 (m, 2H). LCMS: Rt: 1.220 min; MS m/z (ESI): 669.6 [M+H].sup.+.

7.9 Example 9: Preparation of Compound 8

[0510] ##STR00229##

Step 1: Preparation of Compound 8-1

[0511] To a solution of compound A (500 mg, 2.06 mol, 1.0 eq) in EtOH (20 mL) was added 2-(piperidin-1-yl)ethan-1-amine (529 mg, 4.12 mmol, 2.0 eq). The mixture was stirred at room temperature for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure and purified by column chromatography on silica gel (DCM/MeOH=50/1-3011) to give the title compound (480 mg, 63% yield) as yellow oil. LCMS: Rt: 0.730 min; MS m/z (ESI): 371.3 [M+H].sup.+.

Step 2: Preparation of Compound 8

[0512] To a solution of 8-1 (480 mg, 1.30 mmol, 1.0 eq) and compound B (1.1 g, 2.60 mmol, 2.0 eq) in ACN (50 mL) were added K.sub.2CO.sub.3 (539 mg, 3.90 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (98 mg, 0.39 mmol, 0.3 eq) and NaI (58 mg, 0.39 mmol, 0.3 eq). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated and purified by column chromatography on silica gel (DCM/MeOH=100/1-7011) to give the title compound (600 mg, 65% yield) as yellow oil. Then 150 mg product was further purified by prep-HPLC to give the title compound (43 mg) as yellow oil.

[0513] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.25-1.27 (m, 47H), 1.40-1.49 (m, 4H), 1.56-1.73 (m, 8H), 2.30 (t, J=7.6 Hz, 3H), 2.40-2.82 (m, 10H), 3.32-3.38 (m, 1H), 3.43-3.46 (m, 3H), 3.70-3.80 (m, 1H), 3.92-3.97 (m, 2H). LCMS: Rt: 1.090 min; MS m/z (ESI): 709.6 [M+H].sup.+.

7.10 Example 10: Preparation of Compound 9

[0514] ##STR00230##

[0515] To a solution of 8 (120 mg, 0.17 mol, 1.0 eq) and DIPEA (66 mg, 0.51 mmol, 3.0 eq) in DCM (8 mL) at 0° C. was added acetic anhydride (35 mg, 0.34 mmol, 2.0 eq). The mixture was stirred at room temperature for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated under reduced pressure and purified by prep-HPLC to give the title compound (28 mg, 22% yield) as yellow oil.

[0516] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.25-1.27 (m, 47H), 1.41-1.48 (m, 4H), 1.53-1.67 (m, 8H), 2.06 (s, 3H), 2.29 (t, J=7.4 Hz, 2H), 2.45-2.75 (m, 10H), 3.35-3.52 (m, 4H), 3.95-3.97 (m, 2H), 4.98-5.00 (m, 1H). LCMS: Rt: 1.260 min; MS m/z (ESI): 751.6 [M+H].sup.+.

7.11 Example 11: Preparation of Compound 10

[0517] ##STR00231##

Compound 10-1

[0518] LCMS: Rt: 0.880 min; MS m/z (ESI): 357.3 [M+H].sup.+.

Compound 10

[0519] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.83-0.93 (m, 9H), 1.16-1.37 (m, 43H), 1.43-1.69 (m, 8H), 1.92-2.24 (m, 11H), 2.27-2.35 (m, 3H), 2.44-2.65 (m, 4H), 3.35-3.50 (m, 4H), 3.78-3.90 (m, 1H), 3.93-4.01 (m, 2H). LCMS: Rt: 1.660 min; MS m/z (ESI): 695.6 [M+H].sup.+.

7.12 Example 12: Preparation of Compound 11

[0520] ##STR00232##

Compound 11-1

[0521] LCMS: Rt: 0.880 min; MS m/z (ESI): 359.3 [M+H].sup.+.

Compound 11

[0522] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.05 (s, 6H), 1.25 (s, 45H), 1.45-1.65 (m, 7H), 2.28-2.32 (m, 2H), 2.43-2.70 (m, 12H), 3.43-3.44 (m, 4H), 3.75 (s, 1H), 3.96-3.97 (m, 2H). LCMS: Rt: 1.060 min; MS m/z (ESI): 697.6 [M+H].sup.+.

7.13 Example 13: Preparation of Compound 13

[0523] ##STR00233##

[0524] To a solution of compound F (660 mg, 1.65 mmol) in EtOH (20 mL) was added COMPOUND A (200 mg, 0.82 mmol). The reaction was stirred at RT for 10 hours. The reaction mixture was concentrated in vacuo and purified by prep-HPLC to give the target product as yellow oil (20 mg, yield: 4%).

[0525] .sup.1HNMR (400 MHz, CDCl.sub.3): δ: 0.87 (t, J=8 Hz, 9H), 1.35-1.68 (m, 35H), 1.43-1.91 (m, 18H), 2.29-2.33 (m, 2H), 2.51-2.71 (m, 6H), 3.35-3.47 (m, 4H), 3.58-3.63 (m, 2H), 3.83-3.86 (m, 1H), 3.96-3.97 (m, 2H). LCMS: Rt: 1.059 min; MS m/z (ESI): 642.5 [M+H].sup.+.

7.14 Example 14: Preparation of Compound 14

[0526] ##STR00234##

Step 1: Preparation of 14-1

[0527] To a solution of compound G (500 mg, 0.68 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added DIEA (300 mg, 2.05 mmol) and hexanoyl chloride (185 mg, 1.37 mmol). The reaction was stirred at RT for 2 hours. The reaction mixture was poured into NaHCO.sub.3 (aq) (50 ml) and extracted with CH.sub.2Cl.sub.2 (3×50 mL). The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The crude product was purified by flash column chromatography (PE: EtOAc=5:1) to give the target product as yellow oil (280 mg, yield: 48%).

Step 4: Preparation of Compound 14

[0528] To a solution of compound 14-1 (250 mg, 0.3 mmol) in MeOH (10 mL) was added HCl (12N) (0.3 mL) and Pd/C (50 mg). The reaction was stirred at RT for 1 hour under H.sub.2. The reaction mixture was filtrated concentrated in vacuo. The crude product was purified by prep-HPLC to give the target product as yellow oil (30 mg, yield: 13%).

[0529] .sup.1HNMR (400 MHz, CDCl.sub.3): δ: 0.87 (t, J=8 Hz, 12H), 1.11-1.31 (m, 42H), 1.39-1.68 (m, 17H), 2.28-2.69 (m, 11H), 3.39-3.55 (m, 4H), 3.96-3.97 (m, 2H), 5.05-5.08 (m, 1H). LCMS: Rt: 1.932 min; MS m/z (ESI): 740.6 [M+H].sup.+.

7.15 Example 15: Preparation of Compound 15

[0530] ##STR00235##

[0531] compound 15 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.91 (m, 12H), 1.25-1.39 (m, 51H), 1.40-1.47 (m, 3H), 1.50-1.66 (m, 11H), 2.27-2.31 (m, 4H), 2.45-2.65 (m, 10H), 3.35-3.51 (m, 4H), 3.95-3.97 (m, 2H), 4.99-5.04 (m, 1H). LCMS: Rt: 1.540 min; MS m/z (ESI): 807.6 [M+H].sup.+.

7.16 Example 16: Preparation of Compound 16

[0532] ##STR00236##

Step 1: Preparation of Compound 16-1

[0533] To a solution of compound H (400 mg, 0.94 mmol, 1.2 eq) and 1-bromohexane (165 mg, 0.78 mmol, 1.0 eq) in ACN (15 mL) were added K.sub.2CO.sub.3 (323 mg, 2.34 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (76 mg, 0.23 mmol, 0.3 eq) and NaI (35 mg, 0.23 mmol, 0.3 eq). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated and purified by column chromatography on silica gel (DCM/MeOH=70/1) to give the title compound (290 mg, 73% yield) as yellow oil. LCMS: Rt: 1.050 min; MS m/z (ESI): 512.5 [M+H].sup.+

Step 2: Preparation of Compound 16-2

[0534] To a solution of 16-1 (290 mg, 0.57 mmol, 1.0 eq) in EtOH (10 mL) were added 2-bromoethanol (143 mg, 1.14 mmol, 2.0 eq) and DIPEA (221 mg, 1.71 mmol, 3.0 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the desired MS was detected. The reaction mixture was concentrated under reduced pressure. The residue was diluted with DCM and washed with water, brine, dried over Na.sub.2SO.sub.4 and concentrated give the title compound (300 mg, 95%) as yellow oil. It was used in the next step without further purification. LCMS: Rt: 1.220 min; MS m/z (ESI): 556.5 [M+H].sup.+.

Step 3: Preparation of Compound 16-3

[0535] To a solution of 16-2 (300 mg, 0.54 mmol, 1.0 eq) in DCM (10 mL) was added SOCl.sub.2 (193 mg, 1.62 mmol, 3.0 eq). The mixture was stirred at 30° C. for 16 hours. LCMS showed the reaction was complete. The mixture was washed with saturated NaHCO.sub.3 solution, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=10/1) to give the title compound (120 mg, 39% yield) as yellow oil. LCMS: Rt: 0.950 min; MS m/z (ESI): 538.5 [M-Cl].sup.+.

Step 4: Preparation of Compound 16

[0536] To a solution of 16-3 (120 mg, 0.21 mmol, 1.0 eq) and compound I (73 mg, 0.63 mmol, 3.0 eq) in THF (10 mL) were added DIPEA (136 mg, 1.05 mmol, 5.0 eq) and NaI (6 mg, 0.042 mmol, 0.2 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC to give the title compound (27 mg, 20% yield) as yellow oil.

[0537] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.39 (m, 44H), 1.47-1.70 (m, 8H), 1.83-2.19 (m, 6H), 2.41-2.74 (m, 8H), 3.40-3.62 (m, 9H). LCMS: Rt: 1.290 min; MS m/z (ESI): 653.6 [M+H].sup.+.

7.17 Example 17: Preparation of Compound 17

[0538] ##STR00237##

Compound 17-1

[0539] LCMS: Rt: 1.210 min; MS m/z (ESI): 596.6 [M+H].sup.+.

Compound 17-2

[0540] LCMS: Rt: 1.740 min; MS m/z (ESI): 640.5 [M+H].sup.+.

Compound 17

[0541] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 3.61-3.41 (m, 8H), 3.19-3.14 (m, 1H), 2.63-2.45 (m, 9H), 2.03-1.84 (m, 6H), 1.66-1.41 (m, 9H), 1.26 (s, 54H), 0.90-0.86 (m, 9H). LCMS: Rt: 1.860 min; MS m/z (ESI): 737.6 [M+H].sup.+.

7.18 Example 18: Preparation of Compound 18

[0542] ##STR00238##

Step 1: Preparation of Compound 18-1

[0543] To a solution of compound G (500 mg, 0.68 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added DIEA (270 mg, 2.05 mmol) and AcCl (110 mg, 1.37 mmol). The reaction was stirred at RT for 2 hours. The reaction mixture was poured into NaHCO.sub.3 (aq) (50 ml) and extracted with CH.sub.2Cl.sub.2 (3×100 mL). The combined organic layers were washed with brine, dried over anhydrous Na.sub.2SO.sub.4 and concentrated in vacuo. The crude product was purified by flash column chromatography (PE: EtOAc=5:1) to give the target product as yellow oil (300 mg, yield: 57%). LCMS: Rt: 2.360 min; MS m/z (ESI): 774.6 [M+H].sup.+.

Step 2: Preparation of Compound 18

[0544] To a solution of 18-1 (300 mg, 0.39 mmol) in MeOH (10 mL) was added HCl (12N) (0.3 mL) and Pd/C (50 mg). The reaction was stirred at rt for 1 hour under H.sub.2. The reaction mixture was filtrated concentrated in vacuo. The crude product was purified by prep-HPLC to give the target product as yellow oil (30 mg, yield: 11%).

[0545] .sup.1HNMR (400 MHz, CDCl.sub.3): δ: 0.87 (t, J=8 Hz, 9H), 1.11-1.31 (m, 42H), 1.39-1.68 (m, 9H), 2.04 (s, 3H), 2.28-2.32 (m, 2H), 2.48-2.73 (m, 6H), 2.89-2.91 (m, 1H), 3.39-3.55 (m, 6H), 3.96-3.97 (m, 2H), 5.05-5.08 (m, 1H). LCMS: Rt: 1.530 min; MS m/z (ESI): 684.5 [M+H].sup.+.

7.19 Example 19: Preparation of Compound 19

[0546] ##STR00239##

Step 1: Preparation of Compound 19-1

[0547] A mixture of compound G (320 mg, 0.44 mmol, 1 equiv), propionic anhydride (171 mg, 1.32 mmol, 3 equiv) and DIEA (568 mg, 4.40 mmol, 10 equiv) in DCM (5 Ml) was stirred overnight at room temperature. The mixture was concentrated under vacuum. The residual was purified by silica gel column chromatography (PE:EA=60:1 to 10:1 to give the desired product (340 mg, 98.8% yield) as colorless oil. LCMS: Rt: 2.170 min; MS m/z (ESI): 788.6 [M+H].sup.+.

Step 2: Preparation of Compound 19

[0548] A mixture of 19-1 (340 mg, 0.43 mmol, 1.0 equiv), a drop of HCl (36.5%) and Pd/C (60 mg) in MeOH (15 mL) was stirred overnight at room temperature. The mixture was filtered through celite. The filer cake was washed with MeOH (8×10 mL). The filtrate was concentrated under vacuum. The residual was purified by prep-HPLC to give the desired product (34 mg, 11.3% yield) as light brown oil.

[0549] .sup.1HNMR (400 MHz, CDCl.sub.3): δ: 0.83-0.92 (m, 9H), 0.93-0.99 (m, 1H), 1.08-1.18 (m, 3H), 1.19-1.37 (m, 42H), 1.39-1.76 (m, 13H), 2.16-2.42 (m, 4H), 2.44-2.81 (m, 3H), 3.35-3.65 (m, 5H), 3.92-4.03 (m, 2H). LCMS: Rt: 1.480 min; MS m/z (ESI): 698.5 [M+H].sup.+.

7.20 Example 20: Preparation of Compound 20

[0550] ##STR00240##

Step 1: Preparation of Compound 20-1

[0551] To a solution of compound 18 (1.0 g, 1.46 mmol, 1.0 eq) in DCM (30 mL) was added SOCl.sub.2 (521 mg, 4.38 mmol, 3.0 eq). The mixture was stirred at 30° C. for 16 hours. LCMS showed the reaction was complete. The mixture was concentrated under reduced pressure to give the title compound (1.0 g, 97%) as brown oil. LCMS: Rt: 0.960 min; MS m/z (ESI): 666.5 [M-Cl].sup.+.

Step 2: Preparation of Compound 20

[0552] To a solution of 20-1 (250 mg, 0.36 mmol, 1.0 eq) and compound I (124 mg, 1.08 mmol, 3.0 eq) in THE (10 mL) were added DIPEA (233 mg, 1.80 mmol, 3.0 eq) and NaI (11 mg, 0.072 mmol, 0.2 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC to give the title compound (33 mg, 12% yield) as yellow oil.

[0553] .sup.1HNMR (400 MHz, CDCl.sub.3): δ: 0.86-0.90 (m, 9H), 1.26-1.40 (m, 45H), 1.48-1.68 (m, 9H), 1.81-2.07 (m, 8H), 2.31 (t, J=7.4 Hz, 2H), 2.40-2.55 (m, 5H), 2.61-2.75 (m, 3H), 3.07-3.17 (m, 2H), 3.34-3.51 (m, 4H), 3.71-3.80 (m, 1H), 3.96-3.97 (m, 2H), 4.09-4.14 (m, 2H). LCMS: Rt: 1.330 min; MS m/z (ESI): 781.6 [M+H].sup.+.

7.21 Example 21: Preparation of Compound 21

[0554] ##STR00241##

[0555] compound 21 .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 4.09-4.06 (m, 1H), 3.96 (d, J=6 Hz, 2H), 3.75 (s, 1H), 3.46-3.38 (m, 4H), 2.72-2.45 (m, 10H), 2.32-2.28 (m, 2H), 2.05 (s, 3H), 1.78 (d, J=9.2 Hz, 4H), 1.65-1.47 (m, 9H), 1.26 (d, J=3.2 Hz, 51H), 0.90-0.86 (m, 9H). LCMS: Rt: 1.430 min; MS m/z (ESI): 809.6 [M+H].sup.+.

7.22 Example 22: Preparation of Compound 22

[0556] ##STR00242##

Step 1: Preparation of Compound 22-1

[0557] A mixture of compound J (1.0 g, 1.7 mmol, 1 equiv) and Pd/C (100 mg) in MeOH (100 mL) was stirred overnight at room temperature under hydrogen atmosphere. The mixture was filtered with celite; the filtrate was concentrated to give the desired product 22-1 (504 mg, 85.6% yield) as light brown oil. LCMS: Rt: 0.840 min; MS m/z (ESI): 344.3 [M+H].sup.+.

Step 2: Preparation of Compound 22-2

[0558] A mixture of compound 22-1 (504 mg, 1.47 mmol, 1 equiv), potassium carbonate (607 mg, 4.40 mmol, 3 equiv), cesium carbonate (13 mg, 0.04 mmol, 0.03 equiv), compound B (493 mg, 1.18 mmol, 0.8 equiv) and sodium iodide (50 mg, 0.33 mmol, 0.23 equiv) in ACN (35 mL) was stirred overnight at 80° C. The mixture was filtered. The filtrate was concentrated. The residual was purified by silica gel column chromatography (MeOH:DCM=0% to 2.5%) to give the desired product (500 mg, 49.9% yield) as yellow oil. LCMS: Rt: 1.140 min; MS m/z (ESI): 682.6 [M+H].sup.+.

Step 3: Preparation of Compound 22-3

[0559] A mixture of compound 22-2 (500 mg, 0.73 mmol, 1 equiv), 2-bromoethanol (138 mg, 1.10 mmol, 1.5 equiv), DIEA (283 mg, 2.19 mmol, 3 equiv) and sodium iodide (50 mg, 0.33 mmol, 0.45 mmol) in THF (10 mL) was stirred overnight at 70° C. The mixture was washed with water, extracted over EA (3×10 mL), dried and concentrated under vacuum to give the desired product (433 mg, crude) as yellow oil. LCMS: Rt: 1.550 in; MS m/z (ESI): 726.6 [M+H].sup.+.

Step 4: Preparation of Compound 22-4

[0560] A mixture of compound 22-3 (433 mg, 0.60 mmol, 1 equiv) and SOCl.sub.2 (114 mg, 1.8 mmol, 3 equiv) in DCM (8 mL) was stirred overnight at 35° C. The mixture was concentrated, the residual was purified by silica gel column chromatography (MeOH:DCM=0:100 to 1:100) to give the desired product 22-4 (230 mg, 51.8% yield).

Step 5: Preparation of Compound 22

[0561] A mixture of compound 22-4 (115 mg, 0.15 mmol, 1 equiv), compound k (47 mg, 0.46 mmol, 3 equiv), sodium iodide (15 mg, 0.10 mmol, 0.6 equiv) and DIEA (100 mg, 0.78 mmol, 3 equiv) in THF (5 mL) was stirred overnight at 70° C. The mixture was concentrated under vacuum. The residual was purified by prep-HPLC to give the desired product (23 mg, 18.4% yield) as light brown oil.

[0562] .sup.1H NMR (400 MHz, CDCl3): δ: 0.82-0.94 (m, 12H), 1.12-1.38 (m, 53H), 1.49-1.69 (m, 8H), 1.73-2.07 (m, 10H), 2.18-2.25 (m, 1H), 2.26-2.37 (m, 2H), 2.87 (s, 1H), 3.04-3.37 (m, 4H), 3.38-3.53 (m, 4H), 3.54-3.79 (m, 2H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 1.320 min; MS m/z (ESI): 809.7 [M+H].sup.+.

7.23 Example 23: Preparation of Compound 23

[0563] ##STR00243##

[0564] compound 23 .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 0.87-0.90 (m, 12H), 1.26-1.40 (m, 52H), 1.50-1.68 (m, 10H), 2.08-2.23 (m, 7H), 2.31 (t, J=7.6 Hz, 2H), 2.42-2.57 (m, 3H), 2.76-2.84 (m, 3H), 2.98-3.18 (m, 4H), 3.42-3.57 (m, 4H), 3.61-3.65 (m, 2H), 3.79-3.85 (m, 1H), 3.95-3.97 (m, 2H). LCMS: Rt: 1.480 min; MS m/z (ESI): 823.6 [M+H].sup.+.

7.24 Example 24: Preparation of Compound 24

[0565] ##STR00244##

[0566] compound 24 .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 0.82-0.94 (m, 12H), 1.19-1.40 (m, 47H), 1.41-1.69 (m, 13H), 1.73-2.14 (m, 13H), 2.18-2.36 (m, 3H), 2.43-2.84 (m, 4H), 3.01-3.25 (m, 2H), 3.35-3.65 (m, 7H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 1.560 min; MS m/z (ESI):837.7 [M+H].sup.+.

7.25 Example 25: Preparation of Compound 25

[0567] ##STR00245##

[0568] compound 25 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.82-0.95 (m, 12H), 1.16-1.38 (m, 50H), 1.40-1.87 (m, 26H), 1.96-2.07 (m, 2H), 2.18-2.34 (m, 2H), 2.39-2.66 (m, 6H), 3.35-3.75 (m, 6H), 3.96 (d, J=5.6 Hz, 1H). LCMS: Rt: 1.980 min; MS m/z (ESI): 851.7 [M+H].sup.+.

7.26 Example 26: Preparation of Compound 26

[0569] ##STR00246##

[0570] compound 26 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.81-0.93 (m, 12H), 1.15-1.36 (m, 50H), 1.38-1.87 (m, 23H), 1.75-1.87 (m, 2H), 2.18-2.34 (m, 2H), 2.39-2.66 (m, 9H), 3.35-3.52 (m, 7H), 3.53-3.62 (m, 1H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 1.860 min; MS m/z (ESI): 865.7 [M+H].sup.+.

7.27 Example 27: Preparation of Compound 27

[0571] ##STR00247##

Step 1: Preparation of Compound 27-1

[0572] A mixture of compound H (2.0 g, 4.7 mmol, 1 equiv), potassium carbonate (1.9 g mg, 14.1 mmol, 3 equiv), cesium carbonate (460 mg, 1.4 mmol, 0.3 equiv), compound B (2.2 g, 5.1 mmol, 1.1 equiv) and sodium iodide (210 mg, 1.4 mmol, 0.3 equiv) in ACN (70 mL) was stirred overnight at 80° C. The mixture was filtered. The filtrate was concentrated. The residual was purified by silica gel column chromatography (MeOH:DCM=0% to 2.5%) to give the desired product (1.8 g, 50.3% yield) as yellow oil. LCMS: Rt: 1.970 min; MS m/z (ESI): 767.6 [M+H].sup.+.

Step 2: Preparation of Compound 27-2

[0573] A mixture of 27-1 (1.8 g, 2.3 mmol, 1 equiv), 2-bromoethanol (880 mg, 7.0 mmol, 3.0 equiv), DIEA (910 mg, 7.0 mmol, 3 equiv) in ethanol (30 mL) was stirred for overnight at 80° C. The mixture was diluted with EA and washed with water and brine, concentrated. The residue was purified by a column chromatography to give product (1.6 g, 84.4% yield) as yellow oil. LCMS: Rt: 2.180 in; MS m/z (ESI): 810.7 [M+H].sup.+.

Step 3: Preparation of Compound 27-3

[0574] A mixture of 27-2 (1.6 g, 2.0 mmol, 1 equiv) and SOCl.sub.2 (720 mg, 6.0 mmol, 3 equiv) in DCM (20 mL) was stirred overnight at 35° C. The mixture was concentrated, the residue was purified by column chromatography to give the desired product 27-3 (1.4 g, 84.5% yield) as yellow oil.

Step 4: Preparation of Compound 27

[0575] A mixture of 27-3 (200 mg, 0.24 mmol, 1 equiv), compound K (242 mg, 2.4 mmol, 10 equiv), sodium iodide (18 mg, 0.12 mmol, 0.5 equiv) and DIEA (100 mg, 0.72 mmol, 3 equiv) in THF (5 mL) was stirred overnight at 70° C. The mixture was concentrated under vacuum. The residual was purified by prep-HPLC to give the desired product (56 mg, 26% yield) as light brown oil.

[0576] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.43-0.47 (m, 4H), 0.86-0.90 (m, 12H), 1.26 (s, 63H), 1.43-1.67 (m, 11H), 2.28-2.32 (m, 2H), 2.44-2.62 (m, 5H), 2.72-2.79 (m, 4H), 3.40-3.50 (m, 6H), 3.56-3.58 (m, 3H), 3.97 (d, J=6.0 Hz, 2H). LCMS: Rt: 2.270 min; MS m/z (ESI): 893.7 [M+H].sup.+.

7.28 Example 28: Preparation of Compound 28

[0577] ##STR00248##

[0578] compound 28 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.86-0.90 (m, 12H), 1.26 (s, 59H), 1.44-1.68 (m, 15H), 1.86 (m, 2H), 1.99-2.05 (m, 2H), 2.28-2.32 (m, 2H), 2.45-2.57 (m, 10H), 3.12 (m, 1H), 3.40-3.58 (m, 9H), 3.96 (d, J=6.0 Hz, 2H). LCMS: Rt: 2.20 min; MS m/z (ESI): 907.7 [M+H].sup.+.

7.29 Example 29: Preparation of Compound 29

[0579] ##STR00249##

[0580] compound 29 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.86-0.90 (m, 12H), 1.26 (s, 61H), 1.39-1.75 (m, 22H), 2.28-2.32 (m, 2H), 2.47-2.61 (m, 9H), 3.08-3.12 (m, 1H), 3.40-3.60 (m, 9H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 1.690 min; MS m/z (ESI): 921.7 [M+H].sup.+.

7.30 Example 30: Preparation of Compound 30

[0581] ##STR00250##

[0582] compound 30 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.86-0.90 (m, 12H), 1.26 (s, 63H), 1.51-1.77 (m, 20H), 2.28-2.32 (m, 2H), 2.45-2.61 (m, 10H), 3.40-3.56 (m, 9H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 1.590 min; MS m/z (ESI): 935.8 [M+H].sup.+.

7.31 Example 31: Preparation of Compound 31

[0583] ##STR00251##

[0584] compound 31 .sup.1H NMR (400 MHz, CDCl.sub.3): δ: 0.86-0.90 (m, 12H), 1.26 (s, 61H), 1.39-1.79 (m, 24H), 2.28-2.32 (m, 2H), 2.45-2.58 (m, 10H), 3.40-3.58 (m, 9H), 3.96 (d, J=5.6 Hz, 2H). LCMS: Rt: 2.510 min; MS m/z (ESI): 949.8 [M+H].sup.+.

7.32 Example 32: Preparation of Compound 32

[0585] ##STR00252##

Step 1: Preparation of Compound 32-1

[0586] To a solution of 1-1 (1 g, 4.88 mmol, 1.0 eq) in DMF (10 mL) was added NaH (250 mg, 6.25 mmol, 1.3 eq). The mixture was stirred at 0° C. for 1 hour. 2-(bromomethyl)oxirane (868 mg, 6.25 mmol, 1.3 eq) was added in this mixture. The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was extracted by EA (50 ml), washed by brine (50 ml×3) and evaporated under reduced pressure, purified by column chromatography on silica (PE/EA=5/1) to give the title compound (687 mg, 54%) as yellow oil. LCMS: Rt: 0.74 min; MS m/z (ESI): 262 [M+H].sup.+.

Step 2: Preparation of Compound 32-2

[0587] To a solution of 32-1 (500 mg, 1.92 mmol, 1.0 eq) in H2O (5 mL) was added perchloric acid (0.2 ml). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The mixture was purified by prep-HPLC to give the title compound (220 mg, 41%) as yellow oil. LCMS: Rt: 0.735 min; MS m/z (ESI): 280.2 [M+H].sup.+.

Step 3: Preparation of Compound 32-3

[0588] To a stirred solution of 32-2 (188 mg, 0.67 mmol, 1.0 eq), DIPEA (260 mg, 2.01 mmol, 3.0 eq) in DCM (10 mL) were added compound M (578 mg, 2.01 mmol, 3.0 eq) and DMAP (16 mg, 0.134 mmol, 0.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=5/1) to give the title compound (346 mg, 66% yield) as yellow oil.

Step 4: Preparation of Compound 32

[0589] To a solution of 32-3 (320 mg, 0.44 mmol, 1.0 eq) in MeOH (10 mL) were added Pd/C (64 mg) and concentrated HCl (4 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by prep-HPLC to give the title compound (40 mg, 13.2% yield) as yellow oil.

[0590] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.31-1.41 (m, 48H), 2.05-1.61 (m, 10H), 2.19-2.28 (m, 4H), 2.64-2.78 (m, 3H), 3.18-3.21 (m, 1H), 3.22-3.59 (m, 2H), 4.01-4.35 (m, 2H), 5.18-5.22 (m, 1H). LCMS: Rt: 1.240 min; MS m/z (ESI): 694.4 [M+H].sup.+.

7.33 Example 33: Preparation of Compound 33

[0591] ##STR00253##

Step 1: Preparation of Compound 33-1

[0592] To a solution of J-1 (5.5 g, 11.0 mmol, 1.0 eq) in DMF (110 mL) was added NaH (880 mg, 22.0 mmol, 2.0 eq) at room temperature. The mixture was stirred at room temperature for 1 hour. Then 1-bromooctane (4.2 g, 22.0 mmol, 2.0 eq) and NaI (330 mg, 2.2 mmol, 0.2 eq) were added. The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=40/1) to give the title compound (3.5 g, 52% yield) as yellow oil. LCMS: Rt: 1.040 min; MS m/z (ESI): 612.4 [M+H].sup.+.

Step 2: Preparation of Compound 33-2

[0593] To a solution of 33-1 (3.5 g, 5.72 mmol, 1.0 eq) in MeOH (60 mL) was added Pd/C (350 mg). The mixture was stirred at RT under H.sub.2 for 16 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtrate was concentrated and purified by column chromatography on silica gel (DCM/MeOH=30/1-20/1) give the title compound (1.8 g, 86% yield) as light yellow oil. LCMS: Rt: 1.080 min; MS m/z (ESI): 372.3 [M+H].sup.+.

Step 3: Preparation of Compound 33-3

[0594] To a solution of 33-2 (1.8 g, 4.8 mmol, 1.2 eq) and compound B (1.7 g, 4.0 mmol, 1.0 eq) in ACN (50 mL) were added K.sub.2CO.sub.3 (1.6 g, 12.0 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (391 mg, 1.2 mmol, 0.3 eq) and NaI (180 mg, 1.2 mmol, 0.3 eq). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated and purified by column chromatography on silica gel (DCM/MeOH=50/1) to give the title compound (1.7 g, 61% yield) as yellow oil. LCMS: Rt: 1.420 min; MS m/z (ESI): 710.6 [M+H].sup.+.

Step 4: Preparation of Compound 33-4

[0595] To a solution of 33-3 (1.7 g, 2.4 mmol, 1.0 eq) in EtOH (25 mL) were added 2-bromoethanol (600 mg, 4.8 mmol, 2.0 eq) and DIPEA (930 mg, 7.2 mmol, 3.0 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the desired MS was detected. The reaction mixture was concentrated under reduced pressure. The residue was diluted with DCM and washed with water, brine, dried over Na.sub.2SO.sub.4 and concentrated give the title compound (1.8 g, 100%) as yellow oil. It was used in the next step without further purification. LCMS: Rt: 1.980 min; MS m/z (ESI): 754.6 [M+H].sup.+.

Step 5: Preparation of Compound 33-5

[0596] To a solution of 33-4 (1.8 g, 2.39 mmol, 1.0 eq) in DCM (30 mL) was added SOCl.sub.2 (853 mg, 7.17 mmol, 3.0 eq). The mixture was stirred at 30° C. for 16 hours. LCMS showed the reaction was complete. The mixture was washed with saturated NaHCO.sub.3solution, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=10/1) to give the title compound (750 mg, 42% yield) as yellow oil. LCMS: Rt: 1.230 min; MS m/z (ESI): 736.6 [M-Cl].sup.+.

Step 6: Preparation of Compound 33

[0597] To a solution of 33-5 (150 mg, 0.19 mmol, 1.0 eq) and compound I (66 mg, 0.57 mmol, 3.0 eq) in THE (10 mL) were added DIPEA (123 mg, 0.95 mmol, 5.0 eq) and NaI (6 mg, 0.038 mmol, 0.2 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC to give the title compound (65 mg, 40% yield) as yellow oil.

[0598] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.40 (m, 56H), 1.46-1.67 (m, 11H), 1.80-1.92 (m, 1H), 1.96-2.08 (m, 2H), 2.30 (t, J=7.6 Hz, 2H), 2.41-2.73 (m, 10H), 3.15-3.25 (m, 1H), 3.40-3.61 (m, 9H), 3.95-3.97 (m, 2H). LCMS: Rt: 2.250 min; MS m/z (ESI): 851.7 [M+H].sup.+.

7.34 Example 34: Preparation of Compound 34

[0599] ##STR00254##

[0600] compound 34 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.40-0.52 (m, 4H), 0.86-0.90 (m, 12H), 1.26-1.40 (m, 56H), 1.44-1.65 (m, 9H), 1.84-1.92 (m, 1H), 2.30 (t, J=7.6 Hz, 2H), 2.40-2.82 (m, 9H), 3.38-3.63 (m, 9H), 3.95-3.97 (m, 2H). LCMS: Rt: 2.090 min; MS m/z (ESI): 837.7 [M+H].sup.+.

7.35 Example 35: Preparation of Compound 35

[0601] ##STR00255##

[0602] compound 35 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.42 (m, 56H), 1.44-1.55 (m, 9H), 1.56-1.73 (m, 5H), 1.76-1.82 (m, 1H), 2.30 (t, J=7.4 Hz, 2H), 2.47-2.68 (m, 9H), 3.40-3.60 (m, 9H), 3.95-3.97 (m, 2H). LCMS: Rt: 2.520 min; MS m/z (ESI): 865.7 [M+H].sup.+.

7.36 Example 36: Preparation of Compound 36

[0603] ##STR00256##

[0604] compound 36 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.40 (m, 58H), 1.41-1.63 (m, 11H), 1.70-1.85 (m, 4H), 2.05-2.07 (m, 1H), 2.30 (t, J=7.4 Hz, 2H), 2.39-2.65 (m, 9H), 2.85-3.03 (m, 1H), 3.40-3.61 (m, 9H), 3.95-3.97 (m, 2H). LCMS: Rt: 2.480 min; MS m/z (ESI): 879.7 [M+H].sup.+.

7.37 Example 37: Preparation of Compound 37

[0605] ##STR00257##

[0606] compound 37 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.40 (m, 56H), 1.41-1.69 (m, 16H), 1.75-1.84 (m, 2H), 2.07-2.24 (m, 4H), 2.30 (t, J=7.6 Hz, 2H), 2.40-2.61 (m, 5H), 2.74-2.80 (m, 2H), 3.00-3.06 (m, 1H), 3.16-3.23 (m, 1H), 3.40-3.61 (m, 7H), 3.84-3.89 (m, 1H), 3.95-3.97 (m, 2H). LCMS: Rt: 2.520 min; MS m/z (ESI): 893.7 [M+H].sup.+.

7.38 Example 38: Preparation of Compound 38

[0607] ##STR00258##

[0608] compound 38 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 6H), 1.26-1.54 (m, 25H), 1.56-1.70 (m, 6H), 1.91-2.02 (m, 4H), 2.32 (s, 3H), 2.48-2.58 (m, 8H), 3.11-3.23 (m, 1H), 3.41-3.60 (m, 9H). LCMS: Rt: 0.909 min; MS m/z (ESI): 499.4 [M+H].sup.+.

7.39 Example 39: Preparation of Compound 39

[0609] ##STR00259##

[0610] compound 39 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.54 (m, 36H), 1.52-1.67 (m, 6H), 1.91-2.02 (m, 4H), 2.32 (s, 3H), 2.50-2.58 (m, 8H), 3.17-3.21 (m, 1H), 3.43-3.62 (m, 9H). LCMS: Rt: 1.030 min; MS m/z (ESI): 583.5 [M+H].sup.+.

7.40 Example 40: Preparation of Compound 40

[0611] ##STR00260##

Step 1: Preparation of Compound 40-1

[0612] To a solution of 2-(methylamino)ethan-1-ol (7.5 g, 0.10 mol, 1.0 eq) in DCM (300 mL) at 0° C. was added di-tert-butyl dicarbonate (24.0 g, 0.11 mmol, 1.1 eq). The mixture was stirred at room temperature for 1 hour. LCMS showed the reaction was completed. The mixture was washed with water, brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (17.5 g, 100% yield) as colorless oil. LCMS: Rt: 0.820 min; MS m/z (ESI): 120.2 [M-56].sup.+.

Step 2: Preparation of Compound 40-2

[0613] To a stirred solution of 40-1 (12.5 g, 71.2 mol, 1.0 eq) and Benzyl bromide (14.6 g, 85.4 mmol, 1.2 eq) in THE (140 mL) at 0° C. under N.sub.2 was added NaH (3.4 g, 85.4 mmol, 1.2 eq) portionwise. The resulting mixture was stirred at room temperature for 16 hours. TLC showed the reaction was completed. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=15/1) to give the title compound (12.5 g, 60% yield) as colorless oil.

Step 3: Preparation of Compound 40-3

[0614] To a solution of 40-2 (12.5 g, 47.1 mol, 1.0 eq) in DCM (100 mL) under N.sub.2 was added a solution of HCl in 1,4-dioxane (23.6 mL, 94.2 mmol, 2.0 eq, 4.0 M). The resulting mixture was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The mixture was basified to pH=9 with saturated NaHCO.sub.3 aqueous solution. The mixture was extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (5.1 g, 65% yield) as yellow oil. LCMS: Rt: 0.650 min; MS m/z (ESI): 166.2 [M+H].sup.+.

Step 4: Preparation of Compound 40-4

[0615] To a solution of compound A (6.0 g, 25.0 mmol, 1.0 eq) in EtOH (100 mL) was added 40-3 (5.0 g, 30.0 mmol, 1.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was evaporated under reduced pressure and purified by column chromatography on silica (DCM/MeOH=100/1-40/1) to give the title compound (6.1 g, 61%) as yellow oil. LCMS: Rt: 0.890 min; MS m/z (ESI): 408.3 [M+H].sup.+.

Step 5: Preparation of Compound 40-5

[0616] To a stirred solution of 40-4 (300 mg, 0.74 mmol, 1.0 eq), DIPEA (289 mg, 2.22 mmol, 3.0 eq) in DCM (10 mL) were added valeryl chloride (178 mg, 1.48 mmol, 2.0 eq) and DMAP (18 mg, 0.15 mmol, 0.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=100/1) to give the title compound (320 mg, 88% yield) as yellow oil. LCMS: Rt: 0.940 min; MS m/z (ESI): 492.4 [M+H].sup.+.

Step 6: Preparation of Compound 40-6

[0617] To a solution of 40-5 (320 mg, 0.65 mmol, 1.0 eq) in MeOH (15 mL) were added Pd/C (64 mg) and concentrated HCl (4 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by column chromatography on silica gel (DCM/MeOH=75/1) to give the title compound (200 mg, 77% yield) as yellow oil. LCMS: Rt: 0.920 min; MS m/z (ESI): 402.3 [M+H].sup.+.

Step 7: Preparation of Compound 40-7

[0618] To a stirred solution of 40-6 (200 mg, 0.50 mmol, 1.0 eq) and DIPEA (129 mg, 1.0 mmol, 2.0 eq) in DCM (10 mL) at 0° C. was added methane sulfonyl chloride (69 mg, 0.60 mmol, 1.2 eq). The mixture was stirred at RT for 2 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (200 mg, 83% yield) as yellow oil. LCMS: Rt: 1.480 min; MS m/z (ESI): 480.3 [M+H].sup.+.

Step 8: Preparation of Compound 40

[0619] To a solution of 40-7 (200 mg, 0.42 mmol, 1.0 eq) in ACN (10 mL) were added compound I (97 mg, 0.84 mmol, 2.0 eq) and potassium carbonate (174 mg, 1.26 mmol, 3.0 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to give the title compound (200 mg, 83% yield) as yellow oil.

[0620] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.94 (m, 6H), 1.25-1.38 (m, 21H), 1.54-1.62 (m, 6H), 1.87-1.94 (m, 2H), 1.97-2.04 (m, 2H), 2.31 (t, J=7.4 Hz, 3H), 2.39-2.48 (m, 3H), 2.52-2.66 (m, 4H), 2.72-2.76 (m, 3H), 3.13-3.21 (m, 1H), 3.42-3.47 (m, 4H), 3.90-3.97 (m, 1H), 4.11-4.15 (m, 2H). LCMS: Rt: 0.910 min; MS m/z (ESI): 499.4 [M+H].sup.+.

7.41 Example 41: Preparation of Compound 41

[0621] ##STR00261##

Step 1: Preparation of Compound 41-1

[0622] To a stirred solution of 40-4 (600 mg, 1.47 mmol, 1.0 eq), DIPEA (570 mg, 4.41 mmol, 3.0 eq) in DCM (20 mL) were added dodecanoyl chloride (643 mg, 2.94 mmol, 2.0 eq) and DMAP (35 mg, 0.29 mmol, 0.2 eq). The mixture was stirred at RT for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water (20 mL) and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=100/1) to give the title compound (750 mg, 75% yield) as yellow oil. LCMS: Rt: 0.940 min; MS m/z (ESI): 590.4 [M+H].sup.+.

Step 2: Preparation of Compound 41

[0623] To a solution of 41-1 (700 mg, 1.19 mmol, 1.0 eq) in MeOH (15 mL) were added Pd/C (140 mg) and concentrated HCl (4 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by column chromatography on silica gel (DCM/MeOH=75/1) to give the title compound (550 mg, 93% yield) as yellow oil. 250 mg product was then further purified by prep-HPLC to give the title compound (62 mg, 10% yield) as colorless oil.

[0624] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.88 (t, J=6.8 Hz, 6H), 1.26-1.29 (m, 35H), 1.53-1.64 (m, 4H), 2.35 (t, J=7.6 Hz, 2H), 2.44 (s, 3H), 2.66-2.80 (m, 4H), 3.40-3.45 (m, 2H), 3.46-3.59 (m, 2H), 3.64-3.66 (m, 2H), 5.16-5.20 (m, 1H). LCMS: Rt: 1.190 min; MS m/z (ESI): 500.4 [M+H].sup.+.

7.42 Example 42: Preparation of Compound 42

[0625] ##STR00262##

Compound 42-1

[0626] LCMS: Rt: 0.920 min; MS m/z (ESI): 482.5 [M-OMs].sup.+.

Compound 42

[0627] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 6H), 1.25-1.38 (m, 35H), 1.56-1.67 (m, 6H), 1.87-1.96 (m, 2H), 1.97-2.04 (m, 2H), 2.05-2.28 (m, 2H), 2.30-2.32 (m, 3H), 2.42-2.53 (m, 3H), 2.60-2.79 (m, 5H), 3.16-3.21 (m, 1H), 3.40-3.47 (m, 4H), 3.93-4.00 (m, 1H), 4.12-4.14 (m, 2H). LCMS: Rt: 1.080 min; MS m/z (ESI): 597.5 [M+H].sup.+.

7.43 Example 43: Preparation of Compound 43

[0628] ##STR00263##

Step 1: Preparation of Compound 43-1

[0629] A mixture of 3-(methylamino)propane-1,2-diol (5.0 g, 47.6 mmol, 1.0 eq), ((2-bromoethoxy)methyl)benzene (12.3 g, 57.1 mmol, 1.2 eq) and DIPEA (18.5 g, 142.8 mmol, 3.0 eq) in THF (100 mL) was stirred at 70° C. for 16 hours. LCMS showed the reaction was completed. The mixture was poured into water (100 mL) and extracted with EA (50 mL×3). The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=50/1-20/1) to give the title compound (3.7 g, 32% yield) as yellow oil. LCMS: Rt: 0.630 min; MS m/z (ESI): 240.2 [M+H].sup.+.

Step 2: Preparation of Compound 43-2

[0630] To a mixture of 43-1 (2.0 g, 8.36 mmol, 1.0 eq) in methyl acrylate (12 mL) was added NaOH (66 mg, 1.67 mmol, 0.2 eq). The resulting mixture was stirred at 50° C. for 16 hours. TLC showed the reaction was completed. The mixture was poured into water (40 mL) and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep-HPLC to give the title compound (800 mg, 24% yield) as colorless oil. LCMS: Rt: 0.730 min; MS m/z (ESI): 412.2 [M+H].sup.+.

Step 3: Preparation of Compound 43-3

[0631] To a solution of 43-2 (800 mg, 1.94 mol, 1.0 eq) in THF/H2O (8 mL/8 mL) was added Lithium hydroxide monohydrate (326 mg, 7.76 mmol, 4.0 eq). The reaction mixture was stirred at room temperature for 16 hours. LCMS showed the reaction worked completely. The reaction mixture was concentrated under reduced pressure to remove the organic solvent. The aqueous layer was acidified to pH=5 with 2 N HCl and then purified by prep-HPLC to give the title compound (280 mg, 38% yield) as colorless oil. LCMS: Rt: 0.740 min; MS m/z (ESI): 384.2 [M+H].sup.+.

Step 4: Preparation of Compound 43-4

[0632] A mixture of 43-3 (280 mg, 0.73 mmol, 1.0 eq), nonan-1-ol (316 mg, 2.19 mmol, 3.0 eq), EDCI (420 mg, 2.19 mmol, 3.0 eq), DMAP (178 mg, 1.46 mmol, 2.0 eq) and DIPEA (472 mg, 3.65 mmol, 5.0 eq) in DCM (30 mL) was stirred at reflux for 16 hours. LCMS showed the reaction worked completely. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep-HPLC to give the title compound (150 mg, 32% yield) as colorless oil. LCMS: Rt: 0.890 min; MS m/z (ESI): 636.4 [M+H].sup.+.

Step 5: Preparation of Compound 43-5

[0633] To a solution of 43-4 (150 mg, 0.24 mmol, 1.0 eq) in MeOH (10 mL) were added Pd/C (30 mg) and concentrated HCl (5 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and the residue was diluted with DCM washed with saturated NaHCO.sub.3 aqueous solution. The combined organic layers were dried Na.sub.2SO.sub.4 and concentrated to give the title compound (100 mg, 77% yield) as yellow oil. LCMS: Rt: 0.860 min; MS m/z (ESI): 546.4 [M+H].sup.+.

Step 6: Preparation of Compound 43-6

[0634] To a solution of 43-5 (100 mg, 0.18 mmol, 1.0 eq) in DCM (10 mL) was added SOCl.sub.2 (65 mg, 0.54 mmol, 3.0 eq) at RT. The mixture was stirred at 30° C. for 16 hours. LCMS showed the reaction was completed. The mixture was evaporated under reduced pressure to give the title compound (100 mg, 99% yield) as yellow oil. LCMS: Rt: 0.950 min; MS m/z (ESI): 564.3 [M+H].sup.+.

Step 7: Preparation of Compound 43

[0635] To a solution of 43-6 (100 mg, 0.18 mmol, 1.0 eq) and compound B (62 mg, 0.54 mmol, 3.0 eq) in THE (10 mL) were added DIPEA (116 mg, 0.90 mmol, 5.0 eq) and NaI (5 mg, 0.036 mmol, 0.2 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to give the title compound (11 mg, 10% yield) as yellow oil.

[0636] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 6H), 1.27-1.30 (m, 25H), 1.60-1.70 (m, 6H), 1.90-2.06 (m, 4H), 2.28-2.35 (m, 3H), 2.44-2.67 (m, 12H), 3.18-3.26 (m, 1H), 3.47-3.55 (m, 5H), 3.61-3.65 (m, 2H), 3.71-3.77 (m, 1H), 3.80-3.90 (m, 1H), 4.05-4.09 (m, 4H). LCMS: Rt: 0.910 min; MS m/z (ESI): 643.4 [M+H].sup.+.

7.44 Example 44: Preparation of Compound 44

[0637] ##STR00264##

Compound 44-1

[0638] LCMS: Rt: 0.940 min; MS m/z (ESI): 506.3 [M+H].sup.+.

Compound 44-2

[0639] LCMS: Rt: 1.010 min; MS m/z (ESI): 416.3 [M+H].sup.+.

Compound 44-3

[0640] LCMS: Rt: 1.630 min; MS m/z (ESI): 398.1 [M-OMs].sup.+.

Compound 44

[0641] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.94 (m, 6H), 1.25-1.32 (m, 24H), 1.54-1.64 (m, 6H), 1.87-1.93 (m, 2H), 1.97-2.05 (m, 2H), 2.30 (t, J=7.6 Hz, 3H), 2.36-2.39 (m, 3H), 2.46-2.65 (m, 4H), 2.72-2.76 (m, 2H), 3.12-3.20 (m, 1H), 3.40-3.50 (m, 4H), 3.87-3.90 (m, 1H), 4.12-4.16 (m, 2H). LCMS: Rt: 0.890 min; MS m/z (ESI): 513.4 [M+H].sup.+.

7.45 Example 45: Preparation of Compound 45

[0642] ##STR00265##

[0643] compound 45 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.81-0.93 (m, 6H), 1.01-1.14 (m, 1H), 1.16-1.38 (m, 29H), 1.47-1.60 (m, 4H), 1.61-1.70 (m, 1H), 1.76-1.91 (m, 3H), 1.98-2.35 (m, 7H), 2.55-2.97 (m, 6H), 3.35-3.53 (m, 5H), 3.54-3.75 (m, 3H). LCMS: Rt: 1.050 min; MS m/z (ESI): 527.5 [M+H].sup.+.

7.46 Example 46: Preparation of Compound 46

[0644] ##STR00266##

Step 1: Preparation of Compound 46-1

[0645] To a solution of 2-(bromomethyl)oxirane (5.4 g, 20 mmol, 1.0 eq) in THF (50 ml) was added NaH (1.6 g, 40 mmol, 2.0 eq) and stirred for 2 h at RT. Then C.sub.18H.sub.37Br (5.4 g, 40 mmol, 2.0 eq) was added and stirred for 16 h at 70° C. LCMS showed the reaction was completed. H2O was added, exacted with EA, and concentrated and purified by FCC (PE/EA=10/1) to get the 46-1 (1.0 g, 20% yield) as colorless oil.

[0646] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.84-0.89 (s, 3H), 1.30 (s, 32H), 1.50-1.60 (m, 4H), 2.60-2.62 (m, 1H), 2.79-2.80 (m, 1H), 3.14-3.16 (m, 1H), 3.36-3.52 (m, 3H), 3.68-3.73 (m, 1H).

Step 2: Preparation of Compound 46-2

[0647] To a solution of 46-1 (1.0 g, 3.07 mmol, 1.0 eq) and bis (4-methoxybenzyl)amine (617 mg, 3.98 mmol, 1.3 eq) in EtOH (10.0 ml) was stirred for 16 h at RT. LCMS showed the reaction was completed, concentrated and purified by FCC (PE/EA=10/1) to get the 46-2 (1.2 g, 68% yield) as white solid. LCMS: Rt: 1.421 min; MS m/z (ESI): 584.4 [M+H].sup.+.

Step 3: Preparation of Compound 46-3

[0648] To a solution of 46-2 (1.5 g, 2.57 mmol, 1.0 eq) in THF (20 ml) was added NaH (308 mg, 7.72 mmol, 3.0 eq) and stirred for 2 h at RT, then C.sub.6H.sub.13Br (1.26 g, 7.72 mmol, 3.0 eq) was added and stirred for 16 h at 70° C. LCMS showed the reaction was completed, H2O was added, exacted with EA, concentrated and purified by FCC (PE/EA=10/1) to get 46-3 (1.2 g, 70% yield) as yellow oil. LCMS: Rt: 1.585 min; MS m/z (ESI): 669.1 [M+H].sup.+.

Step 4: Preparation of Compound 46-4

[0649] To a solution of 46-3 (1.2 g, 1.8 mmol, 1.0) in EtOH (10 ml) was added Pd/C (120.0 mg) and then was stirred for 16 h at RT under H.sub.2. LCMS showed the reaction was completed, filtered and concentrated to get 46-4 (750.0 mg, 97% yield) as yellow oil. LCMS: Rt: 1.422 min; MS m/z (ESI): 428.4 [M+H].sup.+.

Step 5: Preparation of Compound 46-5

[0650] To a solution of 46-4 (750.0 mg, 1.75 mmol, 1.0 eq) and compound C (469.0 mg, 1.4 mmol, 0.8 eq) in ACN (10 ml) was added K.sub.2CO.sub.3 (724.5 mg, 5.25 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (170 mg, 0.525 mmol, 0.3 eq) and NaI (25 mg, 0.175 mmol, 0.1 eq) was stirred for 16 h at 85° C. LCMS showed the reaction was completed, concentrated and purified by FCC (DCM/MeOH=20/1) to get 46-5 (320 mg, 33% yield) as brown oil. LCMS: Rt: 1.770 min; MS m/z (ESI): 682.6 [M+H].sup.+.

Step 6: Preparation of Compound 46-6

[0651] To a solution of 46-5 (300 mg, 0.44 mmol, 1.0 eq) and 2-Bromoethanol (110 mg, 0.88 mmol, 4.0 eq) in EtOH (5 ml) was added DIEA (170 mg, 1.32 mmol, 3.0 eq) and NaI (25 mg, 0.0176 mmol, 0.1 eq). The mixture was stirred for 48 h at 70° C. LCMS showed the reaction was completed, concentrated to get 46-6 (350 mg, crude) as brown oil. LCMS: Rt: 2.010 min; MS m/z (ESI): 726.6 [M+H].sup.+.

Step 7: Preparation of Compound 46-7

[0652] A mixture of 46-6 (100 mg, 0.137 mmol, 1.0 eq) and SOCl.sub.2 (48 mg, 0.413 mmol, 3.0 eq) in DCM (5 ml) was stirred at 35° C. for 16 h. The reaction was filtered and the filtrate was concentrated. The residue was purified by column chromatography on silica (PE/EA=10/1) to give 46-7 (80 mg, 74% yield) as a colorless oil. LCMS: Rt: 1.030 min; MS m/z (ESI): 708.6 [M-Cl].sup.+.

Step 8: Preparation of Compound 46

[0653] A mixture of 46-7 (100 mg, 0.134 mmol, 1.0 eq), compound I (46 mg, 0.4 mmol, 3.0 eq), DIEA (87 mg, 0.67 mmol, 5.0 eq) in THE (5 ml) was stirred at 70° C. for 16 h. LCMS showed the reaction was complete. After removal of solvent, the residue was purified by prep-HPLC to provide 46 (16.0 mg, 14% yield) as colorless oil.

[0654] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.84-0.89 (s, 9H), 1.14-1.31 (s, 59H), 1.50-1.65 (m, 10H), 1.98-2.09 (m, 2H), 2.28-2.31 (m, 2H), 2.48-2.63 (m, 8H), 3.40-3.61 (m, 10H), 4.03-4.07 (m, 2H). LCMS: Rt: 1.670 min; MS m/z (ESI): 823.7[M+H].sup.+.

7.47 Example 47: Preparation of Compound 47

[0655] ##STR00267##

Step 1: Preparation of Compound 47-1

[0656] To a mixture of NaH (800 mg, 20.01 mmol, 5.0 eq) in THF (50 mL) was added J-1 (2 g, 4.002 mmol, 1.0 eq) at RT under N.sub.2. The reaction mixture was stirred at RT for 4 hours. The C.sub.14H.sub.29Br (1.66 g, 6.004 mmol, 1.5 eq) was added to it. The reaction mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The mixture was poured in water and washed with EA. The combined organic layers were separated and dried over Na.sub.2SO.sub.4. Removal of solvent, FCC to get the compound 47-1 (1.6 g, 57.43%) as yellow oil. LCMS: Rt: 1.660 min; MS m/z (ESI): 696.5 [M+H].sup.+.

Step 2: Preparation of Compound 47-2

[0657] To a solution of 47-1 (1.6 g, 2.299 mmol) in EA (100 mL) was added Pd/C (300 mg). The reaction mixture was stirred at RT for 40 hours under hydrogen. LCMS showed the reaction was complete. The mixture was filtered through diatomite. Removal of solvent to get the compound 47-2 (1.1 g, crude) as yellow oil. LCMS: Rt: 1.100 min; MS m/z (ESI): 456.4 [M+H].sup.+.

Step 3: Preparation of Compound 47-3

[0658] To a solution of compound 47-2 (1.1 g, 2.299 mmol, 1.2 eq) in ACN (30 mL) was added compound C (642 mg, 1.916 mmol, 1.0 eq), K.sub.2CO.sub.3 (794 mg, 5.748 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (187 mg, 0.5748 mmol, 0.3 eq), NaI (29 mg, 0.1916 mmol, 0.1 eq). The reaction mixture was stirred at 80° C. for 88 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 47-3 (500 mg, 36.74%) as yellow oil. LCMS: Rt: 1.540 min; MS m/z (ESI): 710.7 [M+H].sup.+.

Step 4: Preparation of Compound 47-4

[0659] To a mixture of compound 47-3 (500 mg, 0.7040 mmol, 1.0 eq), DIEA (455 mg, 3.520 mmol, 5.0 eq) in EtOH (20 mL) was added 2-bromoethan-1-ol (352 mg, 2.816 mmol, 4.0 eq), NaI (10 mg). The reaction mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound (500 mg, crude) as yellow oil. LCMS: Rt: 2.150 min; MS m/z (ESI): 754.7 [M+H].sup.+.

Step 5: Preparation of Compound 47-5

[0660] To a solution of compound 47-4 (500 mg, 0.6629 mmol, 1.0 eq) in DCM (15 mL) was added SOCl.sub.2 (237 mg, 1.989 mmol, 3.0 eq). The reaction mixture was stirred at 35° C. for 16 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 47-5 (200 mg, 39.05%) as yellow oil. LCMS: Rt: 1.380 min; MS m/z (ESI): 736.7 [M-Cl].sup.+.

Step 6: Preparation of Compound 47

[0661] To a mixture of compound 47-5 (180 mg, 0.2329 mmol, 1.0 eq), DIEA (150 mg, 1.65 mmol, 5.0 eq) in THF (15 mL) was added compound I (81 mg, 0.6988 mmol, 3.0 eq), NaI (20 mg). The reaction mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. After removal of solvent, the residue was purified by prep-HPLC to give the title compound (60 mg, 30.26% yield) as yellow oil.

[0662] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.07-4.03 (m, 2H), 3.60-3.40 (m, 9H), 3.18-3.14 (m, 1H), 2.57-2.45 (m, 10H), 2.31-2.27 (m, 2H), 2.01-1.84 (m, 5H), 1.65-1.48 (m, 15H), 1.26 (s, 53H), 0.90-0.86 (m, 9H). LCMS: Rt: 2.230 min; MS m/z (ESI): 851.8 [M+H].sup.+.

7.48 Example 48: Preparation of Compound 48

[0663] ##STR00268##

Compound 48-1

[0664] LCMS: Rt: 1.040 min; MS m/z (ESI): 612.4 [M+H].sup.+.

Compound 48-2

[0665] LCMS: Rt: 0.930 min; MS m/z (ESI): 372.3 [M+H].sup.+.

Compound 48-3

[0666] LCMS: Rt: 1.010 min; MS m/z (ESI): 626.5 [M+H].sup.+.

Compound 48-4

[0667] LCMS: Rt: 1.270 min; MS m/z (ESI): 670.6 [M+H].sup.+.

Compound 48-5

[0668] LCMS: Rt: 1.030 min; MS m/z (ESI): 652.6 [M-Cl].sup.+.

Compound 48

[0669] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.07-4.03 (m, 2H), 3.61-3.40 (m, 9H), 3.19 (s, 1H), 2.58-2.49 (m, 9H) 2.31-2.27 (m, 2H), 2.01 (s, 2H), 1.89 (s, 1H), 1.65-1.45 (m, 12H), 1.26 (s, 47H), 0.90-0.86 (m, 9H). LCMS: Rt: 1.150 min; MS m/z (ESI): 767.7 [M+H].sup.+.

7.49 Example 49: Preparation of Compound 49

[0670] ##STR00269##

Step 1: Preparation of Compound 49-1

[0671] To a solution of compound 22-1 (800 mg, 2.328 mmol, 1.2 eq) in ACN (20 mL) was added compound C (650 mg, 1.940 mmol, 1.0 eq), K.sub.2CO.sub.3 (804 mg, 5.821 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (190 mg, 0.5821 mmol, 0.3 eq), NaI (29 mg, 0.1940 mmol, 0.1 eq). The reaction mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 49-1 (240 mg, 18.39%) as yellow oil. LCMS: Rt: 1.040 min; MS m/z (ESI): 598.5 [M+H].sup.+.

Step 2: Preparation of Compound 49-2

[0672] To a mixture of compound 49-1 (220 mg, 0.3679 mmol, 1.0 eq), DIEA (238 mg, 1.840 mmol, 5.0 eq) in EtOH (15 mL) was added 2-bromoethan-1-ol (184 mg, 1.472 mmol, 4.0 eq). The reaction mixture was stirred at 70° C. for 30 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound (260 mg, crude) as yellow oil. LCMS: Rt: 1.160 min; MS m/z (ESI): 642.5 [M+H].sup.+.

Step 3: Preparation of Compound 49-3

[0673] To a solution of compound 49-2 (260 mg, 0.3582 mmol, 1.0 eq) in DCM (10 mL) was added SOCl.sub.2 (128 mg, 1.675 mmol, 3.0 eq). The reaction mixture was stirred at 35° C. for 16 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound 49-3 (240 mg, crude) as yellow oil. LCMS: Rt: 2.170 min; MS m/z (ESI): 660.5 [M+H].sup.+.

Step 4: Preparation of Compound 49

[0674] To a mixture of compound 49-3 (220 mg, 0.3331 mmol, 1.0 eq), DIEA (215 mg, 1.666 mmol, 5.0 eq) in THE (20 mL) was added compound I (115 mg, 0.9993 mmol, 3.0 eq), NaI (20 mg). The reaction mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. After removal of solvent, the residue was purified by prep-HPLC to give the title compound (100 mg, 40.6% yield) as yellow oil.

[0675] .sup.1H NMR (400 MHz, CDCl3) δ: 4.07-4.03 (m, 2H), 3.61-3.19 (m, 10H), 2.55 (s, 9H), 2.31-2.27 (m, 2H), 2.04-1.87 (m, 3H), 1.65-1.52 (m, 12H), 1.26 (s, 43H), 0.90-0.86 (m, 9H). LCMS: Rt: 1.220 min; MS m/z (ESI): 739.6 [M+H].sup.+.

7.50 Example 50: Preparation of Compound 50

[0676] ##STR00270##

Step 1: Preparation of Compound 50-1

[0677] To a solution of 43-3 (300 mg, 0.78 mmol, 1.0 eq) and compound N (462 mg, 1.72 mmol, 2.2 eq) in DCM (15 mL) were added HATU (742 mg, 2.19 mmol, 2.5 eq) and DIPEA (504 mg, 3.90 mmol, 3.0 eq). The mixture was stirred at room temperature for 16 hours. LCMS showed the reaction worked completely. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=50/1) to give the title compound (550 mg, 79% yield) as yellow oil. LCMS: Rt: 1.460 min; MS m/z (ESI): 886.7 [M+H].sup.+.

Step 2: Preparation of Compound 50

[0678] To a solution of 50-1 (100 mg, 0.10 mmol, 1.0 eq) in MeOH (10 mL) were added Pd/C (10 mg) and concentrated HCl (3 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated and purified by prep-HPLC to give the title compound (34 mg, 38% yield) as colorless oil.

[0679] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.34 (m, 58H), 1.43-1.53 (m, 2H), 2.38-2.52 (m, 7H), 2.58-2.88 (m, 3H), 3.10-3.18 (m, 4H), 3.54-3.56 (m, 2H), 3.64-3.89 (m, 6H), 3.91-3.95 (m, 1H), 6.13-6.18 (m, 1H), 6.38-6.44 (m, 1H). LCMS: Rt: 1.280 min; MS m/z (ESI): 796.7 [M+H].sup.+.

7.51 Example 51: Preparation of Compound 51

[0680] ##STR00271##

Compound 51-1

[0681] LCMS: Rt: 1.610 min; MS m/z (ESI): 814.7 [M+H].sup.+.

Compound 51

[0682] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.35 (m, 62H), 1.45-1.48 (m, 2H), 1.54-1.66 (m, 2H), 1.98-2.10 (m, 2H), 2.33-2.83 (m, 12H), 3.11-3.23 (m, 5H), 3.46-3.86 (m, 8H), 3.91-3.95 (m, 1H), 6.31-6.37 (m, 1H), 6.64-6.74 (m, 1H). LCMS: Rt: 1.460 min; MS m/z (ESI): 893.7 [M+H].sup.+.

7.52 Example 52: Preparation of Compound 52

[0683] ##STR00272##

[0684] compound 52 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.31 (m, 64H), 1.44-1.48 (m, 2H), 1.54-1.60 (m, 2H), 1.76-1.88 (m, 2H), 2.18-2.80 (m, 16H), 3.09-3.17 (m, 4H), 3.44-3.59 (m, 3H), 3.72-3.86 (m, 4H), 3.89-3.93 (m, 1H), 6.36-6.45 (m, 1H), 6.72-6.78 (m, 1H). LCMS: Rt: 1.410 min; MS m/z (ESI): 921.8 [M+H].sup.+.

7.53 Example 53: Preparation of Compound 53

[0685] ##STR00273## ##STR00274##

Step 1: Preparation of Compound 53-1

[0686] To a mixture of NaH (3.5 g, 87.61 mmol) in THE (200 mL) was added C.sub.16H.sub.33OH (14.16 g, 58.40 mmol) at RT under N.sub.2. The reaction mixture was stirred at RT for 3 hours. 2-(bromomethyl)oxirane (4.0 g, 29.2 mmol) was added to it. The reaction mixture was stirred at RT for 10 hours. TLC showed the reaction was complete. The mixture was poured in water and washed with EA. The combined organic layers were separated and dried over Na.sub.2SO.sub.4. Removal of solvent, FCC to get the compound 53-1 (4.0 g, 46% yield) as colorless oil.

Step 2: Preparation of Compound 53-2

[0687] To a mixture of compound 53-1 (4.0 g, 13.40 mmol), bis(4-methoxybenzyl)amine (6.9 g, 26.8 mmol) in EtOH (200 mL) was stirred at RT for 10 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 53-2 (4.5 g, 67% yield) as colorless oil. LCMS: Rt: 1.088 min; MS m/z (ESI): 556.4 [M+H].sup.+.

Step 3: Preparation of Compound 53-3

[0688] To a mixture of NaH (450 mg, 7.2 mmol) in THF (50 mL) was added 53-3 (1 g, 1.8 mmol) at RT under N.sub.2. The reaction mixture was stirred at RT for 2 hours. The C.sub.6H.sub.13Br (1.04 g, 6.3 mmol) was added to it. The reaction mixture was stirred at 70° C. for 10 hours. LCMS showed the reaction was complete. The mixture was poured in water and washed with EA. The organic layers were separated and dried over Na.sub.2SO.sub.4. Removal of solvent, FCC to get the compound 53-3 (1.0 g, 87% yield) as colorless oil. LCMS: Rt: 1.960 min; MS m/z (ESI): 640.4 [M+H].sup.+.

Step 4: Preparation of Compound 53-4

[0689] To a solution of 53-3 (1.0 g, 1.56 mmol) in EA (100 mL) was added Pd/C (1.0 g). The reaction mixture was stirred at RT for 40 hours under H.sub.2. LCMS showed the reaction was complete. The mixture was filtered through diatomite. Removal of solvent to get the compound 53-4 (0.5 g, 80% yield) as colorless oil. LCMS: Rt: 0.920 min; MS m/z (ESI): 400.3 [M+H].sup.+.

Step 5: Preparation of Compound 53-5

[0690] To a solution of compound 53-5 (0.5 g, 1.25 mmol) in ACN (30 mL) was added C (420 mg, 1.25 mmol), K.sub.2CO.sub.3 (550 mg, 3.75 mmol), Cs.sub.2CO.sub.3 (130 mg, 0.37 mmol), NaI (60 mg, 0.37 mmol). The reaction mixture was stirred at 80° C. for 10 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 53-5 (200 mg, 23% yield) as yellow oil. LCMS: Rt: 1.310 min; MS m/z (ESI): 654.6 [M+H].sup.+.

Step 6: Preparation of Compound 53-6

[0691] To a mixture of 53-5 (200 mg, 0.3 mmol), DIEA (240 mg, 1.83 mmol) in EtOH (20 mL) was added 2-bromoethan-1-ol (150 mg, 1.22 mmol), NaI (45 mg). The reaction mixture was stirred at 70° C. for 20 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound (150 mg, 70% yield) as yellow oil. LCMS: Rt: 1.30 min; MS m/z (ESI): 698.6 [M+H].sup.+.

Step 7: Preparation of Compound 53-7

[0692] To a solution of compound 53-6 (150 mg, 0.21 mmol) in DCM (10 mL) was added SOCl.sub.2 (80 mg, 0.64 mmol). The reaction mixture was stirred at 35° C. for 10 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 53-7 (154 mg, 100% yield) as yellow oil. LCMS: Rt: 0.640 min; MS m/z (ESI): 716.5 [M+H].sup.+.

Step 8: Preparation of Compound 53

[0693] To a mixture of compound 53-7 (150 mg, 0.21 mmol, 1.0 eq), DIEA (80 mg, 0.63 mmol) in THF (10 mL) was added Compound I (50 mg, 0.42 mmol), NaI (30 mg). The reaction mixture was stirred at 70° C. for 10 hours. LCMS showed the reaction was complete. After removal of solvent, the residue was purified by prep-HPLC to give the title compound (20 mg, 12% yield) as yellow oil.

[0694] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.87 (t, J=8 Hz, 9H), 1.20-1.41 (m, 50H), 1.45-2.09 (m, 15H), 2.27-2.55 (m, 12H), 3.21-3.57 (m, 10H), 4.03-4.07 (m, 2H). LCMS: Rt: 1.800 min; MS m/z (ESI): 795.7 [M+H].sup.+.

7.54 Example 54: Preparation of Compound 55

[0695] ##STR00275##

Step 1: Preparation of Compound 55-1

[0696] To a solution of 43-3 (135 mg, 0.35 mmol, 1.0 eq) and 2-octyldecan-1-ol (284 mg, 1.05 mmol, 3.0 eq) in DCM (10 mL) were added EDCI (201 mg, 1.05 mmol, 3.0 eq), DMAP (22 mg, 0.18 mmol, 0.5 eq) and DIPEA (226 mg, 1.75 mmol, 5.0 eq). The mixture was stirred under reflux for 16 hours. LCMS showed the reaction worked completely. The reaction mixture was concentrated and purified by by column chromatography on silica gel (DCM/MeOH=50/1) to give the title compound (150 mg, 48% yield) as colorless oil. LCMS: Rt: 1.510 min; MS m/z (ESI): 888.7 [M+H].sup.+.

Step 2: Preparation of Compound 55-2

[0697] To a solution of 55-1 (150 mg, 0.17 mmol, 1.0 eq) in MeOH (8 mL) were added Pd/C (15 mg) and concentrated HCl (3 drops). The mixture was stirred at RT under H.sub.2 for 2 hours. LCMS showed the reaction was complete. The mixture was filtered through a pad of celite and washed with MeOH. The filtration was concentrated The residue was diluted with DCM and washed with saturated NaHCO.sub.3 aqueous solution. The organic layers were dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (128 mg, 95% yield) as yellow oil. LCMS: Rt: 1.627 min; MS m/z (ESI): 798.5 [M+H].sup.+.

Step 3: Preparation of Compound 55-3

[0698] To a solution of 55-2 (128 mg, 0.16 mmol, 1.0 eq) in DCM (10 mL) was added SOCl.sub.2 (57 mg, 0.48 mmol, 3.0 eq) at RT. The mixture was stirred at 30° C. for 16 hours. LCMS showed the reaction was completed. The mixture was evaporated under reduced pressure to give the title compound (128 mg, 98% yield) as yellow oil. LCMS: Rt: 1.660 min; MS m/z (ESI): 817.6 [M+H].sup.+.

Step 4: Preparation of Compound 55

[0699] To a solution of 55-3 (128 mg, 0.16 mmol, 1.0 eq) and Compound I (55 mg, 0.48 mmol, 3.0 eq) in THF (10 mL) were added DIPEA (103 mg, 0.80 mmol, 5.0 eq) and NaI (7 mg, 0.048 mmol, 0.3 eq). The mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was complete. The mixture was concentrated and purified by prep-HPLC to give the title compound (26 mg, 18% yield) as yellow oil.

[0700] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.32 (m, 58H), 1.55-1.70 (m, 4H), 1.78-2.08 (m, 4H), 2.25-2.33 (m, 3H), 2.40-2.62 (m, 11H), 3.11-3.21 (m, 1H), 3.48-3.63 (m, 5H), 3.71-3.74 (m, 2H), 3.77-3.80 (m, 1H), 3.81-3.90 (m, 1H), 3.96-3.99 (m, 4H). LCMS: Rt: 1.730 min; MS m/z (ESI): 895.7 [M+H].sup.+.

7.55 Example 55: Preparation of Compound 56

[0701] ##STR00276##

Step 1: Preparation of Compound 56-1

[0702] To a mixture of compound 4-bromobutan-1-ol (10 g, 65.79 mmol, 1.0 eq), TsOH (500 mg) in EA (300 mL) was added DHP (6.63 g, 78.94 mmol, 1.2 eq). The reaction mixture was stirred at 24° C. for 3 hours. TLC showed the reaction was complete. The mixture was quenched with water, extracted over EA, concentrated and purified by silica gel column chromatography (PE:EA=50:1) to give the desired product 56-1 (11 g, 70.9% yield) as colorless oil.

Step 2: Preparation of Compound 56-2

[0703] To a mixture of NaH (2 g, 50 mmol, 1.2 eq) in DMF (80 ml) was added dimethyl malonate (6.6 g, 50 mmol 1.2 eq) dropwise. The mixture was stirred at 0° C. for 2 hours. Then compound 56-1 (10 g, 42.37 mmol, 1.0 eq) was added. The reaction mixture was stirred at 90° C. for 4 hours. TLC showed the reaction was complete. The mixture was quenched with water, extracted over EA, concentrated and purified by silica gel column chromatography (PE:EA=4:1) to give the desired product 56-2 (7.9 g, 64.7% yield) as colorless oil.

Step 3: Preparation of Compound 56-3

[0704] To a mixture of LiAlH.sub.4 (2.13 g, 56.15 mmol, 2.1 eq) in THE (150 ml) was added 56-2 (7.7 g, 26.73 mmol 1.0 eq) dropwise in 0° C. The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with water, extracted over EA, concentrated and purified by silica gel column chromatography (PE:EA=1:1) to give the desired product 56-3 (2.4 g, 39.1% yield) as colorless oil.

Step 4: Preparation of Compound 56-4

[0705] To a mixture of Compound P (1.4 g, 4.74 mmol, 2.2 eq) in DCM (40 ml) was added EDCl (1.03 g, 5.37 mmol, 2.5 eq), DIEA (1.39 g, 10.75 mmol, 5 eq). The mixture was stirred at RT for 1 hour. Then 56-3 (500 mg, 2.15 mmol 1.0 eq) and DMAP (100 mg) was added in it. The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with water, extracted over EA, concentrated and purified by silica gel column chromatography (PE:EA=20:1) to give the desired product 56-4 (860 mg, 50.5% yield) as colorless oil.

Step 5: Preparation of Compound 56-5

[0706] To a mixture of 56-4 (860 mg, 1.09 mmol, 1.0 eq) was added HCl/dioxane (4 M, 5 ml). The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with NaHCO.sub.3 solution (50 ml), extracted over EA, concentrated in vacuo. The crude product was used for next step without further purification 56-5 (700 mg, crude).

Step 6: Preparation of Compound 56-6

[0707] To a mixture of 56-5 (650 mg, 0.92 mmol, 1.0 eq) in 20 ml DCM was added DIEA (356 mg, 2.76 mmol, 3.0 eq), MsCl (125 mg, 1.1 mmol, 1.2 eq). The mixture was stirred at RT for 2 hours. TLC showed the reaction was complete. The mixture was quenched with water (50 ml), extracted over EA, concentrated in vacuo. The crude product was used for next step without further purification 56-6 (700 mg, crude).

Step 7: Preparation of Compound 56

[0708] To a mixture of 56-6 (200 mg, 0.25 mmol, 1.0 eq) in 10 ml ACN was added K.sub.2CO.sub.3 (103 mg, 0.75 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (98 mg, 0.08 mmol, 0.3 eq), NaI (20 mg). Then compound I (86.25 mg, 0.75 mmol, 3.0 eq) was added. The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The mixture was quenched with water (50 ml), extracted over EA, concentrated in vacuo. The mixture was concentrated and purified by prep-HPLC to give the title compound (70 mg, 34.7% yield) as yellow oil.

[0709] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.09-4.00 (m, 4H), 3.52-3.51 (m, 2H), 3.15-3.11 (m, 1H), 2.54-2.39 (m, 4H), 2.24-2.23 (m, 4H), 2.02-1.95 (m, 3H), 1.86-1.82 (m, 4H), 1.67-1.58 (m, 2H), 1.42-1.38 (m, 3H), 1.33-1.26 (m. 60H), 0.89-0.86 (m, 12H). LCMS: Rt: 2.21 min; MS m/z (ESI):806.7 [M+H].sup.+.

7.56 Example 56: Preparation of Compound 57

[0710] ##STR00277##

[0711] compound 57 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.33 (m, 58H), 1.37-1.59 (m, 6H), 1.72-1.84 (m, 4H), 2.20-2.46 (m, 10H), 2.54-2.79 (m, 4H), 3.07-3.20 (m, 5H), 3.43-3.58 (m, 5H), 3.63-3.78 (m, 3H), 3.86-3.93 (m, 1H), 6.34-6.41 (m, 1H), 6.70-6.76 (m, 1H). LCMS: Rt: 1.390 min; MS m/z (ESI): 907.8 [M+H].sup.+.

7.57 Example 57: Preparation of Compound 58

[0712] ##STR00278##

Compound 58

[0713] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.32 (m, 58H), 1.35-1.60 (m, 10H), 1.64-1.88 (m, 4H), 2.20-2.29 (m, 3H), 2.35-2.66 (m, 12H), 3.09-3.21 (m, 4H), 3.41-3.62 (m, 5H), 3.71-3.74 (m, 3H), 3.84-3.90 (m, 1H), 6.38-6.42 (m, 1H), 6.71-6.76 (m, 1H). LCMS: Rt: 1.580 min; MS m/z (ESI): 935.8 [M+H].sup.+.

7.58 Example 58: Preparation of Compound 59

[0714] ##STR00279## ##STR00280##

Step 1: Preparation of Compound 59-1

[0715] To a solution of compound Q (2.0 g, 5.11 mmol) in ACN (100 mL) was added 2-(benzyloxy)ethan-1-amine (1.55 g, 10.22 mmol), K.sub.2CO.sub.3 (2.12 g, 15.33 mmol), Cs.sub.2CO.sub.3 (500 mg, 1.53 mmol), NaI (250 mg, 1.53 mmol). The reaction mixture was stirred at 80° C. for 10 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 59-1 (1.8 g, 76% yield) as yellow oil. LCMS: Rt: 0.930 min; MS m/z (ESI): 462.3 [M+H].sup.+.

Step 2: Preparation of Compound 59-2

[0716] To a mixture of compound 59-1 (1.6 g, 3.47 mmol), oxiran-2-ylmethanol (0.4 g, 5.2 mmol) in EtOH (50 mL) was added DIEA (1.34 g, 10.4 mmol). The reaction mixture was stirred at 80° C. for 10 hours. LCMS showed the reaction was complete. Removal of solvent, FCC to get the compound 59-2 (1.1 g, 59% yield) as yellow oil. LCMS: Rt: 0.86 min; MS m/z (ESI): 536.4 [M+H].sup.+.

Step 3: Preparation of Compound 59-3

[0717] To a mixture of 59-2 (500 mg, 0.93 mmol), DIEA (750 mg, 5.6 mmol) in CH.sub.2Cl.sub.2 (20 mL) was added hexanoyl chloride (500 mg, 3.73 mmol). The reaction mixture was stirred at 70° C. for 20 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound 59-3 (500 mg, 73% yield) as yellow oil. LCMS: Rt: 0.60 min; MS m/z (ESI): 732.4 [M+H].sup.+.

Step 4: Preparation of Compound 59-4

[0718] To a solution of 59-3 (500 mg, 0.68 mmol) in EA (50 mL) was added Pd/C (1.0 g). The reaction mixture was stirred at RT for 10 hours under H.sub.2. LCMS showed the reaction was complete. The mixture was filtered through diatomite. Removal of solvent to get the compound 59-4 (400 mg, 91% yield) as yellow oil. LCMS: Rt: 0.480 min; MS m/z (ESI): 642.4 [M+H].sup.+.

Step 5: Preparation of Compound 59-5

[0719] To a solution of 59-4 (200 mg, 0.31 mmol) in DCM (10 mL) was added SOCl.sub.2 (110 mg, 0.93 mmol). The reaction mixture was stirred at 35° C. for 10 hours. LCMS showed the reaction was complete. Removal of solvent to get the compound 59-5 (205 mg, 100% yield) as yellow oil. LCMS: Rt: 0.653 min; MS m/z (ESI): 660.4 [M+H].sup.+.

Step 6: Preparation of Compound 59

[0720] To a mixture of compound 59-5 (200 mg, 0.3 mmol), DIEA (120 mg, 0.9 mmol) in THE (10 mL) was added pyrrolidin-3-ol (53 mg, 0.6 mmol), NaI (45 mg). The reaction mixture was stirred at 70° C. for 10 hours. LCMS showed the reaction was complete. After removal of solvent, the residue was purified by prep-HPLC to give the title compound (70 mg, 32% yield) as yellow oil.

[0721] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.87 (t, J=8 Hz, 12H), 1.20-1.79 (m, 40H), 2.16-2.99 (m, 20H), 3.96-4.11 (m, 3H), 4.34-4.41 (m, 2H), 5.12-5.14 (m, 1H). LCMS: Rt: 1.020 min; MS m/z (ESI): 711.5 [M+H].sup.+.

##STR00281##

7.59 Example 59: Preparation of Compound 60

Step 1: Preparation of Compound 60-1

[0722] To a mixture of compound P (298 mg, 1.0 mmol, 1.0 eq) in DCM (10 ml) was added EDCl (229 mg, 1.2 mmol, 1.2 eq), DIEA (387 mg, 3.0 mmol, 3 eq). The mixture was stirred at RT for 1 hour. Then 56-3 (232 mg, 1.0 mmol 1.0 eq) and DMAP (10 mg) was added in. The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with water, extracted over EA, concentrated and purified by silica gel column chromatography (PE:EA=10:1) to give the desired product 60-1 (330 mg, 64.4% yield) as colorless oil.

Step 2: Preparation of Compound 60-2

[0723] To a mixture of 60-1 (512 mg, 1.0 mmol, 1.0 eq) in 10 ml DCM was added DIEA (387 mg, 3.0 mmol, 3 eq), hexanoyl chloride (160 mg, 1.2 mmol, 1.2 eq). The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with water (30 ml), extracted over DCM, concentrated in vacuo, and purified by silica gel column chromatography (PE:EA=10:1) to give the desired product 60-2 (420 mg, 68%).

Step 3: Preparation of Compound 60-3

[0724] To a mixture of 60-2 (610 mg, 1.0 mmol, 1.0 eq) was added HCl/dioxane (4 M, 5 ml). The mixture was stirred at RT for 16 hours. TLC showed the reaction was complete. The mixture was quenched with NaHCO.sub.3 solution (50 ml), extracted over EA, concentrated in vacuo. The crude product was used for next step without further purification 60-3 (700 mg, crude).

Step 4: Preparation of Compound 60-4

[0725] To a mixture of 60-3 (526 mg, 1.0 mmol, 1.0 eq) in 10 ml DCM was added DIEA (387 mg, 3.0 mmol, 3.0 eq), MsCl (137 mg, 1.2 mmol, 1.2 eq). The mixture was stirred at RT for 2 hours. TLC showed the reaction was complete. The mixture was quenched with water (50 ml), extracted over EA, concentrated in vacuo. The crude product was used for next step without further purification 60-4 (490 mg, crude).

Step 5: Preparation of Compound 60

[0726] To a mixture of 60-4 (151 mg, 0.25 mmol, 1.0 eq) in 10 ml ACN was added K.sub.2CO.sub.3 (103 mg, 0.75 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (98 mg, 0.08 mmol, 0.3 eq), NaI (20 mg). Then compound I (86.25 mg, 0.75 mmol, 3.0 eq) was added. The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The mixture was quenched with water (50 ml), extracted over EA, concentrated in vacuo. The mixture was concentrated and purified by prep-HPLC to give the title compound (33 mg, 21.2% yield) as yellow oil.

[0727] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.09-4.00 (m, 4H), 3.52-3.49 (m, 2H), 3.17-3.09 (m, 1H), 2.56-2.54 (m, 4H), 2.43-2.23 (m, 6H), 2.04-1.80 (m, 6H), 1.67-1.58 (m, 4H), 1.46-1.43 (m, 3H), 1.33-1.26 (m. 36H), 0.89-0.86 (m, 9H). LCMS: Rt: 0.97 min; MS m/z (ESI): 624.5 [M+H].sup.+.

7.60 Example 60: Preparation of Compound 64

[0728] ##STR00282##

Step 1: Preparation of Compound 64-2

[0729] To a solution of compound 64-1 (2.0 g, 16.0 mmol, 1.0 eq.) in DMF (32 mL) was added imidazole (2.2 g, 32.0 mmol, 2.0 eq.) and then TBSCl (3.6 g, 24.0 mmol, 1.5 eq.) dropwise. The mixture was stirred at room temperature for 16 hours. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromotography on silica gel (PE/EA=100/1) to give the title compound (2.6 g, 68% yield) as colorless oil.

Step 2: Preparation of Compound 64-3

[0730] To a solution of compound 64-2 (995 mg, 4.16 mmol, 0.8 eq.) and compound I (600 mg, 5.20 mmol, 1.0 eq.) in ACN (30 mL) was added K.sub.2CO.sub.3 (2.2 g, 15.6 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (508 mg, 1.56 mmol, 0.3 eq.) and NaI (234 mg, 1.56 mmol, 0.3 eq.). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The reaction mixture was concentrated and purified by column chromatography on silica gel (DCM/MeOH=40/1) to give the title compound (670 mg, 61% yield) as yellow oil. LCMS: Rt: 0.770 min; MS m/z (ESI): 274.2 [M+H].sup.+.

Step 3: Preparation of Compound 64-4

[0731] To a solution of compound 64-3 (670 mg, 2.45 mmol, 1.0 eq.) and DIPEA (633 mg, 4.90 mmol, 2.0 eq.) in DCM (25 mL) was added MsCl (337 mg, 2.94 mmol, 1.2 eq.) at 0° C. The mixture was stirred at room temperature for 2 hours. LCMS showed the reaction was completed. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (715 mg, 83% yield) as yellow oil. It was used in the next step without further purification. LCMS: Rt: 0.810 min; MS m/z (ESI): 292.2 [M-OMs+Cl].sup.+.

Step 4: Preparation of Compound 64-5

[0732] To a solution of compound 64-4 (715 mg, 2.04 mmol, 1.0 eq.) and compound W (257 mg, 2.45 mmol, 1.2 eq.) in THE (20 mL) was added DIPEA (791 mg, 6.12 mmol, 3.0 eq.) and NaI (92 mg, 0.612 mmol, 0.3 eq.). The mixture was stirred at at 70° C.; for 16 hours. LCMS showed the reaction was completed. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=20/1) to give the title compound (340 mg, 46% yield) as colorless oil. LCMS: Rt: 0.710 min; MS m/z (ESI): 361.2 [M+H].sup.+.

Step 5: Preparation of Compound 64-6

[0733] To a solution of compound 64-5 (340 mg, 0.94 mmol, 1.0 eq.) in DCM (10 mL) was added compound X (659 mg, 2.35 mmol, 2.5 eq.), EDCI (541 mg, 2.82 mmol, 3.0 eq.), DMAP (57 mg, 0.47 mmol, 0.5 eq.) and DIPEA (607 mg, 4.70 mmol, 5.0 eq.). The mixture was stirred under reflux for 16 hours. LCMS showed the reaction worked completely. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=40/1) to give the title compound (360 mg, 43% yield) as colorless oil. LCMS: Rt: 1.387 min; MS m/z (ESI): 885.5 [M+H].sup.+.

Step 6: Preparation of Compound 64

[0734] To a solution of compound 64-6 (310 mg, 0.35 mmol, 1.0 eq.) in MeOH (5 mL) was added HCl in Dioxane (0.5 mL, 4.0 M). The mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with DCM and washed with saturated NaHCO.sub.3 aqueous solution. The mixture was extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep-HPLC to give the title compound (65 mg, 24% yield) as colorless oil.

[0735] .sup.1H NMR (400 MHz, CDCl3) δ: 0.87-0.91 (m, 6H), 1.30-1.37 (m, 30H), 1.63-1.86 (m, 6H), 1.95-2.07 (m, 12H), 2.30-2.38 (m, 6H), 2.39-2.54 (m, 6H), 2.75-2.79 (m, 6H), 3.10-3.26 (m, 1H), 3.90-4.09 (m, 2H), 4.14-4.20 (m, 3H), 5.29-5.42 (m, 8H). LCMS: Rt: 1.140 min; MS m/z (ESI): 771.5 [M+H].sup.+.

7.61 Example 61: Preparation of Compound 65

[0736] ##STR00283##

Step 1: Preparation of Compound 65-1

[0737] To a mixture of compound 56 (402 mg, 0.5 mmol, 1.0 eq.) in DCM (8 mL) was added DIEA (193.5 mg, 1.5 mmol, 3.0 eq.), MsCl (68 mg, 0.6 mmol, 1.2 eq.). The mixture was stirred at RT for 2 hours. TLC showed the reaction was complete. The mixture was quenched with water (50 mL), extracted over EA, concentrated in vacuo. The crude product was used for next step without further purification (380 mg, crude).

Step 2: Preparation of Compound 65

[0738] To a mixture of compound 65-1 (200 mg, 0.23 mmol, 1.0 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (95.2 mg, 0.69 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (22.9 mg, 0.07 mmol, 0.3 eq.), NaI (10 mg). Then compound Y (51.75 mg, 0.69 mmol, 3.0 eq.) was added. The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was complete. The mixture was quenched with wather (50 mL), extracted over EA, concentrated in vacuo. The mixture was concentrated and purified by prep-HPLC to give the title compound (15 mg, 7.5% yield) as yellow oil.

[0739] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 0.79-0.83 (m, 12H), 1.23-1.27 (m, 62H), 1.29-1.37 (m, 2H), 1.51-1.61 (m, 2H), 1.76-1.93 (m, 7H), 2.13-2.16 (m, 4H), 2.17-2.25 (m, 3H), 2.41-2.51 (m, 7H), 3.05-3.06 (m, 1H), 3.52-3.54 (m. 2H), 3.92-4.03 (m, 4H). LCMS: Rt: 0.588 min; MS m/z (ESI):863.6 [M+H].sup.+.

[0740] The following compounds were prepared in analogous fashion as Compound 65, using corresponding starting material.

TABLE-US-00005 Compound Characterization [00284] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 0.86-0.89 (m, 12H), 1.26-1.32 (m, 58H), 1.33-1.51 (m, 8H), 1.82-2.02 (m, 7H), 2.22-2.25 (m, 7H), 2.40-2.57 (m, 7H), 3.04-3.20 (m, 1H), 3.55-3.57 (m, 2H), 3.99-4.09 (m, 4H). LCMS: Rt: 0.588 min; MS m/z (ESI): 863.6 [M + H].sup.+. [00285] .sup.1H NMR (400 MHz, CCl.sub.3D): δ: 0.86-0.89 (m, 12H), 1.25-1.29 (m, 62H), 1.39-1.42 (m, 4H), 1.57- 1.69 (m, 3H), 1.77-1.85 (m, 2H), 1.96-2.10 (m, 4H), 2.23-2.24 (m, 4H), 2.50-3.18 (m, 11H), 3.61-3.75 (m, 2H), 4.02-4.07 (m, 4H). LCMS: Rt: 0.903 min; MS m/z (ESI): 877.6 [M + H].sup.+.

7.62 Example 62: Preparation of Compound 68

[0741] ##STR00286##

Step 1: Preparation of Compound 68-1

[0742] To a solution of compound SM2 (14.0 g, 16.8 mmol, 1.0 eq.) and compound S (13.6 g, 50.4 mmol, 3.0 eq.) in DCM (100 mL) were added DIEA (10.8 g, 83.9 mmol, 5.0 eq.), EDCI (9.7 g, 50.4 mmol, 3.0 eq.) and DMAP (1.0 g, 8.4 mmol, 0.5 eq.). The mixture was stirred at 40° C.; for 16 hours. TLC showed the reaction was completed. The reaction mixture was concentrated and purified by column chromatography on silica gel (PE/EA=50:1) to give the title compound 68-1 (10.1 g, crude) as yellow oil.

Step 2: Preparation of Compound 68-2

[0743] To a mixture of compound 68-1 (10.1 g, 13.6 mmol, 1.0 eq.) and Pd/C (1 g, 10%) in the MeOH (50 mL) and THE (50 mL) was added compound T (199 mg, 14.9 mmol, 1.1 eq.) at room temperature. The mixture was stirred for 24 h at room temperature under hydrogen atmosphere. TLC showed the reaction was completed. The mixture was filtered, the filtrate was concentrated and then purified by column chromatography on silica gel (PE/EA=10:1) to give the title compound 68-2 (9.1 g, crude) as colorless oil. .sup.1H NMR (400 MHz, CDCl3) δ: 0.86-0.90 (m, 12H), 1.26-1.46 (m, 53H), 1.56-1.62 (m, 2H), 1.83 (s, 2H), 1.96-2.02 (m, 1H), 2.23-2.24 (m, 4H), 3.64 (s, 2H), 4.02-4.11 (m, 4H).

Step 3: Preparation of Compound 68-3

[0744] A mixture of compound 68-2 (150 mg, 0.23 mmol, 1.0 eq.), MsCl (32 mg, 0.28 mmol, 1.2 eq.), DIEA (89 mg, 0.69 mmol, 3.0 eq.) in DCM (5 mL) was stirred for an hour at ambient temperature. The mixture was quenched with water, extracted over EA, dried and concentrated to give the desired product 68-3 (212 mg, crude) as yellow oil.

Step 4: Preparation of Compound 68

[0745] A mixture of compound 68-3 (212 mg, 0.29 mmol, 1.0 eq.), compound E (50 mg, 0.35 mmol, 1.2 eq.), K.sub.2CO.sub.3 (120 mg, 0.87 mmol, 3.0 eq.) and Cs.sub.2CO.sub.3 (3 mg, 0.01 mmol, 0.03 eq.) in ACN (3 mL) was stirred overnight at 80° C. The mixture was concentrated, purified by Prep-HPLC to give the desired product 68 (18 mg, 7.98% yield) as light yellow oil.

[0746] .sup.1H NMR (400 MHz, CDCl3) δ: 0.83-0.92 (m, 12H), 1.17-1.37 (m, 56H), 1.38-1.45 (m, 2H), 1.64-1.67 (m, 2H), 1.70-1.86 (m, 6H), 1.92-2.04 (m, 2H), 2.19-2.26 (m, 4H), 2.40-2.49 (m, 3H), 2.57-2.65 (m, 2H), 3.41-3.51 (m, 2H), 3.97-4.12 (m, 4H). LCMS: Rt: 0.080 min; MS m/z (ESI): 778.5 [M+H].sup.+.

7.63 Example 63: Preparation of Compound 69

[0747] ##STR00287##

Step 1: Preparation of Compound 69-1

[0748] A mixture of compound 68-3 (361 mg, 0.49 mmol, 1.0 eq.), compound I (68 mg, 0.59 mmol, 1.2 eq.), K.sub.2CO.sub.3 (204 mg, 1.48 mmol, 3.0 eq.) and Cs.sub.2CO.sub.3 (5 mg, 0.01 mmol, 0.03 eq.) in ACN (7 mL) was stirred overnight at 80° C. The mixture was concentrated, purified by FCC to give the desired product 69-1 (78 mg, 21.06% yield) as colorless oil. LCMS: Rt: 1.290 min; MS m/z (ESI): 750.7 [M+H].sup.+.

Step 2: Preparation of Compound 69-2

[0749] A mixture of compound 69-1 (78 mg, 0.10 mmol, 1.0 eq.), MsCl (14 mg, 0.12 mmol, 1.2 eq.), DIEA (40 mg, 0.31 mmol, 3.0 eq.) in DCM (2 mL) was stirred for an hour at ambient temperature. The mixture was quenched with water, extracted over EA, dried and concentrated to give the desired product 69-2 (98 mg, crude) as yellow oil.

Step 3: Preparation of Compound 69

[0750] A mixture of compound 69-2 (78 mg, 0.09 mmol, 1.0 eq.), compound U (10 mg, 0.11 mmol, 1.2 eq.), K.sub.2CO.sub.3 (39 mg, 0.28 mmol, 3.0 eq.) and Cs.sub.2CO.sub.3 (1 mg, 0.003 mmol, 0.03 eq.) in ACN (3 mL) was stirred overnight at 80° C. The mixture was concentrated, purified by Prep-HPLC to give the desired product 69 (23 mg, 8.75% yield) as light yellow oil.

[0751] .sup.1H NMR (400 MHz, CDCl3) δ: 0.83-0.92 (m, 12H), 0.98-1.06 (m, 3H), 1.17-1.47 (m, 52H), 1.54-1.72 (m, 5H), 1.78-2.06 (m, 8H), 2.20-2.27 (m, 4H), 2.37-2.46 (m, 4H), 2.49-2.66 (m, 5H), 3.01-3.12 (m, 1H), 3.52-3.59 (m, 2H), 3.98-4.11 (m, 4H). LCMS: Rt: 0.093 min; MS m/z (ESI): 821.6 [M+H].sup.+.

[0752] The following compounds were prepared in analogous fashion as Compound 69, using corresponding starting material.

TABLE-US-00006 Compound Characterizaton [00288] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.95 (m, 15H), 1.26-1.38 (m, 56H), 1.63-1.66 (m, 4H), 1.80-1.86 (m, 2H), 1.96-2.09 (m, 4H), 2.23-2.24 (m, 4H), 2.64 (s, 4H), 2.85-2.92 (m, 6H), 3.09 (s, 1H), 3.32-3.38 (m, 1H), 3.72-3.78 (m, 1H), 3.99-4.11 (m, 4H). LCMS: Rt: 1.950 min; MS m/z (ESI): 835.7 [M + H].sup.+. [00289] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.89 (m, 12H), 1.25-1.38 (m, 52H), 1.71-1.80 (m, 12H), 2.06-2.24 (m, 4H), 2.82-2.93 (m, 6H), 3.11-4.00 (m, 12H), 4.01-4.05 (m, 4H), 5.18-5.34 (m, 2H). LCMS: Rt: 1.575 min; MS m/z (ESI): 861.7 [M + H].sup.+. [00290] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.47 (m, 55H), 1.58-1.70 (m, 4H), 1.82-2.01 (m, 11H), 2.22-2.24 (m, 4H), 2.43-2.58 (m, 8H), 3.13-3.26 (m, 2H), 3.58 (s, 2H), 3.99-4.11 (m, 4H). LCMS: Rt: 1.870 min; MS m/z (ESI): 847.7 [M + H].sup.+. [00291] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.91 (m, 15H), 1.26-1.48 (m, 60H), 1.82-2.02 (m, 8H), 2.22-2.24 (m, 4H), 2.49-2.65 (m, 8H), 3.10-3.15 (m, 1H), 3.60 (s, 2H), 3.99-4.11 (m, 4H). LCSM: Rt: 1.850 min; MS m/z (ESI): 835.7 [M + H].sup.+. [00292] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.14-3.98 (m, 4H), 3.58 (s, 2H), 3.21 (s, 1H), 2.72-2.36 (m, 8H), 2.23 (d, J = 6.9 Hz, 4H), 2.10-2.93 (m, 4H), 1.83 (s, 2H), 1.74-1.00 (m, 64H), 0.97-0.86 (m, 15H). LCMS: Rt: 1.970 min; MS m/z (ESI): 849.8 [M + H].sup.+. [00293] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.93 (m, 15H), 1.26-1.35 (m, 56H), 1.54-1.65 (m, 3H), 1.72-2.04 (m, 9H), 2.23 (d, J = 6.8 Hz, 4H), 2.37-2.63 (m, 10H), 3.00-3.12 (m, 1H), 3.52-3.58 (m, 2H), 3.99-4.13 (m, 4H). LCMS: Rt: 2.207 min; MS m/z (ESI): 849.9 [M + H].sup.+. [00294] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.91 (m, 15H), 1.26-1.68 (m, 63H), 1.83-1.99 (m, 7H), 2.22-2.24 (m, 4H), 2.39-2.61 (m, 10H), 3.05- 3.09 (m, 1H), 3.53-3.55 (m, 2H), 3.99-4.09 (m, 4H). LCMS: Rt: 2.127 min; MS m/z (ESI): 863.9 [M + H].sup.+. [00295] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 15H), 1.26-1.44 (m, 63H), 1.54-1.68 (m, 2H), 1.79-2.01 (m, 7H), 2.23-2.24 (m, 4H), 2.36-2.63 (m, 10H), 3.01-3.12 (m, 1H), 3.51-3.58 (m, 2H), 3.99-4.11 (m, 4 H). LCMS: Rt: 2.397 min; MS m/z (ESI): 877.9 [M + H].sup.+. [00296] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-2.03 (m, 72H), 2.22-2.24 (m, 4H), 2.37-2.46 (m, 4H), 2.56-2.66 (m, 4H), 3.02-3.14 (m, 2H), 3.52-3.59 (m, 2H), 3.99-4.09 (m, 4H). LCMS: Rt: 2.187 min; MS m/z (ESI): 861.9 [M + H].sup.+. [00297] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.07-1.43 (m, 58H), 1.56-1.97 (m, 15H), 2.22-2.63 (m, 13H), 3.03- 3.10 (m, 1H), 3.50-3.57 (m, 2H), 3.99-4.09 (m, 4H), 5.09-5.30 (m, 1H). LCMS: Rt: 2.217 min; MS m/z (ESI): 875.9 [M + H].sup.+. [00298] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.48 (m, 60H), 1.56-1.61 (m, 3H), 1.68-1.73 (m, 3H), 1.81-2.02 (m, 10H), 2.23 (d, J = 6.8 Hz, 4H), 2.34-2.42 (m, 4H), 2.49- 2.61 (m, 5H), 3.02-3.13 (m, 1H), 3.47-3.54 (m, 2H), 3.98-4.11 (m, 4H). LCMS: Rt: 2.367 min; MS m/z (ESI): 890.0 [M + H].sup.+.

7.64 Example 64: Preparation of Compound 70

[0753] ##STR00299##

Step 1: Preparation of Compound 70-1

[0754] A mixture of compound 43-3 (410 mg, 1.06 mmol, 1.0 eq.), compound Z (720 mg, 2.66 mmol, 2.5 eq.), TsOH (20 mg) in toluene (50 mL) was stirred for 2 h at 180° C. TLC showed the reaction was completed. The mixture was concentrated and purified by column chromatography silica gel (EA:PE=0% to 5%) to give the compound 70-1 (700 mg, 78% yield) as colorless oil. LCMS: Rt: 1.750 min; MS m/z (ESI): 888.7 [M+H].sup.+.

Step 2: Preparation of Compound 70-2

[0755] A mixture of compound 70-1 (500 mg, 0.56 mmol, 1.0 eq.) and Pd/C (110 mg, 0.56 mmol, 1.0 eq.) in solution of MeOH (10 mL) and concentrated HCl (5 drops) was stirred for 16 h under H2 at RT. TLC showed the reaction was completed. The mixture was concentrated to give the desired product 70-2 (310 mg, 70% yield) as colorless oil. LCMS: Rt: 1.490 min; MS m/z (ESI): 798.7 [M+H].sup.+

Step 3: Preparation of Compound 70-3

[0756] To a solution of compound 70-2 (150 mg, 0.19 mmol, 1.0 eq.) was dissolved in DCM (10 mL) was added SOCl.sub.2 (113 mg, 0.95 mmol, 5.0 eq.) at RT. The mixture was stirred for 16 hours at 35° C. TLC showed the reaction was completed, the mixture was evaporated under reduced pressure to provide 70-3 (150 mg, crude) as yellow oil.

Step 4: Preparation of Compound 70

[0757] A mixture of compound K (93 mg, 0.92 mmol, 1.2 eq.), compound 70-3 (150 mg, 0.38 mmol, 1.0 eq.), DIEA (70 mg, 0.54 mmol, 3.0 eq.), NaI (7 mg, 0.05 mmol, 0.3 eq.) in THF (10 mL) was stirred overnight at 75° C. The mixture was concentrated under vacuum. The residual was purified by Prep-HPLC to give the desired product 70 (16 mg, 10% yield) as yellow oil.

[0758] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.37-0.55 (m, 2H), 0.56-0.77 (m, 1H), 0.78-0.96 (m, 12H), 1.13-1.33 (m, 54H), 1.51-1.70 (m, 2H), 1.84-2.34 (m, 10H), 2.50-2.65 (m, 5H), 2.75-2.93 (m, 4H), 2.94-3.07 (m, 1H), 3.19-3.43 (m, 1H), 3.44-3.67 (m, 3H), 3.68-3.82 (m, 4H), 3.83-3.93 (m, 1H), 3.94-4.05 (m, 4H). LCMS: Rt: 1.540 min; MS m/z (ESI): 881.8 [M+H].sup.+.

[0759] The following compounds were prepared in analogous fashion as Compound 70, using corresponding starting material.

TABLE-US-00007 Compound Characterization [00300] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.73-0.95 (m, 12H), 1.05-1.35 (m, 56H), 1.36-1.47 (m, 1H), 1.48-1.69 (m, 6H), 1.70-1.88 (m, 3H), 1.89-2.19 (m, 3H), 2.20-2.36 (m, 3H), 2.37-2.56 (m, 3H), 2.57-2.71 (m ,7H), 2.96-3.22 (m, 2H), 3.42-3.65 (m, 3H), 3.66-3.75 (m, 1H), 3.76-3.82 (m, 2H), 3.83-3.89 (m, 1H), 3.90-3.91 (m, 1H), 3.92-4.04 (m, 4H). LCMS: Rt: 2.100 min; MS m/z (ESI): 909.8 [M + H].sup.+. [00301] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.30 (m, 66H), 1.53-1.87 (m ,6H), 2.49-2.56 (m, 6H), 2.57-2.60 (m, 8H), 3.46-3.60 (m, 4H), 3.72-3.91 (m, 4H), 3.96-3.98 (m, 4H). LCMS: Rt: 0.617 min; MS m/z (ESI): 923.8 [M + H].sup.+. [00302] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.72-0.97 (m, 12H), 1.10-1.35 (m, 55H), 1.36-1.48 (m, 6H), 1.50-1.65 (m, 5H), 1.66-1.77 (m, 2H), 1.78-1.90 (m, 2H), 1.91-2.16 (m, 5H), 2.17-2.34 (m, 3H), 2.35-2.71 (m, 10H), 3.37-3.64 (m, 3H), 3.65- 3.91 (m, 5H), 3.92-4.09 (m, 4H). LCMS: Rt: 1.050 min; MS m/z (ESI): 937.8 [M + H].sup.+. [00303] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.72-0.96 (m, 12H), 1.02-1.31 (m, 55H), 1.32-1.53 (m, 6H), 1.54-1.69 (m, 8H), 1.70-1.85 (m, 3H), 1.86-2.21 (m, 9H), 2.22-2.35 (m, 1H), 2.36-2.67 (m, 8H), 2.90-3.16 (m, 2H), 3.44-3.58 (m, 2H), 3.64-3.90 (m, 5H), 3.91-4.05 (m, 3H). LCMS: Rt: 2.070 min; MS m/z (ESI): 951.8 [M + H].sup.+.

7.65 Example 66: Preparation of Compound 77

[0760] ##STR00304##

Compound 77

[0761] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.89 (m, 12H), 1.18-1.55 (m, 40H), 1.73-1.79 (m, 14H), 1.86-1.96 (m, 10H), 2.23-2.25 (m, 4H), 2.96-2.99 (m, 4H), 3.74-3.94 (m, 2H), 3.95-3.98 (m, 5H), 4.01-4.03 (m, 1H), 4.06-4.09 (m, 1H). LCMS: Rt: 1.345 min; MS m/z (ESI): 764.6 [M+H].sup.+.

7.66 Example 67: Preparation of Compound 81

[0762] ##STR00305##

Step 1: Preparation of Compound 81-1

[0763] To a solution of compound S (5.2 g, 19.2 mmol, 1.0 eq.) and compound SM2 (5.5 g, 23.0 mmol, 1.2 eq.) in DCM (100 mL) were added EDCI (11.0 g, 57.6 mmol, 3.0 eq.), DMAP (1.2 g, 9.6 mmol, 0.3 eq.) and DIPEA (12.4 g, 9.6 mmol, 0.3 eq.). The mixture was stirred under 50° C. for 16 hours. After which the TLC showed complete disappearance of starting compound S, the reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dryied over Na.sub.2SO.sub.4 and concentrated. The crude product was purified by column chromatography silica gel (PE/EA=20/1) to give the title compound 81-1 (3.8 g, 40.43% yield) as yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.96 (m, 6H), 1.26-1.49 (m, 28H), 1.59-1.66 (m, 2H), 1.73-2.84 (m, 2H), 1.99-2.08 (m, 1H), 2.20-2.31 (m, 2H), 3.42-3.64 (m, 4H), 4.02-4.24 (m, 2H), 4.46-4.57 (m, 2H), 7.26-7.37 (m, 5H).

Step 2: Preparation of Compound 81-2

[0764] To a solution of compound 81-1 (900 mg, 1.83 mmol, 1.0 eq.) and compound V (682 mg, 3.66 mmol, 2.0 eq.) in DCM (20 mL) were added EDCI (1.05 g, 5.49 mmol, 3.0 eq.), DMAP (67 mg, 0.55 mmol, 0.3 eq.) and DIPEA (71 mg, 0.55 mmol, 0.3 eq.). The mixture was stirred at 50° C. for 16 hours. TLC showed the reaction was completed. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography silica gel (0-1% EA in PE) to give the title compound 81-2 (1.15 mg, 94.96% yield) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.89 (m, 9H), 1.25-1.40 (m, 42H), 1.59-1.62 (m, 4H), 1.77-1.83 (m, 1H), 1.97-2.03 (m, 1H), 2.20-2.33 (m, 4H), 3.45-3.48 (m, 2H), 4.00-4.10 (m, 4H), 4.47-4.53 (m, 2H), 7.26-7.40 (m, 5H).

Step 3: Preparation of Compound 81-3

[0765] To a solution of compound 81-2 (1.15 g, 1.74 mmol, 1.0 eq.) in MeOH (20 mL) were added Pd/C (288 mg) and CHCl.sub.2CH.sub.2Cl (279 mg, 2.09 mmol, 1.2 eq.). The mixture was stirred at room temperature under H.sub.2 for 2 hours. TLC showed the reaction was completed. The reaction mixture was filtrated by a pad of Celite and washed with MeOH. The filtrate was concentrated and purified by column chromatography silica gel (PE/EA=10/1) to give the title compound 81-3 (859 mg, 86.8% yield) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.36 (m, 38H), 1.37-1.44 (m, 4H), 1.54-1.64 (m, 5H), 1.64-1.95 (m, 1H), 1.99-2.06 (m, 1H), 2.20-2.34 (m, 4H), 3.63-3.69 (m, 2H), 4.02-4.13 (m, 4H).

Step 4: Preparation of Compound 81-4

[0766] To a solution of compound 81-3 (850 mg, 1.49 mmol, 1.0 eq.) in DCM (15 mL) were added SOCl.sub.2 (532 mg, 4.47 mol, 3.0 eq.) and pyridine (236 mg, 2.98 mmol, 2.0 eq.). The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was completed. The reaction mixture was extracted with DCM and water. The combined organic layers were washed with brine dried over Na.sub.2SO.sub.4 and concentrated to give the title compound 81-4 (850 mg, 97.14% yield) as yellow oil.

Step 5: Preparation of Compound 81

[0767] To a solution of compound 81-4 (850 mg, 1.45 mmol, 1.0 eq.) in THE (10 mL) were added compound I (334 mg, 2.9 mmol, 2.0 eq.), DIPEA (562 mg, 4.35 mmol, 3.0 eq.) and NaI (66 mg, 0.44 mmol, 0.3 eq.). The reaction mixture was stirred at 70° C. for 16 hours. LCMS showed the reaction was completed. The reaction mixture was poured into water and extracted with EA. The organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography silica gel (DCM/MeOH=70/1-50/1) to give crude title compound 81 (418 mg, 43.27% yield). Crude compound 81 (100 mg) was purified by prep-HPLC to give the title compound (28 mg, 28.0% yield) as colorless oil.

[0768] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.16-1.39 (m, 45H), 1.55-1.66 (m, 5H), 1.75-1.87 (m, 2H), 1.94-2.11 (m, 4H), 2.23-2.32 (m, 4H), 2.48-2.69 (m, 3H), 3.09-3.26 (m, 1H), 3.46-3.68 (m, 2H), 4.01-4.11 (m, 4H). LCMS: Rt: 1.185 min; MS m/z (ESI): 666.5 [M+H].sup.+.

[0769] The following compounds were prepared in analogous fashion as Compound 81, using corresponding starting material.

TABLE-US-00008 Compound Characterization [00306] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.12-3.94 (m, 4H), 2.36-2.19 (m, 4H), 1.98 (s, 2H), 1.83 (s, 3H), 1.59-1.51 (m, 6H), 1.45-1.26 (m, 53H), 0.88 (t, J = 6.8 Hz, 9H). LCMS: Rt: 1.205 min; MS m/z (ESI): 680.6 [M + H].sup.+. [00307] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.11-3.92 (m, 5H), 2.33-2.19 (m, 5H), 2.03-1.92 (m, 2H), 1.88-1.88 (m, 3H), 1.60-1.51 (d, 5H), 1.45- 1.26 (m, 54H), 0.88 (t, J = 6.8 Hz, 9H). LCMS: Rt: 22.575 min; MS m/z (ESI): 694.6 [M + H].sup.+. [00308] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.89 (m, 9H), 1.25-1.73 (m, 55H), 1.75-1.82 (m, 6H), 1.93-1.97 (m, 4H), 1.98-2.04 (m, 4H), 2.28- 2.29 (m, 5H), 2.30-2.70 (m, 3H), 2.51-4.00 (m, 2H), 4.04-4.09 (m, 5H). LCMS: Rt: 1.600 min; MS m/z (ESI): 764.8 [M + H].sup.+. [00309] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.89 (m, 9H), 1.25-1.68 (m, 51H), 1.69-1.86 (m, 13H), 1.95-2.04 (m, 4H), 2.23-2.30 (m, 2H), 2.31- 2.41 (m, 6H), 2.85-3.28 (m, 3H), 3.31-3.47 (m, 2H), 4.03-4.09 (m, 5H). LCMS: Rt: 1.900 min; MS m/z (ESI): 778.8 [M + H].sup.+. [00310] .sup.1H NMR (40 0MHz, CDCl.sub.3) δ: 0.82-0.90 (m, 9H), 1.15-1.47 (m, 62H), 1.55-1.87 (m, 12H), 1.93-2.01 (m, 1H), 2.19-2.30 (m, 4H), 2.35-2.40 (m, 1H), 2.43-2.58 (m, 1H), 2.69-2.82 (m, 1H), 3.36-3.57 (m, 2H), 4.03-4.11 (m, 4H). LCMS: Rt: 1.325 min; MS m/z (ESI): 792.7 [M + H].sup.+. [00311] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.44 (m, 48H), 1.53-1.83 (m, 12H), 1.95-1.99 (m, 1H), 2.23-2.32 (m, 4H), 2.39- 2.43 (m, 2H), 2.56-2.63 (m, 3H), 3.46-3.48 (m, 2H), 4.00-4.10 (m, 4H). LCMS: Rt: 0.093 min; MS m/z (ESI): 708.5 [M + H].sup.+. [00312] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26 (s, 39H), 1.35-1.48 (m, 11H), 1.59- 1.83 (m, 12H), 1.95-2.01 (m, 1H), 2.23-2.32 (m, 4H), 2.37-2.41 (m, 2H), 2.54-2.56 (m, 2H), 2.75 (s, 1H), 3.46-3.48 (m, 2H), 4.00-4.10 (m, 4H). LCMS: Rt: 0.080 min; MS m/z (ESI): 722.5 [M + H].sup.+. [00313] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.38 (m, 43H), 1.51-1.63 (m, 7H), 1.77-1.86 (m, 4H), 1.95-2.00 (m, 1H), 2.23-2.32 (m, 4H), 2.52-2.77 (m, 4H), 3.18-3.32 (m, 2H), 3.60-3.71 (m, 2H), 4.00- 4.10 (m, 4H), 5.22-5.30 (m, 1H). LCMS: Rt: 1.370 min; MS m/z (ESI): 680.8 [M + H].sup.+. [00314] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.11-3.99 (m, 4H), 3.59-3.54 (m, 2H), 3.21-3.10 (m, 1H), 2.57 (s, 2H), 2.46 (s, 2H), 2.30 (t, J = 7.6 Hz, 2H), 2.24 (d, J = 6.9 Hz, 2H), 2.10-1.80 (m, 6H), 1.70-1.56 (m, 4H), 1.45 (s, 2H), 1.34-1.26 (m, 53H), 0.88 (t, J = 6.8 Hz, 9H). LCMS: Rt: 2.320 min; MS m/z (ESI): 758.7 [M + Na].sup.+. [00315] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.10-4.00 (m, 4H), 3.74-3.59 (m, 1H), 2.91-2.63 (m, 2H), 2.30-2.23 (m, 4H), 2.03-1.59 (m 14H), 1.37- 1.26 (m, 57H), 0.87 (d, J = 6.8 Hz, 9H). LCMS: Rt: 1.680 min; MS m/z (ESI): 750.8 [M + H].sup.+.

7.67 Example 68: Preparation of Compound 82

[0770] ##STR00316##

Compound 82-1

[0771] LCMS: Rt: 1.487 min; MS m/z (ESI): 684.5 [M+H].sup.+.

Compound 82

[0772] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.26-1.39 (m, 45H), 1.56-1.75 (m, 6H), 1.78-2.09 (m, 10H), 2.23-2.36 (m, 5H), 2.46-2.68 (m, 7H), 3.08-3.26 (m, 2H), 3.53-3.65 (m, 2H), 4.00-4.11 (m, 4H). LCMS: Rt: 1.275 min; MS m/z (ESI): 763.5 [M+H].sup.+.

[0773] The following compounds were prepared in analogous fashion as Compound 82, using corresponding starting material.

TABLE-US-00009 Compound Characterization [00317] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.93 (m, 12H), 1.26-1.99 (m, 59H), 2.22-2.61 (m, 14H), 3.03-3.10 (m, 1H), 3.53-3.56 (m, 2H), 4.02-4.07 (m, 4H). LCMS: Rt: 1.227 min; MS m/z (ESI): 765.8 [M + H].sup.+. [00318] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.91 (m, 12H), 1.26-2.00 (m, 61H), 2.22-2.61 (m, 14H), 3.04-3.09 (m, 1H), 3.53-3.56 (m, 2H), 4.01-4.07 (m, 4H). LCMS: Rt: 1.227 min; MS m/z (ESI): 779.8 [M + H].sup.+. [00319] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.26-1.44 (m, 53H), 1.51-1.66 (m, 4H), 1.70-2.03 (m, 6H), 2.22-2.24 (m, 2H), 2.28-2.30 (m, 2H), 2.35-2.66 (m, 10H), 3.02-3.12 (m, 1H), 3.51-3.59 (m, 2H), 3.99-4.10 (m, 4H). LCMS: Rt: 1.177 min; MS m/z (ESI): 793.9 [M + H].sup.+. [00320] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.10-3.99 (m, 4H), 3.59-3.525 (m, 2H), 3.15-3.01 (m, 2H), 2.66-2.62 (m, 4H), 2.52 (s, 4H), 2.31-2.23 (m, 2H), 2.24-2.22 (m, 2H), 2.01-1.95 (m, 5H), 1.63-1.30 (m, 58H), 0.88 (t, J = 6.8 Hz, 9H). LCMS: Rt: 2.075 min; MS m/z (ESI): 777.7 [M + H].sup.+. [00321] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.89-1.18 (m, 9H), 1.20-1.41 (m, 51H), 1.42-1.57 (m, 5H), 1.43-1.60 (m, 4H), 1.89-1.94 (m, 4H), 2.22-2.36 (m, 10H), 2.27-2.53 (m, 4H), 2.61-3.06 (m, 1H), 3.06-3.99 (m, 2H), 4.02-4.06 (m, 4H). LCMS: Rt: 1.377 min; MS m/z (ESI): 791.9 [M + H].sup.+. [00322] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.89-1.18 (m, 9H), 1.20-1.41 (m, 51H), 1.42-1.57 (m, 8H), 1.43-1.60 (m, 8H), 1.89-1.94 (m, 8H), 2.22-2.36 (m, 5H), 2.27-2.57 (m, 1H), 3.06-3.99 (m, 2H), 4.02-4.06 (m, 4H). LCMS: Rt: 1.297 min; MS m/z (ESI): 805.8 [M + H].sup.+. [00323] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.89-0.92 (m, 12H), 1.35-1.45 (m, 59H), 1.42-1.45 (m, 4H), 1.58-1.62 (m, 6H), 1.99-2.24 (m, 4H), 2.27-2.31 (m, 10H), 2.51-2.53 (m, 1H), 2.55-4.01 (m, 2H), 4.03-4.06 (m, 4H). LCMS: Rt: 2.257 min; MS m/z (ESI): 835.9 [M + H].sup.+. [00324] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.92 (m, 12H), 1.25-1.35 (m, 60H), 1.38-1.47 (m, 4H), 1.56-1.69 (m, 6H), 1.82-2.00 (m, 4H), 2.22-2.32 (m, 10H), 2.59-3.08 (m, 1H), 3.53-3.57 (m, 2H), 3.98-4.10 (m, 4H). LCMS: Rt: 1.827 min; MS m/z (ESI): 849.7 [M + H].sup.+. [00325] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.25-1.44 (m, 62H), 1.58-1.66 (m, 4H), 1.88-2.00 (m, 6H), 2.22-2.32 (m, 4H), 2.39-2.62 (m, 10H), 3.06-3.56 (m, 1H), 3.99-4.02 (m, 2H), 4.01-4.10 (m, 4H). LCMS: Rt: 0.493 min; MS m/z (ESI): 863.7 [M + H].sup.+. [00326] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.89-1.31 (m, 9H), 1.35-1.45 (m, 58H), 1.46-1.53 (m, 8H), 1.55-1.63 (m, 7H), 1.96-1.97 (m, 4H), 1.98-1.99 (m, 4H), 2.22-2.27 (m, 4H), 12.59-2.63 (m, 2H), 3.54-3.57 (m, 2H), 4.01-4.03 (m, 4H). LCMS: Rt: 2.377 min; MS m/z (ESI): 847.9 [M + H].sup.+. [00327] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.88-1.05 (m, 9H), 1.07-1.43 (m, 60H), 1.45-1.68 (m, 10H), 1.75-1.80 (m, 5H), 1.90-2.32 (m, 9H), 2.37-2.63 (m, 4H), 3.05-3.10 (m, 1H), 3.49-4.00 (m, 2H), 4.01-4.10 (m, 4H). LCMS: Rt: 0.480 min; MS m/z (ESI): 861.8 [M + H].sup.+. [00328] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.25-1.70 (m, 68H), 1.78-1.99 (m, 9H), 2.22-2.51 (m, 8H), 2.52-2.60 (m, 5H), 3.00-3.15 (m, 2H), 3.49-3.52 (m, 2H), 4.01-4.09 (m, 4H). LCMS: Rt: 0.480 min; MS m/z (ESI): 875.8 [M + H].sup.+. [00329] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.12-3.99 (m, 4H), 3.57-3.53 (m, 2H), 2.63 (m, 2H), 2.60- 2.54 (m, 4H), 2.49 (s, 3H), 2.31-2.27 (m, 2H), 2.24-2.22 (m, 2H), 1.99-1.95 (m, 3H), 1.89- 1.29 (m, 72H), 0.91-0.86 (m, 12H). LCMS: Rt: 1.165 min; MS m/z (ESI): 863.7 [M + H].sup.+. [00330] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.12-3.98 (m, 4H), 3.58-3.51 (m, 2H), 3.13-3.03 (m, 1H), 2.64-2.37 (m, 10H), 2.33-2.20 (m, 4H), 1.83- 1.25 (m, 75H), 0.90-0.86 (m, 12H). LCMS: Rt: 1.445 min; MS m/z (ESI): 877.8 [M + H].sup.+. [00331] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.13-3.96 (m, 4H), 3.60-3.49 (m, 2H), 3.11-3.01 (m, 1H), 2.64-2.39 (m, 10H), 2.35-2.20 (m, 4H), 1.76- 1.13 (m, 77H), 0.90-0.86 (m, 12H). LCMS: Rt: 1.555 min; MS m/z (ESI): 891.8 [M + H].sup.+. [00332] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.06-4.01 (m, 4H), 3.55 (s, 2H), 3.12-3.00 (s, 2H), 2.67-2.52 (m, 4H), 2.45-2.41 (m, 3H), 2.36-2.22 (m, 4H), 2.02-1.97 (m, 5H), 1.62-1.33 (m, 73H), 0.89-0.86 (m, 9H). LCMS: Rt: 1.575 min; MS m/z (ESI): 875.8 [M + H].sup.+. [00333] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.13-3.97 (m, 4H), 3.55-3.46 (m, 2H), 2.66-2.51 (m, 4H), 2.42-2.36 (m, 4H), 2.31-2.18 (m, 4H), 2.01- 1.89 (m, 4H), 1.77-1.20 (m, 77 H), 0.89-0.86 (m, 9H). LCMS: Rt: 1.795 min; MS m/z (ESI): 889.8 [M + H].sup.+. [00334] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.10-3.99 (m, 4H), 3.52-3.49 (m, 2H), 3.11-3.02 (m, 1H), 2.59-2.56 (m, 2H), 2.53-2.51 (m, 2H), 2.41- 2.37 (m, 4H), 2.29-2.24 (m, 2H), 2.24-2.22 (m, 2H), 2.01-1.88 (m, 6H), 1.82-1.67 (m, 3H), 1.62-1.33 (m, 73H), 0.89-0.86 (m, 9H). LCMS: Rt: 2.315 min; MS m/z (ESI): 903.9 [M + H].sup.+.

7.68 Example 69: Preparation of Compound 89

[0774] ##STR00335##

Step 1: Preparation of Compound 89-1

[0775] To a solution of compound 81-1 (3.8 g, 7.7 mmol, 1.0 eq.) and compound X (4.3 g, 15.4 mmol, 2.0 eq.) in DCM (30 mL) were added EDCI (4.4 g, 23.1 mmol, 3.0 eq.), DMAP (470 mg, 3.9 mmol, 0.5 eq.) and DIPEA (5.0 g, 38.5 mmol, 5.0 eq.). The mixture was stirred under 50° C. for 16 hours. After which the TLC showed complete disappearance of starting compound 81-1, the reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dryied over Na.sub.2SO.sub.4 and concentrated. The crude product was purified by column chromatography silica gel (PE/EA=50/1) to give the title compound 89-1 (5.1 g, 87.9% yield) as yellow oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.17-1.47 (m, 42H), 1.55-1.60 (m, 4H), 1.62-1.79 (m, 1H), 1.96-2.10 (m, 5H), 2.22-2.32 (m, 4H), 2.70-2.82 (m, 2H), 3.39-3.50 (m, 2H), 3.94-3.12 (m, 4H), 4.44-4.54 (m, 2H), 5.24-5.46 (m, 4H), 7.26-7.37 (m, 5H).

Step 2: Preparation of Compound 89-2

[0776] To a stirred solution of compound 89-1 (5.0 g, 6.6 mmol, 1.0 eq.) in DCM (60 mL) were added BCl.sub.3 (9.3 g, 79.2 mmol, 12.0 eq.) at −78° C. The mixture was stirred at −78° C. for 2 hours. After which the TLC showed complete disappearance of starting compound 89-1, the reaction mixture was poured into NaHCO.sub.3 and extracted with DCM. The organic layer was washed with brine and dryied over Na.sub.2SO.sub.4 and concentrated. The crude product was purified by column chromatography on silica gel (PE/EA=20/1) to give the title compound 89-2 (2.5 g, 56.8% yield) as yellow solid. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.17-1.48 (m, 44H), 1.49-1.59 (m, 5H), 1.63-1.74 (m, 2H), 1.89-2.07 (m, 4H), 2.20-2.32 (m, 4H), 2.73-2.79 (m, 1H), 3.56-3.71 (m, 2H), 4.00-4.17 (m, 4H), 5.31-5.47 (m, 3H).

Step 3: Preparation of Compound 89-3

[0777] To a solution of compound 89-2 (400 mg, 0.6 mmol, 1.0 eq.) and DIPEA (155 mg, 1.2 mmol, 2.0 eq.) in DCM (20 mL) was added MsCl (83 mg, 0.7 mmol, 1.2 eq.) at 0° C. The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was completed. The reaction mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine dried over Na.sub.2SO.sub.4 and concentrated to give the title compound 89-3 (440 g, 98.9% yield) as yellow oil.

Step 4: Preparation of Compound 89

[0778] To a solution of compound 89-3 (440 mg, 0.6 mmol, 1.0 eq.) and compound I (115 mg, 1.2 mmol, 2.0 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (111 mg, 1.8 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (65 mg, 0.2 mmol, 0.3 eq.) and NaI (30 mg, 0.2 mmol, 0.3 eq.). The reaction mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was completed. The reaction mixture was poured into water and extracted with EA. The organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep-HPLC to give the title compound (107 mg, 23.8% yield) as colorless oil.

[0779] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.19-1.53 (m, 46H), 1.58-1.84 (m, 7H), 1.86-2.10 (m, 7H), 2.14-2.34 (m, 5H), 2.38-2.71 (m, 4H), 2.75-2.81 (m, 1H), 3.09-3.27 (m, 1H), 3.33-3.73 (m, 2H), 4.00-4.13 (m, 4H), 5.24-5.59 (m, 3H). LCMS: Rt: 1.395 min; MS m/z (ESI): 760.6 [M+H].sup.+.

[0780] The following compounds were prepared in analogous fashion as Compound 89, using corresponding starting material.

TABLE-US-00010 Compound Characterization [00336] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.91 (m, 9H), 1.26-1.53 (m, 47H), 1.59-1.64 (m, 5H), 1.74-1.77 (m, 4H), 2.02-2.08 (m, 5H), 2.22-2.25 (m, 2H), 2.28-2.32 (m, 2H), 2.47-2.51 (m, 2H), 2.60-2.63 (m, 2H), 2.75-2.79 (m, 2H), 3.01-3.25 (m, 1H), 3.50-3.53 (m, 2H), 4.01-4.07 (m, 4H), 5.32-5.38 (m, 4H). LCMS: Rt: 1.480 min; MS m/z (ESI): 774.7 [M + H].sup.+. [00337] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 9H), 1.21-1.41 (m, 48H), 1.54-1.67 (m, 10H), 1.75-1.82 (m, 3H), 1.96-1.99 (m, 1H), 2.02-2.10 (m, 4H), 2.20-2.30 (m, 4H), 2.38-2.40 (m, 2H), 2.44-2.47 (m, 2H), 2.54-2.61 (m, 2H), 2.64-2.80 (m, 2H), 3.43-3.52 (m, 4H), 5.28-5.44 (m, 4H). LCMS: Rt: 1.155 min; MS m/z (ESI): 802.7 [M + H].sup.+. [00338] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 5.52-5.31 (m, 4H), 4.10-4.03 (m, 5H), 3.86-3.47 (m, 2H), 2.77-2.42 (m, 7H), 2.32-2.30 (m, 2H), 2.28-2.23 (m, 2H), 2.06-1.96 (m, 6H), 1.82-1.72 (m, 6H), 1.36-1.26 (m, 50H), 0.91-0.87 (m, 9H). LCMS: Rt: 1.370 min; MS m/z (ESI): 788.7 [M + H].sup.+. [00339] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.83-0.89 (m, 9H), 1.06-1.90 (m, 65H), 1.96-2.07 (m, 5H), 2.18-2.65 (m, 7H), 2.73-2.81 (m, 2H), 3.43-3.55 (m, 1H), 4.01-4.10 (m, 4H), 5.27-5.43 (m, 4H). LCMS: Rt: 1.445 min; MS m/z (ESI): 816.8 [M + H].sup.+.

7.69 Example 70: Preparation of Compound 95

[0781] ##STR00340## ##STR00341##

Step 1: Preparation of Compound 95-2

[0782] To a solution of compound 95-1 (12.1 g, 66.4 mmol, 1.0 eq.) and compound SM3 (17.6 g, 662 mmol, 5.0 eq.) in DCM (330 mL) at 0° C. was added TRITON B (2.2 g, 13.3 mmol, 0.2 eq.) dropwise. The mixture was stirred at RT for 16 hours. TLC showed the reaction was completed. The mixture was concentrated and purified by column chromatography on silica gel (PE/EA=10/1-4/1) to give the title compound (16.7 g, 87% yield) as colorless oil.

Step 2: Preparation of Compound 95-3

[0783] To a solution of compound 95-2 (15.8 g, 54.8 mmol, 1.0 eq.) in EtOH (250 mL) was added concentrated H.sub.2SO.sub.4 (30 mL). The mixture was stirred at 90° C. for 16 hours. TLC showed the reaction was completed. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel concentrated and purified by column chromatography on silica gel (PE/EA=10/1-5/1) to give the title compound (8.9 g, 43% yield) as colorless oil.

Step 3: Preparation of Compound 95-4

[0784] To a solution of compound 95-3 (8.9 g, 23.3 mol, 1.0 eq.) in THF/H.sub.2O (120 mL/40 mL) was added lithium hydroxide monohydrate (7.8 g, 186.4 mmol, 8.0 eq.). The reaction mixture was stirred at room temperature for 16 hours. TLC showed the reaction worked completely. The reaction mixture was concentrated under reduced pressure to remove the organic solvent. The aqueous layer was acidified to pH=3-4 with 2 N HCl and then extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (6.7 g, 88% yield) as colorless oil.

Step 4: Preparation of Compound 95-5

[0785] A mixture of compound 95-4 (3.26 g, 10 mmol, 1.0 eq.), compound SM4 (7.3 g, 30 mmol, 3.0 eq.), EDCI (5.8 g, 30 mmol, 3.0 eq.), DMAP (611 mg, 5 mmol, 0.5 eq.) and DIPEA (6.5 g, 50 mmol, 5.0 eq.) in DCM (100 mL) was stirred at reflux overnight. TLC showed the reaction worked completely. The reaction mixture was concentrated and purified by column chromatography on silica gel (PE/EA=100/1-50/1-20/1) to give the title compound (5.7 g, 73% yield) as colorless oil.

Step 5: Preparation of Compound 95-6

[0786] To a solution of compound 95-5 (5.7 g, 7.4 mmol, 1.0 eq.) in MeOH (100 mL) were added Pd/C (570 mg) and trichloroethane (1.1 g, 8.1 mmol, 1.1 eq.). The mixture was stirred at RT under H.sub.2 for 2 hours. TLC showed the reaction was complete. The mixture was filtered through a pad of Celite and washed with EA. The filtration was concentrated and concentrated and by column chromatography on silica gel (PE/EA=10/1-4/1) to give the title compound (3.8 g, 76% yield) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 12H), 1.27-1.30 (m, 48H), 1.59-1.68 (m, 2H), 2.51-2.67 (m, 4H), 2.79-2.84 (m, 1H), 3.48-3.58 (m, 4H), 3.69-3.74 (m, 3H), 3.82-3.89 (m, 2H), 3.98-4.01 (m, 4H).

Step 6: Preparation of Compound 95-7

[0787] To a solution of compound 95-6 (1.5 g, 2.2 mmol, 1.0 eq.) and TEA (445 mg, 4.4 mmol, 2.0 eq.) in DCM (22 mL) was added MsCl (302 mg, 2.64 mmol, 1.2 eq.). The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was complete. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (1.6 g, 96% yield) as yellow oil. It was used in the next step without further purification.

Step 7: Preparation of Compound 108

[0788] To a solution of compound 95-7 (300 mg, 0.39 mmol, 1.0 eq.) and compound I (91 mg, 0.79 mmol, 2.0 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (163 mg, 1.18 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (39 mg, 0.12 mmol, 0.3 eq.) and NaI (18 mg, 0.12 mmol, 0.3 eq.). The mixture was stirred at 80° C.; for 10 hours. LCMS showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC to give the title compound (18 mg, 6% yield) as colorless oil. .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.80-0.83 (m, 12H), 1.19-1.31 (m, 50H), 1.51-1.94 (m, 8H), 2.41-2.52 (m, 6H), 2.96-2.98 (m, 1H), 3.06-3.16 (m, 1H), 3.39-3.92 (m, 13H). LCMS: Rt:1.607 min; MS m/z (ESI): 782.7 [M+H].sup.+.

Step 8: Preparation of Compound 95-9

[0789] To a solution of compound 108 (110 mg, 0.14 mmol, 1.0 eq.) in DCM (10 mL) were added SOCl.sub.2 (50 mg, 0.42 mmol, 3.0 eq.). The mixture was stirred at 35° C.; for 10 hours. TLC showed the reaction mixture was complete. The reaction mixture was concentrated to give the title compound (112 mg, 100% yield) as yellow oil.

Step 9: Preparation of Compound 95

[0790] To a solution of compound 95-9 (110 mg, 0.14 mmol, 1.0 eq.) and compound I (32 mg, 0.27 mmol, 2.0 eq.) in THE (10 mL) were added DIEA (54 mg, 0.42 mmol, 3.0 eq.) and NaI (21 mg, 0.14 mmol, 1.0 eq.). The mixture was stirred at 70° C.; for 10 hours. LCMS showed the reaction was completed. The mixture was concentrated and purified by prep-HPLC to give the title compound (25 mg, 20% yield) as colorless oil.

[0791] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.85-0.90 (m, 12H), 1.26-1.43 (m, 52H), 1.56-1.99 (m, 14H), 2.51-2.59 (m, 10H), 3.13-3.15 (m, 2H), 3.46-3.56 (m, 4H), 3.70-3.99 (m, 8H). LCMS: Rt:1.477 min; MS m/z (ESI): 879.9 [M+H].sup.+.

[0792] The following compounds were prepared in analogous fashion as Compound 95, using corresponding starting material.

TABLE-US-00011 Compound Characterization [00342] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.87-0.90 (m, 12H), 0.99-1.03 (m, 3H), 1.19-1.26 (m, 55H), 1.61-1.77 (m, 6H), 2.47-2.62 (m, 15H), 3.46- 3.56 (m, 5H), 3.71-3.98 (m, 8H). LCMS: Rt: 1.057 min; MS m/z (ESI): 881.9 [M + H].sup.+. [00343] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 4.01-3.96 (m, 4H), 3.88-3.76 (m, 5H), 3.68-3.43 (m, 4H), 2.62-2.56 (m, 8H), 2.39-2.37 (m, 3H), 1.83- 1.78 (m, 4H), 1.72 (s, 3H), 1.35-1.15 (m, 60H), 0.89-0.86 (m, 15H). LCMS: Rt: 1.985 min; MS m/z (ESI): 895.8 [M + H].sup.+. [00344] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 3.98-3.96 (m, 4H), 3.84-3.79 (m, 5H), 3.54-3.46 (m, 5H), 2.61-2.54 (m, 7H), 2.52-2.50 (m, 3H), 2.40 (s, 2H), 1.61-1.60 (m, 2H), 1.55-1.53 (m, 4H), 1.48-1.42 (m, 7H), 1.35-1.15 (m, 54H), 0.89-0.84 (m, 15H). LCMS: Rt: 1.875 min; MS m/z (ESI): 909.8 [M + H].sup.+. [00345] .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.86-0.90 (m, 15H), 1.27-1.32 (m, 56H), 1.38-1.58 (m, 14H), 1.71-1.83 (m, 2H), 2.39-2.59 (m, 14H), 3.43-3.56 (m, 5H), 3.70-3.83 (m, 4H), 3.89-3.99 (m, 4H). LCMS: Rt: 1.997 min; MS m/z (ESI): 952.0 [M + H].sup.+.

7.70 Example 71: Preparation of Compound 104

[0793] ##STR00346## ##STR00347##

Step 1: Preparation of Compound 104-2

[0794] To a solution of compound 104-1 (2.4 g, 7.4 mmol, 1.0 eq.) and compound compound X (4.3 g, 16.3 mmol, 2.2 eq.) in DCM (40 mL) were added EDCI (4.3 g, 22.2 mmol, 3.0 eq.), DMAP (452 mg, 3.7 mmol, 0.5 eq.) and DIPEA (4.8 g, 37 mmol, 5.0 eq.). The mixture was stirred under reflux for 16 hours. TLC showed the reaction was complete. The reaction mixture was concentrated and purified by column chromatography silica gel (PE/EA=20/1) to give the title compound 104-2

(3.8 g, 62% yield) as colorless oil. (MC22-163-118)

Step 2: Preparation of Compound 104-3

[0795] To a stirred solution of compound 104-2 (3.7 g, 4.4 mmol, 1.0 eq.) in DCM (30 mL) were added BCl.sub.3 (44 mL, 44 mmol, 10.0 eq.) at −78° C. The mixture was stirred at −78° C. for 1 hours. TLC showed the reaction was complete. The reaction was quenched with NaHCO.sub.3(aq) and extracted with DCM. The organic layer was washed with brine and dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=10/1-4/1) to give the title compound 104-3 (2.0 g, 61% yield) as colorless oil. (MC22-163-119)

Step 3: Preparation of Compound 104-4

[0796] To a solution of compound 104-3 (2.0 g, 2.7 mmol, 1.0 eq.) and DIPEA (698 mg, 5.4 mmol, 2.0 eq.) in DCM (25 mL) was added MsCl (371 mg, 3.2 mmol, 1.2 eq.). The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was complete. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (2.2 g, 100% yield) as yellow oil. It was used in the next step without further purification. (MC22-163-120)

Step 4: Preparation of Compound 104-5

[0797] To a solution of compound 104-4 (2.0 g, 2.5 mmol, 1.0 eq.) and compound Y (376 mg, 5.0 mmol, 2.0 eq.) in ACN (30 mL) were added K.sub.2CO.sub.3 (1.0 g, 7.5 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (244 mg, 0.75 mmol, 0.3 eq.) and NaI (112 mg, 0.75 mmol, 0.3 eq.). The mixture was stirred at 80° C.; for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=50/1) to give the title compound (1.1 g, 58% yield) as colorless oil. (MC22-163-123) LCMS: Rt: 1.247 min; MS m/z (ESI): 790.6 [M+H].sup.+.

Step 5: Preparation of Compound 104-6

[0798] To a solution of compound 104-5 (1.1 g, 1.4 mmol, 1.0 eq.) and DIPEA (362 mg, 2.8 mmol, 2.0 eq.) in DCM (15 mL) was added MsCl (192 mg, 1.68 mmol, 1.2 eq.). The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was complete. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (1.1 g, 91% yield) as yellow oil. It was used in the next step without further purification. (MC22-163-125)

Step 6: Preparation of Compound 104

[0799] To a solution of compound 104-6 (300 mg, 0.34 mmol, 1.0 eq.) and compound K (69 mg, 0.68 mmol, 2.0 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (141 mg, 1.02 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (33 mg, 0.102 mmol, 0.3 eq.) and NaI (15 mg, 0.102 mmol, 0.3 eq.). The mixture was stirred at 80° C.; for 16 hours. LCMS showed the reaction was completed. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep HPLC to give the title compound (15 mg, 5% yield) as colorless oil. (MC22-163-126)

[0800] .sup.1H NMR (400 MHz, CDCl3) δ: 0.40-0.69 (m, 4H), 0.87-0.90 (m, 6H), 1.26-1.34 (m, 36H), 1.56-1.78 (m, 6H), 1.99-2.04 (m, 6H), 2.06-2.19 (m, 4H), 2.28-2.58 (m, 8H), 2.70-2.81 (m, 6H), 3.45-3.60 (m, 4H), 3.71-3.92 (m, 4H), 4.04-4.09 (m, 4H), 5.30-5.55 (m, 8H). LCMS: Rt: 0.093 min; MS m/z (ESI): 873.7 [M+H].sup.+.

[0801] The following compounds were prepared in analogous fashion as Compound 104, using corresponding starting material.

TABLE-US-00012 Compound Characterization [00348] .sup.1H NMR (400 MHz, CDCl3) δ: 0.87-0.90 (m, 6 H), 1.26-1.83 (m, 55 H), 2.02-2.06 (m, 6 H), 2.19-2.26 (m, 3 H), 2.38- 2.59 (m, 12 H), 2.76-2.79 (m, 2 H), 3.43-3.61 (m, 4 H), 3.72-3.92 (m, 4 H), 4.05-4.08 (m, 4 H), 5.31-5.56 (m, 8H). LCMS: Rt: 0.093 min; MS m/z (ESI): 929.8 [M + H].sup.+.

7.71 Example 72: Preparation of Compound 108

[0802] ##STR00349##

[0803] compound 108 .sup.1H NMR (400 MHz, CDCl.sub.3) δ: 0.80-0.83 (m, 12H), 1.19-1.31 (m, 50H), 1.51-1.94 (m, 8H), 2.41-2.52 (m, 6H), 2.96-2.98 (m, 1H), 3.06-3.16 (m, 1H), 3.39-3.92 (m, 13H). LCMS: Rt:1.607 min; MS m/z (ESI): 782.7 [M+H].sup.+.

7.72 Example 73: Preparation of Compound 133

[0804] ##STR00350##

Step 1: Preparation of Compound 133-1

[0805] To a solution of compound 89 (2.0 g, 2.6 mmol, 1.0 eq.) and DIPEA (672 mg, 5.2 mmol, 2.0 eq.) in DCM (20 mL) was added MsCl (357 mg, 3.1 mmol, 1.2 eq.). The mixture was stirred at room temperature for 2 hours. TLC showed the reaction was complete. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give the title compound (2.2 g, 100% yield) as yellow oil. It was used in the next step without further purification. (MC22-163-115)

Step 2: Preparation of Compound 133

[0806] To a solution of compound 133-1 (300 mg, 0.36 mmol, 1.0 eq.) and compound SM8 (84 mg, 0.72 mmol, 2.0 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (149 mg, 1.08 mmol, 3.0 eq.), Cs.sub.2CO.sub.3 (35 mg, 0.108 mmol, 0.3 eq.) and NaI (16 mg, 0.108 mmol, 0.3 eq.). The mixture was stirred at 80° C. for 16 hours. LCMS showed the reaction was completed. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by prep HPLC to give the title compound (115 mg, 37% yield) as colorless oil. (MC22-163-116)

[0807] .sup.1H NMR (400 MHz, CDCl3) δ: 0.86-0.90 (m, 12H), 1.26-1.47 (m, 51H), 1.53-1.77 (m, 4H), 1.89-2.07 (m, 8H), 2.22-2.32 (m, 4H), 2.37-2.64 (m, 10H), 2.75-2.79 (m, 2H), 3.02-3.14 (m, 1H), 3.54-3.59 (m, 2H), 3.99-4.10 (m, 4H), 5.29-5.43 (m, 4H). LCMS: Rt: 0.107 min; MS m/z (ESI): 859.7 [M+H].sup.+.

[0808] The following compounds were prepared in analogous fashion as Compound 133, using corresponding starting material.

TABLE-US-00013 Compound Characterization [00351] .sup.1H NMR (400 MHz, CDCl3) δ: 0.88-0.91 (m, 12H), 1.26-2.07 (m, 63H), 2.22-2.79 (m, 18H), 3.02-3.14 (m, 1H), 3.48-3.60 (m, 2H), 4.01-4.07 (m, 4H), 5.32-5.38 (m, 4H). LCMS: Rt: 0.507 min; MS m/z (ESI): 873.8 [M + H].sup.+. [00352] .sup.1H NMR (400 MHz, CDCl3) δ: 0.86-0.90 (m, 12 H), 1.26-1.37 (m, 54 H), 1.59-1.76 (m, 4 H), 1.75- 2.06 (m, 8 H), 2.22-2.32 (m, 4 H), 2.37-2.61 (m, 10 H), 2.75-2.80 (m, 2 H), 3.03-3.14 (m, 2 H), 3.54-3.58 (m, 2 H), 3.99-4.07 (m, 4 H), 5.30-5.42 (m, 4 H). LCMS: Rt: 0.093 min; MS m/z (ESI): 887.7 [M + H].sup.+. [00353] .sup.1H NMR (400 MHz, CDCl3) δ: 5.35-5.34 (m, 4H), 4.03-4.02 (m, 4H), 3.61-3.51 (m, 2H), 3.16- 3.08 (m, 2H), 2.98-2.77 (m, 6H), 2.05-1.28 (m, 75H), 0.90-0.88 (m, 9H). LCMS: Rt: 0.427 min; MS m/z (ESI): 871.8 [M + H].sup.+. [00354] .sup.1H NMR (400 MHz, CDCl3) δ: 5.36-5.34 (m, 4H), 4.06-4.02 (m, 5H), 3.54-3.46 (m, 2H), 2.77- 2.64 (m, 2H), 2.64-2.55 (m, 4H), 2.41-2.22 (m, 10H), 2.05-1.96 (m, 5H), 1.79-1.75 (m, 8H), 1.62- 1.61 (m, 4H), 1.37-1.18 (m, 51H), 0.89-0.88 (m, 9H). LCMS: Rt: 0.107 min; MS m/z (ESI): 885.7 [M + H].sup.+. [00355] .sup.1H NMR (400 MHz, CDCl3) δ: 0.86-0.93 (m, 12H), 1.26-2.02 (m, 69H), 2.22-2.63 (m, 14H), 3.04-3.17 (m, 1H), 3.52-3.59 (m, 2H), 4.01-4.09 (m, 4H), 5.33-5.36 (m, 2H). LCMS: Rt: 0.453 min; MS m/z (ESI): 861.7 [M + H].sup.+. [00356] .sup.1H NMR (400 MHz, CDCl3) δ: 0.77-0.84 (m, 12H), 1.19-1.95 (m, 71H), 2.16-2.60 (m, 14H), 2.97-3.07 (m, 1H), 3.47-3.55 (m, 2H), 3.92-4.03 (m, 4H), 5.26-5.29 (m, 2H). LCMS: Rt: 0.480 min; MS m/z (ESI): 875.8 [M + H].sup.+. [00357] .sup.1H NMR (400 MHz, CDCl3) δ: 0.86-0.90 (m, 12 H), 1.26-1.44 (m, 59 H), 1.59-1.70 (m, 4 H), 1.82- 2.02 (m, 10 H), 2.22-2.32 (m, 4 H), 2.36-2.65 (m, 10 H), 3.01-3.13 (m, 1 H), 3.49-3.62 (m, 2 H), 3.99-4.10 (m, 4 H), 5.32-5.39 (m, 2 H). LCMS: Rt: 0.093 min; MS m/z (ESI): 889.8 [M + H].sup.+. [00358] .sup.1H NMR (400 MHz, CDCl3) δ: 5.36-5.34 (m, 2H), 4.06-4.01 (m, 4H), 3.66-3.55 (m, 1H), 3.18- 3.04 (m, 2H), 2.74-2.22 (m, 16H), 1.66-1.25 (m, 70H), 0.89-0.86 (m, 9H). LCMS: Rt: 0.107 min; MS m/z (ESI): 873.8 [M + H].sup.+. [00359] 1H NMR (400 MHz, CDCl3) δ: 5.36-5.34 (m, 2H), 4.06-4.02 (m, 4H), 3.59-3.25 (m, 2H), 3.13- 2.98 (m, 1H), 2.68-2.22 (m, 12H), 2.01-1.21 (m, 76H), 0.89-0.86 (m, 9H). LCMS: Rt: 0.107 min; MS m/z (ESI): 887.8 [M + H].sup.+. [00360] .sup.1H NMR (400 MHz, CDCl3) δ: 0.06-1.03 (m, 9H), 1.19-1.56 (m, 64H), 1.57-1.67 (m, 12H), 1.78-1.92 (m, 9H), 1.94-2.27 (m, 5H), 2.42-2.53 (m, 1H), 3.50-3.52 (m, 2H), 3.99-4.07 (m, 4H), 5.34-5.35 (m, 2H). LCMS: Rt: 0.107 min; MS m/z (ESI): 901.8 [M + H].sup.+.

7.73 Example 74: Preparation of Compound 145

[0809] ##STR00361##

Step 1: Preparation of Compound 145-1

[0810] To a solution of compound SM2 (800 mg, 3.36 mmol, 1.0 eq.) and compound SM7 (144 mg, 8.40 mmol, 2.5 eq.) in toluene (50 mL) was added TsOH (20 mg). The mixture was stirred at 180° C. for 16 hours. TLC showed the reaction was completed. The mixture was diluted with EA and washed with Saturated NaHCO.sub.3 aqueous solution. The organic layer was washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (PE/EA=100/1) to give 145-1 (850 mg, 49% yield) as colorless oil. (MC21-151-081)

Step 2: Preparation of Compound 145-2

[0811] To a solution of compound 145-1 (850 mg, 1.73 mmol, 1.0 eq.) in MeOH (10 mL) was added Pd/C (20 mg). The mixture was stirred at RT under H.sub.2 for 16 hours. TLC showed the reaction was completed. The mixture was filtered through a pad of Celite and washed with EA. The filtration was concentrated and purified by column chromatography on silica gel (PE/EA=5/1) to give 145-2 (610 mg, 63% yield) as colorless oil. (MC21-151-082)

Step 3: Preparation of Compound 145-3

[0812] To a solution of compound 145-2 (400 mg, 1.00 mmol, 1.0 eq.) and DIPEA (129 mg, 1.00 mmol, 1.0 eq.) in DCM (10 mL) at 0° C.; was added MsCl (114 mg, 1.00 mmol, 1.0 eq.). The mixture was stirred at room temperature for 1 hour. The mixture was poured into water and extracted with DCM. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated to give 145-3 (478 mg) as yellow oil. It was used in the next step without further purification. (MC21-151-086) LCMS: Rt: 0.900 min; MS m/z (ESI): 428.4 [M+H].sup.+.

Step 4: Preparation of Compound 145

[0813] To a solution of compound 145-3 (478 mg, 1.0 mmol, 1.0 eq) and compound K (121 mg, 1.2 mmol, 1.2 eq.) in ACN (10 mL) were added K.sub.2CO.sub.3 (414 mg, 3.0 mmol, 3.0 eq), Cs.sub.2CO.sub.3 (98 mg, 0.3 mmol, 0.3 eq) and NaI (14 mg, 0.1 mmol, 0.1 eq). The mixture was stirred at 80° C.; for 16 hours. LCMS showed the reaction was complete. The mixture was poured into water and extracted with EA. The combined organic layers were washed with brine, dried over Na.sub.2SO.sub.4 and concentrated. The residue was purified by column chromatography on silica gel (DCM/MeOH=10/1) to give 145 (300 mg, 62% yield) as yellow oil. (MC21-151-087) LCMS: Rt: 0.810 min; MS m/z (ESI): 484.4 [M+H].sup.+.

[0814] The following compounds were prepared in analogous fashion as Compound 145, using corresponding starting material.

TABLE-US-00014 Compound Characterization [00362] LCMS: Rt: 0.830 min; MS m/z (ESI): 498.4 [M + H].sup.+. [00363] LCMS: Rt: 0.800 min; MS m/z (ESI): 512.4 [M + H].sup.+. [00364] LCMS: Rt: 0.850 min; MS m/z (ESI): 526.5 [M + H].sup.+. [00365] LCMS: Rt: 0.847 min; MS m/z (ESI): 540.5 [M + H].sup.+. [00366] LCMS: Rt: 0.840 min; MS m/z (ESI): 554.4 [M + H].sup.+. [00367] LCMS: Rt: 0.870 min; MS m/z (ESI): 512.3 [M + H].sup.+. [00368] LCMS: Rt: 0.770 min; MS m/z (ESI): 414.2 [M + H].sup.+. [00369] LCMS: Rt: 0.770 min; MS m/z (ESI): 442.3 [M + H].sup.+. [00370] LCMS: Rt: 0.773 min; MS m/z (ESI): 470.3 [M + H].sup.+. [00371] LCMS: Rt: 0.820 min; MS m/z (ESI): 526.4 [M + H].sup.+. [00372] LCMS: Rt: 1.160 min; MS m/z (ESI): 554.4 [M + H].sup.+. [00373] LCMS: Rt: 1.220 min; MS m/z (ESI): 582.4 [M + H].sup.+. [00374] LCMS: Rt: 1.460 min; MS m/z (ESI): 610.4 [M + H].sup.+. [00375] LCMS: Rt: 1.380 min; MS m/z (ESI): 798.6 [M + H].sup.+. [00376] LCMS: Rt: 1.050 min; MS m/z (ESI): 593.5 [M + H].sup.+. [00377] LCMS: Rt: 0.870 min; MS m/z (ESI): 606.6 [M + H].sup.+. [00378] LCMS: Rt: 1.135 min; MS m/z (ESI): 794.7 [M + H].sup.+. [00379] LCMS: Rt: 0.850 min; MS m/z (ESI): 498.6 [M + H].sup.+. [00380] LCMS: Rt: 0.900 min; MS m/z (ESI): 554.5 [M + H].sup.+.

7.74 Example 75: Preparation and Characterization of Lipid Nanoparticles

[0815] Briefly, a cationic lipid provided herein, DSPC, cholesterol, and PEG-lipid were solubilized in ethanol at a molar ratio of 50:10:38.5:1.5, and mRNA were diluted in 10 to 50 mM citrate buffer, pH=4. The LNPs were prepared at a total lipid to mRNA weight ratio of approximately 10:1 to 30:1 by mixing the ethanolic lipid solution with the aqueous mRNA solution at a volume ratio of 1:3 using a microfluidic apparatus, total flow rate ranging from 9-30 mL/min. Ethanol were thereby removed and replaced by DPBS using dialysis. Finally, the lipid nanoparticles were filtered through a 0.2 μm sterile filter.

[0816] Lipid nanoparticle size were determined by dynamic light scattering using a Malvern Zetasizer Nano ZS (Malvern UK) using a 173° backscatter detection mode. The encapsulation efficiency of lipid nanoparticles were determined using a Quant-it Ribogreen RNA quantification assay kit (Thermo Fisher Scientific, UK) according to the manufacturer's instructions.

[0817] As reported in literature, the apparent pKa of LNP formulations correlates with the delivery efficiency of LNPs for nucleic acids in vivo. The apparent pKa of each formulation was determined using an assay based on fluorescence of 2-(p-toluidino)-6-napthalene sulfonic acid (TNS). LNP formulations comprising of cationic lipid/DSPC/cholesterol/DMG-PEG (50/10/38.5/1.5 mol %) in PBS were prepared as described above. TNS was prepared as a 300 uM stock solution in distilled water. LNP formulations were diluted to 0.1 mg/ml total lipid in 3 mL of buffered solutions containing 50 mM sodium citrate, 50 mM sodium phosphate, 50 mM sodium borate, and 30 mM sodium chloride where the pH ranged from 3 to 9. An aliquot of the TNS solution was added to give a final concentration of 0.1 mg/ml and following vortex mixing fluorescence intensity was measured at room temperature in a Molecular Devices Spectramax iD3 spectrometer using excitation and mission wavelengths of 325 nm and 435 nm. A sigmoidal best fit analysis was applied to the fluorescence data and the pKa value was measured as the pH giving rise to half-maximal fluorescent intensity.

7.75 Example 61: Animal Study

[0818] Lipid nanoparticles comprising compounds in the following table encapsulating human erythropoietin (hEPO) mRNA were systemically administered to 6-8 week old female ICR mice (Xipuer-Bikai, Shanghai) at 0.5 mg/kg dose by tail vein injection and mice blood were sampled at specific time points (e.g., 6 hours) post administration. In addition to the aforementioned tested groups, lipid nanoparticles comprising dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA, usually abbreviated to MC3) encapsulating hEPO mRNA were similarly administered at the same dose to age and gender comparative groups of mice as a positive control.

[0819] Mice were euthanized by CO.sub.2 overdoses after the last sampling time point. Serum were separated from total blood by centrifugation at 5000 g for 10 minutes at 4° C., snap-frozen and stored at −80° C. for analysis. ELSA assay were carried out using a commercial kit (DEP00, R&D systems) according to manufacturer's instructions.

[0820] Characteristics of tested lipid nanoparticles, including expression levels over MC3 measured from the tested group are listed the table below.

TABLE-US-00015 TABLE 4 Encapsulation Expression Apparant Lipid size (nm) polydispersity Efficiency over MC3 Pka  1 72.4 0.12  96.3% D 5.902  2 69.05 0.094  94.5% D 5.177  3 105.6 0.124  92.8% D  4 54.52 0.2  94.6% D 5.083  5 78.36 0.384  92.4% D 7.315  6 126.4 0.241  93.4% D 7.035  7 85.46 0.063  96.7% D 8.7  8 102.9 0.156  88.2% D 7.327  9 70.97 0.144  90.8% C  10 84.22 0.356  94.8% D  11 77.98 0.196  82.2% D 7.701  13 61.3 0.128  94.2% D 7.089  14 56.31 0.4  86.1% D 4.536  15 62.11 0.139  93.5% C 5.875  16 74.31 0.060  89.8% C  17 79.43 0.186  92.8% C 5.835  18 65.06 0.17  90.6% D 4.916  19 117.5 0.156  88.5% D 4.957  20 67.7 0.145  90.7% D 5.96  21 74.88 0.221  85.5% D 5.49  22 68.67 0.206  83.5% D 5.457  23 74.73 0.076  88.5% B 5.993  24 56.39 0.094  93.0% C 6.005  25 58.01 0.112 94.70% C  26 55.8 0.153 93.30% C 5.696  27 65.62 0.139  73.3% D 5.351  28 54.96 0.2  86.9% D 5.905  29 53.73 0.110  80.1% C 5.699  30 49.64 0.107  92.7% D 5.409  31 51.35 0.127  80.1% D 5.704  32 59.1 0.232  44.6% D 3.519  33 63.54 0.06 94.30% C 5.943  34 58.33 0.103 89.60% D 5.482  35 56.71 0.1 96.80% C 5.940  36 59.91 0.078 94.20% C 5.774  37 51.77 0.156 92.70% C 5.726  38 97.62 0.194  76.2% D 6.971  39 111.2 0.162  94.7% D 6.712  40 230 0.326  36.4% D  41 82.81 0.136  83.5% D 5.746  42 82.81 0.155  94.4% D 6.283  43 138.6 0.362  78.4% D 7.227  44 112.8 0.178  85.2% D 6.65  45 68.33 0.289  86.8% D 7.466  46 103.3 0.039  95.9% C 6.047  47 82.89 0.082  91.3% C 6.120  48 79.39 0.113  91.7% C 6.263  49 99.66 0.031  83.7% A 6.221  50 58.14 0.099  90.4% C 8.298  51 75.11 0.057  94.7% C 7.727  52 66.03 0.14  96.5% D  53 90.06 0.014 89.79% C 6.069  55 75.43 0.034  89.5% C 6.851  56 53.32 0.107  91.0% B 6.403  57 70.05 0.055  97.0% C 7.73  58 56.37 0.032  97.3% D  59 94.89 0.049  96.1% D  60 73.31 0.068  92.6% C 6.985  61 70.83 0.055  96.2% C 7.219  62 58.44 0.075  94.1% D 7.062  63 150.2 0.076  95.9% C 6.928  64 73.8 0.075  91.6% D  65 73.61 0.082  97.0% C 6.428  66 72.85 0.044  96.6% B 6.749  67 130.6 0.116 91.83% D 6.444  68 55.01 0.084 89.42% C 6.22  69 110.30 0.013 91.81% C 6.789  70 78.45 0.049 93.32% A 6.351  71 76.45 0.077 94.62% B 6.727  72 81.89 0.055 95.22% A  73 76.26 0.088 94.69% A 6.616  74 101.00 0.031 93.97% A 6.539  77 56.88 0.046 89.92% B 6.439  78 159.72 0.017 93.77% C 6.166  79 103.64 0.01 91.29% A 5.927  80 98.95 0.027 92.20% A 6.063  81 72.78 0.023 88.28% B 6.79  82 84.46 0.079 92.38% A 6.621  83 94.04 0.019 92.39% A 6.182  84 83.05 0.01 93.44% B 6.785  85 74.14 0.066 92.17% B 6.438  86 73.58 0.03 92.81% A  87 64.08 0.044 92.47% B 6.372  88 59.54 0.063 92.91% B 6.309  89 69.36 0.04 92.56% B 6.926  90 70.38 0.043 93.02% C 7.024  91 63.89 0.111 92.11% B 6.608  92 70.85 0.051 90.25% A  93 63.32 0.111 85.30% C 6.377  94 77.15 0.04 93.93% A 6.793  98 77.16 0.01 93.58% B 6.997  99 77.25 0.025 94.14% B 7.059 100 64.54 0.055 91.24% B 6.666 101 59.51 0.111 92.79% B 6.488 106 65.31 0.046 93.17% A 6.035 107 61.18 0.067 93.66% B 108 47.87 0.097 92.04% D 109 74 0.011 92.59% A 6.204 110 75.71 0.038 92.71% A 6.147 111 62.77 0.048 92.54% B 6.072 A: ≥2 B: ≥1 and <2 C: ≥0.1 and <1 D: <0.1