Novel Process for Preparing Nucleotide P(V) Monomers

Abstract

The present invention relates to an optimized method for preparing a P(V) monomer of formula (IIIa) or (IIIb) from a nucleoside using DBU as a base at about 0.8 equivalent to nucleoside.

##STR00001##

Claims

1. A method for preparing a compound of formula (IIIa) or (IIIb) comprising the step of reacting a modified nucleoside of formula (I): ##STR00040## wherein PG is a hydroxyl protecting group, R.sup.1 is H and R.sup.2 is —O(CH.sub.2).sub.2OCH.sub.3 or, R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH.sub.2—O— or —CH(CH.sub.3)O—, wherein the oxygen is attached via position R.sup.2; with a compound of formula (IIa) or (IIb): ##STR00041## wherein R.sup.3 is a C.sub.6-C.sub.10 aryl substituted by halo and halo is selected from F, Cl and Br and R.sup.4 is a linear or branched C.sub.1-C.sub.6-alkyl; with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as base, wherein DBU is used in an amount of about 0.8 equivalent to nucleoside of formula (I); to produce a monomer of formula (IIIa) or (IIIb): ##STR00042##

2. The method of claim 1, wherein the nucleoside of formula I has the following formula (Ia): ##STR00043##

3. The method of claim 1, wherein the nucleoside of formula I has the following formula (Ib): ##STR00044##

4. The method of claim 1, wherein the nucleoside of formula I has the following formula (Ic): ##STR00045##

5. The method of claim 1 wherein the nucleobase is selected from the group consisting of A, T, G, C, and 5-methyl cytosine.

6. The method claim 1 wherein the nucleobase is selected from the group consisting of A, T and G.

7. The method claim 5 wherein in A, G, C or 5-methyl cytosine the exocyclic amino moiety is protected by an amino protecting group.

8. The method of claim 7, wherein the amino protecting group is selected from the group consisting of DMF or iBu.

9. The method of claim 1 further comprising the step of coupling compounds of formula (IIIa) or (IIIb) as P(V) monomers for preparing an antisense oligonucleotide.

10. The method of claim 1, wherein R.sup.1 is H and R.sup.2 is —O(CH.sub.2).sub.2OCH.sub.3.

11. The method of claim 1, wherein R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH.sub.2—O— wherein the oxygen is attached via position R.sup.2.

12. The method of claim 1, wherein R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH(CH.sub.3)O—, wherein the oxygen is attached via position R.sup.2.

13. The method of claim 1, wherein R.sup.3 is a phenyl substituted by 1 to 5 F.

14. The method of claim 1, wherein R.sup.4 is methyl or ethyl.

15. Use of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as base in the production of a compound of formula (IIIa) or (IIIb) starting from a nucleoside of formula (I) as claimed in claim 1, wherein DBU is used in an amount of about 0.8 equivalent to nucleoside.

16. A compound of formula (IIIa) or (IIIb): ##STR00046## wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are as defined in claim 1.

17. The compound of formula (IIIa) or (IIIb) of claim 16 prepared according to the process of claim 1.

18. An oligonucleotide manufactured according to a method of claim 1.

19. (canceled)

Description

DETAILED DESCRIPTION OF THE INVENTION

[0100] In one aspect, the invention relates to a method for preparing a P(V) monomer of formula (IIIa) or (IIIb) comprising the step of reacting a modified nucleoside of formula (I):

##STR00009##

wherein PG is a hydroxyl protecting group,
R.sup.1 is H and R.sup.2 is —O(CH.sub.2).sub.2OCH.sub.3 or,
R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH.sub.2—O— or —CH(CH.sub.3)O—, wherein the oxygen is attached via position R.sup.2;
with a compound of formula (IIa) or (IIb):

##STR00010##

wherein R.sup.3 is a C.sub.6-C.sub.10 aryl substituted by halo and halo is selected from F, Cl and Br and R.sup.4 is a linear or branched C.sub.1-C.sub.6-alkyl;
with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as base, wherein DBU is used in an amount of about 0.8 equivalent to nucleoside of formula (I);
to produce a monomer of formula (IIIa) or (IIIb):

##STR00011##

[0101] In one or all embodiments of the invention, the compound of formula (IIa) or (IIb) is:

##STR00012##

[0102] In one or all embodiments of the invention, the nucleoside of formula I has the following formula (Ia):

##STR00013##

[0103] In one or all embodiments of the invention, the nucleoside of formula I has the following formula (Ib):

##STR00014##

[0104] In one or all embodiments of the invention, the nucleoside of formula I has the following formula (Ic):

##STR00015##

[0105] In one or all embodiments of the invention, the nucleobase is selected from the group consisting of A, T, G, C, and 5-methyl cytosine.

[0106] In one or all embodiments of the invention, the nucleobase is selected from the group consisting of A, T and G.

[0107] In one or all embodiments of the invention, the nucleobase is A, G, C or 5-methyl cytosine wherein the exocyclic amino moiety is protected by an amino protecting group.

[0108] The amino protecting group can be selected from the group consisting of DMF and iBu.

[0109] In one or all embodiments, the method of the invention further comprises the step of coupling compounds of formula (IIIa) or (IIIb) as P(V) monomers for preparing an antisense oligonucleotide. This can be done according to processes known in the art.

[0110] In one or all embodiments of the invention, R.sup.1 is H and R.sup.2 is —O(CH.sub.2).sub.2OCH.sub.3.

[0111] In one or all embodiments of the invention, R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH.sub.2—O— wherein the oxygen is attached via position R.sup.2.

[0112] In one or all embodiments of the invention R.sup.1 and R.sup.2 together form a 2′-4′ bridge selected from —CH(CH.sub.3)O—, wherein the oxygen is attached via position R.sup.2.

[0113] In one or all embodiments of the invention R.sup.3 is a phenyl substituted by 1 to 5 F, for example phenyl substituted by 1, 2, 3, 4 or 5 F, for example substituted by 4 or 5 F, for example substituted by 5 F.

[0114] In one or all embodiments of the invention R.sup.4 is methyl or ethyl for example methyl.

[0115] In one embodiment of the invention the method of the invention is a method for preparing a compound of formula (IIIa1) or (IIIb1) comprising the step of reacting a modified nucleoside of formula (I):

##STR00016##

wherein PG is a hydroxyl protecting group,
with a compound of formula (IIa1) or (IIb1):

##STR00017##

with 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as base in MeCN, wherein DBU is used in an amount of about 0.8 equivalent to nucleoside of formula (Ia);
to produce a monomer of formula (IIIa1) or (IIIb1):

##STR00018##

[0116] In another aspect, the invention relates to the use of 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU) as base in the production of a compound of formula (IIIa) or (IIIb) starting from a nucleoside of formula (I) as described herein, wherein DBU is used in an amount of about 0.8 equivalent to nucleoside.

[0117] In another aspect, the invention relates to a compound of formula (IIIa) or (IIIb):

##STR00019##

[0118] wherein R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are as defined herein.

[0119] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are of formula (IIIa1) or (IIIb1):

##STR00020##

[0120] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are selected from the group consisting of the following compounds:

##STR00021## ##STR00022## ##STR00023##

[0121] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are the following compounds:

##STR00024##

[0122] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are the following compounds:

##STR00025##

[0123] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are the following compounds:

##STR00026##

[0124] In an embodiment, the compounds of formula (IIIa) or (IIIb) according to the invention are the following compounds:

##STR00027##

[0125] An other aspect of the invention is a compound of formula (IIIa) or (IIIb) prepared according to the process of the invention. In one embodiment, it is a compound of formula (IIIa) or (IIIb) directly obtained by the process of the invention.

[0126] In another aspect, the invention relates to an oligonucleotide manufactured according to the method of the invention.

[0127] In one or all aspects or embodiments of the invention, DBU can be reacted in solution in MeCN. Other suitable solvents can also be contemplated by the person skilled in the art.

[0128] In one or all aspects or embodiments of the invention, the reaction can be conducted in solution in MeCN. Other suitable solvents can also be contemplated by the person skilled in the art.

[0129] In an embodiment, the compound of formula (I) is selected from the group consisting of:

##STR00028##

[0130] As it can be evidenced from table 6 hereinbelow, the inventors found a new process for preparing LNA monomers through a P(V) reagent based system with surprisingly optimized conditions, resulting in a cheapest and higher yield process than described in the art. For instance, when DBU is used as 0.8 equivalents in the preparation of LNA-T, LNA-A and LNA-G, the isolated yield of compounds III is significantly higher than when 1.3 DBU equivalents are used as recommended in the prior art. As for the preparation of LNA-C, the same or a better yield is obtained when using 0.8 equivalents of DBU compared to when using 1.3 DBU equivalents. This means that the process can be significantly cheaper because less DBU equivalents are needed.

EXAMPLES

[0131] ##STR00029##

General Synthesis for P(V) LNA a Monomers Using Either 0.8 Eq DBU or 1.3 Eq DBU

[0132] To suspension of 5′-ODMTr-LNA-A (500 mg 0.73 mmol) and (+)-1 (423 mg, 0.95 mmol) in MeCN (9 mL) solution of DBU (87 μL, 0.58 mmol, 0.8 eq) in MeCN (2 ml) was added at room temperature, then stirred at room temperature for 40 min (TLC-full conv.). Reaction mixture was filtered through silicagel plug, rinsed with EtOAc (35 mL). Filtrate was washed with water (15 mL) 10% Na.sub.2HPO.sub.4 (20 mL), water (20 mL), sat. NaHCO.sub.3 (20 mL), dried over Na.sub.2SO.sub.4 and evaporated. Residue was purified by column chromatography (eluent EtOAc in hexanes from 50% to 70%).

TABLE-US-00001 TABLE 1 Isolated yield DBU eq. (+)-1 (−)-1 LNA-A 0.8 89-91% 80% 1.3 26-65% 76%

General Synthesis for P(V) LNA T Monomers Using Either 0.8 Eq DBU or 1.3 Eq DBU

[0133] To suspension of 5′-ODMTr-LNA-T (500 mg 0.87 mmol) and (−)-1 (507 mg 1.13 mmol) in MeCN (9 mL) solution of DBU (104 μL 0.58 mmol 0.8 eq) in MeCN (2 ml) was added at room temperature, and then stirred at room temperature for 40 min (TLC-full conv.). Reaction mixture was filtered through silicagel plug, rinsed with EtOAc (35 mL). Filtrate was washed with 10% Na.sub.2HPO.sub.4 (20 mL), water (20 mL), sat. NaHCO.sub.3 (20 mL), dried over Na.sub.2SO.sub.4 and evaporated. Residue was purified by column chromatography (eluent EtOAc/hexanes+1/1).

TABLE-US-00002 TABLE 2 Isolated yield DBU eq. (+)-1 (−)-1 LNA-T 0.8 83% 88-84% 1.3 62% 79%

General Synthesis for P(V) LNA C Monomers Using Either 0.8 Eq DBU or 1.3 Eq DBU

[0134] To solution of 5′-ODMTr-LNA-C (588 mg 0.87 mmol) and (+)-1 (505 mg 1.13 mmol) in MeCN (11 mL) solution of DBU (DBU 106 μL 0.58 mmol 0.8 eq) in MeCN (2 ml) was added at room temperature, and then stirred at room temperature for 40 min (TLC-full conv.). Reaction mixture was filtered through silicagel plug, rinsed with EtOAc (35 mL). Filtrate was washed with 10% Na.sub.2HPO.sub.4 (20 mL), water (20 mL), sat. NaHCO.sub.3 (20 mL), dried over Na.sub.2SO.sub.4 and evaporated. Residue was purified by column chromatography (eluent EtOAc/hexanes+1/2 then 1/1).

TABLE-US-00003 TABLE 3 Isolated yield DBU eq. (+)-1 (−)-1 LNA-C 0.8 81% 84% 1.3 88% 80%

General Synthesis for P(V) LNA G-iBu Monomers Using Either 0.8 Eq DBU or 1.3 Eq DBU

[0135] To solution of 5′-ODMTr-LNA-G.sup.iBu (500 mg, 0.75 mmol) and (+)-1 (434 mg, 0.97 mmol) in MeCN (11 mL) solution of DBU (91.2 μL 0.60 mmol 0.8 eq) in MeCN (1 mL) was added at room temperature, and then stirred at room temperature for 40 min (TLC-full conv.). Reaction mixture was filtered through silicagel plug, rinsed with EtOAc (35 mL). Filtrate was washed with 10% Na.sub.2HPO.sub.4 (20 mL), water (20 mL), sat. NaHCO.sub.3 (20 mL), dried over Na.sub.2SO.sub.4 and evaporated. Residue was purified by column chromatography (eluent EtOAc in hexanes from 50% to 75%).

TABLE-US-00004 TABLE 4 Isolated yield DBU eq. (+)-1 (−)-1 LNA-G.sup.iBu 0.8 79% 83% 1.3 49% 66%

General Synthesis for P(V) LNA G-DMF Monomers Using Either 0.8 Eq DBU or 1.3 Eq DBU

[0136] To suspension of 5′-ODMTr-LNA-G.sup.DMF (500 mg 0.77 mmol) in MeCN (6 mL) and THF (13 mL) a solution of (+)-1 (444 mg 1.00 mmol) in MeCN (5 mL) was added, followed with solution of DBU (DBU 91.5 μL 0.61 mmol 0.8 eq) in MeCN (1 mL). The reaction mixture stirred at room temperature for 1 h (gradually LNA-G.sup.DMF is dissolved). Reaction mixture was filtered through silicagel plug, product was washed out from plug with EtOAc (100 mL), EtOAc/THF (1/2 80 mL), THF (50 mL). Filtrate was washed with 10% Na.sub.2HPO.sub.4 (100 mL), water (100 mL), sat. NaHCO.sub.3 (100 mL), brine (100 mL) dried over Na.sub.2SO.sub.4 and evaporated. Residue was purified by column chromatography (eluent THF in EtOAc from 10% to 30%).

TABLE-US-00005 TABLE 5 Isolated yield DBU eq. (+)-1 (−)-1 LNA-G.sup.DMF 0.8 75% 84% 1.3 43% 54%

TABLE-US-00006 TABLE 6 yield overview Isolated yield DBU eq. (+)-1 (−)-1 LNA-T 0.8 83% 88-84% 1.3 62% 79% LNA-C- 0.8 81% 84% pg 1.3 88% 80% LNA-A- 0.8 89-91% 80% pg 1.3 26-65% 76% LNA- 0.8 79% 83% G.sup.iBu 1.3 49% 66% LNA- 0.8 75% 84% G.sup.DMF 1.3 43% 54%

[0137] 1.1 Analytical Data

##STR00030##

[0138] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 8.71 (1H, s), 7.64 (1H, d, J=1.2 Hz), 7.46-7.41 (2H, m), 7.36-7.26 (6H, m), 7.25-7.20 (1H, m) 6.87-7.60 (4H, m), 5.70 (1H, s), 5.16 (1H, d, J=7.3 Hz), 4.95 (1H, s), 4.85 (1H, s), 4.66 (1H, s), 4.39 (1H, dt, J=12.8 3.5 Hz), 3.81 (2H, s), 3.78 (6H, s), 3.47 (1H, d, J=11.1 Hz), 3.42 (1H, d, J=11.1 Hz), 2.60-2.52 (1H, m), 2.35-2.25 (1H, m), 2.09-1.62 (5H, m), 1.76 (3H, s), 1.67-1.64 (6H, m)

[0139] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm) 163.7, 158.8, 149.9, 145.0, 144.2, 135.3, 135.2, 134.2, 130.3, 128.3, 128.2, 127.2, 113.5, 111.8, 111.0, 87.7, 87.6, 87.2, 86.9, 85.6, 78.4, 74.3, 74.2, 72.4, 66.6, 57.7, 55.4, 39.0, 33.8, 27.7, 27.6, 23.6, 22.6, 22.0, 12.7.

[0140] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm) 101.4.

[0141] [α].sub.D.sup.20=−37.0 (c.1, DCM).

[0142] LCMS ESI (m/z): 817.3 [M−H].sup.−

##STR00031##

[0143] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 8.8-8.6 (1H, br. s), 7.64 (1H, d, J=1.0 Hz), 7.52-7.46 (2H, m), 7.40-7.33 (4H, m), 7.33-7.26 (2H, m), 7.26-7.19 (1H, m) 6.87-7.79 (4H, m), 5.69 (1H, s), 5.25 (1H, d, J=11.0 Hz), 4.90-4.88 (1H, m), 4.57 (1H, s), 4.60 (1H, s), 4.29 (1H, dt, J=12.6 3.1 Hz), 3.87 (1H, d, J=8.3 Hz), 3.79-3.76 (1H, m), 3.78 (3H, s), 3.78 (3H, s), 3.59 (1H, d, J=11.0 Hz), 3.42 (1H, d, J=11.0 Hz), 2.57-2.48 (1H, m), 2.08-1.65 (6H, m), 1.65 (6H, s), 1.58 (3H, d, J=1.0 Hz)

[0144] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 163.7, 158.8, 149.9, 144.9, 144.2, 135.3, 135.2, 134.2, 130.5, 130.4, 128.5, 128.1, 127.2, 113.4, 113.4, 111.9, 111.1, 87.4, 87.1, 86.1, 78.3, 78.3, 74.6, 74.6, 72.2, 65.9, 57.9, 55.3, 39.0, 33.8, 33.7, 27.8, 27.6, 23.5, 22.7, 21.8, 12.6.

[0145] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 100.9.

[0146] [α].sub.D.sup.20=+94.9 (c. 1, DCM).

[0147] LCMS ESI (m/z): 817.25 [M−H].sup.−

##STR00032##

[0148] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 13.5-13.2 (1H, br. s), 8.35-8.28 (2H, m), 7.82 (1H, J=1.0 Hz), 7.55-7.40 (5H, m), 7.39-7.28 (6H, m), 7.27-7.22 (1H, m), 6.86-6.83 (4H, m), 5.75 (1H, s), 5.19 (1H, d, J=7.5 Hz), 4.95 (1H, s), 4.86 (1H, s), 4.71 (1H, s), 4.40 (1H, dt, J=12.7, 3.5 Hz), 3.84-3.79 (7H, m), 3.50 (1H, d, J=11.1 Hz), 3.44 (1H, d, J=11.1 Hz), 2.62-2.52 (1H, m), 2.36-2.25 (1H, m), 2.09-1.66 (6H, m), 1.85 (3H, d, J=1.0 Hz), 1.76 (3H, s), 1.65 (3H, s).

[0149] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 159.8, 158.8, 147.7, 145.0, 144.2, 137.3, 135.4, 135.3, 135.2, 132.6, 130.3, 130.3, 130.1, 128.3, 128.4, 127.2, 113.5, 112.2, 111.8, 87.9, 87.8, 87.5, 87.6, 85.6, 78.3, 74.1, 74.0, 72.4, 66.7, 57.7, 55.4, 39.0, 33.8, 33.7, 27.7, 27.6, 23.6, 22.7, 22.0, 13.8.

[0150] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm) 101.4

[0151] [α].sub.D.sup.20=+15.9 (c 0.1, DCM).

[0152] LCMS ESI (m/z): ): 922.81 [M+H].sup.+

##STR00033##

[0153] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 13.5-13.3 (1H, br. s), 8.33-8.28 (2H, m), 7.82 (1H, d, J=1.0 Hz), 7.55-7.48 (3H, m), 7.46-7.37 (6H, m), 7.35-7.29 (2H, m), 7.27-7.22 (1H, m), 6.89-6.81 (4H, m), 5.74 (1H, s), 5.28 (1H, d, J=11.2 Hz), 4.92-4.88 (1H, m), 4.86 (1H, s), 4.65 (1H, s), 4.30 (1H, dt, J=12.7, 3.2 Hz), 3.89 (1H, d, J=8.2 Hz), 3.81-3.77 (1H, m), 3.79 (3H, s), 3.79 (3H, s), 3.62 (1H, d, J=11.0 Hz), 3.44 (1H, d, J=11.0 Hz), 2.57-2.48 (1H, m), 2.08-1.62 (6H, m), 1.78 (3H, d, J=1.0 Hz), 1.66 (3H, s), 1.65 (3H, s)

[0154] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 159.8, 158.9, 158.8, 147.7, 144.9, 144.2, 137.2, 135.4, 135.3, 135.2, 132.6, 130.5, 130.1, 128.5, 128.3, 128.2, 127.2, 113.4, 113.4, 112.2, 111.9, 87.6, 87.5, 87.4, 87.1, 86.1, 78.2, 78.1, 74.5, 74.4, 72.3, 65.9, 57.9, 55.4, 38.9, 33.8, 33.7, 27.8, 27.6, 23.5, 22.7, 21.8, 13.7.

[0155] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 100.9.

[0156] [α].sub.D.sup.20=+152.3 (c. 1, DCM).

[0157] LCMS ESI (m/z): 922.79 [M+H].sup.+

##STR00034##

[0158] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 9.04 (1H, s), 8.82 (1H, s), 8.38 (1H, s), 8.06-7.97 (2H, m), 7.63-7.55 (1H, m), 7.55-7.47 (2H, m), 7.46-7.39 (2H, m), 7.36-7.25 (1H, m), 7.24-7.17 (1H, m) 6.88-6.78 (4H, m), 6.22 (1H, s), 5.32 (1H, d, J 6.75 Hz), 4.98 (1H, s), 4.96 (1H, s), 4.88 (1H, s), 4.42 (1H, dt, J=12.7 3.5 Hz), 4.05 (1H, d, J 8.4 Hz), 3.96 (1H, d, J=8.4 Hz), 3.78 (6H, s), 3.49 (2H, s), 2.62-2.53 (1H, m), 2.35-1.2.25 (1H, m), 2.11-1.64 (5H, m), 1.78 (3H, s), 1.63 (3H, s)

[0159] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 164.6, 158.7, 153.2, 151.2, 149.7, 145.0, 144.3, 140.2, 135.4, 135.3, 133.7, 132.9, 130.2, 129.0, 128.2, 128.1, 128.0, 127.1, 123.4, 113.4, 111.8, 87.5, 87.4, 86.7, 85.9, 85.6, 78.7, 75.7, 75.6, 73.0, 66.6, 58.5, 55.4, 39.0, 33.8, 33.7, 27.7, 27.6, 23.5, 22.6, 22.0.

[0160] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm) 101.6.

[0161] [α].sub.D.sup.20=−71.7 (c. 1, DCM).

[0162] LCMS ESI (m/z): 932.7 [M+H].sup.+

##STR00035##

[0163] .sup.1H NMR (300 MHz, CDCl.sub.3, ppm): 9.1-9.0 (1H, br, s), 8.80 (1H, s), 8.33 (1H, s), 8.04-7.98 (2H, m), 7.62-7.56 (1H, m) 7.54-7.43 (4H, m), 7.37-7.26 (6H, m), 7.24-7.18 (1H, m) 6.85-6.80 (4H, m), 6.20 (1H, s), 5.42 (1H, d, J=10.4 Hz), 4.95 (1H, s), 4.84 (1H, s), 4.66 (1H, s), 4.31 (1H, dt, J=12.6 3.3 Hz), 4.14 (1H, d, J=8.4 Hz), 3.95 (1H, d, J=8.4 Hz), 3.77 (6H, s), 3.59 (1H, d, J=11.0 Hz), 3.53 (1H, d, J=11.0 Hz), 2.56-2.47 (1H, m), 2.10-1.60 (6H, m), 1.64-1.60 (6H, m).

[0164] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 164.6, 158.7, 153.0, 151.0, 149.6, 144.8, 144.2, 140.3, 135.4, 135.3, 133.6, 132.9, 130.2, 129.9, 128.2, 128.0, 127.9, 127.0, 123.4, 113.3, 111.8, 87.2, 87.1, 86.7, 86.1, 85.9, 78.6, 78.5, 76.0, 75.9, 72.8, 65.8, 58.9, 55.2, 38.0, 33.6, 33.6, 27.6, 27.5, 23.3, 22.5, 21.6.

[0165] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 101.1.

[0166] [α].sub.D.sup.20=+48.5 (c 0.1, DCM).

[0167] LCMS ESI (m/z): 932.7 [M+H].sup.+

##STR00036##

[0168] .sup.1H NMR (400 MHz, CDCl.sub.3, ppm): 12.02 (1H, s), 8.97 (1H, s), 7.79 (1H, s), 7.44-7.38 (2H, m), 7.34-7.23 (6H, m), 7.22-7.17 (1H, m), 6.84-6.77 (4H, m), 5.99 (1H, d, J=10.3 Hz), 5.83 (1H, s), 4.97 (1H, s), 4.91-4.88 (1H, m), 4.73 (1H, s), 4.40 (1H, dt, J=12.6, 3.2 Hz), 4.15 (1H, d, J=8.7 Hz), 3.95 (1H, d, J=8.7 Hz), 3.78 (3H, s), 3.78 (3H, s), 3.55-3.49 (2H, m), 2.64 (1H, septet, J=6.8 Hz), 2.60-2.53 (1H, m), 2.20-2.11 (1H, m), 2.10-1.65 (5H, m), 1.70 (3H, s), 1.69 (3H, s), 1.27 (3H, d, J=6.8 Hz), 1.25 (3H, d, J=6.8 Hz).

[0169] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 178.7, 158.7, 155.5, 147.5, 147.2, 144.8, 144.4, 138.6, 135.4, 130.3, 128.3, 128.0, 127.0, 122.3, 113.30, 111.9, 87.1, 87.0, 86.6, 86.5, 78.4, 73.0, 66.5, 59.3, 55.3, 38.9, 36.7, 33.9, 33.8, 27.8, 27.7, 23.5, 22.7, 21.8, 19.2, 19.0.

[0170] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 100.3.

[0171] [α].sub.D.sup.20=+85.3 (c 0.1, DCM).

[0172] LCMS ESI (m/z): 914.73 [M+H].sup.+

##STR00037##

[0173] .sup.1H NMR (400 MHz, CDCl.sub.3, ppm): 11.99 (1H, s), 8.89 (1H, s), 7.81 (1H, s), 7.43-7.37 (2H, m), 7.33-7.23 (6H, m), 7.22-7.17 (1H, m), 6.84-6.78 (4H, m), 5.97 (1H, J=7.7 Hz), 5.84 (1H, s), 5.01-4.87 (1H, m), 4.93-4.87 (2H, m), 4.51 (1H, dt, J=12.7 3.5 Hz), 4.07 (1H, J=8.5 Hz), 4.00 (1H, J=68.5 Hz), 3.78 (6H, s), 3.48-3.41 (2H, m), 2.63-2.56 (1H, m), 2.57 (1H, septet, J=6.9 Hz), 2.36-2.25 (1H, m), 2.14-1.64 (5H, m), 1.78 (3H, s), 1.68 (3H, s), 1.24 (3H, d, J=6.9 Hz), 1.22 (3H, d, J=6.9 Hz).

[0174] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 178.5, 158.7, 155.5, 147.5, 147.3, 144.8, 144.4, 138.5, 135.4, 130.2, 128.2, 128.0, 127.0, 122.2, 113.3, 111.9, 87.2, 87.1, 86.6, 86.5, 86.1, 79.2, 73.0, 66.6, 58.9, 55.3, 39.0, 36.6, 33.7, 33.6, 27.9, 27.8, 23.5, 22.7, 22.0, 19.1, 18.9.

[0175] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 103.3.

[0176] [α].sub.D.sup.20=−14.6 (c. 1, DCM).

[0177] LCMS ESI (m/z): 914.77 [M+H].sup.+

##STR00038##

[0178] .sup.1H NMR (400 MHz, CDCl.sub.3, ppm): 9.2-9.0 (1H, br. s), 8.59 (1H, s), 8.0-7.8 (1H, br. s), 7.44-7.39 (2H, m), 7.34-7.23 (6H, m), 7.22-7.16 (1H, m), 6.75-6.77 (4H, m), 5.97 (1H, s), 5.39 (1H, d, J=6.5 Hz), 4.98-4.92 (2H, m), 4.88-4.85 (1H, m), 4.41 (1H, dt, J=12.5 3.5 Hz), 4.03 (1H, d, J=8.2 Hz), 3.93 (1H, d, J=8.2 Hz), 3.77 (6H, s), 3.51 (1H, d, J=10.9 Hz), 3.43 (1H, d, J=10.9 Hz), 3.10 (3H, s), 3.05 (3H, s), 2.59-2.52 (1H, m), 2.32-2.22 (1H, m), 2.07-1.65 (5H, m), 1.76 (3H, s), 1.62 (3H, s).

[0179] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 158.7, 158.5, 157.1, 149.6, 145.1, 144.3, 135.5, 135.4, 130.2, 128.3, 128.1, 127.0, 113.4, 111.8, 87.1, 87.1, 86.6, 86.1, 85.7, 79.1, 75.9, 75.8, 73.0, 66.3, 58.8, 55.4, 41.4, 39.0, 35.3, 33.8, 33.7, 27.8, 27.6, 23.5, 22.7, 21.9, 21.3.

[0180] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 100.6.

[0181] [α].sub.D.sup.20=−37.4 (c. 1, THF).

[0182] LCMS ESI (m/z): 899.71 [M+H].sup.+

##STR00039##

[0183] .sup.1H NMR (400 MHz, CDCl.sub.3, ppm): 9.2-9.0 (1H, br. s), 8.55 (1H, s), 8.0-7.8 (1H, br. s), 7.48-7.41 (2H, m), 7.37-7.34 (6H, m), 7.22-7.16 (1H, m), 6.84-6.74 (4H, m), 5.96 (1H, s), 5.39 (1H, d, J=9.5 Hz), 4.96-4.75 (2H, m), 4.85-4.81 (1H, m), 4.27 (1H, dt, J=12.7 3.1 Hz), 4.14 (1H, d, J=8.3 Hz), 3.92 (1H, d, J=8.3 Hz), 3.76 (6H, s), 3.64-3.50 (2H, m), 3.12 (3H, s), 3.06 (3H, s), 2.55-2.46 (1H, m), 2.09-1.58 (6H, m), 1.53 (3H, s), 1.61 (3H, s).

[0184] .sup.13C NMR (100 MHz, CDCl.sub.3, ppm): 158.7, 158.3, 157.0, 144.9, 144.4, 135.5, 135.4, 130.3, 130.2, 128.3, 128.1, 127.5, 127.1, 113.4, 113.4, 111.8, 87.0, 86.7, 86.2, 79.0, 76.3, 76.2, 73.0, 65.8, 59.4, 55.3, 41.5, 38.9, 35.3, 33.7, 33.7, 27.7, 27.5, 23.4, 22.6, 21.7.

[0185] .sup.31P NMR (160 MHz, CDCl.sub.3, ppm): 100.0.

[0186] [α].sub.D.sup.20=+70.8 (c 0.1, THF).

[0187] LCMS ESI (m/z): 899.74 [M+H].sup.+