SEL ONIUM DE TAXO-DITERPENOIDE SOLUBLE DANS L'EAU

WATER SOLUBLE ONIUM SALT OF TAXO-DITERPENOID

Abrégé français

Cette invention concerne des sels onium de taxo-diterpénoïde-C?n¿, 2-0-aza-arènes qu'on utilise comme promédicaments solubles dans l'eau. Par exemple, le taxol-2'-méthylpyridinium toxylate se caractérise par une solubilité aqueuse élevée, une activation rapide par la protéine sérique, une bonne stabilité dans la plupart des autres solutions aqueuses, la formation d'une protéine:un intermédiaire de taxol et une bonne rétention dans le système circulatoire. La toxicité de la forme activée est comparable ou supérieure à celle du taxol non dérivé. En outre on peut synthétiser du taxol-2'-MPT en n'effectuant qu'une seule étape de réaction simple entre du taxol et du 2-fluoro-1-MPT. Cette invention peut s'appliquer au taxol et aux substances mimétiques du taxol comprenant des groupes hydroxyle qui réagissent avec les sels onium de 2-halogéné-aza-arènes. Par exemple, le taxol, les analgoues à substitution C-2 de taxol, et le Taxotere comprennent chacun des groupes hydroxyle réactifs situés en positions 2' et 7. Cette invention s'applique également à une gamme étendue de 2-halogéné-aza-arènes.

Abrégé anglais

Onium salts of taxo-diterpenoid-Cn, 2-0-aza-arenes are employed as water
soluble prodrugs. For example, taxol-2'-methylpyridinium tosylate (MPT) is
characterized by an elevated aqueous solubility, rapid activation by serum
protein, good stability in most other aqueous solutions, formation of a
protein:taxol intermediate and good retension within the circulatory system.
The toxicity of the activated form is comparable or greater than underivatized
taxol. Furthermore, taxol-2'-MPT can be synthesized by a simple one step
reaction between taxol and 2-fluoro-1-MPT. The invention is applicable to
taxol and taxol mimetics having hydroxyls that are reactive with onium salts
of 2-halogenated-aza-arenes. For example, taxol, C-2 substituted analogs of
taxol, and Taxotere each have reactive hydroxyls at the 2' and 7 positions.
The invention is also applicable to a wide array of 2-halogenated-aza-arenes.

Revendications

99
CLAIMS:
1. A taxo-diterpenoid represented by the following formula:
<IMG>
wherein:
R x is selected from the group consisting of Ph and tB u O;
R10 is selected from the group consisting of OAc and OH;
R y is selected from the group consisting of phenyl and
C-2 substituents represented by the following structures:
<IMG>
R2' and R7 are each selected from the group consisting of OH and
an opium salt of a 2-O-aza-arene, with the proviso that at least
one of R2' and R7 is said opium salt of the 2-O-aza-arene, said
onium salt of the 2-O-aza-arene being selected from the group

-100-
consisting of opium salt I and opium salt
II represented by the following formulas:
<IMG>
wherein:
Z1 and Z2 are each selected from the group
consisting of C and N;
Z3 is selected from the group consisting of S
and O;
R1 is selected from the group consisting of C1-
C6 alkyl, allyl, propargyl, and
fused C6-aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl,
propargyl, and fused C6-aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl,
propargyl,, C1-C6 o-alkyl, OH,
halogen, and fused C6-aryl;
if Z1 is N, then R3 is absent;
R4 and R8 are each selected from the group
consisting of H, C1-C6 alkyl, allyl,
propargyl, C1-C6 O-alkyl, OH,
halogen, and fused C6-aryl; and
if Z2 is C, then R~ is selected from the group
consisting of H, C1-C6 alkyl, allyl,
propargyl, C1-C6 O-alkyl, OH,
halogen, and fused C6-aryl;
if Z2 is N, then R5 is absent; and

101
S is a counter ion.
2. A taxo-diterpenoid as described in claim 1
wherein:
S- is selected from the group of counter ions
consisting of C1-, Br , I-, CH3COO', BF4-,
C104-, ArSO3', and C1-C6 AlkylSO3-.
3. A taxo-diterpenoid as described in claim 1
wherein:
R2' is said opium salt and
R7 is OH.
4. A taxo-diterpenoid as described in claim 3
wherein:
R x is Ph;
R10 is AcO; and
said onium salt is onium salt I wherein:
Z1 and Z2 are both C;
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
s- is selected from the group of counter ions
consisting of Cl-, Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 Alky1SO3-.
5. A taxo-diterpenoid as described in claim 4
wherein:
R1 is methyl; and
S- is ArSO3- or CH3COO-.
6. A taxo-diterpenoid as described in claim 3
wherein:
R x is tBuo;
R10 is OH; and
said onium salt is opium salt I wherein:
Z1 and Z2 are both C;

102
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
s' is selected from the group of counter ions
consisting of C1-, Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 AlkylSO3-.
7. A taxo-diterpenoid as described in claim 6
wherein:
R1 is methyl; and
S- is ArsO3- or CH3COO-.
8. A taxo-diterpenoid as described in claim 1
wherein:
R2' and R7 are both said onium salt.
9. A taxo-diterpenoid as described in claim 8
wherein:
R x is Ph;
R10 is AcO; and
said onium salt is onium salt I wherein:
Z1 and Z2 are both C;
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
S- is selected from the group of counter ions
consisting of Cl-, Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 AlkylSO3-.
10. A taxo-diterpenoid as described in claim 9
wherein:
R1 is methyl; and
S- is ArSO3- or CH3COO-.

103
11. A taxo-diterpenoid as described in claim S
wherein:
R x is tBuO;
R10 is OH; and
said onium salt is onium salt I wherein:
Z1 and Z2 are both C;
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
S- is selected from the group of counter ions
consisting of Cl-, Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 AlkylSO3-.
12. A taxo-diterperloid as described in claim 11
wherein:
R1 is methyl; and
S- is ArSO3- or CH~COO .
13. A taxo-diterpenoid as described in claim 1
wherein:
R7 is said opium salt and
R2' is OH.
14. A taxo-diterpenoid as described in claim 13
wherein:
R x is Ph;
R10 is AcO; and
said onium salt is onium salt I wherein:
Z1 and Z2 are both C;
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
S- is selected from the group of counter ions
consisting of Cl-, Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 AlkylSO3-.

104
15. A taxo-diterpenoid as described in claim 14
wherein:
R1 is methyl; and
S- is ArSO3-, or CH3COO-.
16. A taxo-diterpenoid as described in claim 13
wherein:
R x is tBuo;
R10 is OH; and
said onium salt is onium salt I wherein:
Z1 and Z2 are both C;
R1 is C1-C6 alkyl;
R2, R3, R4, and R4 are each H; and
S- is selected from the group of counter ions
consisting of Cl-; Br-, I-, CH3COO-, BF4-,
ClO4-, ArSO3-, and C1-C6 AlkylSO3-.
-.
17. A taxo-diterpenoid as described in claim 16
wherein:
R1 is methyl; and
S- is ArSO3- or CH3COO-.

105
18. Use of a taxo-diterpenoid as defined in any one of
claims 1 to 17 in treatment of a tumor or other cancer in a
mammal.
19. Use of an aqueous solution of a taxoid onium salt
prodrug in a form administrable to a mammal by a route
selected from the group consisting of injection and infusion,
wherein the taxoid onium salt prodrug is represented by the
following structure:
<IMG>
wherein S- is selected from the group consisting of OAc-,
Cl-, Br-, I-, BF4-, ClO4-, ArSO3-, and AlkylSO3-, R1 is
selected from the group consisting of phenyl and tBuO, R2 is
selected from the group consisting of OAc and OH, and R3 is
selected from the group consisting of R3 1 and R3 2'
where R3 1 is represented by the following structure:
<IMG>

106
wherein:
P is selected from the group consisting of N and
S,
Z is selected from the group consisting of C, N,
S, and O,
R4 is selected from the group consisting of H,
alkyl, allyl, propargyl, bridging aryl
and bridging alkyl,
R5 is selected from the group consisting of H,
alkyl, allyl, propargyl, bridging aryl
and bridging alkyl,
if Z is C, then R6 is selected from the group
consisting of H, alkyl, allyl, aryl,
propargyl, O-alkyl, and OH,
R7 is selected from the group consisting of H,
alkyl, allyl, aryl, propargyl, O-alkyl,
and OH,
if Z is C, then R8 is selected from the group
consisting of H, alkyl, allyl, aryl,
propargyl, O-alkyl, and OH, and
where R3 2 is represented by the following structures
<IMG>

107
wherein
P is selected from the group consisting of N and
S,
Z is selected from the group consisting of C, N,
S, and O,
R9 is selected from the group consist ing of H,
alkyl, allyl, propargyl, bridging aryl
and bridging alkyl,
R10 is selected from the group consisting of H,
alkyl, allyl, propargyl, bridging aryl
and bridging alkyl,
R11 is selected from the group consisting of H,
alkyl, allyl, aryl, propargyl, O-alkyl,
OH, halogen, bridging aryl, and bridging alkyl,
if Z is C, then R12 is selected from the group
consisting of H, alkyl, allyl, aryl,
propargyl, O-alkyl, and OH, halogen, bridging
aryl, and bridging alkyl,
in treatment of a tumor or other cancer in a mammal.
20. A use as described in claim 19 wherein the mammal is
a human tumor or cancer patient.
21. A use as described in claim 19 wherein the mammal is
sensitive or intolerant to a side effect of taxol.

108
22. A use as described in claim 19 wherein tire solution
is in a form suitable for a dosage range of 50 -
300 mg/m2.
23. A use as described in claim 19 wherein the
solution is in a form suitable for a dosage range of
50 and 4,900 mg/m2.
24. A use as described in claim 19 wherein the
solution is in a form suitable for a dosage range of
50 and 21,000 mg/m2.
25. A use as described in claim 19 wherein the
solution is in a form suitable for a dosage range of
50 and 28,000 mg/m2.
26. A use as described in claim 25 wherein the tumor or
other cancer is refractory or insensitive to taxol.
27. A use as described in claim 19 wherein S- is any
counter ion classified as generally regarded as safe (GRAS) by
the U.S. Food and Drug Administration.
28. A use as described in claim 19 wherein the taxoid
onium salt prodrug is taxol-2'-MPA represented by the
following structure:

109
<IMG>
29. A use as described in claim 19 wherein the taxoid
onium salt prodrug is taxotere-2'-MPA represented by the
following structure:
<IMG>
30. A use as described in claim 19 wherein the solution
is in a form suitable for administration by infection of a
bolus.
31. A use as described in claim 19 wherein the solution
is in a form suitable for administration by infusion.

110
32. A use as described in claim 19 wherein the solution
is suitable for intravenous administration.
33. A use as described in claim 19 wherein the solution
is suitable for intraperitoneal administration.

Description

CA 02180444 1996-07-03
218044
WO 95/18804 PGT/US95/00478
WATER SOLUBLE ONIUM SALT OF TARO-DITERPENOID
S ~chnic~, Field:
The invention relates to taxol prodrugs. More particularly,
the invention relates to taxol and taxol memetics derivatized with
onium salts of aza-arenes to form water soluble prodrugs which
are activated upon contact with serum protein.
Background:
Taxol, an antineoplastic agent originally isolated from ~ Taxus
brevifolia, is approved for usage in the treatment of~ ovarian
cancer and is expected to see usage in breast, lung, and skin
cancers as well. However, since Taxol possesses an extremely low
water solubility, i.e., less than 1.5 x 10'6 M, it has been necessary
to formulate Taxol in a mixture of CremaphorT"', a
polyoxyethylated castor oil, and ethanol in order to achieve a
therapeutic concentration. This formulation can induce a variety
2 0 of significant side effects including hypersensitivity reactions.
While premedication and slow administration of the drug
can circumvent these problems in the clinic, the entire protocol is
quite cumbersome and requires extensive close monitoring of
patients. Although taxol's dramatic efficacy has driven clinical
2 5 usage forward despite these problems, a water soluble form of
taxol could completely obviate the need for this troublesome
protocol.
One approach to bypassing these formulation difficulties,
. previously attempted by several groups including our own, is the
3 0 introduction of solubilizing functionality that normal metabolic
a pathways could remove in vivo. Compounds of this type, termed
prodrugs, consist, in the case of taxol, primarily of ester
derivatives at the 2' and 7 positions. Currently none of these
' protaxols have given success in the clinic. In each case, the
3 5 prodrug is rapidly cleared from circulation by the kidneys.
RECTIFIED SHEET (1~~E 9'1)
ISA/EP

21804
CA 02180444 1996-07-03
WO 95118804 PCT/US95/00478
-2-
Taxol is only one of a class of taxo-diterpenoids having
bioactivity. Another preferred taxo-diterpenoid having clinically
significant activity is TaxotereT"''. C-2 substituted taxol analogs are
also known to have bioactivity. Unfortunately, all known
bioactive taxo-diterpenoids have a low aqueous solubility.
What is needed is a prodrug which is activated by contact
with serum and which is retained for a clinically significant period
after administration.
Summary:
The invention is an opium salt of a taxo-diterpenoid-Cn,2-O-
aza-arene. The opium salt of aza-arene includes a delocalized
charge which imparts aqueous solubility. The invention herein
teaches that opium salts of taxo-diterpenoid-Cn,2-O-aza-arenes
are soluble and stable in aqueous media; that, with the addition of
serum protein, 2-O-aza-arene is displaced and a soluble
protein:taxo-diterpenoid intermediate is formed; and that the
protein:taxo-diterpenoid intermediate dissociates over time to
provide a bioactive taxo-diterpenoid. Preferred taxo-diterpenoids
2 0 include taxol, C-2 substituted analogs of taxol, and TaxotereTM.
Taxo-diterpenoid-Cn,2-O-aza-arene may be produced in a one
step synthesis by reacting opium salts of 2-halogenated aza-
arenes with reactive hydroxyls on the taxo-diterpenoid. Reactive
hydroxyls on taxol and TaxotereTM are located at C2~ and C7. A
2 5 preferred opium salt of 2-halogenated aza-arene is 2-fluoro-1-
methylpyridinium tosylate. Other employable opium salts of 2-
halogenated aza-arenes are disclosed by T.Mukaiyama,
Angewandte Chemie 1979, 18(18), 707-808, incorporated herein
by reference.
3 0 More particularly, the invention is directed to water soluble
taxo-diterpenoids represented by the following formula:
SUBSTITUTE SHEET (RULE 26)

CA 02180444 2004-09-24
28395-34
-3-
O
18 ~°R O
lgR~
Rx NH O 11 10 g
12 6
2'
Ph 3' 1' 0~~~~~ A 15 B 8 C
- 13 \ 3 5
14 1 _2~H4_ ~'0
HR p OAc 20
O
wherein RX is Ph or tBuO; R10 is OAc or OH; Ry is a C-2
substituent defined below; and R2~ and R~ are each selected from
the group consisting of OH and an opium salt of a 2-O-aza-arene,
with the proviso that at least one of R2~ and R~ is the opium salt
of the 2-O-aza-arene. The opium salt of the 2-O-aza-arene can be
represented by either of the following formulas for opium salt I
or opium salt II:
R4
3R~Z~~ZZ.RS eR Z3
~ ~i ~~ O
2 R ~1V ~0 sR
SO R~ S R
opium salt ~ opium salt II
1 5 wherein Z 1 and Z 2 are each either C or N; Z 3 is S or 0; Rl is
selected from the group consisting of Cl-C6 alkyl, allyl, arenyl,
propargyl, and fused aryl; R2 and R6 are each selected trom the
group consisting of H, C1-C6 alkyl, allyl, arenyl, propargyl, and
fused aryl; if Z 1 is C, then R3 is selected from the group consisting
of H, Cl-C6 alkyl, allyl, arenyl, propargyl, Cl-C6 0-alkyl, OH,
halogen, and fused aryl; if Z 1 is N, then R3 is absent; R4 and R$
are each selected from the group consisting of H, Cl-C6 alkyl, allyl,

CA 02180444 2004-09-24
28395-34
-4-
arenxyl, propargyl, C1-C6 O-alkyl, OH, halogen, and fused aryl; and
if ZZ is C, then R5 is selected from the group consisting of H, Cl-C6
alkyl, allyl, arenyl, propargyl, C1-C6 O-alkyl, OH, halogen, and
fused aryl; if Z2 is N, then RS is absent; and S- is a counter ion.
R y is phenyl or a C-2 substituent selected from the group
represented by the following structures:
O ~ S S N~ N
\ / \ / \ / ~ , ~ i
Me2N ~ ~ ~ I % \ ~SPh
Brief Description of the DrawinQS:
Figure 1 illustrates the kinetics of taxol release from taxol
2'-MPT (2) in various aqueous solutions at 25 °C (horizontal line).
Although stable in water and aqueous buffer solutions, methylene
chloride extraction of plasma treated with compound 2 showed
complete conversion of 2 into taxol (1) within 10 minutes (curve,
20% of total recovered).
Figure 2 A illustrates a transmission electron micrograph
2 0 (TEM) of self-assembled helical fibrous nanostructures of taxol-2'
MPT,. i.e., compound 2 in buffered solutions (2,1 mM in 100 mM
PBS) above the critical aggregation concentration (CAC) of this
compound using a negative phosphotungstate stain and a
magnification of x25000. The inset shows a portion of one of the
2 5 fibrils further magnified to illustrate the helical nature of the
structure.

CA 02180444 2004-09-24
28395-34
Figure 28 illustrates a transmission electron
micrograph (TEM) of self-assembled spherical nanostructures of
taxol-2'-MPT (Compound 2) in unbuffered solutions (2,1 mM
in H20) above the critical aggregation concentration (CAC) of
this compound using a negative uranyl acetate stain and a
magnification of x45000.
Figure 3 illustrates a tubulin polymerization-
depolymerization measurements with negative control
(triangles), positive taxo.l control (diamonds), and taxol-
2'-MPT, i.e., compound 2 (dots). Calcium chloride promoted
depolymerization is suppressed by taxol but not by taxol-2'-
MPT.
Figure 4 illustrates the relative cytotoxicities of
taxol-2'-MPT (compound 2) and taxol against a variety of cell
lines.
Figure 5 illustrates the efficacy of taxol-2'-MPT in
lung tumor xenograft nude mouse models 5% dext rose
(triangles), taxol (diamonds), and taxol-2'-MPT (dots).
Detailed Description
The synthesis, physical properties, and
pharmaceutical profiles of water soluble opium salts of
taxo-diterpenoid-Cn,2-O-aza-arenes are described.
Synthesis of Taxo1-2'-MPTe
Taxol-2'-MPT (methylpyridinium tosylate), compound
2, was synthesized according to the method of T. Mukaiyama,
Anctewandte Chemie 1979, 18(18), 707-808, Taxol (10 mg,

CA 02180444 1996-07-03
21~~44~ ...
WO 95118804 PCT/US95/00478
- 6 -
0.012 mM), from NaPro Biochemicals, Boulder CO, USA,
was dried by azeotropic distillation with toluene
(2x1.0 mL) and then dissolved in methylene chloride
(0.4 mL) and treated sequentially under an atmosphere
of dry argon, with freshly distilled triethylamine (5
microL, 0.04 mM, 3 equivalents) and 2-fluoro-1-
methylpyridinium tosylate (5 mg, 0.018 mM, 1.5
equivalents) Aldrich Chemicals, and allowed to stir
at ambient temperature for 30 minutes. The clear
colorless solution rapidly turned to a clear pale
yellow. The course of the reaction was monitored
through thin layer chromatography (TLC) (E. Merck
RP-18 silica, 65 tetrahydrofuran . 35 water,
UV/phosphomolybdic acid) and after thirty minutes of
stirring at ambient temperature, judged complete as
no taxol remained and only one compound was apparent
by TLC (Rf 0.8). Purification via reverse phase
high pressure liquid chromatography (HPLC) (C~8
column, imM NH40Ac pH 6.5 buffer / methanol gradient,
1.5 mL / min. W) to give, after removal of solvent
in vaccuo, pure taxol-2'-MPT (2) (12 mg, 93% yield)
as a white amorphous solid. All spectroscopic data
('H NMR and HRMS) were in accord with the structure
assigned to 2. ~H NMR (CDC13, 125 MHz) d: 1.055 (s,
3 H, C17-H), 1.083 (s, 3 H, C19-8), 1.724 (s, 3 H,
C19-H), 1.858 (m, 1 H, C6-68), 1.913 (s, 3 H, CH3-Ph),
2.193 (s, 3 H, C10-OC(O)C83), 2.514 (m, 1 H, C6-a8),
3.663 (d, 1 H, J = 7.0 Hz, C3-8), 4.110 (d, 1 H, J =
8.5, C20-8H, A of AB), 4.133 (s, 3 H, N-CH3), 4.230
(d, 1 H, J = 8.5, C20-a8, B of AB), 4.315 (dd, 1 H, J
- 8.7, 10.7, C7-H), 4.901 (dd, 1 H, J = 1.0, 7.7,
C5-H), 5.501 (d, 1 H, C2-8, J = 7.0), 5.702 (bt, 1 H,
C2'-Ii, J = 8.0), 5.951 (dd, 1 H, C13-H, J = 1.0,
8.0), 6.120 (bd, 1 H, C3'-8, J = 10.0), 6.181 (s, 1
H, C10-H) 7.702 (t, 1 H, N-H, J = 7.5), 7.33 - 7.45
(m, 8 H, Ar-H), 7.56 - 7.62 (m, 4 H, Ar-H), 7.56 -

CA 02180444 1996-07-03
WO 95/18804
PGTIUS95I00478
-
7.62 (m, 4 H, Ar-8), 7.68 - 7.75 (m, 4 H, Ar-8), 8.00
- 8.05 (m, 1 H Ar-H), 8.23 - 8.28 (m, 1 H, Pyr-B),
8.41 (m, 1 H, Pyr-H). IR (neat, RCl plate) Gait: 3640
- 3120 (bm), 3030-2870 (bm), 2320 (m), 1720 (~), 1630
(m), 1560 (m), 1500 (m), 1360 (s), 1160 (m), 1070
(m), 700 (m). UV/Vis (CHC13) nm: 254, 280. FAB HRMS:
calc for C53H5~0~4N2: 945.3810; found: 945.3810
The molecular structures of taxol, compound i,
and of taxol-2'-MPT, compound 2, are illustated in
to Scheme lA. The synthesis of taxol-2'-MPT is
illustrated in Scheme iB.
Ac0 HO O
.. O OH
OH
O O ' O v0' ''~~
H H
Ph N~, '' wOH HO : H. O Ph~N,~ w0 HO gz0 H. : O
Bz0 pAe ~O Ph ,\ ~/ OAc
O Ph ~ ~ N
TsD~
2: taxol-2'-methylpyrldlnium tosylate
1: Taxol ~ taxol H (taxol-2'-MP'?]
b
Ta 00
. \
N-' Ts00
taxol H \ / ~ ~ / N
O
(CH~CH~N, CHZCI= 2 O taxol
90'.:
Schemes lA and 18
-7-
The synthesis of taxol-7-MPT differed only
slightly from the synthesis of Taxol-2'-MPT. Taxol
(10 mg, 0.012 mM), from NaPro Biochemicals, Boulder
CO, USA, was dissolved in methylene chloride (2.0 mL)
and treated sequentially with triethylamine (67

CA 02180444 1996-07-03
WO 95/18804 ~ ~ PCT/US95I00478
- 8 -
microL, 0.48 mM, 40 equivalents) and 2-fluoro-1-
methylpyridinium tosylate (34 mg, 0.12 mM, 10
equivalents) Aldrich Chemicals, and allowed to stir
at ambient temperature for 5 minutes. Purification
via reverse phase high pressure liquid chromatography
(HPLC) gave pure taxol-2'-MPT (2) (12 mg, 93% yield)
as a white amorphous solid. The Rf of taxol-7-MPT is
about 0.3 minutes less than the Rf of taxol-2'-MPT.
The yield was 11 mg or 85%. Spectroscopic data ('H
NMR and HRMS) were as expected.
~nthesis of Taxol-bis-2' 7-MPT:
The synthesis of taxol-bis-2',7-MPT differed
from the synthesis of Taxol-7-MPT only with respect
to reaction time. Taxol (10 mg, 0.012 mM), from
NaPro Biochemicals, Boulder CO, USA, was dissolved in
methylene chloride (2.0 mL) and treated sequentially
with triethylamine (67 microL, 0.48 mM, 40
equivalents) and 2-fluoro-1- methylpyridinium
tosylate (34 mg, 0.12 mM, 10 equivalents) Aldrich
Chemicals, and allowed to stir at ambient temperature
for 18 hours. Purification via reverse phase high
pressure liquid chromatography (HPLC) gave pure
taxol-2'-MPT (2) (12 mg, 93% yield) as a white
amorphous solid. The Rf of taxol-bis-2',7-MPT is
about 0.3 minutes less than the Rf of taxol-2'-MPT.
The yield was 13 mg or 85%. Spectroscopic data ('H
NMR and HRMS) were as expected.
Alternative synthetic schemes based upon the
method of T. Mukaiyama (Angewandte Chemie 1979,
X8(18), 707-808) using a variety of opium salts of 2-
halogenated aza-arenes for derivatizing either the 2'
or the 7 positions of taxol are illustrated in the
following scheme:

CA 02180444 1996-07-03
21~04~~
wo 9mssoa
~ 9
Acp 0 pH
BzNH O
~I
Ph~o-~ ~ _ ~)T~5on.26-mrdine,
off ' = r p cH; ~:,:
haioonium car., pH - H ' 2)haloonlum cal
O8: O
26.iutidine, 2.5-i~idinc, Chi, O!=
' CH?~;= Taxol (7)
Aeo O pH Ac0 O
°_:NH 0 i ExhIH 0 r ~R
Pt~~p ~» Ph~G ~»
OR ~ ~ 0 O T F.S ~ i 0
OH = H OAc pH pe: OAc
GE.
S .~ o . ~. '°~~
F s X O (> Alley! X ~ ~ Alley! X O ~ AJicyl t X O CAlky
X = SF,; TsO; hstides
Due to a difference in retention time using our
standard HPLC conditions (see Fig. 1) and differing
ultraviolet absorption maxima ( 120/ 1254 = 1. 6 for 2 and
0.3 for 1), the stability of 2 was easily assayed by
HPLC (Fig. 1). In all of the ensuing studies, the
only degradation products detected were taxol and the
pyridinone that results from hydrolysis of pyridinium
salts (Fig. ic.). Taxol-2'-MPT appears completely
stable in the solid state in a temperature range of
'80 C to 25 C regardless of the presence of an inert
atmosphere. In water, 5% dextrose, and 1.5% saline 2
' is stable for several days but begins to exhibit slow
degradation after 4 days. In phosphate buffered
saline (PBS) or ammonium acetate - phosphate buffer
systems of pH 6.0 to 7.3, 2 is stable at 25 C for

CA 02180444 1996-07-03
WO 95/18804 21 ~ 0 ~~ ~ PCTIUS95/00478
- 10 -
over 21 days. Taxol-2'-MPT (2) is, however, unstable
in 5% HC1 (pH 1.1) and brine. Most significantly, 2
breaks down rapidly when incubated at 37 C with
human plasma. This result suggests the presence of
factors within plasma that initiate the degradation
of 2 to taxol. Since taxol has been shown to bind to
albumin to the extent of ca. 85% in sera, it is
suspected that basic lysine residues on this protein
may initiate breakdown.
The kinetics of taxol release from taxol-2'-MPT
(2) in various aqueous solutions at 25°C is shown in
Figure 1. In sterile water, pH 6.2 phosphate
buffered saline, or 5% dextrose, no taxol release is
observed over a period of 11 minutes, as shown by the
horizontal line. Although stable in water and
aqueous buffer solutions, methylene chloride
extraction of plasma treated with compound 2 showed
complete conversion of 2 into taxol (1) within 10
minutes, as shown by the curved line. Under these
conditions 20% of total is recovered. More
particularly, Taxol-2'-MPT (2) was dissolved in the
aqueous system with the aid of sonication for five
minutes. Aliquots were then removed at the times
shown and partitioned into methylene chloride to
quench the reaction. Samples were then analyzed
using a Waters Maxima HPLC instrument equipped with
an autoinjector (3.9x300 mm C~8 column equipped with a
precolumn. The flow rate was 1.5 mL/minute. The
eluent gradient A-B extended over 30 minutes. "A"
was 80% 80mM ammonium acetate, pH 6Ø "B" was 100%
methanol. An ultraviolet diode array detector was
employed. The ratio of compound 2 (Rf 16.2 min.) to
taxol (compound 1, Rf 16.8 min.) remaining was
determined from the relative areas of the peaks after
normalization with previously determined calibration
curves.

CA 02180444 1996-07-03
wo ~snsso4 21 ~ ~'~'~'~' ~ pcrnUS9siooa's
-ii_
~,QlubilitY Me~sur~men~:
The solubility and partition coefficient data
for 2 and taxol were determined using an HPLC method.
10
Solubility was found by forming a solution in
water with the aid of sonication for five minutes,
centrifugation of the samples, and injection of the
supernatant. The values reported were normalized
using calibration curves constructed far both
compounds by preparing known concentrations in the
range 1 x 10'6 to 1 x 10'3 M in methylene chloride and
subjecting to HPLC analysis under the same
conditions. One should note that the solubility of
taxol is at the detection limit of the instrument and
thus represents an upper limit. The solubility of 2
was found at a concentration at which the solution
was clear (see below) and thus represents a lower
limit. Compound 2 exhibited similar solubilities for
various buffer systems in the pH range 6.2 to 7.4.
Partition coefficients were determined by dissolving
the compound in the organic phase, shaking the
resulting solution with water for ten minutes, and
analyzing each phase by HPLC as above. No
degradation of 2 was noted during these studies.

CA 02180444 1996-07-03
WO 95/18804 ~ ~ ~ ~ PCT/US95/00478
- 12 -
These data clearly show that 2 is significantly more
soluble in water than the parent taxol. The
solubility demonstrated in:a range of aqueous systems
is higher than the clinically relevant dosages (3 to
3 0 mM ) .
Self-Assembling Structures:
While the water solutions of 2 were optically
clear at all concentrations examined, buffered
solutions of concentrations greater than 1 x 10-3 M
exhibited a haze to the naked eye and diffuse
scattering of monochromatic light. Ultraviolet
spectroscopic absorption measurements at 340 nm (Fig.
3) showed an exponential increase in optical density
(OD) above a critical concentration of 4 x 10~ M, a
result characteristic of macromolecular structure in
solution. Transmission electron microscopy (TEM)
confirmed the presence of supramolecular structures
in these solutions. Uniform aggregates of fibrillar
structure (Fig. 2a) with helical conformations were
observed. These structures exhibited varying (up to
800 ~1) lengths but consistent diameter of ca. 80 A
with a helical twist of about 7. Additionally,
freshly sonicated solutions of 2 showed the presence
of spherical structures (Fig. 2b) with diameters of
about 50 A. It is likely that the long term
stability of these solutions is due, at least in
part, to stabilization provided by this structured
environment.
Microtubule polymerization-depolvmerization
measurements:
Microtubule polymerization-depolymerization
measurements (Fig. 3) with taxol-2'-MPT (2) were very
similar to GTP-saline controls and drastically
different from taxol. Compound 2 does not appear to

CA 02180444 1996-07-03
,~~.. 218 a ~ ~ 4
wo 9s~m rcrms~srooa~8
- 13 -
bind to tubulin in the manner of taxol. In the
buffered aqueous environment of this assay, 2 is not
converted to taxol and thus does not affect the
tubulin-microtubule equilibria. Taxol, recovered
from human plasma treated with 2, exhibited the
expected microtubule stabilization, indicating that Z
does act as a prodrug for taxol.
Tubulin polymerization-depolymerization
measurements are illustrate in Figure 3. Negative
controls are shown with triangles; positive taxol
controls are shown with diamonds; and taxol-2'-MPT,
i.e., compound 2 is shown with dots. The
measurements indicate that calcium chloride promoted
depolymerization is suppressed by taxol but not by
taxol-2'-MPT.
More particularly, measurement were performed in
96 well plates at 37°C following the protocol of R.
Merlock and W. Wrasidlo (Analytical Biochemistry
1993, in press). Calcium chloride additian is
indicated by the arrow. In each case, 1.0 mM GTP was
used to promote the initial polymerization of
tubulin. Negative control employed tubulin (1.0
mg/mLj lone, CaClz (0,25 mM) added after 20 minutes.
Positi _ taxol control employed tubulin (1.0 mg/mL)
with taxol (10'~M) and CaCl2 (0.25mM) added after 20
minute:. The experimental taxol-2'-MPT employed
tubulin (1.0 mg/mL) with taxol-2'-MPT (10'x) and CaClZ
(0.25 mM) added after 20 minutes. Turbidity was
measured as optical density at 340 nm using a
microplate reader (Molecular Devices Thermomax).
Toxicity Meas~pe:~~ntS
Compound 2 was tested for its cytotoxicity
against a cell line panel including leukemia,
ovarian, lung, and breast carcinoma cells (Fig. 4).
The differential cytotoxicity profiles for 2 and

CA 02180444 1996-07-03
WO 95118804 2 1$ PCT/US95I00478
- 14 -
taxol were similar, although ome differences were
noted. Both compounds exhibited ICso values ranging
from 10'S to 10'12 M with means close to one nanomolar.
Normal cells had cytotoxicity levels three to four
orders of magnitude below mean values. Extremely
high cytotoxicity levels were recorded for human
leukemia, metastatic melanoma and cervical carcinoma.
As expected for a prodrug of taxol in the cellular
environment, 2 shows the same remarkable tumor cell
selectivity and cell line specificity as taxol.
The relative cytotoxicities of taxol-2'-MPT
(compound 2) and taxol against a variety of cell
lines are illustrated in Figure 4. More
particularly, cells were plated on 96 well plates
with the following controls: no cells and toxic
control (1 x 10'3 M SDS). The drug was added to the
first set of wells and diluted via standard dilution
method from the stock. Plates were incubated at
37°C, 5% COz in sterile air in an humidified incubator
for 72 hours. An aliquot of 50 ~cL of a solution of
2,3-bis(methoxy-4-nitro-5-sulfophenyl)-5-
[(phenylamino)carbonyl]-1H-tetrazolium hydroxide
(XTT), 1 mg mL'', in phosphate buffered saline (PBS,
100mM) was added to the wells. In the presence of
viable cells, this colorless clear media is
enzymatically altered to give a pink coloration. The
plates were read at 450 nm using a plate reader.
Percentage cytotoxicity was calculated using the
formula: %C = 1 - (OD toxin)(OD growth
control)'' (100) .
Efficacy of taxol-2~-MPT in lung tumor xenoaraft:
The encouraging in vitro data obtained with
taxol-2'-MPT (2) prompted us to study its in vivo
action using nude mice inflicted with human lung
carcinoma xenografts (Fig. 1). The samples of 2 used

CA 02180444 1996-07-03
Wo 95~~~~
21$049
- 15 -
for this study were formulated in sterile PBS without
CremaphorT", indicating the suitability of this
compound for simple bolus administration.
Preliminary data shows that the control of tumor
growth exhibited by 2 is at least comparable to that
of taxol and significantly (0.001 p-value, multiple
linear regression model) different from controls.
These results provide a reasonable indication that 2
is converted rapidly to taxol in vivo and should thus
exhibit pharmacology similar to taxol. Indeed, in
metabolic study using tritiated 2, only 5% of the
compound was excreted through the kidneys, a result
that is completely in accord with the behavior of
taxol.
The efficacy of taxol-2'-MPT in lung tumor
xenograft nude mouse model is illustrated in Figure
5. The tumor model was generated from an ATCC A549
non-small cell lung adenocarcinoma cell line that was
maintained under the standard cell proliferation
conditions (37°C, 5% carbon dioxide in sterile air).
Hemocytometer counted cells suspended in Hanks medium
(Gibco, Grand Island NY) were implanted S.C. (106
cells in 0.4 mL per tumor volume determined using the
equation (length)(width)Z/2. The test compounds (1.0
microM) were injected I.P. on day 1,3, and 7 using
the following media: control. 5% dextrose in water
(D5W), triangles; taxol, suspended in Cremaphor/D5W
(5/95, 18.0 mg/kg of animal weight), diamonds; and
taxol-2'-MPT, dissolved in D5W (23.9 mg/kg of animal
wight), dots. The procedures used for the
maintenance of tumors and the experimental details
were according to protocols set forth by the
Developmental Therapeutics Program, National Cancer
Institute, viz., National Cancer Institute Cancer
Chemotherapy Reports, 3 (1972).

CA 02180444 1996-07-03
WO 95/18804 ~ ~ ~ ~ ~ PCT/US95/00478
- 16 -
Mechanisms of taxol release:
The mechanism of acid catalyzed taxol (i)
release from taxol-2'-MPT is illustrated in Scheme 2.
C
~ TsO~ TsO~
/ O "~"' ~ ~ --~-- taxol H +
v
H~O p iaxol HO ~pO ;axol p
- 2 ~' ~ 1-methylpyridfnone
Scheme 2
However, the release of taxol from taxol-2~-MPT
can also be catalyzed by serum protein and by
proteins having nucleophilic groups. When contacted
with serum protein, taxol-2'-MPT is observed to
displace its MPT group and form a protein:taxol
intermediate. Dialysis of the protein:taxol
intermediate indicates a dissociation period of hours
or days. Displacement of the MPT group by serum
proteins seems to be specific for such serum
proteins. Tested non-serum proteins seemed to lack
this activity. In particular, immunoglobulins and
serum albumen seem to be particularly effective
displacing the MPT group and forming protein:taxol
intermediates. The precise nature of the bonding
between the protein and taxol has not been
characterized. Scheme 2 illustrates alternative
pathways for MPT release.
Discussion
Taxol-2'-MPT has proven to be a remarkably
stable compound in most aqueous media. It is
probable that this stability is conferred upon 2 by
the facile formation of supramolecular aggregates, a
process that is probably driven by the amphiphillic

CA 02180444 1996-07-03
218~4~~
wo msso4 rcr~s9siooa~s
- i~ -
nature of the compound. The stability, water
solubility, and lack of cytotoxicity of taxol-2'-MPT
makes this class of compound an ideal a prodrug for
taxol and rnemetics of taxol. While essentially
completely stable in aqueous media at physiological
pH and ion strength, the compound rapidly discharges
taxol in sera. This profile is ideal for a
clinically useful prodrug to taxol. It is possible
that these properties should allow the formulation of
taxol-2'-MPT (2) without the use of Cremaphor~' or
ethanol.

CA 02180444 1996-07-03
wo 9s~i8so4 2 :~ 8 ~ t~ ~
PCT/US95/00478
~ 18 ~
SCR 1432
Ae0 0 OH
BzNH O
Scheme 3 Ph~O
OH ! 0 1)T~SDTt, 2,6-lutidine,
~ MFPT, OH ~ H ' Cl~i Cl
26-luiidine, OEz OAC 2)MP'PT 22.~lulidine
CHI~i~ Taxol (7) Ch~CIZ ,
Ts0
Ac0 0 OH Ac0 p
Ph N"r ' BzNH p ~ ' O
p °~ . Ph~p...
~ w
I ~ OH : H~O OTcS ~ O
TsO~ ~ ~ 08z OAc OH H OA
O8z c
Nu ' Nu
EtNH 1' O Ac0 O OH ' ezNH p Ac° O Nu
~,,~ ~I
Ph~ ~ Ph~O'
O I
N'u OH \ H ° OT~S , ~ I
Ubz OAc OH : H v
O~: OAS
H?n,~ Hh~~~2 H!'~~OH l2 f;~h'~.~'SH
Nu ~ O U
i
R~O. Sn !O FO NH3 H2S
NH2
Ac0 O OH Ac0 O
EzNH O ~ QzNH O ~ / I
I O "_ 1 J
Ph~ ! Ph~p_
I : O : I'~ I
OT'c5
OH O3~ H pAc OH : H : O
OEz OAc
N~ ~ Nu
r r
~~o
azNH ° v ° o~ Ago o Nu
1 ) 1 B~NH o ~ ~ ~ I
Ph l_ °_,
I ~ Pr~''o_ ~~~'''~
Nu : ° '
ON H : OT~S : °
03z OAS pH : N
p" OAc
0..

CA 02180444 1996-07-03
218l~~~~
"~..~
WO 95/18804 PGTIUS95I~00478
~ 19
Synthetic Methods
Preparation of 7-TES deacetylbaccatin ITI (4)
HO O OH _ HO O QTES
H O~,.. ~ K I I --w'. H 0..,.
~H ~'O ~ H ~O
HO pgz OAc HO OgZ OAc
10-deacetylbaccatin III (3)
S
7-TES deacetylbaccatin III (4). To a solution of 10-
deacetylbaccatin III ( 3 , 3.0 g, S .S 1 mmol, Indena Corpation,
Italy) in pyridine (250 mL) was added chlorotriethylsilane
( 18.5 mL, 110 mmol) dropwise. The resulting solution was
stirred at 25 °C for 17 hours. After dilution with diethylether
(750 mL), the solution was washed with aqueous CuS04 (3 x
200 mL) and brine ( 200 mL) . The organic layer was dried
(MgS04), concentrated, and purified by flash chromatography
(silica, 35 -~509~o ethylacetate in petroleum ether) to give
alcohol 4 (3.39 g, 9l~Yo) as a white solid.
Physical Data for 7-TES deacetylbaccatin III (4).
Rf= 0.32 (silica, 5090 ethylacetate in hexanes); IR (thin film)
vma,; 3464, 2954, 2282, 1710, 1453, 1362, 1271, 1242, 1105,
994 cm-1; 1H NMR (500 MHz, CDC13) 8 8.06 (dd, J= 8.0, 0.9 Hz, 2
H,Bz),7.57(t,J=7.9 Hz, lH,Bz),7.44(t,J=7.9Hz,2H,Bz),
S.Sfi(d,J=7.0Hz,1H,2-H),5.14(d,J=l.9Hz,1H,10-H),4.92
(d, J= 9.5 Hz, 1 H, S-H), 4.84-4.78 (m, 1 H, 13-H), 4.37 (dd, J=

CA 02180444 1996-07-03
WO 95/18804 ~ PCT/US95/00478
- 20 -
10.6, 7.0 Hz, 1 H, 7-H), 4.27 (d, J = 8.5 Hz, 1 H, 20-H), 4.25 (d, J
= 1.9 Hz, 1 H, 10-OH ) , 4.12 ( d, J =. 8 :5 Hz, 1 H, 2 0-H ) , 3 . 91 ( d, J
=
7.0 Hz, 1 H, 3-H), 2.48-2.40 (m, 1 H, 6-H), 2.25 (s, 3 H, Me),
2.25-2.17 (m, 2 H, 14-CH2); 2.04 (s, 3 H, Me), 1:90-1.82 (m, 1 H,
6-H), 1.70 (s, 3 H, Me), 1.03 (s, 6 H, Me, Me), 0.90 (t, J= 8 Hz, 9
H, Si(CH2C~j3)3), 0.58-0.42 (band, 6 H, Si(C~2CH3)3); 13C NMR
(125 MHz, CDC13) 8 210.3, 170.8, 167.0, 141.8, 135.1, 133.6,
130.1, 129.4, 128.6, 84.2, 80.7, 78.8, 76.5, 74.8, 74.6, 72.9, 67.9,
57.9, 47.0, 42.7, 38.6, 37.2, 26.8, 22.6, 19.5, 15.2, 9.9, 6.7, 5.1;
FAB HRMS (NBA / CsI) m / a 791.2251, M + Cs+ calcd for
C35HSOO10Si 791.2228.
Preparation of enone 5
HO O OTES ' HO /O- pTES
HO~~w( K I I --
O ~ H ~-O
HO pBZ pAc HO OBz OAc
4 5
Enone 5. To a solution of 7-TES deacetylbaccatin III (4,
1.5 g, 2.28 mmol) and 4-methylmorpholine N-oxide (NMO, 240
mg, 2.OS mmol) in CH?C1? ( S mL) was added 4 A molecular
sieves (200 mg) and the suspension was stirred at 2S °C for 10
minutesutes. A catalytic amount of tetrapropylammonium
perruthenate from Aldrich Chemical Company Inc. (TPAP, 40
mg, 0.11 mmol) was added by portions and the reaction

CA 02180444 1996-07-03
~.. 218 A4~4
WO 95118804 PGT/US95100498
- 21 -
mixture was stirred at 2 S °C for 0.5 hours. Small amounts of 4-
methylmorpholine N-oxide and TPAP were added alternatively
at 0.5 hour intervals until the starting material was consumed
to the extent of ca. 9596 by TLC. The reaction mixture was
filtered through silica gel, eluted with CH2C12 ( 100 mL), and
concentrated to give enone S ( 1.44 g, 9696) as a white solid.
Physical Data for Enone 5 . R f = 0.5 ( silica, S 096
ethylacetate in hexanes); IR (thin film) vma,; 3446, 2957, 2882,
1726, 1672, 1456, 1367, 1243, 1106 cm-1; 1H NMR (500 MHz,
CDCa3) 8 8.05 (dd, J= 8.0, 1.0 Hz, 2 H, Bz), 7.61 (t, J= 7.S Hz, 1 H,
Bz), 7.45 (t, J = 7.S Hz, 2 H, Bz), 5.63 (d, J= 7.S Hz, 1 H, 2-H),
5.30 (d,J=2.OHz, 1 H, 10-H),4.90(d,J=8.O Hz, 1H,5-H),4.36
(dd, J = 10.5, 7.0 Hz, 1 H, 7-H), 4.31 (d, J = 8.S Hz, 1 H, 20-H),
4.30 (d, J = 2.0 Hz, 1 H, 10-OH), 4.11 (d, J = 8.S Hz, 1 H, 20-H),
3.93 (d, J = 7.5 Hz, 1 H, 3-H), 2.92 (d, J = 19.5 Hz, 1 H, 14-H),
2.62 (d, J= 19.5 Hz, 1 H, 14-H), 2.50-2.42 (m, 1 H, 6-H), 2.17 (s,
3 H, Me), 2.08 (s, 3 H, Me), 1.90-1.82 (m, 1 H, 6-H), 1.77 (s, 1 H,
1-4H), 1.70 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.14 (s, 3 H, Me), 0.90
( t, J = 8.0 Hz, 9 H, Si(CH? C~,3 ) 3 ), 0.60-0.42 ( band, 6 H,
Si(c~CH3)3); 13C NMR ( 125 MHz, CDC13) s 208.2, 198.1, 170.2,
166.8, 156.6, 139.1, 134.0, 130.0, 128.8, 128.8, 84.0, 80.4, 78.5,
76.2, 75.7, 72.9, 72.8, 58.8, 45.9, 43.4, 42.5, 37.2, 33.0, 21.7,
17.5, 13.6, 9.6, 6.7, 5.1; FAB HRMS (NBA / NaI) m / a
657.3070, M + Na+ calcd for C3SH:~gOlpSi 657.3095.

CA 02180444 1996-07-03
WO 95/18804 ~ ~ PGT/US95/l10478
Preparation of triol 6
H O O pTES
- 22 -
O~C X I I ---.-
HO H =v0 -
OBa OAc '~" OH OAc
6
S Triol 6. To a solution of enone 5 ( 1.44 g, 2.19 mmol) in
MeOH (300 mL) at 0 °C was slowly added an aqueous solution
of K2C03 (3.0 g in 32 mL of H20). The solution was stirred at 0
°C for 2.5 hours. The reaction was then quenched with aqueous
NH4C1 ( 150 mL) and the resulting mixture was extracted with
CHZC12 (2 x 200 mL). The organic layer was dried (Na?S04),
concentrated, and purified by flash chromatography ( silica, 3 S
-~ 509'o ethylacetate in petroleum ether) to give enone S (270
mg, 199'0) and triol 6 (912 mg, 93% based on 8l~Yo conversion).
Physical Data for Triol 6. R f = 0.24 ( silica, S 09'0
ethylacetate in hexanes); IR (thin film) vma,; 3414, 2957, 2881,
1727, 1664, 1370 cm-1; 1H NMR (500 MHz, CDC13) 8 5.23 (d, J=
9.5 Hz, 1 H, 10-H), 4.89 (d, J= 9.5 Hz, 1 H, S-H), 4.63 (d, J= 9.S
Hz, 1 H, 20-H), 4.56 (d, J= 9.5 Hz, 1 H, 20-H), 4.32 (dd, J= 11.0,
7.0 Hz, 1 H, 7-H), 4.28 (d, J = 2.5 Hz, 1 H, 10-OH), 3.89 (dd, J =
.. 20 6.5, 4.0 Hz, 1 H, 2-H), 3.57 (d, J= 6.5 Hz, 1 H, 3-H), 2.78 (d, J=
19.SHz,1H,14-H),2.58(d,4.OHz,lH,2-OH),2.~2(d,J=19.5
Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, 6-H), 2.03 (s, 3 H, Me), 1.92-
1.84 (m, 1 H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3

CA 02180444 1996-07-03
WO 95/18804 PCTlUS95/00478
- 23
H, Me), 0.90 (t, J~ 8.0 Hz, 9 H, Si(CHzC$3)3), 0.60-0.40 (band, 6
H, Si(C~CH3)3); 13C NMR ( 125 MHz, CDC13) 8 208.9, 198.5, 170.1,
156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1,
42.8, 37.3, 32.7, 21.6, 17.5, 13.6, 9.7, 6.7, 5.1; FAB HRMS (NBA /
S NaI) m / a 575.2648, M + Na+ calcd for CZgH4~09Si 575.2652.
Preparation of Carbonate 7
HO 0 OTES HO O pTES
~ i~
H O H ~O O~ H ~.
O H OAc L,.O OAc
O 7
Carbonate 7. A solution of triol 6 ( 60.0 mg, 0.109
mmol) in THF ( 2 mL) was treated with carbonyldiimidazole
( 110.0 mg, 0.678 mmol) and stirred at 40 °C for 0.5 hour The
reaction mixture was concentrated and redisolved in THF ( 5
mL). TLC analysis confirmed total consumption of starting
material. 1 N aqueous HCl ( 5 mL) was added and the resulting
solution was allowed to stir for 15 minutes at 25 °C.
diethylether ( 25 mL) was added, the organic layer was
separated, washed with aqueous NaHC03 ( 10 mL) and brine ( 10
mL), dried (MgSO~), and concentrated to give carbonate 7 (58
mg, 939'0) as a white foam.
Physical Data for Carbonate 7. Rf= 0.50 (silica, 3590
ethylacetate in hexanes); IR (thin film) vna,; 3438, 2957, 2882,

CA 02180444 1996-07-03
WO 95/18804 ~ ~ PCT/US95/00478
- ~4 -
1820, 1731, 1685, 1370, 1236 cm-1; 1H NMR (500 MHz, CDC13) 8
5.27 (d, J= 2.5 Hz, 1 H, 10-H), 4:89 (d, J= 9.0 Hz, 1 H, 5-H), 4.60
(d, J = 9.0 Hz, 1 H, 20-H), 4.45 (d, J= 9.0 Hz, 1 H, 20-H), 4.43 (d,
J= 6.0 Hz, 1 H, 2-H), 4.33 (dd, J= 10.0, 7.5 Hz, 1 H, 7-H), 4.28
(d, J= 2.5 Hz, 1 H, 10-OH), 3.54 (d, J= 6.0 Hz, 1 H, 3-H), 2.88 (d,
J= 20.0 Hz, 1 H, 14-H), 2.75 (d, J = 20.0 Hz, 1 H, 14-H), 2.54-
2.47 (m, 1 H, 6-H), 2.08 (s, 3 H, Me), 2.06 (s, 3 H, Me), 1.92-1.84
(m, 1 H, 6-H), 1.77 (s, 3 H, Me), 1.31 (s, 3 H, Me), 1.15 (s, 3 H,
Me), 0.88 (t, J= 8.5 Hz, 9 H, Si(CH?C$3)3), 0.55-0.45 (band, 6 H,
Si(C~j2CH3)3); ~3C NMR ( 125 MHz, CDC13) b 208.4, 195.5, 170.5,
154.0, 152.0, 141.2, 88.4, 83.9, 79.8, 79.0, 76.7, 75.7, 71.9, 60.3,
43.0, 41.6, 39.8, 37.7, 31.6, 21.5, 17.8, 14.4, 9.7, 6.6, 5.0; FAB
HRMS (NBA) m / a 579.2652, M + H+ calcd for C?9H4?OlpSi
579.2626.
Preparation of nButyl-C-2 ester derivative (Alcohol 8)
HO O OTES HO O OTES
nBuLi
O '~ O
' THF
O H ' O HO H ' O
''p OAc O OAc
O 7 O 8
Alcohol 8. A solution of carbonate 7 ( 10 mg,
0.0173 mmol) in tetrahydrofuran ( 1 mL) at -78 °C was treated
with n-Butyllithium from Aldrich Chemical Company, Inc.
(0.087 mL of a 1.6 M solution in hexanes, 0.139 mmol) and

CA 02180444 1996-07-03
2184444
WO 95/18804 PCT/US95/00478
- 25 -
stirred for 1.0 hour The reaction mixture was poured into a
mixture of diethylether ( 10 mL) and aqueous NH~CI (S mL).
The organic layer was separated and the aqueous layer was
extracted with diethylether (2 x 5 mL). The combined organic
S layer was washed with a saturated solution of brine ( S mL),
dried (MgS04), concentrated, and purified by flash
chromatography (silica, 35 -~ 509~o ethylacetate in hexanes) to
give 8 (7.9 mg, 729'0) as an amorphous solid.
Physical Data for Alcohol 8. R f = 0.3 G ( silica, 3 590
ethylacetate in petroleum ether); IR (filin) v,~,~,a,; 3437, 2962,
2865, 1726, 1671, 1367, 1239, 1105 cm-1; 1H NMR (500 MHz,
CDC13) 8 5.36 (d, J= 6.5 Hz, 1H, 2-H), 5.26 (d, J= 2.5 Hz, 1H, 10-
H), 4.89 (br d, J= 8.0 Hz, 1H, 5-H), 4.47 (d, J= 8.0 Hz, 1H, 20-H),
4.32 (dd, J= 10.5, 6.S Hz, 1H, 7-H), 4.26 (d, J= 2.5 Hz, 1H, 10-
OH), 4.1S (d, J= 8.0 Hz, 1H, 20-H), 3.81 (d, J= 6.5 Hz, 1H, 3-H),
2.73 (d, J= 20.0 Hz, 1H, 14-H), 2.57 (d, J= 20.0 Hz, 1H, 14-H),
2.49-2.41 (m, 1H, 6-H), 2.38-2.23 (m, 2H, OCC~?(CH2)?CH3), 2.06
(s, 3H, Me), 2.04 (s, 3H, Me), 1.90-1.82 (m, 1H, G-H), 1.67 (s, 1H,
OH), 1.64 (s, 3H, Me), 1.68-1.52 (m, 2H, C~CCH?C~~CH~CH3), 1.41-
1.30 (m, 2H, OC(CH~)~C~,?CH3), 1.19 (s, 3H, Me), 1.07 (s, 3 H,
Me), 0.94-0.86 (band, 12H, CH3 of Bu, OSi(CH?C~3)3), 0.58-0.45
(band, 6H, OSi(C~?CH3)3); FAB HRMS (NBA) m / a 637.3421, M
+ H+ calcd for C33H~~OlpSi 637.3408.

CA 02180444 1996-07-03
WO 95/18804 ~ PCT/US95/00478
- 26
Preparation of vinyl-C-2 ester derivative (Alcohols 9
and 10 )
HO O OTES ~ HO .O OTES
U
O '~ O
~~O or _ _ 0
O p H OA~c ~ HO = H
MgBr _ ,O OAc
O
O
HO O OTES
O
H O H~'O
p OH
O
S 10
Alcohols 9 and 10. A solution of carbonate 7 ( 111.3
mg, 0.192 mmol) in tetrahydrofuran ( 2 mL) at -7 8 °C was
treated with vinyllithium (3.7 mL of a 0.52 M solution in
diethylether, 1.92 mmol, prepared from tetravinyltin and
nButyllithium: methodology from Wakefield, B.J. Organolithium
Methods, Academic Press: London, 1988, p. 46) and stirred for
2.25 hour The reaction mixture was poured into a mixture of
CH2C12 (20 mL) and aqueous NH~CI (10 mL), the organic layer
was separated, and the aqueous layer was extracted with
CHZC1? (3 x 10 mL). The combined organic layer was washed
with brine (IS mL), dried (MgSO.~), concentrated, and purified
by flash chromatography (silica, 30 -~ SO% ethylacetate in

CA 02180444 1996-07-03
218!~~44
WO 95/18804 PGT/US95I00478
- Z7
petroleum ether) to give 9 (b0.0 mg, 529'0), and 10 (25.7 mg,
2496 ) as white foams.
Physical Data for Alcohol 9. Rf = 0.5 2 ( silica, S 096
ethylacetate in hexanes); IR (film) v~"a,; 3442, ~295b, 2882,
1727, 1672, 1407, 13b8, 1243, 1182, 1110, 1050, 986, 826,
73 b cm-1; 1 H NMR ( S 00 MHz, CDC13 ) 8 6.51 ( dd, J = 17 .0, 1.0 Hz,
1H, vinyl H), b.13 (dd, J= 17.0, 10.5 Hz, 1H, vinyl H), b.00 (dd, J
= 10.5, 1.0 Hz, 1H, vinyl H), S.45 (br d, J= b.5 Hz, 1H, 2-H), 5.30
(d, J = 2.5 Hz, 1H, 10-H), 4.91 ( br d, J = 9.5 Hz, 1H, 5-H), 4.44 ( d,
J= 8.5 Hz, 1H, 20-H), 4.35 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.30
(d, J= 2.5 Hz, 1H, 10-OH), 4.14 (d, J= 8.5 Hz, 1H, 20-H), 3.88 (d,
J= b.5 Hz, 1H, 3-H), 2.79 (d, J= 20.0 Hz, 1H, 14-H), 2.G1 (d, J=
20.0 Hz, 1H, 14-H), 2.48 (ddd, J = 14.5, 9.5, b.5 Hz, 1H, b-H),
2.09 (s, 3H, Me), 2.08 (s, 3H, Me), 1.89 (ddd, J= 14.5, 10.5, 2.0
Hz, 1H, b-H), 1.72 (s, 1H, OH), 1.b8 (s, 3H, Me), 1.22 (s, 3H, Me),
1.12 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.62-0.4G
(band, 6H, OSi(C~? C H 3 ) 3 ); FAB HRMS ( NBA / CsI) m / a
739.1925, M + Cs+ calcd for C31H.~~OIpSi 739.1915.
Physical Data for Alcohol 10. Rf = 0.24 (silica, 509%
ethylacetate in hexanes); IR (film) vma,; 3439, 2955, 2881,
1711, 1b71, 1409, 13b5, 1188, 1115, 980, 833, 735 cm-1; 1H
NMR (500 MHz, CDCl3) 8 6.48 (br d, J = 17.0 Hz, 1H, vinyl H),
b.10 (dd, J= 17.0, 10.5 Hz, 1H, vinyl H), 5.97 (br d, J= 10.5 Hz,
1H, vinyl H), 5.47 (br d, J= b.0 Hz, 1H, 2-H), 5.25 (d, J= 2.5 Hz,
1H, 10-H), 4.75 (dd, J= 9.5, 3.5 Hz, 1H, S-H), 4.38 (d, J= 8.5 Hz,
1H, 20-H), 4.30 (d, J= 2.5 Hz, 1H, 10-OH), 4.24 (d, J= 8.5 Hz, 1H,
20-H), 3.90 (dd, J = 11.5, 6.0 Hz, 1H, 7-H), 3.28 (d, J = 19.5 Hz,

CA 02180444 1996-07-03
WO 95/18804 2 1 PCT/US95/00478
- 28 -
1H, 14-H), 3.24 (d, J= 6.0 Hz, 1H, 3-H), 3.06 (br s, 1H, OH), 2.58
(d, J= 19.5 Hz, 1H, 14-H), 2.38 (ddd, J= 14.5, 9.5, 6.0 Hz, 1H, 6-
H), 2.07 (s, 3H, Me), 1.98 (ddd, J= 14.5, 11.5, 3.5 Hz, 1H, 6-H),
1.87 (s, 1H, OH), 1.61 (s, 3H, Me), 1.23 (s, 3H, Mej, 1.13 (s, 3H,
Me), 0.90 (t, J= 8.0 Hz, 9H, OSi(CH?C,j~3)3), 0.59-0.45 (band, 6H,
OSi(C~j2CH3)3); FAB HRMS (NBA / CsI) m / a 697.1802, M + Cs+
calcd for C?gH4.~OgSi 697.1809.
Preparation of 2-Furyl-C-2 ester derivative (Alcohol
11)
HO O OTES O V HO O OTES
O ~ O
w
O H 'A O HO H O
O c O p OAc
O
7 11
Alcohol 11. A solution of carbonate 7 (46 mg, 0.0795
1S mmol) in tetrahydrofuran (3 mL) at -78 °C was treated with 2-
furyllithium (4 mL of a 0.47 M suspension in diethylether, 1.88
mmol, prepared from furan (Aldrich Chemical Company, Inc.)
and n-Butyllithium (Aldrich Chemical Company, Inc.);
methodology from Ramanathan, V.; Levine, R. J. Org. Chem.
1962, 27, 1216) and stirred for 10 minutesutes The reaction
mixture was poured into a mixture of CH? C1? ( 15 mL) and
aqueous NH4C1 (20 mL). The organic layer was separated and

CA 02180444 1996-07-03
Wo 9~~s$~ 218 Q 4-~.4 p~~s~g
- a9 -
the aqueous layer was extracted with CH~C12 (2 x 10 mL). The
combined organic layer was washed with brine ( 10 mL), dried
(MgS04) and concentrated to give 11 which was taken into the
next step without further purification. '
S Physical Data for Alcohol 11. R f = 0.3 8 ( silica, 209
ethylacetate in petroleum ether); IR (film) vI,r,a,; 3442, 2956,
2882, 1727, 1672, 1468, 1300, 1240, 1110, 1007, 733 cm-1; 1H
NMR (S00 MHz, CDC13) 8 7.GG-7.G4 (m, 1H, furan), 7.24 (br d, J=
3.S Hz, 1H, furan), 6.58 (dd, J= 3.5, 1.S Hz, 1H, furan), 5.55 (d, J
= 6.5 Hz, 1H, 2-H), 5.31 (d, J= 2.0 Hz, 1H, 10-H), 4.92 (br d, J=
9.0 Hz, 1H, S-H), 4.43 (d, J = 8.5 Hz, 1H, 20-H), 4.37 (dd, J =
10.5, 6.5 Hz, 1H, 7-H), 4.32 (d, J= 2.0 Hz, 1H, 10-OH), 4.18 (d, J
= 8.5 Hz, 1H, 20-H), 3.93 (d, J = G.S Hz, 1H, 3-H), 2.88 (d, J =
20.0 Hz, 1H, 14-H), 2.G3 (d, J= 20.0 Hz, 1H, 14-H), 2.55-2.37 (m,
1H, 6-H), 2.15 (s, 3H, Me), 2.09 (s, 3H, Me), 1.93-1.87 (m, 1H, G-
H), 1.81 (s, 1H, OH), 1.71 (s, 3H, Me), 1.23 (s, 3H, Me), 1.15 (s,
3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), O.G2-0.42 (band,
6H, OSi(C~CH3)3); FAB HRMS (NBA / NaI) m / a GG9.2717, M +
Na+ calcd for C33H.~~0~ 1Si GG9.2707.

CA 02180444 1996-07-03
218~~4~
WO 95/18804 PCT/US95/00478
Preparation of 2-thiophenyl-C-2 ester derivative
(Alcohol 12 )
HO O OTES 5 V HO O OTES
o -~ 'C 1 ~- o ~~ ~' 1
0 H ~ I-t W--O
p OAc
S p OAc
O ~ ~ O
12
5
Alcohol 12. A solution of carbonate 7 (50.0 mg,
0.0864 mmol) in tetrahydrofuran (5 mL) at -78 °C was treated
with 2-thienyllithium from Aldrich Chemical Company, inc.
( 1.30 mL of a 1.0 M solution in tetrahydrofuran, 1.30 mmol)
10 and stirred for 0.5 hour The reaction mixture was poured into
a mixture of diethylether ( 10 mL) and aqueous NH~CI ( 5 mL) .
The organic layer was separated and the aqueous layer was
extracted with diethylether (2 x 10 mL). The combined organic
layer was washed with brine ( 10 mL), dried (MgSO~),
15 concentrated, and purified by flash chromatography ( silica, 10
--~ 359~o ethylacetate in hexanes) to give 7 ( 1G.5 mg, 33%), 12
(36.8 mg, 969 based on 679~o conversion) as an amorphous
solid.
Physical Data for Alcohol 12. Rf = 0.5G (silica, 50%
20 ethylacetate in hexanes); IR (film) vma~ 3403, 2945, 2881,
1717, 1669, 1520, 1413, 1360, 1248, 107$; 1H NMR (500 MHz,
CDC13) 8 7.84 (dd, J= 3.5, 1.0 Hz, 1H, thiophene), 7.64 (d, J= I.O,

CA 02180444 1996-07-03
wo 9srissoa rcrms9s~~a
~ 31 ~
5.0 Hz, 1H, thiophene), 7.14 (dd, J= 5.0, 3.5 Hz, 1H, thiophene),
5.53 (br d, J = 6.5 Hz, 1H, 2-H), 5.29 (d, J = 2.5 Hz, 1H, 10-H),
4.90 (br d, J = 7.5 Hz, 1H, 5-H), 4.44 (d, J = 8.5 Hz, 1H, 20-H),
4.35 (dd, J = 10.5 Hz, 6.5 Hz, 1H, 7-H), 4.29 (d, J = 2.5 Hz, 1H,
10-OH), 4.19 (d, J= 8.5 Hz, 1H, 20-H), 3.90 (d, J= 6.5 Hz, 1H, 3-
H), 2.89 (d, J= 19.5 Hz, 1H, 14-H), 2.62 (d, J= 19.5 Hz, 1H, 14-
H), 2.49-2.43 (m, 1H, 6-H), 2.15 (s, 3H, Me), 2.07 (s, 3H, Me),
1.92-1.84 (m, 1H, 6-H), 1.73 (s, 1H, OH), 1.71 (s, 3H, Me), 1.21
( s, 3H, Me), 1.13 ( s, 3H, Me), 0.91 ( t, J = 8.0 Hz, 9H,
OSi(CH2C~3)3), 0.56-0.49 (band, 6H, OSi(C~,~CH3)3); FAB HRMS
(NBA) m / a 663.2655, M + H+ calcd for C33H~~OIpSSi 663.2659.
Preparatioa of 3-thiophenyl-C-2 ester derivatives
(Alcohol 13 and 14)
a
HO O OTES ~ ~ HO O OTES
S
O .:~. 0
p H ~O HO ~ H WO
OAc ~ O OAc
O S O
13

CA 02180444 1996-07-03
218~4~~.
WO 95/18804 PCT/US95/00478
- 32 -
HO O pTES
O
~H
HO _ O
p OH
S O
14
Alcohols 13 and 14. A solution of carbonate 7 ( 107.9
mg, 0.186 mmol) in tetrahydrofuran (G.2 mL) at -78 °C was
treated with 3-thienyllithium ( 2.7 G mL of a 0.41 M solution in
diethylether : tetrahydrofuran : hexanes (4.5 : 1 : 2), 1.13
mmol, prepared from 3-bromothiophene and n-Butyllithium;
methodology from Camici, L.; Ricci, A.; Taddei, M. Tetrahedron
Lett. 19 8 6, 2 7, 515 S ) and stirred for 1.5 hour The rear tion
mixture was poured into a mixture of CH?C12 ( 15 mL) and
aqueous NH4C1 ( 20 mL), the organic layer was separated, and
the aqueous layer was extracted with CH?Cl? (2 x 10 mL). The
combined organic layer was washed with brine ( 10 mL) , dried
(MgS04), concentrated, and purified by flash chromatography
(silica, 20 -~ 30% ethylacetate in hexanes) to give 7 ( 1G.9 mg,
1G~'o), 13 (87.0 mg, 83% based on 84% conversion), and
hydrolyzed C4 acetate 14 (C4-hydrolyzed side product, 9.7 mg,
10~'o based on 84% conversion) as amorphous solids.
Physical Data for Alcohol 13. Rf = 0.74 (silica, 50%
ethylacetate in hexanes), 0.41 (silica, 10% ethylacetate in
benzene, 3 elutions); IR (thin film) v,T,a~; 3442, 3110, 2956,
2882, 1725, 1672, 1410, 1368, 1244, 1198, 1101, 988, 825,

CA 02180444 1996-07-03
WO 95/18804 ~ PCTlIJS95lOp478
- 33 -
744 cm-1; 1 H NMR ( 500 MHz, CDC13 ) 8 8.18 ( dd, J = 3 .0, 1.2 Hz,
1H, thiophene), 7.54 (dd, J = 5.0, 1.2 Hz, 1H, thiophene), 7.37
(dd, J= 5.0, 3.0 Hz, 1H, thiophene), 5.56 (dd, J= 6.5, 1.0 Hz, 1H,
2-H), 5.31 (d, J= 2.5 Hz, 1H, 10-H), 4.92 (dd, J= 7.5, 2.0 Hz, 1H,
S S-H), 4.40-4.34 (m, 2H, 20-H, 7-H), 4.31 (d, J = 2.5 Hz, 1H, 10-
OH), 4.15 (d, J= 8.S Hz, 1H, 20-H), 3.93 (d, J= G.S Hz, 1H, 3-H),
2.88 (d, J= 20 Hz, 1H, 14-H), 2.63 (dd, J= 20.0, 1.0 Hz, 1H, 14-
H), 2.47 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, G-H), 2.18 (s, 3H, Me),
2.10 (s, 3H, Me), 1.89 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, G-H), 1.81
(br s, 1H, OH), 1.72 (s, 3H, Me), 1.23 (s, 3H, Me), 1.15 (s, 3H,
Me), 0.93 ( t, J = 8.0 Hz, 9H, OSi( CH?C$3 ) 3 ), O.G2-0.48 ( band, GH,
Si(C$?CH3)3); FAB HRMS (NBA / CsI) m / a 795.1640, M + Cs+
calcd for C33H4601oSSi 795.1635.
Physical Data for Alcohol 14: Rf = 0.54 (silica,
5096 ethylacetate in hexanes); IR (thin film) vna,; 3437, 3108,
2955, 2880, 1709, 1674, 1605, 1520, 1410, 1360, 1258, 1194,
1103, 1004, 829, 744 cm-1; iH NMR (500 MHz, CDC13) S 8.15
(dd, J= 3.0, 1.0 Hz, 1H, thiophene), 7.49 (dd, J= 5.0, 1.0 Hz, 1H,
thiophene), 7.35 (dd, J= 5.0, 3.0 Hz, 1H, thiophene), 5.59 (d, J=
6.0 Hz, 1H, 2-H), 5.27 (d, J= 2.5 Hz, 1H, 10-H), 4.73 (dd, J= 9.5,
3.5 Hz, 1H, 5-H), 4.40 (d, J = 8.5 Hz, 1H, 20-H), 4.3 2 (d, J = 2.5
Hz, 1H, 10-OH), 4.15 (d, J= 8.5 Hz, 1H, 20-H), 3.92 (dd, J= 11.5,
6.0 Hz, 1H, 7-H), 3.44 (d, J= 19.5 Hz, 1H, 14-H), 3.30 (d, J= G.0
Hz, 1H, 3-H), 2.91 (br s, 1H, OH), 2.61 (d, J= 19.5 Hz, 1H, 14-H),
2.38 (ddd, J= 14.5, 9.5, G.0 Hz, 1H, G-H), 2.09 (s, 3H, Me), 1.99
(ddd, J= 14.5, 11.5, 3.5 Hz, 1H, G-H), 1.81 (br s, 1H, OH), 1.65 (s,
3H,Me),1.24(s,3H,Me),1.1G(s,3H,Me),0.91(t,J=8.O Hz,

CA 02180444 1996-07-03
2 ~ 8 ~ 4.~-~.
WO 95/18804 PCTIUS95I00478
34
9H, OSi(CH2C~3)3), 0.60-0.46 (band, 6H, OSi(C~2CH3)3); FAB
HRM~ (NBA / CsI) m / a 753.1530, M + Cs+ calcd for
C31H4409SSi 753.1530.
Preparation of 2-pyridinyl-C-2 ester derivatives
( Alcohol 1 S , 16 and triol 6 )
HO O OTES I N~ V HO O OTES
O -''' O
H ~-O ~! H ~ O
O p OAc N O p OAc
O
/ O
7 15
HO O OTES HO O pTES
O O
H~ O = O
HO ~ OH HO
N' pHH OAc
N
/ O
16 6
Alcohol 15 and 16, and triol 6. A solution of
carbonate 7 ( 62.6 mg, 0.108 mmol) in tetrahydrofuran ( S .~
mL) at -78 °C was treated with 2-lithiopyridine ( 1.15 mL of a
0.44 M solution in diethylether-pentane 1 : 1, 0.506 mmol,

CA 02180444 1996-07-03
218a9-
wo 9sr~sgoa pcr~s9s~ooa~s
prepared from 2-bromopyridine and r-Butyllithium;
methodology from Malmberg, H.; Nilsson, M. Tetrahedron,
1986, 42, 3 981 ) and stirred for 1.3 hour The reaction mixture
was poured into a mixture of ethylacetate (~10 mL) and
5 aqueous NH4C1 ( S mL), the organic layer was separated, and the
aqueous layer was extracted with ethylacetate (2 x 10 mL).
The combined organic layer was washed with brine ( 5 mL),
dried (MgS04), concentrated, and purified by flash
chromatography ( silica, 70 --~ 1009~o ethylacetate in petroleum
10 ether) to give 6 (16.3 mg, 2790), ~5 (28.0 mg, 3990), and 16 (8.4
mg, 139'0) as amorphous solids.
Physical Data for Alcohol 15. Rf = 0.60 (silica,
ethylacetate); 1H NMR (500 MHz, CDC13) 8 8.77 (ddd, J= 4.5, 1.7,
1.0 Hz, 1H, pyridine), 8.05 (br d, J= 7.5 Hz, 1H, pyridine), 7.89
15 (ddd, J= 7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J= 7.5, 4.5,
1.0 Hz, 1H, pyridine), 5,61 (dd, J= 6.5, 1.0 Hz, 1H, 2-H), 5.33 (d,
J = 2.5 Hz, 1H, 10-H), 4.92 (dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.39
(dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.36 (d, J = 9.0 Hz, 1H, 20-H),
4.33 (d, J = 2.5 Hz, 1H, 10-OH), 4.28 (d, J = 9.0 Hz, 1H, 20-H),
20 3.96 (d, J= 6.5 Hz, 1H, 3-H), 2.98 (d; J= 20.0 Hz, 1H, 14-H), 2.71
(dd, J= 20.0, 1.0 Hz, 1H, 14-H), 2.50 (s, 1H, OH), 2.48 (ddd, J=
14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.15 (s, 3H, Me), 2.11 (s, 3H, Me),
1.90 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.76 (s, 3H, Me), 1.24
(s, 3H, Me), 1.16 (s, 3H, Me), 0.93 (t, J = 8.0 Hz, 9H,
25 OSi(CH2C~3)3), 0.63-0.47 (band, 6H, OSi(C,~7CH3)3); FAB HRMS
(NBA / CsI) m / a 790.2060, M + Cs+ calcd for C3~H.~~OIpNSi
790.2024.

CA 02180444 1996-07-03
WO 95/18804 ~ ~ PCT/US95/00478
- 36 -
Physical Data for Alcohol 16. Rf = 0.45 (silica,
ethylacetate); IR (film) vma,; 335, 2954, 2879, 1732, 1674,
1589, 1362, 1305, I24I, lIlG, 998, 829, 741 cm-I; IH NMR
(S00 MHz, CDC13) 8 8.73 (br d, J= 4.5 Hz, 1H, pyridine), 8.15 (br
d, J= 7.5 Hz, 1H, pyridine), 7.90 (ddd, J= 7.5, 7.5, 1.7 Hz, 1H,
pyridine), 7.56 (ddd, J= 7.5, 4.5, 1.0 Hz, 1H, pyridine), 5.53 (dd,
J= 7.5, 1.0, 1H, 2-H), 5.30 (d, J= 2.5 Hz, 1H, 10-H), 4.84 (dd, J=
9.5, 3.0 Hz, 1H, S-H), 4.81 (br s, 1H, OH), 4.31 (d, J= 2.5 Hz, 1H,
10-OH), 4.25 (s, 2H, 20-CH2), 3.97 (dd, J= 11.5, 6.5 Hz, 1H, 7-H),
3.31 (d, J= 19.5 Hz, 1H, 14-H), 3.23 (d, J= 7.5 Hz, 1H, 3-H), 2.57
(br d, J= 19.5 Hz, 1H, 14-H), 2.43 (ddd, J= 14.5, 9.5, G.S Hz, 1H,
6-H), 2.11 (s, 3H, Me), 1.95 (ddd, J= 14.5, 11.5, 3.0 Hz, 1H, 6-H),
1.92 (br s, 1H, OH), 1.70. (s, 3H, Me), 1.24 (s, 3H, Me), 1.17 (s,
3H, Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH?C_H3)3), 0.60-0.46 (band,
6H, OSi(C~2CH3)3).
Physical Data for Triol 6. Rf = 0.24 (silica, 50%
ethylacetate in hexanes); IR (thin film) vma~ 3414, 2957, 2881,
1727, 1664, I370 cm-1; 1H NMR (500 MHz, CDCI3) 8 5.23 (d, J=
9.5 Hz, 1 H, 10-H), 4.89 (d, J= 9.5 Hz, 1 H, 5-H), 4.63 (d, J= 9.5
Hz, I H, 20-H), 4.56 (d, J= 9.5 Hz, 1 H, 20-H), 4.32 (dd, J= 11.0,
7.0 Hz, 1 H, 7-H), 4.28 (d, J= 2.5 Hz, 1 H, 10-OH), 3.89 (dd, J=
6.5, 4.O Hz, 1 H, 2-H), 3.57 (d, J = 6.S Hz, 1 H, 3-H), 2.78 (d, J=
19.SHz,1H,14-H),2.58(d,4.OHz,lH,2-OH),2.52(d,J=19.5
Hz, 1 H, 14-H), 2.49-2.42 (m, 1 H, G-H), 2.03 (s, 3 H, Me), 1.92-
1.84 (m, 1 H, 6-H), 1.G8 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04 (s, 3
H, Me), 0.90 (t, J= 8.0 Hz, 9 H, Si(CH?C,~3)3), 0.60-0.40 (band, 6
H, Si(C~T?CH3)3); 13C NMR ( 125 MHz, CDC13) b 208.9, 198.5, 170.1,

CA 02180444 1996-07-03
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WO 95118804 PGT/US95I00478
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156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8, 72.5, 58.8, 45.8, 43.1,
42.8, 37.3, 32.7, 21.6, 17.5, 13.6, 9.7, G.7, 5.1; FAB HRMS (NBA /
NaI) m / a 575.2648, M + Na+ calcd for CZgH~09Si 575.2652.
S Preparation of 3-pyridinyl-C-2 ester derivative
(Alcohol 17 )
N
HO O OTES ' ~ HO O pTES
I I ""-'~ O K I I
H ~O p H ~O
OAc ~ ' p OAc
O/~ N ~-
O
Alcohol 17. To a solution of 3-lithiopyridine ( 1.15
mmol) in tetrahydrofuran (7 mL), prepared from 3-
bromopyridine (Aldrich Chemical Company Inc.) and n-
Butyllithium (Aldrich Chemical Company Inc.) at -100 °C
1S (methodology from Parham, W.E.; Piccirilli, ft. M. J. Org. Chem.
1977, 42, 257), was added a solution of carbonate 7 ( 133.1
mg, 0.230 mmol) in tetrahydrofuran (2 mL) via cannula. The
resulting solution was stirred for 1 h, allowed to warm to -78
°C, stirred for 1 h, and poured into a mixture of ethylacetate
( 10 mL) and aqueous NH:~Cl ( 10 mL). The organic layer was
separated and the aqueous layer was extracted with

CA 02180444 1996-07-03
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WO 95/18804 PCT/US95/00478
- 3$ -
ethylacetate (2 x 10 mL). The combined organic layer was
washed with brine ( 10 mL), dried (MgS04), concentrated, and
purified by flash chromatography (silica, 70 -~ 9590
ethylacetate in petroleum ether) to give 7 (64.8 ring, 499%) and
S 17 (43.9 mg, 579~o based on S 19'o conversion) as an amorphous
solid.
Physical Data for Alcohol 17. R f = 0.5 G ( silica,
ethylacetate); IR (film) vma,; 3435, 2956, 2882, 1731, 1671,
1592, 1366, 1280, 1240, 1109, 991, 824, 739 cm-1; 1H NMR
( S 00 MHz, CDC13 ) 8 9 . 2 4 ( br s, 1 H, pyridine ) , 8 . 81 ( d, J = 1.0,
4.5
Hz, 1H, pyridine), 8.30 (ddd, J= 8.0, 2.0, 2.0 Hz, 1H, pyridine),
7.44 (dd, J= 8.0, 4.5 Hz, 1H, pyridine), 5.6G (d, J= G.5 Hz, 1H, 2-
H), 5.32 (s, 1H, 10-H), 4.92 (dd, J= 9.5, Z.0 Hz, 1H, 5-H), 4.38
(dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.32 (br s, 1H, OH), 4.30 (d, J=
8.5 Hz, 1H, 20-H), 4.13 (d, J= 8.5 Hz, 1H, 20-H), 3.9G (d, J= G.S
Hz, 1H, 3-H), 2.92 (d, J= 19.5 Hz, 1H, 14-H), 2.GG (d, J= 19.5 Hz,
1H, 14-H), 2.48 (ddd, J= 15.5, 9.5, G.5 Hz, 1H, G-H), 2.18 (s, 3H,
Me), 2.10 (s, 3H, Me), 2.03 (s, 1H, OH), 1.89 (ddd, J= 14.5, 10.5,
2.0 Hz, 1H, 6-H), 1.72 (s, 3H, Me), 1.23 (s, 3H, Me), 1.1G (s, 3H,
Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH~C~-3)3), O.G2-0.48 (band, GH,
OSi(C~?CH3)3); FAB HRMS (NBA / CSI) m / a 790.2030, M + Cs+
calcd for C34H4~010NSi 790.2024.

CA 02180444 1996-07-03
2184
WO 95!18804 PCT/US95I00478
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Preparation of 4-N,N-dimethylaniline-C-2 ester
derivative (Alcohol 18)
HO O pTES Me N ~ ~ ~ HO' O pTES
2
O ~ -~- O
O -~'' H V O
p OAc O p OAc
Me2N
O
O ~
S
Alcohol 18. A solution of carbonate ? ( 150 mg,
0.259 mmol) in tetrahydrofuran (20 mL) at -78 °C was treated
with 4-lithio-N,N dimethylaniline (6.5 mL of a 0.39 M solution
in diethylether : pentane (3 : 1), 2.54 mmol, prepared from 4-
bromo-N,N dimethylaniline and t Butyllithium; methodology
from Jones, F.N.; Hauser, C.R. J. Org. Chem. 1962, 27, 4389) and
stirred for 15 minutesutes The reaction mixture was poured
into a mixture of CH?Cl? (35 mL) and aqueous NH~CI (20 mL),
the organic layer was separated, and the aqueous layer was
1 S extracted with CH?Ch ( 2 x 20 mL) . The combined organic layer
was washed with brine (20 mL), dried (MgSO.~), concentrated,
and purified by flash chromatography ( silica, 10 -~ 3 S%
ethylacetate in petroleum ether) to give 18 (SS.O mg, 30%) as
an amorphous solid.
Physical Data for Alcohol 18. R f = 0.26 ( silica, 3 590
ethylacetate in hexanes); IR (film) vr"a~; 3414, 2924, 1706,
1669, 1605, 15 30, 1094; 1 H NMR ( 500 MHz, CDCl3 ) 8 7.90 ( d, J =

CA 02180444 1996-07-03
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9.0 Hz, 2H, Ar), 6.64 ( d, J = 9.0 Hz, 2H, Ar), 5.60 ( br d, J = 7 .0 Hz,
1H, 2-H), 5.29 (d, J = 2.5 Hz, 1H, 10-H), 4.89 (br d, J = 9.5 Hz,
1H, 5-H), 4.37 (d, J = 8.5 Hz,. 1H, 20-H), 4.36 (dd, J= 10.5, 6.5
Hz, 1H, 7-H), 4.31 (d, J= 2.5 Hz, 1H, 10-OH), 4.13 (br d, J= 8.5
Hz, 1H, 20-H), 3.90 (d, J= 7.0 Hz, 1H, 3-H), 3.05 (s, 6H, NMe?),
2.93 (s, 1H, OH), 2.90 (d, J= 20.0 Hz, 1H, 14-H), 2.61 (br d, J=
20.0 Hz, 1H, 14-H), 2.49-2.40 (m, 1H, 6-H), 2.16 (s, 3H, Me),
2.08 (s, 3H, Me), 1.90-1.83 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.20
(s, 3H, Me), 1.13 (s, 3H, Me), 0.90 (t, J = 8.0 Hz, 9H,
OSi(CH?C~3)3), 0.56-0.49 (band, 6H, OSi(CH~CH3)3); FAB HRMS
(NBA / NaI) m / a 722.3354, M + Na+ calcd for C3~H~301pNSi
722.3336.
Preparation of 1-naphthalene-C-2 ester derivative
(Alcoho119)
HO O OTES HO O OTES
O " O
O H 'A O / HO H 'A O
O c ~ O O c
O ~ ~ ,O
7 19
Alcohol 19. A solution of carbonate 7 (47 mg, 0.0812
mmol) in tetrahydrofuran (2 mL) at -78 °C was treated with 1-
lithionaphthalene (6.3 mL of a 0.32 M solution in diethylether,
a

CA 02180444 1996-07-03
218~4~4
wo mssoa rcr~s9s~ood~s
- 41 -
2.03 mmol, prepared from 1-bromonaphthalene from Aldrich
Chemical Company Inc. and tButyllithium; methodology from
Gilman, H.; Moore, F.W. J. Am. Chem. Soc. 1940, 62, 1843) and
stirred for S minutesutes The reaction mixture was poured
S into a mixture of CH2C12 ( 1S mL) and aqueous NH~CI ( 20 mL),
the organic layer was separated, and the aqueous layer was
extracted with CH?C12 (2 x 10 mL). The combined organic layer
was washed with brine (10 mL), dried (MgSO~) and
concentrated to give alcohol 19 which was taken into the next
step without further purification.
Physical Data for Alcohol 19. R f = 0.27 ( 209
ethylacetate in petroleum ether); IR (film) vr"a,; 3442, 2954,
2882, 1724, 1671, 1461, 1362, 1279, 1228, 1195, 1092, 987,
826, 73fi cm-1; 1H NMR (500 MHz, CDC13) 8 8.66 (s, 1H,
naphthalene), 8.07 (dd, J= 9.0, 2.0 Hz, 1H, naphthalene), 7.97-
7.89 (m, 3H, naphthalene), 7.68-7.57 (m, 2H, naphthalene),
5.71 (br d, J = 6.5 Hz, 1H, 2-H), 5.35 (d, J = 2.5 Hz, 1H, 10-H),
4.94 (br d, J= 8.0 Hz, 1H, S-H), 4.41 (dd, J= 11.0, 7.0 Hz, 1H, 7-
H), 4.37 (d, J= 8.5 Hz, 1H, 20-H), 4.35 (d, J= 2.0 Hz, 1H, 10-OH),
4.18 (d, J = 8.5 Hz, 1H, 20-H), 4.00 (d, J = 6.5 Hz, 1H, 3-H), 3.02
(d, J= 19.5 Hz, 1H, 14-H), 2.69 (d, J= 19.5 Hz, 1H, 14-H), 2.54-
2.45 (m, 1H, 6-H), 2.27 (s, 3H, Me), 2.13 (s, 3H, Me), 1.94-1.87
(m, 1H, 6-H), 1.86 (s, 1H, OH), 1.75 (s, 3H, Me), 1.25 (s, 3H, Me),
1.20 (s, 3H, Me), 0.94 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.63-0.49
(band, 6H, OSi(Cj~~CH3)3); FAB HRMS (NBA) m /e 707.3270, M
+ H+ calcd for C39H~pOlpSi 707.3252.

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Preparation of phenylacetylide-C-2 ester derivative
(Alcohol 20)
HO O OTES HO O pTES
/ - Li
O ----.. O
H~O HO H ' '' O
\'p OAc p OAc
O/~
O
5
Alcohol 20. A solution of carbonate 7 (5.0 mg, 0.00864
mmol) in tetrahydrofuran (0.5 mL) at -78 °C was treated with
lithium phenylacetylide from Aldrich Chemical Company Inc.
(0.13 mL of a 1.0 M solution in tetrahydrofuran, 0.13 mmol)
10 and stirred for 0.5 hour The reaction mixture was treated with
aqueous NH~Cl (0.5 mL), allowed to warm to 25 °C, and diluted
with H20 ( 5 mL) and diethylether ( 5 mL) . The organic layer
was separated, dried, and concentrated to give a 9 : 1 mixture
of carbonate 7 and alcohol 20 (5.0 mg, 95%) as a film.
15 Physical Data for Alcohol 20. Rf = 0.59 (50%
ethylacetate in hexanes); 1H NMR (300 MHz, CDCl3) 8 7.63-7.57
(m, 2 H, Ar), 7.53-7.27 (m, 3 H, Ar), 5.43 (d, J= 6.5 Hz, 1H, 2-H),
5.28 (d, J = 2.5 Hz, 1H, 10-H), 4.90 (br d, J = 7.5 Hz, 1H, S-H),
4.67 (d, J = 8.5 Hz, 1H, 20-H), 4.44 (d, J = 8.5 Hz, 1H, 20-H),
20 4.37-4.30 (m, 1 H, 7-H), 4.28 (d, J= 2.5 Hz, 1H, 10-OH), 3.88 (d,
J= 6.5 Hz, 1H, 3-H), 2.85 (d, J= 20.2 Hz, 1H, 14-H), 2.63 (d, J=
20.2 Hz, 1H, 14-H), 2.55-2.47 (m, 1 H, 6-H), 2.11 (s, 3 H, OAc),

CA 02180444 1996-07-03
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2.08 (s, 3 H, 18-Me), 1.94-1.8~ (m, 1 H, 6-H), 1.67 (s, 3 H, Me),
1.41 (s,3H,Me), 1.21 (s,3 H,Me),0.91 (t,J=8.OHz,9H,
OSi(CH2C~3)3), 0.59-0.42 (band, 6H, OSi(C~2CH3)3).
S Preparation of Hydroxycarbamate-C-2 ester derivative
( Alcohol 21 )
HO O pTES HO O OTES
~ NH2
I I "'' O K I I
p H ~O HO H ~O
OAC ~~ O OAc
/j N
O
H O
7 21
Alcohol 21. A solution of carbonate 7 (S .0 mg, 0.00864
mmol) in MeOH (0.5 mL) at 25 °C was treated with n-Butyl-NHS
from Aldrich Chemical Company Inc. ( 0.05 mL, 0.5 06 mmol)
and stirred for 10 minutes. The reaction mixture was
concentrated and purified by flash chromatography (silica, 30
-~ 509o ethylacetate in petroleum ether) to give 21 ( 5.2 mg,
9290) as an~amorphous solid.
Physical Data for Alcohol 21. .Rf = 0.13 (silica, 30~Yo
ethylacetate in petroleum ether); IR (film) v~,a~ 3434, 2957,
2881, 1711, 1671, 1368, 1243, 1108, 987, 829 cm-1; 1H NMR
(500 MHz, CDC13) 8 x.27 (d, J= 2.0 Hz, 1H, 10-H), 5.23 (d, J= 6.5
Hz, 1H, 2-H), 4.91 (br d, J= 8.0 Hz, 1H, 5-H), 4.79 (t, J= 6.0 Hz,
1H, NH), 4.47 (d, J= 8.5 Hz, 1H, 20-H), 4,34 (dd, J= 11.0, 7.0 Hz,

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1H, 7-H), 4.30 (d, J= 2.S Hz, 1H, 10-OH), 4.28 (d, J= 8.5 Hz, 1H,
20-H), 3.78 (d, J = G.S Hz, 1H, 3-H), 3.29-3.12 (m, 2H, NHC~?),
2.70 (d, J= 20.0 Hz, 1H, 14-H), 2.60 (d, J= 20.0 Hz, 1H, 14-H),
2.51-2.42 (m, 1H, 6-H), 2.24 (s, 1H, OH), 2.OG (s, 3H, Me), 2.05
S (s, 3H, Me), 1.94-1.86 (m, 1H, G-H), 1.69 (s, 3H, Me), 1.55-1.46
(m, 2H, NHCH2C~i_?), 1.40-1.30 (m, 2H, NHCH?CH?C~?), 1.21 (s,
3H, Me), 1.09 (s, 3H, Me), 0.95-0.80 (m, 12H, CH3 of Bu,
OSi(CHzC~3)3), O.G1-0.47 (band, 6H, OSi(C,~-I~CH3)3); FAB HRMS
(NBA / NaI) m / a 674.3336, M + Na+ calcd for C33H~301pNSi
674.3336.
Preparation of NN methyl-phenyl-hydroxycarbamate-
C-2 ester derivative (Alcohol 22)
HO O OTES HO O OTES
NLiMe
-'~"' O ?C
O H ~.O HO H~O
OAc p OAc
O \ / M O
1S 7 a 22
Alcohol 22. A solution of carbonate 7 (5.0 mg, 0.00864
mmol) in tetrahydrofuran (0.5 mL) at -78 °C was treated with
LiNMePh ( 0.2 mL of a 0.47 M solution in diethylether, 0.094
mmol, prepared from N-methylaniline (Aldrich) and n-
Butyllithium) and stirred for 1.25 hour The reaction mixture
was poured into a mixture of diethylether ( S mL) and aqueous

CA 02180444 1996-07-03
~~.8n44~
wo 9mssoa pcrnrs~s
~ 45 ~
NH4C1 (5 mL), the organic layer was separated, and the aqueous
layer was extracted with diethylether (2 x S mL). The
combined organic layer was washed with brine ( S mL) , dried
(MgS04), concentrated, and purified by flash chromatography
S (silica, 15 -~ 3S9~o ethylacetate in hexanes) to give 7 (2.5 mg,
5096) and 22 (2.8 mg, 939'o based on 509'o conversion) as an
amorphous solid.
Physical Data for Alcohol 2 2. R f = 0.2 2 ( sili ca 3 S 9'0
ethylacetate in petroleum ether); 1H NMR (500 MHz, CDC13) 8
7.45-7.18 (band, SH), S.2S (br d, J= 6.S Hz, 1H, 2-H), 5.20 (d, J
= 2.5 Hz, 1H, 10-H), 4.70 (br d, J= 8.0 Hz, 1H, S-H), 4.26 (d, J=
2.5 Hz, 1H, 10-OH), 4.22 (dd, J= 10.5, G.S Hz, 1H, 7-H), 4.19 (d, J
= 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.S Hz, 1H, 20-H), 3.58 (d, J=
7.0 Hz, 1H, 3-H), 3.27 (s, 3H, MeN), 2.52 (d, J= 20.0 Hz, 1H, 14-
H), 2.35 (d, J= 20.0 Hz, 1H, 14-H), 2.40-2.31 (m, 1H, 6-H), 2.03
(s, 1H, OH), 1.97 (s, 3H, Me), 1.85-1.76 (m, 1H, 6-H), 1.GG (s, 3H,
Me), 1.57 (s, 3H, Me), 1.18 (s, 3H, Me), 1.08 (s, 3H, Me), 0.87 (t,
J - 8.0 Hz, 9H, OSi(CH?C~3)3), 0.55-0.43 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NBA) m/ a 686.3358, M + H+ calcd
for C3~H~1010IVSi G86.33G1.

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Preparation of Thioether-C-2 ester derivative (Alcohol
23)
HO O pTES HO O OTES
- ~ .~ ..
O~( X I I ~" O
HO H ' "' O ~H~O
p OAc HO p OAc
SPh-' "
O 9 O
23
Alcohol 2 3 . A solution of vinyl ester 9 ( S S . G mg,
0.0916 mmol) and 4-dimethylaminutesopyridine from Aldrich
Chemical Company Inc. (DMAP, 1.8 mg, 0.0147 rnmol) in CH?Cl?
(4.3 mL) at 25 °C was treated with PhSH from Aldrich Chemical
Company Inc. (0.030 mL, 0.292 mmol) and stirred for 1.5 hour
The reaction mixture was concentrated and purified by flash
chromatography (silica, 30% ethylacetate in petroleum ether)
to give 23 (58.1, 889%) as a white solid.
Physical Data for Alcohol 23. Rf = 0.37 (silica, 30%
ethylacetate in hexanes), 0.34 ( 10% ethylacetate in PhH, 2
elutions); IR (film) v,T,a,; 3441, 3057, 2956, 2883, 1732, 1672,
1~00, 1367, 1238, 1111, 988, 825, 739 cm -1; 1H NMR (500
MHz, CDCl3) 8 7.39-7.24 (band, SH), 5.44 (d, J= G.5 Hz, 1H, 2-H),
5.28 (d, J= 2.5 Hz, 1H, 10-H), 4.90 (dd, J= 9.5, 2.0 Hz, 1H, S-H),
4.38 (d, J= 8.0 Hz, 1H, 20-H), 4.33 (dd, J= 10.5, G.SHz, 1H, 7-H),
4.29 (d, J = 2.5 Hz, 1H, 10-OH), 4.18 (d, J = 8.0 Hz, 1H, 20-H),
3.83 (d, J= 6.5 Hz, 1H, 3-H), 3.24-3.13 (m, 2H, C~?SPh), 2.7G (d,

CA 02180444 1996-07-03
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WO 95/18$04 PCT/US951~478
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J = 19.5 Hz, 1H, 14-H), 2.72-2.58 (m, 3H, 14-H, CH?C~?SPh),
2.47 (ddd, J~ 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.39 (s, 1H, OH), 2.07
(s, 3H, Me), 2.05 (s, 3H, Me), 1.89 (ddd, J= 14.5, 10.5, 2.0 Hz,
1H, 6-H), 1.68 (s, 3H, Me), 1.23 (s, 3H, Me), 1.12~(s, 3H, Me),
S 0.92 (t, J = 8.0 Hz, 9H, OSi(CH?C~,3)3), 0.61-0.47 (band, 6H,
OSi(C~2CH3)3); FAB HRMS (NBA / CsI) m / a 849.2085, M + Cs+
calcd for C37H$?OIpSSi 849.2105.
Preparation of intermediates 2 5-2 7 and 2-furanyl-C-
2-taxoid ( 28 )
HO O pTES . Ac0 O OTES
a
O
H~H ;VO . HO H =1i0
O p OAc O p OAc
TESO Ph
~. ~ ~ / 25
O ~ 1 ~N, O
0 2e~ BZ ~ b
Ac0 O OTES
Ac0 O O R
BzNH
.", ~ HO~
Ph O~~w
OR H ; O HO - H : 0
OHO p OAc O 0 OAc
27 : R - TES ~ / 26
O d O
28: R=H
Acetate 25. A solution of alcohol 11 and 4-
dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CH?Cl?
( 3 mL) at 2 S °C was treated with acetic anhydride ( 0.5 0 mL,

CA 02180444 1996-07-03
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WO 95/18804 PCTIUS95/00478
_ 48 _
5.30 mmol) and stirred for 3 h. The reaction mixture was
diluted with CH?Cl~ ( 5 mL), treated .with aqueous NaHC03 ( 7
mL), and stirred vigorously for 25 min. The organic layer was
separated and the aqueous layer was--'-extracted with CH?Cl? (2
S x 10 mL). The combined organic layer was washed with brine
(S mL), dried (MgSO~), concentrated, and purified by
preparative TLC (silica, 109% ethylacetate in benzene, 3
elutions) to give 25 (36 mg, 66% from carbonate 7) as a white
foam.
Physical Data for Acetate 2 S . R f = 0.3 8 ( 20~Yo
ethylacetate in petroleum ether); IR (film) v,T,a,; 3509, 2956,
2881, 172 i, 1674, 1469, 1371, 1299, 1227, 1108, 746 cm-1; 1H
NMR (S00 MHz, CDC13) s 7.65 (br s, 1H, furan), 7.24 ( br d, J=
3.0 Hz, 1H, furan), 6.58-6.54 (m, 2H, 10-H, furan), S.59 (d, J=
6.5 Hz, 1H, 2-H), 4.92 (br d, J = 7.S Hz, 1H, S-H), 4.46 (dd, J =
10.5, 6.5 Hz, 1H, 7-H), 4.42 (d, J= 8.5 Hz, 1H, 20-H), 4.16 (d, J=
8.5 Hz, 1H, 20-H), 3.87 (d, J= 6.5 Hz, 1H, 3-H), 2.89 (d, J= 20.0
Hz, 1H, 14-H), 2.63 (d, J= 20.0 Hz, 1H, 14-H), 2.59-2.48 (m, 1H,
6-H), 2.22 (s, 3H, Me), 2.17 (s, 3H, Me), 2.14 (s, 3H, Me), 1.90-
1.83 (m, 1H, 6-H), 1.65 (s, 3H, Me), 1.25 (s, 3H, Me), 1.18 (s, 3H,
Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.64-0.52 (band, 6H,
OSi(C~-i2CH3)3); FAB HRMS (NBA / CsI) m / a 821.1966, M + Cs+
calcd for C35H4g01?Si 821.1969.
Alcohol 2 6. A solution of enone 2 S ( 3 6 mg, 0.0 ~ 2 3
mmol) in MeOH (3 mL) containing two drops of CH3COOH at 0 °C
was treated with NaBH~. (200 mg, 5.29 mmol, added by

CA 02180444 1996-07-03
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- 49 -
portions) and stirred for 6 h. The reaction mixture was diluted
with CH2C12 (10 mL), treated with aqueous NH4C1 (5 mL), and
stirred for 10 min. The organic layer was separated and the
aqueous layer was extracted with CHZC12 (2 x 10 mL). The
combined organic layer was washed with brine ( S mL), dried
( MgS04), concentrated, and purified by preparative TLC ( silica,
509'6 ethylacetate in petroleum ether) to give 26 (30 mg, 830 )
as an amorphous solid.
Physical Data for Alcohol 26. Rf = 0.42 (silica, 509'0
ethylacetate in petroleum ether); 1 H NMR ( 300 MHz, CDC13 ) b
7.62 (br s, 1H, furan), 7.25 (d, J= 3.5 Hz, 1H, furan), 6.58 (d, J=
3.5 Hz, 1H, furan), 6.43 (s, 1 H, 10-H), 5.51 (d, J= 7.0 Hz, 1H, 2-
H), 4.96 (d, J= 7.5 Hz, 1H, S-H), 4.85-4.79 (m, 1 H, 13-H), 4.48
(dd, J = 10.5, 7.5 Hz, 1H, 7-H), 4.38 (d, J ~ 8.0 Hz, 1H, 20-H),
4.15 (d, J= 8.0 Hz, 1H, 20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 2.61-
2.48 (m, 2 H, 6-H and 14-H), 2.28 (s, 3 H, OAc), 2.20-2.10 (m, 1
H, 14-H), 2.18 (s, 6 H, OAc and 18-Me), 1.98-1.80 (m, 1 H, 6-H),
1.18 ( s, 3 H, 16-Me), 1.04 ( s, 3 H, 17-Me), 0.90 ( t, J = 8.0 Hz, 9H,
OSi(CH2Cj~3)3), 0.65-0.50 (band, 6H, OSi(C~?CH3)3).
DiTES taxoid 27. To a solution of alcohol 26 (30.0 mg,
0.0434 mmol, previously azeotroped twice with benzene) and
~-lactam 24 (28.0 mg, 0.0734 mmol, previously azeotroped
twice with benzene) in THF ( 2 mL), prepared from the Ojima-
Holton protocol (Holton, R.A. Chem Abstr. 1990, X14, 1G45G8q;
Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe, L. R. J. Org.
Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L; Zhao, M.;

CA 02180444 1996-07-03
218044.x.
WO 95/18804 PCT/US95/00478
- 50 -
Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992,
48, 6985-7012), at 0 °C was added NaN(SiMe3)~ (0.130 mL of a
1.0 M solution in THF, 0.130mmo1) dropwise. The resulting
solution was stirred for 5 min and poured into a mixture of
S CH2C12 (10 mL) and aqueous NH4C1 (5 mL). The organic layer
was separated and the aqueous layer was extracted with
CHZC12 (2 x 5 mL). The combined organic layer was washed
with brine (5 mL), dried (MgSO~), concentrated, and purified
by preparative TLC ( silica, 60% ethylacetate in petroleum
ether) to give 27 ( 12 mg, 2690) as an amorphous solid which
was taken directly into the next step.
Taxoid 28. A solution of silyl ether 27 (6 mg, 0.00560
mmol) in THF ( 1 mL) at 25 °C was treated with HF~pyridine ( 1
mL) and stirred for 1 h. The reaction mixture was poured into
a mixture of ethylacetate ( 10 mL) and aqueous NaHC03 ( 10 mL)
and the resulting mixture was stirred for 10 min. The organic
layer was separated and the aqueous layer was extracted with
ethylacetate (2 x 10 mL). The combined organic layer was
washed with brine (5 mL), dried (MgSO~), concentrated, and
purified by preparative TLC (silica, 60~o ethylacetate in
petroleum ether) to give 28 (3 mg, 64%) as a colorless film.
Physical Data for Taxoid 2 8. R f = 0.1 ( S 0%
ethylacetate in petroleum ether); IR (film) vma~ 3383, 2933,
2898, 1729, 1649, 1519, 1242, 1110, 1071 cm-1; 1H NMR (500
MHz, CDCl3) 8 7.77-7.73 (m, 2H), 7.68-7.66 (m, 1H, fura.n), 7.55-
7.33 (band, 9H), 6.98 (d, J= 9.0 Hz; 1H, NH), 6.58 (dd, J= 3.5,

CA 02180444 1996-07-03
2i$04~4
WO 95118804 PCT/IJS95I00478
- 51 -
1.5 Hz, 1H, furan), 6.27-6.21 (m, 2H, 10-H, 13-H), 5.80 (dd, J=
9.0, 2.0 Hz, 1H, 3'-H), 5.57 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (dd, J=
10.0, 2.0 Hz, 1H, 5-H), 4.80 (d, J= 2.0 Hz, 1H, 2'-H), 4.43-4.37
(m, 2H, 7-H, 20-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 3.77 (d, J= 7.0
Hz,l H, 3-H), 2.60-2.52 (m, 1H, 6-H), 2.47 (d, J= 4.0 Hz, 1H, OH),
2.38 (s, 3H, Me), 2.35-2.21 (m, 2H, 14-CH2), 2.25 (s, 3H, Me),
1.94-1.86 (m, 1H, 6-H), 1.81 (br s, 3 H, Me), 1.76 (s, 1H, OH),
1.68 (s, 3H, Me), 1.25 (s, 3H, Me), 1.13 (s, 3H, Me).
Preparation of 2-thiophenyl-C-2 taxol ( 3 2 )
HO O pTES _ Ac0 Q OTES
O~ ~ ~ T ~ a-~- O
H~H~O HD H~O
OAC OAc
'S / TESO~, Ph \S ~ 29
O ~N O
12
24 Bz ~, b
Ac0 O OR Ac0 O pTES
BzNH O
c
P h p.,.. ..,~-- H 0~...
w
OR , HO H = O HO H
S O OAc S OAc
31 : R w TES _ d ~ ~ O 30
O 32: R=H
Acetate 29. A solution of alcohol 12 (36.0 mg, 0.0543
mmol) and 4-dimethylaminopyridine (DMAP, 33.0 mg, 0.270

CA 02180444 1996-07-03
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mmol) in CH2C12 (3.0 mL) at 25 °C was treated with acetic
anhydride (0.50 mL, 5.30 mmol) and stirred for 1 h. The
reaction mixture was diluted with CH?Cl2 ( 10 mL), treated with
aqueous NaHC03 (7 mL), aid stirred vigorously for 0.5 h. The
S organic layer was separated and the aqueous layer was
extracted with CH?Cl? (2 x 10 mL). The combined organic layer
was washed with brine (5 mL), dried (MgSO.~), concentrated,
and purified by flash chromatography ( silica, 10 --~ 3 S %
ethylacetate in hexanes) to give 29 (29.5 mg, 7790) as an
amorphous solid.
Physical Data for Acetate 29. Rf = 0.56 (silica, 509
ethylacetate in petroleum ether); IR (film) vn,a,; 3457, 2956,
1712, 1669, 1525, 1413, 1376, 1264, 1227, 1073; 1H NMR (500
MHz, CDCI 3 ) b 7 .84 ( dd, J = 4.0, 1.5 Hz, 1 H, thiophene) , 7 .6 3 ( dd,
J = 5.0, 1.5 Hz, 1H, thiophene), 7.13 (dd, J = 5.0, 4.0 Hz, 1H,
thiophene), 6.56 (s, IH, 10-H), 5.58 (br d, J= 6.5 Hz, IH, 2-H),
4.90 (br d, J= 8.0 Hz, 1H, S-H), 4.44 (dd, J= 10.5, 7.0 Hz, 1H, 7-
H), 4.42 (d, J= 8.5 Hz, 1H, 20-H), 4.18 (d, J= 8.5 Hz, 1H, 20-H),
3.85 (d, J= 6.5 Hz, 1H, 3-H), 2.91 (d, J= 19.5 Hz, 1H, 14-H), 2.64
(dd, J= I9.5, I.0 Hz, 1H, 14-H), 2.55-2.48 (m, 1H, 6-H), 2.20 (s,
3H, Me), 2.15 (s, 3H, Me), 2.14 (s, 3H, Me), 1.89-1.82 (m, 1H, 6
H), 1.65 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), o.ss (t, J
= 8.0 Hz, 9H, OSi(CH?C~-13)3), 0.59-0.53 (band, 6H, OSi(CF_i~CH3)3);
FAB HRMS (NBA / CsI) m / a 837.1736, M + Cs+ calcd for
C3$H.~gUIISSi 837.1741.
Alcohol 30. A solution of enone 29 (29.0 mg, 0.0411

CA 02180444 1996-07-03
218 fl .4~~-
WO 95/18804 PGT/US95100478
~ 53 ~
mmol) in MeOH ( S mL) at 0 °C was treated with NaBH.~ ( 30.2
mg, 0.80 mmol, added by portions) and stirred for 2.5 h. The
reaction mixture was diluted with CH2C12 ( 15 mL), treated with
aqueous NH4C1 ( 5 mL), and stirred for 10 min. The organic
S layer was separated and the aqueous layer was extracted with
CH2C12 (2 x 10 mL). The combined organic layer was washed
with brine (5 mL), dried (MgSO.~), concentrated, and purified
by flash chromatography (silica, 25 --a 5090 ethylacetate in
petroleum ether) to give 29 {4.0 mg, 149'0) and 30 ( 14.7 mg,
5996 based on 8696 conversion) as an amorphous solid.
Physical Data for Alcohol 3 0. R f = 0.3 4 ( silica, 5 0%
ethylacetate in petroleum ether); IR (film) vma~ 3478, 2946,
2892, 1717, 1520, 13GS, 1238, 1083; 1H NMR (500 MHz, CDC13)
8 7.85 (dd, J = 3.5, 1.5 Hz, 1H, thiophene), 7.G 1 (dd, J = 5.0, 1.5
Hz, 1H, thiophene), 7.12 {dd, J= S.O,, 3.5 Hz, 1H, thiophene), G.43
(s, 1H, 10-H), 5.51 (d, J= 7.0 Hz, 1H, 2-H), 4.94 (br d, J= 7.5 Hz,
1H, S-H), 4.83-4.77 (m, 1H, 13-H), 4.45 (dd, J= 10.5, 7.S Hz, 1H,
7-H), 4.41 (d, J= 8.0 Hz, 1H, 20-H), 4.19 (br d, J= 8.0 Hz, 1H,
20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 2.55-2.48 (m, 1H, G-H), 2.24
(s, 3H, Me), 2.2G-2.21.(m, 2H, J.4-CH?), 2.1G (d, J= 1.0 Hz, 3H,
18-Me), 2.15 (s, 3H, Me), 2.00 (d, J= 5.0 Hz, 1H, OH), 1.90-1.82
(m, 1H, G-H), 1.GG (s, 3H, Me), 1.58 (s, 1H, OH), 1.15 (s, 3H, Me),
1.02 (s, 3H, Me), 0.90 (t, J= 8.0 Hz, 9H, OSiCH?C,~3)3), 0.59-0.55
(band, GH, OSi(CH?C H 3 ) 3 ); FAB HRMS (NBA / CsI) m / a
839.1893, M + Cs+ calcd for C3~H~pOIISSi 839.1897.
DiTES taxoid 31. To a solution of alcohol 30 ( 14.5 mg,

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0.0205 mmol, previously azeotroped twice with benzene) and
~i-lactam 24 ( 16.0 mg, 0.0420 mmol, previously azeotroped
twice with benzene) in THF (.1:0 mL), prepared from the Ojima-
Holton protocol (Holton, R.A. Chem Abstr: 1990, 114, 164568q;
Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe, L. R. J. Org.
Chem. 1991, 56, 1681-1683; Ojima, L; Habus, I.; Zhao, M.;
Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992,
48, 6985-7012), at -78 °C was added NaN(SiMe3)? (0.051 mL of
a 1.0 M solution in THF, 0.051 mmol) dropwise. The resulting
solution was stirred for 0.5 h and poured into a mixture of
diethylether (10 mL) and aqueous NH~CI (5 mL). The organic
layer was separated and the aqueous layer was extracted with
diethylether (2 x 5 mL). The combined organic layer was
washed with brine (5 mL), dried (MgSO~), concentrated, and
purified by flash chromatography ( silica, 10 ~ 3 590
ethylacetate in hexanes) followed by preparative TLC ( silica,
159~o ethylacetate in benzene) to give 30 (3.0 mg, 21%) and 31
(7.6 mg, 439~o based on 799~o conversion) as a white solid.
Physical Data for DiTES taxoid 31. Rf = 0.48 ( silica,
509~o ethylacetate in hexanes); IR (film) v",a~ 3382, 2913, 2850,
1722, 1653,1461,1243,1083, 1014; 1H NMR (500 MHz, CDCl;) 8
7.90 ( br d, J = 4.0 Hz, 1 H, thiophene) , 7.74 ( d, J = 8.0 Hz, 2 H,
NBz), 7.62 (br d, J =5.0 Hz, 1H, thiophene), 7.48 ( t, J = 7.0 Hz,
1H, Ar), 7.42-7.28 (band, 7H, Ar), 7.14 (dd, J = 5.0, 4.0 Hz, 1H,
thiophene), 7.10 (d, J= 9.0 Hz, 1H, NH), 6.42 (s, 1H, 10-H), G.20
(br t, J= 9:0 Hz, 1H, 13-H), 5.65 (br d, J= 9.0 Hz, 1H, 3'-H), 5.57
(d, J=7.0 Hz, 1H, 2-H), 4.94 (br d, J = 8.5 Hz, 1H, 5-H), 4.67 (d, J

CA 02180444 1996-07-03
2i80~4~
wo 9snssoa rcr~s9sioo4~s
- ss -
= 1.5 Hz, 1H, 2'-H), 4.44 rdd, J= 11.0, 6.5 Hz, 1H, 7-H), 4.43 (d, J
= 8.5 Hz, 1H, 20-H), 4.26 (d, J= 8.5 Hz, 1H, 20-H), 3.77 (d, J=
7.O ~Hz, 1H, 3-H), 2.51 (s, 3H, Me), 2.54-2.47 (m, 1H, 6-H), 2.34
(dd, J= 15.0, 9.5 Hz, 1H, 14-H), 2.15 (s, 3H, Me), 2.10 (dd, J=
15.0, 9.0, 1H, 14-H), 1.99 (s, 3H, Me), 1.93-1.86 (m, 1H, 6-H),
1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.18 (s, 3H, Me), 1.16 (s, 3H,
Me), 0.90 (t, J= 8.0 Hz, 9H, Si(CH?C~3)3), 0.79 (t, J= 8.0 Hz, 9H,
Si(CHZC~,3)3), O.S7-0.55 (band, 6H, Si(C~?CH3)3), 0.45-0.40
( band, 6H, Si( C~2 C H 3 ) 3 ); FAB HRMS ( NBA / CsI) m / a
1220.3685 M + Cs+ calcd for C5~H7~O1.~NSSi? 1220.3658.
Taxoid 32. A solution of silyl.ether 31 (7.5 mg, 0.00689
mmol) in THF (0.8 mL) at 2S °C was treated with HF~pyridine
( 0.1 S 0 mL) and stirred for 1 h. The rear tion mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 ( 5 mL) and the resulting mixture was stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO.~),
concentrated, and purified by flash chromatography ( silica, SO
--~ 10096 ethylacetate in petroleum ether) to give 32 (4.2 mg,
7190) as a.colorless film.
Physical Data for Taxoid ~ 3 2 . R f = 0.44 ( silica, 7 S 9'0
ethylacetate in petroleum ether); IR (film) vma~ 3417, 2929,
1716, 1649, 1521, 1460, 1417, 1368, 1247, 1076; 1 H NMR ( 500
MHz, CDC13) S 7.90 (dd, J= 4.0, 1.0 Hz, 1H, thiophene), 7.73 (d, J
= 7.0 Hz, 2H, NBz), 7.63 (dd, J = 5.0, 1.0 Hz, 1H, thiophene),

CA 02180444 1996-07-03
WO 95/18804
PCT/US95100478
~ 56 ~
7.51-7.32 (band, 8H, Ar), 7.14 (dd, J = 5.0, 4.0 Hz, 1H,
thiophene), 6.96 (d, J= 9.0 Hz, 1H, NH), G.24 (s, 1H, 10-H), G.19
(br t, J= 9.0 Hz, 1H, 13-H), 5.7S (dd, J= 9.0, 2.5 Hz, 1H, 3'-H),
5.55 (d, J= 7.0 Hz, 1H, 2-H), 4.94 (br d, J= 8.0 Hz, 1H, 5-H), 4.7G
S ( dd, J = S .0, 2.5 Hz, 1 H, 2'-H ) , 4.41 ( d, J = 8 . 5 Hz, 1 H, 2 0-H ) ,
4.40-4.33 (m, 1H, 7-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 3.73 (d, J
= 7.0 Hz, 1H, 3-H), 3.52 (d, J= S.0 Hz, 1H, 2'-OH), 2.58-2.49 (m,
1H, G-H), 2.44 (d, J= 4.0 Hz, 1H, 7-OH), 2.35 (s, 3H, Me), 2.29 (d,
J= 9.0 Hz, 2H, 14-CH?), 2.22 (s, 3H, Me), 1.91-1.83 (m, 1H, G-H),
1.7G (s, 3H, Me), 1.GG (s, 3H, Me), 1.23 (s, 3H, Me), 1.10 (s, 3H,
Me); FAB HRMS (NBA / CsI) m / a 992.1252, M + Cs+ calcd for
C45H49N014S 992.1928.
Preparation of 3-thiophenyl-C-2 taxol (36)
HO O pTES , ACO O OTES
o~-k r ~ ~ o
HO H = VO 'O
HO H
O OAC TESO, Ph / O OAC
S~ 33
,
13 0 24 BZ ~ b
ACO O OR ACO O pTES
BzNH O
Ph~O~,.. ...,~ HO~...
W w
OR H ' O HO - H _ O
HO 0 OAC p OAC
i~
35 : R = TES S 34
S
O 36 : R = H ."~ d O

CA 02180444 1996-07-03
Wo9~~~~ 2180~~~
PCT/US95/004?$
- 57
Acetate 33. A solution of alcohol 13 (68.4 mg, 0.103
mmol) and 4-dimethylaminapyridine (DMAP, 37.8 mg, 0.309
mmol) in CH2C12 (4.4 mL) at 25 °C was treated with acetic
anhydride (0.370 mL, 3.92 mmol) and stirred for 2 h. The
reaction mixture was diluted with CH2Cl2 (5 mL), treated with
aqueous NaHC03 ( 7 mL), and stirred vigorausly for 25 min.
The organic layer was separated and the aqueous layer was
extracted with CH?C12 (2 x 10 mL). The combined organic layer
was washed with brine (5 mL), dried (MgSO.~), concentrated,
and purified by flash chromatography (silica, 309~o ethylacetate
in hexanes) to give 33 (66.0 mg, 9190) as an amorphous solid.
Physical Data for Acetate 33. Rf = 0.48 {silica, 109'0
ethylacetate in benzene, 3 elutions); IR (film) vn,a,; 3518, 2956,
2881, 1727, 1676, 152,0, 1460, 1371, 1236, 1098, 985, 824,
744 cm 1; 1H NMR (S00 MHz, CDCl3) 8 8.19 (dd, J= 3.0, 1.1 Hz,
1H, thiophene), 7.55 (dd, J = 5.0, 1.1 Hz, 1H, thiophene), 7.38
(dd, J= 5.0, 3.0 Hz, 1H, thiophene), 6.S8 (s, 1H, 10-H), 5.61 (dd,
J= 6.5, 0.7 Hz, 1H, 2-H), 4.92 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.47
(dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.38 (d, J = 8.5 Hz, 1H, 20-H),
4.14 (d, J= 8.5 Hz, 1H, 20-H), 3:88 (d, J= 6.5 Hz, 1H, 3-H), 2.89
(d, J= 20 Hz, 1H, 14-H); 2.64 (br d, J= 20 Hz, 1H, 14-H), 2.54
(ddd, J= 14.5, 9.5, 6.5 Hz; lH, 6-H), 2.23 (s, 3H, Me), 2.18 (s, 3H,
Me), 2.17 (s, 3H, Me), 1.87 (ddd; J = 14.5, 10.5, 2.0, 1H, 6-H),
1.85 (s, 1H, OH), 1.66 (s; 3H, Me)., 1.26 (s, 3H, Me), 1.19 (s, 3H,
Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.65-0.54 (band, 6H,
OSi(C~2CH3)3); FAB HRMS (NBA / CsI) m / a 837.1760, M+Cs+
calcd for C35H~g011SSi 837.1741.

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Alcohol 34. A solution of enone 3 3 ( S 7.3 mg, 0.0813
mmol) in MeOH-THF ( S : 1, 4.1 mL) at 0 °C was treated with
NaBH4 (69.1 mg, 1.83 mmol, added by portions) and stirred for
2.5 h. The reaction mixture was diluted with CH?Cl? ( 10 mL),
S treated with aqueous NH~CI (5 mL), and stirred for 10 min.
The organic layer was separated and the aqueous layer was
extracted with CH?Cl? (2 x 10 mL). The combined organic layer
was washed with brine (S mL), dried (MgSO:~), concentrated,
and purified by flash chromatography (silica, 3096 ethylacetate
in hexanes) to give 33 (G.8 mg, 129'0) and 34 (45.2 mg, 8996
based on 8896 conversion) as an amorphous solid.
Physical Data for Alcohol 34. Rf = 0.48 (silica, SO9'o
ethylacetate in hexanes); IR (film) vma,; 3520, 2953, 2881,
1719, 1520, 1370, 1238, 1100, 979, 823, 74G cm-1; 1H NMR
( 500 MHz, CDC13 ) 8 8.20 ( dd, J = 3 .0, 1.0 Hz, 1H, thiophene), 7 .5 7
( dd, J = 5 .0, 1.0 Hz, 1 H, thiophene) , 7 .3 5 ( dd, J = 5 .0, 3 .0 Hz, 1 H,
thiophene), G.45 (s, 1H, 10-H), 5.54 (d, J= 7.0 Hz, IH, 2-H), 4.9G
(br d, J= 8.5 Hz, IH, S-H), 4.82 (br dd, J= 12.0, $.0 Hz, 1H, 13-
H), 4.48 (dd, J= 10.5, G.5 Hz, 1H, 7-H), 4.3G (d, J= 8.5 Hz, 1H,
20-H), 4.15 (d, J = 8.5 Hz, 1H, 20-H), 3.85 (d, J= 7.0 Hz, 1H, 3-
H), 2.53 (ddd, J= 14.5, 9.5, G.S., 1H, G-H), 2.27 (s, 3H, Me), 2.28-
2.21 (m, 2H, 14-CH2), 2.18 (s, GH, Me, Me), 2.03 (s, IH, OH), 1.87
(ddd, J= 14.5, I0.5, 2.0 Hz, IH, G-H), I.G7 (s, 3H, Me), I.GS (s,
1H, OH), 1.18 (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H,
OSi(CH2C~3)3), 0.G4-0.50 (band, GH, OSi(C~?CH3)3); FAB HRMS
NBA / CsI) m / a 839.1908 M + Cs+ calcd for C3~H;pOIISSi
839.1897.

CA 02180444 1996-07-03
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DiTES taxoid 35. To a solution of alcohol 34 ( 19.5 mg,
0.0276 mrilol, previously azeotroped twice with benzene) and
~i-lactam 24 (27.5 mg, 0.0721 mmol, previously azeotroped
twice with benzene) in THF ( 1.4 mL), prepared from the Ojima-
Holton protocol (Holton, R.A. Chem Abstr. 1990, 114, 1G45G8q;
Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe, L. R. J. Org.
Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L; Zhao, M.;
Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T. Tetrahedron 1992,
48, 6985-7012), at 0 °C was added NaN(SiMe3)? (0.066 mL of a
1.0 M solution in THF, 0.066. mmol) dropwise. The resulting
solution was stirred for 0.5 h and poured into a mixture of
CHZC12 ( 10 mL) and aqueous NH.~CI ( 5 mL) . The organic layer
was separated and the aqueous layer was extracted with
CH2C12 (2 x 5 mL). The combined organic layer was washed
with brine (5 mL), dried (MgSO.~), concentrated, and purified
by flash chromatography (silica, 20 ~ 30 9~o ethylacetate in
hexanes) to give 34 ( 1.1 mg, 69'0) and 35 ( 17.3 mg, G 19'o based
on 949ro conversion) as a white solid.
Physical Data for DiTES taxoid 3 5 . Rf = 0.8 G ( silica,
509'o ethylacetate in hexanes); IR (film) vma~; 3519, 3437, 2953,
2879, 1726, 1GGG, 1515, 1483, 1369, 1240, 1100, 979, 825,
74G cm-1; 1H NMR (500 MHz, CDCl3) 8 8.32 (dd, J= 3.0, 1.2 Hz,
1H, thiophene), 7.7G-7.73 (m, 2H), 7.G0 (dd, J= 5.0, 1.2 Hz, 1H,
thiophene), 7.52-7.29 (band, 9H), 7.10 (d, J= 9.0 Hz, 1H, NH),
6.44 (s, 1H, 10-H), G.2G (br t, J= 9.0 Hz, 1H, 13-H), 5.72 (dd, J=
9.0, 2.0 Hz, 1H, 3'-H), 5.G1 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J=

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WO 95/18804 ~ ~ PCT/US95/00478
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9.5, 2.0 Hz, 1H, S-H), 4.70 (d, J= 2.0 Hz, 1H, 2'-H), 4.48 (dd, J=
10.5, 6.S Hz, 1H, 7-H), 4.37 (d, J= 8.S Hz; 1H, 20-H), 4.23 (d, J=
8.S Hz, 1H, 20-H), 3.81 (d, J= 7.0 Hz, 1H, 3-H), 2.56-2.49 (m, 1H,
6-H), 2.54 (s, 3H, Me), 2.35 (dd, J= 15.5, 9.0 Hz, 1H~, 14-H), 2.17
S (s, 3H, Me), 2.07 (dd, J= 1S.S, 9.0 Hz, 1H, 14-H), 2.03 (d, J= 1.0
Hz, 3H, 18-Me), 1.94-1.87 (m, 1H, 6-H), 1.69 (s, 3H, Me), 1.68 (s,
1H, OH), 1.20 (s, 3H, Me), 1.18 (s, 3H, Me), 0.93 (t, J= 8.0 Hz, 9H,
OSi(CH2C,~3)3), 0.81 (t, J= 8.0 Hz, 9H, OSi(CH~C~3)3), 0.63-0.53
(band, 6H, OSi(C,~?CH3)3), 0.S2-0.36 (band, 6H, OSi(C~-I?CH3)3);
FAB HRMS (NBA / CsI) m / a 1220.3675, M + Cs+ calcd for
C57H~~014SSi2N 1220.3658.
Taxoid 3 6. A solution of silyl ether 3 5 ( 17.3 mg, 0.015 9
mmol) in THF (0.6 mL) at 25 °C was treated with HF~pyridine
1S (0.150 mL) and stirred for 2 h. The reaction mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 (S mL) and the resulting mixture was stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate ( 2 x 10 mL). The combined
organic layer was washed with brine ( S mL), dried ( MgSO.~),
concentrated, and purified by preparative TLC ( silica, 2 S%
ethylacetate in petroleum ether) to give 36 (7.7 mg, 56%) as a
colorless film.
Preparation of Taxoid 36. Rf = 0.11 (silica, 50%
ethylacetate in hexanes); IR (film) vma~ 3496, 3434, 2940,
1723, 1648, 1519, 1370, 1243, 1071, 975 cm-1; 1H NMR (500
MHz, CDC13) b 8.32 (dd, J= 3.0, 1.0 Hz, 1H, thiophene), 7.75-7.72

CA 02180444 1996-07-03
~ ~ s ~~~~
WO 95/18804 PCT/US95/00478
- 61 -
(m, 2H), 7.60 (dd, J = 5.0, 1.0 Hz, 1H, thiophene), 7.53-7.33
(band, 9H), 6.95 (d, J= 9.0 Hz, 1H, NH), 6.28-G.23 (m, 2H, 10-H,
13-H), 5.81 (dd, J= 9.0, 2.O Hz, 1H, 3'-H), 5.58 (d, J= 7.0 Hz, 1H,
2-H), 4.95 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.80 (dd, J= 4.5, 2.0 Hz,
1H, 2'-H), 4.41 (br t, J= 7.5 Hz, 1H, 7-H), 4.3G (d, J= 8.5 Hz, 1H,
20-H), 4.22 (d, J= 8.5 Hz, 1H, 20-H), 3.78 (d, J= 7.0 Hz, 1H, 3-
H), 3.49 (d, J= 4.S Hz, 1H, 2'-0H), 2.55 (ddd, J= 14.5, 9.5, G.5 Hz,
1H, 6-H), 2.45 (br s, 1H, OH), 2.40 (s, 3H, Me), 2.34 (dd, J= 15.5,
9.0 Hz, 1H, 14-H), 2.25 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.24 (s,
3H, Me), 1.89 (ddd, J= 14.5, 11.0, 2.0 Hz, 1H, G-H), 1.81 (d, J=
2.0 Hz, 3H, 18-Me), 1.74 (br s, 1H, OH), 1.G7 (s, 3H, Me), 1.24 (s,
3H, Me), 1.13 (s, 3H, Me); FAB HRMS (NBA / CsI) m / a
992.1940, M + Cs+ calcd for C~~H.~901,~NS 992.1928.

CA 02180444 1996-07-03
Wo 9~~8~4
PCT/US95I00478
~ 62 ~
Preparation of 2-pyridinyl-C-2 taxol (40)
HO O pTES ~ Ac0 O OTES
-'~' O
HO H ~''O HO H 'O
N' p OAc TESO, , Ph N' O OAc
O ~ ~ ~N, ~ / O 37
o 24 Bz
b
Ac0 O OR Ac0 O OTES
B=N H O
Ph~O~... ~ HO....
-
OR HO H ; O HO H ' 0
N p OAc N p OAc
w
O 40: R=HE~ d ~ / ,O 38
S
Acetate 3 7. A solution of alcohol 15 ( 2 3 .2 mg, 0.03 5 3
mmol) and 4-dimethylaminop~yridine (DMAP, 12.9 mg, 0.106
mmol) in CH2C1? ( 1.5 mL) at 25 °C was treated with acetic
anhydride (0.126 mL, 1.34 mmol) and stirred for 2 h. The
reaction mixture was diluted with ethylacetate (S mL), treated
with aqueous NaHC03 (7 mL), and stirred vigorously for 2S
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO:~),
concentrated, and purified by flash chromatography ( silica, 70
--~ 1009~o ethylacetate in petroleum ether) to give 3 7 ( 19.0 mg,

CA 02180444 1996-07-03
WO 95/18804 ~ PCTIUS95100478
- 63 -
7790) as an amorphous solid.
Preparation of Acetate 37. Rf = 0.58 (silica,
ethylacetate); IR (film) vI"a,; 3482, 2954, 2881, 1730, 1675,
1370, 1304, 1231, 1118, 987, 823, 739 cm-l; 1H NMR (S00
S MHz, CDC13) b 8.77 (ddd, J= 4.5, 1.7, 1.0 Hz, 1H, pyridine), B.OS
(ddd, J = 8.0, 1.0, 1.0 Hz, 1H, pyridine), 7.89 (ddd, J = 8.0, 8.0,
1.7 Hz, 1 H, pyridine) , 7 .5 3 ( ddd, J = 8.0, 4.5 , 1.0 Hz, 1 H,
pyridine), 6.59 (s, 1H, 10-H), 5.65 (dd, J= 6.6, 1.0 Hz, 1H, 2-H),
4.92 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.48 (dd, J= 10.5, 7.0 Hz, 1H,
7-H), 4.35 (d, J= 8.5 Hz, 1H, 20-H), 4.26 (dd, J= 8.5, 1.0 Hz, 1H,
20-H), 3.91 (d, J= 6.5 Hz, 1H, 3-H), 3.00 (d, J= 20.0 Hz, 1H, 14-
H), 2.71 (dd, J= 20.0, 1.0 Hz, 1H, 14-H), 2.54 (ddd, J= 14.5, 9.5,
?.0 Hz, 1H, 6-H), 2.53 (s, 1H, OH), 2.23 (s, 3H, Me), 2.18 (s, 3H,
Me), 2.14 (s, 3H, Me), 1.88 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H),
1S 1.70 (s, 3H, Me), 1.27 (s, 3H, Me),.1.20 (s, 3H, Me), 0.92 (t, 9H, J
= 8.0 Hz, OSi(CH2C~3)3), 0.64-0.52 (band, 6H, OSi{C~?CH3)3); FAB
HRMS (NBA / CsI) m / a 832.2139, M + Cs+ calcd for
C36H49011NSi 832.2129.
Alcohol 38. A solution of enone 37 {47.6 mg, 0.0680
mmol) in MeOH-THF ( 5 : 1, 3.8 mL) at 0 °C was treated with
NaBH4 (46.0 mg, 1.22 mmol, added by portions) and stirred for
1.5 h. The reaction mixture was diluted with ethylacetate { 10
mL), treated with aqueous NH.~CI (5 mL), and stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO~),

CA 02180444 1996-07-03
2~.$~44~
WO 95/18804 PCT/US95/00478
- s~ -
concentrated, and purified by flash chromatography ( basic
alumina, ethylacetate --~ 109% MeOH in ethylacetate) to give 27f
(28.0 mg, 59~Yo) as an amorphous solid.
Physical Data for Alcohol 3 8. R f = 0.3 G ( silica,
S ethylacetate); IR (film) vma,; 3487, 2951, 2880, 1736, 1583,
1369, 1307, 1236, 1132, 983, 824, 739 cm-1; 1H NMR (500
MHz, CDC13 ) 8 8 . 7 9 ( dm, J = 4 .5 Hz, 1 H, pyridine ) , 8 .13 ( br d, J =
7.5 Hz, 1H, pyridine), 7.88 (ddd, J = 7.5, 7.5, 1.7 Hz, 1H,
pyridine), 7.51 (ddd, J = 7.5, 4.5, 1.0 Hz, 1H, pyridine), 6.46 (s,
1H, 10-H), 5.64 (d, J= 7.0 Hz, 1H, 2-H), 4.96 (dd, J= 9.5, 2.0 Hz,
1H, S-H), 4.85 (br t, J= 8.0 Hz, 1H, 13-H), 4.49 (dd, J= 10.5, 6.5
Hz, 1H, 7-H), 4.31 (d, J= 8.0 Hz, 1H, 20-H), 4.25 (d, J= 8.0 Hz,
1H, 20-H), 3.89 (d, J= 7.0 Hz, 1H, 3-H), 2.53 (ddd, J= 14.5, 9.5,
6.5 Hz, 1H, 6-H), 2.36-2.11 (m, 2H, 14-CH?), 2.25 (s, 3H, Me),
2.19 (d, J= 1.0 Hz, 3H, 18-Me), 2.18 (s, 3H, Me), 1.88 (ddd, J=
14.0, 10.5, 2.5 Hz, 1H, 6-H), 1.70 (s, 3H, Me), 1.20 (s, 3H, Me),
1.05 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.65-0.51
(band, 6H, OSi(C~~ C H 3 ) 3 ) ; FAB HRMS ( NBA / CsI) m / a
834.2311, M + Cs+ calcd for C3~H~ 1 O11 NSi 834.2286.
DiTES taxoid 3 9. To a solution of alcohol 3 8 ( 10.3 mg,
0.0147 mmol, previously azeotroped twice with benzene) and
~i-lactam 24 ( 17.0 mg, 0.0446 mmol, previously azeotroped
twice with benzene) in THF (0.75 mL) at 0 °C, prepared from
the Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114,
164568q; Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe,
L. R. J. Org. Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L;

CA 02180444 1996-07-03
wo 9snsso4 218 ~ 4 4 4
PCT/US95/00478
- 65 -
Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.; Brigand, T.
Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)2
(0.038 mL of a 1.0 M solution in THF, 0.038 mmol) dropwise.
The resulting solution was stirred for 20 min and poured into a
mixture of ethylacetate ( 10 mL) and aqueous NH4C1 ( S mL) .
The organic layer was separated and the aqueous layer was
extracted with ethylacetate (2 x 5 mL). The combined organic
layer was washed with brine ( 5 mL), dried ( MgSO.~),
concentrated, and purified by preparative TLC ( silica, 6090
ethylacetate in petroleum ether) to give 39 (2.7 mg, l7~Yo) as a
film.
Physical Data for DiTES taxoid 3 9. Rf = 0.2 8 ( silica,
5096 ethylacetate in hexane); IR (film) vr"~,; 3429, 2952, 2927,
2878, 1728, 1662, 1585, 1369, .1236, 1124, 1016, 984, 742 cm-
1; 1H NMR (500 MHz, CDC13) 8 8.78 (br d, J = 4.S Hz, 1H,
pyridine), 8.21 (d, J = 8.0 Hz, 1H, pyridine), 7.95 (ddd, J = 8.0,
8.0, 1.7 Hz, 1H, pyridine), 7.75-7.70 (m, 2H), 7.54-7.22 (band,
9H), 7.12 (d, J= 9.0 Hz, 1H, NH), 6.45 (s, 1H, 10-H), 6.27 (br t, J
= 9.0 Hz, 1H, 13-H), 5.73-5.67 (m, 2H, 2-H, 3'-H), 4.95 (dd, J=
9.5, 2.0 Hz, 1H, 5-H); 4.70 (d, J= 2.0 Hz, 1H, 2'-H), 4.48 (dd, J=
10.5, 6.5; 1H, 7-H), 4:32 (br s, 2H, 20-CH?), 3.85 (d, J= 7.0 Hz,
1H, 3-H), 2.56-2.48 (m, 1H, 6-H), 2.52 (s, 3H, Me), 2.40 (dd, J=
15.0 Hz, 9.~ Hz, 1H,'14-H), 2.20-2.12 (m, 2H, 14-H, OH), 2.18 (s,
3H, Me), 2.04 (s, 3H, Me); 1.92 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H,
G-H), 1.72 (s, 3H, Me), 1.22 (s, 3H, Me), 1.19 (s, 3H, Me), 0.93 (t,
J = 8.0 Hz, 9H, OSi(CH? C~,3 ) 3 ), 0.81 ( t, J = 8.0 Hz, 9H,
OSi(CH2C~3)3), 0.64-0.34 (band, 12H, OSi(C~?CH3)3); FAB HRMS

CA 02180444 1996-07-03
WO 95/18804
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~ ~8a44~
- 66 -
(NBA / CsI) m / a I2I5.40G5, M + Cs-~ calcd for CSgH~gOI.:~N?Si?
1215 .4046.
Taxoid 40. A solution of silyl ether 3 9 ( 2.7 mg, 0.00249
S mmol) in THF (O.4 mL) at 25 °C was treated with HF~pyridine
(0.170 mL) and stirred for 3 h. The reaction mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 (5 mL) and the resulting mixture was stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine ( 5 mL), dried ( MgSO.~),
concentrated, and purified by preparative TLC ( silica,
ethylacetate) to give 40 (0.8 mg, 3890) as a colorless film.
Physical Data for Taxoid 40. R f = 0.5 4 ( silica,
ethylacetate); 1H NMR (500 MHz, CDC13) 8 8.80 (br d, J = 4.5 Hz,
1H, pyridine), 8.22 (d, J= 7.5 Hz, 1H, pyridine), 7.93 (ddd, J=
7.5, 7.5, 1.5 Hz, 1H, pyridine), 7.75-7.71 (m, 2H), 7.54-7.30
(band, 9H), G.98 (d, J= 9.0 Hz, 1H, NH), 6.30-G.24 (m, 2H, 10-H,
13-H), 5.82 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), 5.G7 (d, J= 7.0 Hz, 1H,
2-H), 4.95 (dd, J = 10.0, 2.0 Hz, 1H, S-H), 4.81 (dd, J = 4.5, 2.5
Hz, 1H, 2'-H), 4.41 (ddd, J= 11.0, 7.0, 4.5 Hz, 1H, 7-H), 4.31 (s,
2H, 20-CH2), 3.81 (d, J = 7.0 Hz, 1H, 3-H), 3.52 (br s, 1H, OH),
3.50 (d, J= 4.5 Hz, 1H, 2'-OH), 2.5G (ddd, J= 14.5, 9.5, 7.0 Hz,
1H, 6-H), 2.4G (d, J = 4.0 Hz, 1H, 7-OH), 2.43-2.30 (m, 2H, 14-
CHI), 2.38 (s, 3H, OAc), 2.25 (s, 3H, OAc), 1.90 (ddd, J= 14.5,
11.0, 2.0 Hz, 1H, G-H), 1.81 (s, 3H, Me), 1.71 (s, 3H, Me), 1.2G (s,
3H, Me), 1.15 (s, 3H, Me).

CA 02180444 1996-07-03
WO 95/18804
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~ 67
Prepartion of 3-pyridinyl-C-2-taxol (44)
.
HO O pTES AcO O pTES
a
O
p H 'O ~ H ~0
OAc TESO~. , Ph / ~ O OAC
N~ ~N N~ 41
17 0 24 ~Bz
~b
Ac0 O OR AcO O OTES
BzNH O
Ph~O~~.. .."~
H0~~
p H .
OR ' - O
/ ,O p OAc / O p OAc
N~~ 43 :.R = TES d. N~;~"''~ 42
O 44: R=H ~ O
Acetate 41. A solution of alcohol 17 (42.9 mg, O.OG52
mmol) and 4-dimethylaminopyridine (DMAP, 23.9 mg, 0.196
mmol) in CH2C12 (2.8 mL) at 25 °C was treated with acetic
anhydride (0.235 mL, 2.49. mmol) and stirred for 2 h. The
reaction mixture was diluted with ethylacetate (5 mL), treated
with aqueous NaHC03 (7. mL), and stirred vigorously for 25
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate ( 2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO.~),
concentrated, and purified by flash chromatography ( silica,

CA 02180444 1996-07-03
218p~-~4
WO 95/18804 PCT/US95/00478
- 68 -
ethylacetate) to give 41 (43.5 mg, 95°Yo) as a white solid.
Physical Data for Acetate 41. R f = O.G 1 ( silica,
ethylacetate); IR (film) vma,; 3470, 3327, 2955, 2881, 1731,
1675, 1592, 1370, 1279, 1229, 1108, 822, 738 cm-1; 1H NMR
S ( 500 MHz, CDC13 ) 8 9.2 3 ( br s, 1 H, pyridine ) , 8 .7 9 ( br s, 1 H,
pyridine), 8.30 (ddd, J= 8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.43 (dd,
J = 8.0, 5.0 Hz, 1H, pyridine), G.58 (s, 1H, 10-H), 5.70 (dd, J =
6.5, 1.0 Hz, 1H, 2-H), 4.91 (dd, J= 9.5, 2.0 Hz, 1H, 5-H), 4.47 (dd,
J= 10.5, 7.0 Hz, 1H, 7-H), 4.28 (d, J= 8.0 Hz, 1H, 20-H), 4.11 (d,
J = 8.0 Hz, 1H, 20-H), 3.91 (d, J = G.5 Hz, 1H, 3-H), 2.93 (d, J =
20.0 Hz, 1H, 14-H), 2.G8 (dd, J = 20.0, ,1.0 Hz, 1H, 14-H), 2.53
(ddd, J= 14.5, 9.5, 7.0 Hz, 1H, G-H), 2.24 (br s, 1H, OH), 2.22 (s,
3H, Me), 2.18 (s, 3H, Me), 2.17 (s, 3H, Me), 1.85 (ddd, J = 14.5,
10.5, 2.0 Hz, 1H, G-H), 1.G6 (s, 3H, Me), 1.26 (s, 3H, Me), 1.18 (s,
3H, Me), 0.90 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), O.G3-0.51 (band,
6H, OSi(C~2CH3)3); FAB HRMS (NBA / CsI) m / a 832.2145, M +
Cs+ calcd for C3~H.~gOI INSi 83.2.2129.
Alcohol 42. A solution of enone 41 (39.8 mg, 0.0569
mmol) in MeOH-THF ( S : 1, 3.1 mL) at 0 °C was treated with
NaBH4 (G5.0 mg, 1.72 mmol, added by portions) and stirred for
1.5 h. The reaction mixture was diluted with ethylacetate ( 10
mL), treated with aqueous NH~CI (5 mL), and stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO~),
concentrated, and purified by flash chromatography (silica,

CA 02180444 1996-07-03
~1~'a4.4~
WO 95/18804 PCT/US95100478
69
ethylacetate) to give 41 ( 3.7 mg, 996) and 42 ( 24.3 mg, G79~o
based on 9196 conversion) as an amorphous solid.
Physical Data for Alcohol 42. Rf = 0.42 ( silica,
ethylacetate); IR (film) vma,; 3490, 2953, 2881, 1727, 1592,
1369, 1235, 1110, 822, 740 cm-1; 1H NMR (500 MHz, CDCl3) 8
9.30 (d, J= 2.0 Hz, 1H, pyridine), 8.81 (dd, J= 5.0, 2.0 Hz, 1H,
pyridine), 8.35 (ddd, J= 8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.44 (dd,
J= 8.0, S.0 Hz, 1H, pyridine), G.4G (s, 1H, 10-H), 5.64 (d, J= 7.0
Hz, 1H, 2-H), 4.9~ (dd, J= 9.5, 1.5 Hz, 1H, S-H), 4.83 (br dd, J=
12.5, 7.5 Hz, 1H, 13-H), 4.49 .(dd, J=,10.5, G.5 Hz, 1H, 7-H), 4.28
(d, J= 8.0 Hz, 1H, 20-H), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 3.89 (d, J
= 7.0 Hz, 1H, 3-H), 2.53 (ddd, J = 14,5, 9.5, G.5 Hz, 1H, G-H),
2.30-2.20 (m, 2H, 14-CH2), 2.28 (s, 3H, Me), 2.19 (d, J= 1.0 Hz,
3H, 18-Me), 2.18 (s, 3H, Me), 1.87 (ddd, J= 14.5, 10.5, 2.0 Hz,
1H, 6-H), 1.68 (s, 3H, Me), 1.63 (br s, 2H, OH, OH), 1.19 (s, 3H,
Me), 1.04 (s, 3H, Me), 0.92 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3),
0.64-0.51 (band, GH, OSi(C~?CH3)3); FAB HRMS (NBA / CsI) m /
a 834.2270, M + Cs+ calcd for C3~H~lOI INSi 834.2286.
DiTES taxoid 43. To a solution of alcohol 42 ( 12.6 mg,
0.018 mmol, previously azeotroped twice with benzene) and ~3-
lactam 24 ( 17.0 mg, 0.044 mmol, previously azeotroped twice
with benzene) in THF (0.97 mL) at 0 °C, prepared from the
Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114,
1G45G8q; Ojima, L; Habus, L; Zhao, M.Georg, G. L; Jayasinghe,
L. R. J. Org. Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L;
Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.; Brigand, T.

CA 02180444 1996-07-03
21'8~~~~:
WO 95/18804 PCT/US95/00478
- 70 -
Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)?
(0.054 mL of a 1.0 M solution in THF, ,0,.054 mmol) dropwise.
The resulting solution was stirred for 0.5 h and poured into a
mixture of ethylacetate ( 10 mL) and aqueous NH~Cl ( 5 mL) .
The organic layer was separated and the aqueous layer was
extracted with ethylacetate (2 x S mL). The combined organic
layer was washed with brine ( S mL), dried ( MgSO.~),
concentrated, and purified by flash chromatography (silica, SO
-~ 95f6 ethylacetate in hexanes) to give 42 ( 1.0 mg, 890) and 4 3
(8.6 mg, 489'o based on 92% conversion) as a white solid.
Physical Data for DiTES taxoid 43. Rf = 0.40 (silica ,
5096 ethylacetate in hexanes); IR (film) vma,; 3433, 2955, 2880,
1730, 1662, 1370; 1238, 1112, 1018, 985, 824, 740 cm-1; 1H
NMR (500 MHz, CDC13) s 9.34 (d, J= 2.0 Hz, 1H, pyridine), 8.82
(dd, J= 5.0, 2.0 Hz, 1H, pyridine), 8.42 (ddd, J= 8.0, 2.0, 2.0 Hz,
1H, pyridine), 7.74-7.69 (m, 2H), 7.51-7.20 (band, 9H), 7.08 (d,
J= 9.0 Hz, 1H, NH), 6.46 (s, 1H, 10-H), G.22 (br t, J= 9.0 Hz, 1H,
13-H), 5.74-5.6G (m, 2H, 2-H, 3'-H), 4.95 (dd, J= 9.5, 2.0 Hz, 1H,
5-H), 4.70 (d, J= 2.0 Hz, 1H, 2'-H), 4.48 (dd, J= 10.5,6.5 Hz, 1H,
7-H), 4'.30 (d, J= 8.0 Hz, 1H, 20-H), 4.21 (d, J= 8.0 Hz, 1H, 20-
H), 3.86 (d, J= 7.0 Hz, IH, 3-H), 2.58-2.48 (m, 1H, 6-H), 2.54 (s,
3H, Me), 2.40 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.17 (s, 3H, Me),
2.14 (dd, J = 15.5, 9.0 Hz, 1H, 14-H), 2.03 (br s, 3H, Me), 1.95-
1.86 (m, 1H, 6-H), 1.73 (s, 4H, Me,,OH); I:22 (s, 3H, Me), 1.I8 (s,
2S 3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH?C~- 3)3), 0.82 (t, J= 8.0 Hz,
9H, OSi(CH2C~3)3), 0.65-0.37 (band, 12H, OSi(C,~IpCH3)3,
OSi(C~?CH3)3); FAB HRMS (NBA / CsI) m / a I21S.406G, M +

CA 02180444 1996-07-03
2i8~.4.44
WO 95118804 PCT/US95/00478
- 91 -
Cs+ calcd for CSgH7gO1:~N2Si? 1215.4046.
Taxoid 44. A solution of silyl ether 43 ( 6.4 mg, 0.005 9
mmol) in THF (0.4 mL) at 25 °C was treated with HF~pyridine
(0.160 mL) and stirred for 1.25 h. The reaction mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 (5 mL) and the resulting mixture was stirred for 10
min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate ( 2 x 10 mL). The combined
organic layer was washed with brine ( 5 mL), dried ( MgSO.~),
concentrated, and purified by preparative TLC ( silica,
ethylacetate) to give 44 (3.8 mg, 7590 as a colorless film.
Physical Data for Taxoid 44. R f = 0.5 9 ( silica,
ethylacetate); IR (film) vma,; 3396, 2928, 1728, 1644, 1371,
1273, 1241, 1111, 1071 cm-1; 1H NMR (500 MHz, CDC13) S 9.34
(br s, 1H, pyridine), 8.83 (br d, J = 3.5 Hz, 1H, pyridine), 8.41
(br d, J = 8.0 Hz, 1H, pyridine), 7.75-7.68 (m, 2H), 7.53-7.34
(band, 9H), 6.91 (d, J= 9.0 Hz, 1H, NH), 6.27 (s, 1H, 10-H), 6.23
(br t, J= 9.0 Hz, 1H, 13-H), 5.78 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H),
5.69 (d, J= 7.0 Hz, 1H, 2-H), 4.95 (dd, J= 9.5, 2.0 Hz, 1H, 5-H),
4.79 (dd, J = 5.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J = 11.0, 6.5, 4.0
Hz, 1H, 7-H), 4.29 (d, J= 8.5 Hz, 1H, 20-H), 4.20 (d, J= 8.5 Hz,
1H, 20-H), 3.82 (d, J= 7.0 Hz, 1H, 3-H), 3.54 (d, J= 5.5 Hz, 1H,
2'-OH), 2.56 (ddd, J= 14.5, 9.5, G.5 Hz, 1H, 6-H), 2.49 (d, J= 4.0
Hz, 1H, 7-OH), 2.43-2.26 (m, 2H, 14-CH?), 2.38 (s, 3H, Me), 2.24
(s, 3H, Me), 1.89 (ddd, J= 14.5, 11.0, 2.0 Hz, 1H, 6-H), 1.83 (s,
1H, OH), 1.82 (s, 3H, Me), 1.69 (s, 3H, Me), 1.25 (s, 3H, Me), 1.14

CA 02180444 1996-07-03
21804.4~-
WO 95/18804 PCT/US95/00478
- 72
(s; 3H, Me); FAB HRMS (NBA / Csl)~ m / a 987.2325, M + Cs+
calcd for C4~H~OD1.~N2 987.2316.
Preparation of 4-N,N dimethylaniline-C-2 taxol ( 48 )
HO O pTES Ac0 O OTES
I I a~ O
0 H ~ O '~'~!' H ~ O
p OAc O p OAc
Me2N ~ ~ Me2N
p 4~
TESO,. Ph
~b
Ac0 O pR o 24 Bz Ac0 O pTES
BzNH O
0".. ~ .~.~. ~ H 0.,..
_ w
OR = H : O
H O ' OAc
HO ~! OAc O O
Me N
Me2N ~ / O d7 : R = TES d2 ~ ~ O 46
'48: R=H
Acetate 45. A solution of alcohol 18 (50.0 mg, 0.0714
mmol) and 4-dimethylaminopyridine (DMAP, 26.0 mg, 0.213
mmol) in CH?C1~ (3.0 mL) at 25 °C was treated with acetic
anhydride (0.250 mL, 2.65 mmol) and stirred for 2.5 h. The
reaction mixture was diluted with CH?Cl? ( 10 mL), treated with
aqueous NaHC03 ( 7 mL) , and stirred vigorously for 2 ~ min.
The organic layer was separated and the aqueous layer was
extracted with CH?Cl? (2 x 10 mL). The combined organic layer

CA 02180444 1996-07-03
,..~ 218 0 4 ~.~.
WO 95/18804 PCT/US95100478
~ 73 -
was washed with brine (S mL), dried (MgSO:~), concentrated,
and purified by flash cf~romatography (silica, 1090 ethylacetate
in benzene) to give 45 (41.0 mg, 7790) as an amorphous solid.
Physical Data for Acetate 45. Rf = 0.27 (silica, 359%
S ethylacetate in hexanes); IR (film) vma,; 3425, 2945, 1722,
1674, 1605, 1365, 1275, 1232, 1179, 1094; 1H NMR (500 MHz,
CDC13) 8 7.$9 (d, J= 9.O Hz, 2H, Ar), G.64 (d, J= 9.O Hz, 2H, Ar),
6.56 (s, 1H, 10-H), S.G4 (d, J= G.5 Hz, 1H, 2-H), 4.90 (br d, J=
8.0 Hz, 1H, 5-H), 4.45 (dd, J= 10.5, 7.0 Hz, 1H, 7-H), 4.3G (d, J=
9.0 Hz, 1H, 20-H), 4.11 (d, J= 9.0 Hz, 1H, 20-H), 3.85 (d, J= 6.5
Hz, 1H, 3-H), 3.05 (s, 6H, NMe?), 2.90 (d, J= 20.0 Hz, 1H, 14-H),
2.62 (d, J= 20.0 Hz, l.I~, 14-H), 2.51 (ddd, J= 14.0, 8.0, 7.0, 1H,
6-H), 2.20 (s, 3H, Me), 2..1G (s, 3H, Me), 2.15 (s, 3H, Me), 2.04 (s,
1H, OH), 1.84 (ddd, J= 14.0, 10.5, 2.0 Hz, 1H, G-H), 1.G3 (s, 3H,
Me), 1.23 (s, 3H, Me), 1.1G (s, 3H, Me), 0.89 (t, J= 8.0 Hz, 9H,
OSi(CH2C~3)3), 0.58-0.53 (band, GH, OSi(C~?CH3)3); FAB HRMS
(NBA / CsI) m / a 874.8589, M + Cs+ calcd for C39H~>OlINSi
874.8594.
Alcohol 46. A solution of _enone 45 ( 40.0 mg, 0.05 3 9
mmol) in MeOH-THF (5.8 : 1, 4.1 mL) at 0 °C was treated with
NaBH.~ (30.2 mg, 0.80 mmol, added by portions), stirred for 1 h,
allowed to warm to 25 °C and stirred for 1.5 h. The reaction
mixture was diluted with CH?Cl~ ( 15 mL), treated with aqueous
NH4C1 (S mL), and stirred for 10 min. The organic layer was
separated and the aqueous layer was extracted with CHaCI? ( 2
x 10 mL). The combined organic layer was washed with brine

CA 02180444 1996-07-03
WO 95/18804 ~ ~ ~ PCT/US95/00478
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(5 mL), dried (MgSO.~), concentrated, and purified by flash
chromatography (silica, 25 -~ 509'0 'ethylacetate in petroleum
ether) to give 45 (6.0 ~rng, 159'0) and 46 (30.0 mg, 88~'o based on
859'o conversion) as an amorphous solid.
Physical Data for Alcohol 46. Rf = 0.30 (silica, 50%
ethylacetate in petroleum ether); 1H NMR (500 MHz, CDC13) s
7.93 (d, J= 9.0 Hz, 2H, Ar), 6.64 (d, J= 9.0 Hz, 2H, Ar), 6.42 (s,
1H, 10-H), 5.57 (d, J = 7.0 Hz, 1H, 2-H), 4.94 ( br d, J = 8.0 Hz,
1H, 5-H), 4.83-4.75 (m, 1H, 13-H), 4.46 (dd, J= 10.5, 6.5 Hz, 1H,
7-H), 4.34 (d, J= 8.5 Hz, 1H, 20-H), 4.13 (d, J= 8.5 Hz, 1H, 20-
H), 3.82 (d, J = 7.0 Hz, 1H, 3-H), 3.04 (s, 6H, Me?N), 2.54-2.44
(m, 1H, 6-H), 2.26 (s, 3H, Me), 2.23 (d, J= 7.S Hz, 2H, 14-CH?),
2.16 (s, 6H, Me, Me), 2.08 (d, J= 4.5 Hz, 1H, OH), 1.89-1.80 (m,
2H, 6-H, OH), 1.64 (s, 3H, Me), 1.16 (s, 3H, Me), 1.01 (s, 3H, Me),
0.89 (t, J = 8.5 Hz, 9H, OSi(CH?C -~i3)3), 0.62-0.48 (band, 6H,
OSi(C~?CH3) 3) .
DiTES taxoid 47. ~1 o a solution of alcohol 46 ( 14.0 mg,
0.0188 mmol, previously azeotroped twice with benzene) and
~-lactam 24 (25.0 mg, 0.0656 mmol, previously azeotroped
twice with benzene) in THF (0.75 mL) at 0 °C, prepared from
the Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114,
164568q; Ojima, I.; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe,
L. R. J. Org. Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L;
Zhao, M.; Zucco, M.; Parl:, Y.H.; Sun, C. M.; Brigaud, T.
Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe;)~
(0.056 mL of a 1.0 M solution in THF, 0.056 mmol) dropwise.

CA 02180444 1996-07-03
218 ~ 4-~.~
WO 95/18804 PCT/IJS95/00478
- ?5' -
The resulting solution was stirred for 20 min and poured into a
mixture of CH2C12 ( 10 mL) and aqueous NH:~CI (5 mL). The
organic layer was separated and the aqueous layer was
extracted with CH2Cl2 (2 x S mL). The combined organic layer
was washed with brine (S mL), dried (MgSO~), concentrated,
and purified by flash chromatography (silica, 10 -~ lS9~o
ethylacetate in benzene, then 5090 ethylacetate in petroleum
ether) to give 47 ( 12.0 mg, 5 79'0) as a white solid.
Physical Data for DiTES ~taxflid ~ 47. Rf = 0.2 G ( silica,
1596 ethylacetate in PhH); IR (film) vma~ 3425, 2946, 2882,
1722, 1669,1600, 1365, 1275, 1238, 1179, 1094 cm-1; 1H NMR
(500 MHz, CDC13) 8 7.9G (d, J = 9.0 Hz, 2H, Ar), 7.77-7.72 (m,
2H), 7.54-7.26 (band, 8H), 7.12 (d, J= 8.5 Hz, 1H, NH), G.G9 (d, J
= 9.0 Hz, 2H), 6.43 (s, 1H, 10-H), G.23 (br t, J= 9.0 Hz, 1H, 13-H),
5.68-5.63 (m, 2H, 2-H, 3'-H), 4.93 (br d, J = 8.0 Hz, 1H, 5-H),
4.67 (d, J= 2.0 Hz, 1H, 2'-H), 4.45 (dd, J= 10.5 Hz, 6.5 Hz, 1H, 7-
H), 4.36 (d, J= 8.5 Hz, 1H, 20-H), 4.20 (d, J= 8.5 Hz, 1H, 20-H),
3.78 (d, J= 7.0 Hz, 1H, 3-H); ,3.a4 (s,;. GHQ Me?N), 2.55-2.4G (m,
iH, G-H), 2.53 (s, 3H, OAc), 2.3G (dd, J = 15.5, 9.0 Hz, 1H, 14-H),
2.15 (s, 3H~,~Me), 2.09 (dd, J= 15.S, 9.0 Hz, iH, 14-H), 2.00 (d, J
= 1.0 Hz, 3H, Me), 1.92-1.84 (m, 2H, G-H, OH), 1.G7 (s, 3H, Me),
1.20 (s, 3H, Me), 1.1G (s, 3H, Me), 0.90 (t, J = 8.0 Hz, 9H,
OSi(CH~C~3)3), 0.79 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), O.G3-0.35
(band, 12 H, OSi(C~?CH3)3); FAB HRMS (NBA / CsI) m / a
1257.4503, M + Cs~ calcd for C~IHg~OI:~N?Si~ 1257.4515.
Taxoid 48. A solution of :silyl ether 47 ( 12.0 mg, 0.0107

CA 02180444 1996-07-03
wo 9siissoa 21 g p ~ ~
rcT~rs9srooa~s
- ~s -
mmol) in THF ( 1.0 mL) at 25 °C was treated with HF~pyridine
(0.05 mL) and stirred for 1.5 h. The reaction mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 (5 mL) and the resulting mixture was stirred for 10
S min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgSO:~),
concentrated, and purified by flash chromatography ( silica, 50
~ 759o ethylacetate in petroleum ether) to give 48 (8.0 mg,
8490) as a colorless film.
Physical Data for Taxoid 48. Rf = 0.44 (silica, 75%
ethylacetate in petroleum ether); IR (film) vma~ 3414, 2914,
2850, 1722, 1664, 1660, 1371, 1275, 1243, 1179 cm-1; 1H NMR
(500 MHz, CDC13) 8 7.95 (d, J= 9.0 Hz, 2H), 7.77-7.72 (m, 2H),
I5 7.55-7.30 (band, 8H), 7.03 (d, J= 9.0 Hz, IH, NH), 6.67 (d, J=
9.0 Hz, 2H), 6.24 (s, 1H, 10-H), 6.20 (br t, J= 9.0 Hz, 1H, 13-H),
5.76 (dd, J= 9.0, 2.5 Hz, 1H, 3'-H), S.G2 (d, J= 7.0 Hz, 1H, 2-H),
4.93 (br d, J= 7.5 Hz, 1H, 5-H), 4.76 (dd, J= 5.0, 2.5 Hz, 1H, 2'-
H), 4.37 (ddd, J= 11.5, G.S, 4.0 Hz, 1H, 7-H), 4.34 (d, J= 8.5 Hz,
1H, 20-H), 4.18 (d, J= $.5 Hz, 1H, 20-H), 3.73 (d, J= 7.0 Hz, 1H,
3-H), 3.57 (d, J= S.0 Hz, 1H, 2'-OH), 3.04 {s, GH, Me?N), 2.58-
2.48 {m, 1H, 6-H), 2.44 (d, J = 4.0 Hz, 1H, 7-OH), 2.37 (s, 3H,
Me), 2.30-2.25 (m, 2H, 14-CH?), 2.22 (s, 3H, Me), 1.95 (s, 1H,
OH), 1.88-1.81 (m, 1H, G-H), 1.74 (d, J = 1.0 Hz, 3H, Me), 1.65
(s, 3H, Me), 1.21 (s, 3H, Me), 1.11 (s, 3H, Me); FAB HRMS (NBA /
CsI) m / a 1029.2760, M + Cs+ calcd for C.~~H~~,N?Ol~
1029.2786.

CA 02180444 1996-07-03
WO 95/18804 PCT/US95/00478
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Preparation of 1-naphthalene-C-2-taxol ( 5 2 )
HO O pTES Ac0 O OTES
O a O
"~ _
O H : O
/ t HO 0 OAc / , HO p OAc
1
~,,. ' ,O 49
TESO~, , Ph
~b
Ac0 O
Ac0 O OR ~ 24 ~BZ OTES
BzNH
c H 0....
P h p.,..
/ 0 H
OR HO H' : / HO pAc 0
O OAc I O
O 51 : R = TE~ d ~ ~p 5 0
52 : R = .~JH
Acetate 49. A solution of previous alcohol 19 and 4-
dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CH?Cl?
(3 mL) at 25 °C was treated with acetic anhydride (0.50 mL,
5.30 mmol) and stirred for 3 h. The reaction mixture was
diluted with CH2C1? ( S mL), treated with aqueous NaHC03 ( 7
mL), and stirred vigorously for 25 min. The organic layer was
separated and the aqueous layer was extracted with CH?C1 ~ ( 2
x 10 mL). The combined organic layer was washed with brine
(S mL), dried (MgSO.~), concentrated, and purified by
preparative TLC (silica, 10% ethylacetate in benzene) to give

CA 02180444 1996-07-03
218044-
PCT/US95/00478
98 -
49 (54.1 mg, 899% from carbonate 7) as an amorphous solid.
Physical Data 'for Acetate 49. Rf = 0.27 (209'0
ethylacetate in petroleum ether); IR (film) vmaa 3416, 2953,
2879, 1726; 1676, 1370, 1224, 1089 cm-1; 1H NMR (500 MHz,
S CDC13) S 8.66 (s, 1H, naphthalene), 8.OG (dd, 1H, J= 9.0, 2.0 Hz,
naphthalene), 7.98-7.89 (m, 3H, naphthalene), 7.68-7.55 (m,
2H, naphthalene), G.G1 (s, 1H, 10-H), 5.75 (d, J= 7.0 Hz, 1H, 2-
H), 4.95 (br d, J= 8.0 Hz, 1H, S-H), 4.50 (dd, J= 10.5, 7.0 Hz, 1H,
7-H), 4.35 (d, J= 8.S Hz, 1H, 20-H), 4.16 (d, J= 8.5 Hz, 1H, 20-
H), 3.96 (d, J= 8.S Hz, 1H, 20-H), 3.96 (d, J= 7.0 Hz, 1H, 3-H),
3.03 (d, J= 20.0 Hz, 1H, 14-H), 2.70 (d, J= 20.0 Hz, 1H, 14-H),
2.61-2.50 (m, 2H, G-H, OH), 2.27 (s, 3H, Me), 2.24 (s, 3H, Me),
2.21 (s, 3H, Me), 1.91-1.83 (m, 1H, G-H), 1.70 (s, 3H, Me), 1.30
(s, 3H, Me), 1.20 ,(s, 3H, Me), 0.93 (t, J = 8.0 Hz, 9H,
OSi(CH2CI~3)3), O.GG-0.57 (band, 6H, OSi(C~?CH3)3); FAB HRMS
(NBA / CsI) m / a 881.2326, M + Cs+ calcd for C.~1H~~OlISi
881.2333.
Alcohol S 0. A solu tion of enone 49 ( S 4.1 mg, 0.07 2 2
mmol) in MeOH (10 mL) at 25 °C was treated with NaBH.~ (54.5
mg, 1.44 mmol, added by portions) and stirred for 2.0 h. The
reaction mixture was diluted with CH?C1? ( 10 mL), treated with
aqueous NH4C1 (S mL), and stirred for 10 min. The organic
layer was separated and the aqueous layer was extracted with
CH?Cl? (2 x 10 mL). The combined organic layer was washed
with brine (5 mL), dried (MgSO:~), concentrated, and purified
by preparative TLC ( silica, 209~o ethylacetate in petroleum

CA 02180444 1996-07-03
~1~04~~-
WO 95/18804 ~ PCT/US95100478
- ?9 -
ether) to give 50 (2G mg, 4890 ) as an amorphous solid.
Physical Data for Alcohol S 0. R f = 0.12 ( 209%
ethylacetate in petroleum ether); IR (film) vma,; 3 S 24, 295 3,
1719, 1369, 1231, 1093, 829 cm-1; iH NMR (500 MHz, CDC13) 8
S 8.70 (s, 1H, naphthalene), 8.11 (dd, J - 8.5, 1.5 Hz, 1H,
naphthalene), 7.9G-7.8G (m, 3H, naphthalene), 7.GS-7.54 (m,
2H, naphthalene), G.45 (s, 1H, 10-H), S.G8 (d, J= 7.0 Hz, 1H, 2-
H), 4.98 (br d, J = 8.0 Hz, 1H, S-H), 4.88-4.81 (m, 1H, 13-H),
4.51 (dd, J = 10.5, 7.0 Hz, 1H, 7-H), 4.34 (d, J = 8.5 Hz, 1H, 20-
H), 4.19 (d, J= 8.5 Hz, 1H, 20-H), 3.93 (d, J= 7.0 Hz, 1H, 3-H),
2.58-2.50 (m, 1H, G-H), 2.41-2.14 (r~w:,13H, 14-CH2, 13-OH), 2.37
,:;
(s, 3H, Me), 2.21 (br s, 3H, Me), 2.19 (s, 3H, Me), 1.92-1.84 (m,
1H, G-H), 1.72 (s, 1H, OH) .1.71 (s, 3H, Me), 1.22 (s, 3H, Me), 1.05
(s, 3H, Me), 0.93 (t, J= 8.0 Hz, 9H, OSi(CH?C~3)3), 0.65-0.51
(band, 6H, OSi(C~CH3)3); FAB HRMS (NBA / CsI) m / a
883.2484, M + Cs+ calcd for C.~IHs:~OI lSi 883.2490.
DiTES taxoid 51. To a solution of alcohol SO (20.0 mg,
0.0266 mmol, previously azeotroped twice with benzene) and
~i-lactam 24 (20.0 mg, Ot0525 mmol, previously azeotroped
twice with benzene) in THF ( 1.1 mL) at -78 °C, prepared from
the Ojima-Holton protocol (Holton, R.A. Chem Abstr. 1990, 114,
1645G8q; Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe,
L. R. J. Org. Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L;
Zhao, M.; Zucco, M.; Part, Y.H.; Sun, C. M.; Brigaud, T.
Tetrahedron 1992, 48, 6985-7012), was added NaN(SiMe3)?
(0.065 mL of a 1.0 M solution in THF, 0.065mmo1) dropwise.

CA 02180444 1996-07-03
~ 1.80444
WO 95/18804 PGT/US95/004?8
- 80 -
The resulting solution was stirred for 10 min and poured into a
mixture of CH2C12 ( 10 mL) and aqueous NH.~CI ( 5 mL) . The
organic layer was separated and the aqueous layer was
extracted with CH2C1? (2 x 5 mL). The combined organic Iayer
was washed with brine (5 mL), dried (MgSO~), concentrated,
and purified by preparative TLC (silica, 206 ethylacetate in
petroleum ether) to give S 1 ( 18.7 mg, 6290) as a white solid.
Taxoid 5 2. A solution of silyl ether 51 ( 18.7 mg, 0.0165
mmol) in THF (2 mL) at 25 °C was treated with HF~pyridine (1
mL) and stirred for 1 h. The reaction mixture was poured into
a mixture of ethylacetate ( 10 mL) and aqueous NaHC03 ( S mL)
and the resulting mixture was stirred for 10 min. The organic
layer was separated and the aqueous layer was extracted with
1 S ethylacetate ( 2 x 10 mL). The combined organic layer was
washed with brine (5 mL), dried (MgSO:~), concentrated, and
purified by preparative TLC (silica, 50% ethylacetate in
petroleum ether) to give 5 2 { 12.8 mg, 8690) as a colorless film.
Physical Data for Taxoid 5 2. R f = 0.16 ( silica, 5090
ethylacetate in petroleum ether); IR (film) vm~~ 3420, 2967,
2896, 1721, 1652, 1519, 1370, 1233, 1073, 776 cm-1; 1H NMR
{500 MHz, CDC13) 8 8.67 (s, 1H, naphthalene), 8.04 (dd, J= 8.5,
1.5 Hz, 1H, naphthalene), 7.95 (br d, J - 8.5 Hz, 1H,
naphthalene), 7.87 (br d, J= 9.0 Hz, 1H), 7.81 (br d, J= 8.5 Hz,
1H), 7.65-7.61 (m, 2H), 7.56-7.51 (m, 1H), 7.49-7.22 (band, 9H),
6.94 {d, J= 9.0 Hz, 1H, NH), 6.23-6.16 (m, 2H, 10-H, 13-H), 5.78
(dd, J = 9.0, 2.0 Hz, 1H, 3'-H), 5.64 (br d, J = 7.0 Hz, 1H, 2-H),

CA 02180444 1996-07-03
WO 95/18804 21 $ ~ ~ ~ ~ PCT/US95/00478
- 81 -
4.87 (br d, J = 8.0 Hz, 1H, 5-H), 4.78-4.72 (m, 1H, 2'-H), 4.38-
4.31 (m, 1H, 7-H), 4.24 (d, J= 8.5 Hz, 1H, 20-H), 4.16 (d, J= 8.5
Hz, 1H, 20-H), 3.76 (d, J= 7.0 Hz, 1H, 3-H), 3.53 (br s, 1H, OH),
2.52-2.43 (m, 1H, 6-H), 2.42 (d, J= 4.0 Hz, 1H, OH), 2.40 (s, 3H,
S Me), 2.36 (dd, J= 15.5, 9.0 Hz, 1H, 14-H), 2.25 (dd, J= 15.5, 9.0
Hz, 1H, 14-H), 2.17 (s, 3H, Me), 1.85-1.77 (m, 2H, 6-H, OH), 1.74
(br s, 3H, Me), 1.63 (s, 3H, Me), 1.17 (s, 3H, Me), 1.09 (s, 3H,
Me); FAB HRMS (NBA / CsI) m / a 1036.2505, M + Cs+ calcd for
C~1H~3N014 1036.2520
Preparation of thioether-C-2 taxol ( 5 f )
HO ,O OTES _ Ac0 O pTES
o~-k r ~ ~ o
O '~"~ H ~-O
HO pAc HO OAc
SPh O SPh O 53
23 TESO,, Ph
~b
/~N
Ac0 O OR ~ 24 BZ Ac0 O pTES
BzNH O
Ph p~,.. -~~. H0~~..
OR = H : O
H .. O
HO ~ ~Ac HO OAc
~54
SPh 0 55 : R ~ TE~ d SPh O
56 : R = .~..JH
1 S Acetate 5 3 . A solu lion of alcohol 2 3 ( 2 5 . 2 mg, 0.03 51
mmol) and 4-dimethylaminopyridine (DMAP, 12.2 mg, 0.0999

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- 62 -
mmol) in CH2C12 ( 1.5 mL) at 25 °C was treated with acetic
anhydride (0.120 mL, 1.27 mmol) and stirred for 1.5 h. The
reaction mixture was diluted with CH2C1~ (5 mL), treated with
aqueous NaHC03 (7 mL), and stirred vigorously for 25 min.
S The organic layer was separated and the aqueous layer was
extracted with CH2C1? (2 x 10 mL). The combined organic layer
was washed with brine (5 mL), dried (MgSO:~), concentrated,
and purified by flash chromatography (silica, 30% ethylacetate
in petroleum ether) to give 53 (25.3 mg, 95%) as a colorless oil.
Physical Data for Acetate 53. Rf = 0.41 (silica, 109'0
ethylacetate in benzene, 2 elutions); IR (film) vma,; 3471, 2954,
2881, 1729, 1675, 1370, 1226, 986, 824, 738 cmn; 1H NMR
(S00 MHz, CDCl3) 8 7.38-7.25 (band, 5H, SPh), 6.54 (s, 1H, 10-
H), 5.49 (br d, J= 6.5 Hz, 1H, 2-H), 4.90 (dd, J= 9.5, 2.0 Hz, 1H,
S-H), 4.42 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J= 8.0 Hz, 1H,
20-H), 4.17 (d, J= 8.0 Hz, 1H, 20-H), 3.78 (d, J= 6.5 Hz, 1H, 3-
H), 3.23-3.13 (m, 2H, C~i~SPh), 2.78 (d, J = 20.0 Hz, 1H, 14-H),
2.72-2.58 (m, 3H, C~-I?CH?SPh, 14-H), 2.52 (ddd, J= 14.5, 9.5,
6.5, 1H, 6-H), 2.45 (s, 1H, OH), 2.21 (s, 3H, Me), 2.15 (s, 3H, Me),
2.04 (s, 3H, Me), 1.86 (ddd, J= 14.5, 10.5, 2.0 Hz, 1H, 6-H), 1.62
(s, 3H, Me), I.23 (s, 3H, Me), 1.19 (s, 3H, Me), 0.91 (t, J= 8.0 Hz,
9H, OSi(CH?C~3)3), 0.64-0 52 (band, 6H, OSi(C -~i?CH3)3); FAB
HRMS (NBA / CsI) m / a 891.2225, M + Cs+ calcd for
C39H54011SSi 891.2210
Alcohol 5 4. A solution of enone 5 3 ( 24.4 mg, 0.03 2
mmol) in MeOH-THF (5 : 1, 1.9 mL) at 0 °C was treated with

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NaBH4 ( 18.1 mg, 0.48 mmol, added by portions) and stirred for
1.25 h. The reaction mixture was diluted with CH?Cl? (5 mL),
treated with aqueous NH~CI ( 5 mL), and stirred for 10 min.
The organic layer was separated and the aqueous layer was
extracted with CH?C12 ( 2 x 5 mL). The combined organic layer
was washed with brine ( 5 mL), dried ( MgSO:~), concentrated,
and purified by flash chromatography (silica, 309~o ethylacetate
in hexanes) to give 54 ( 14.6 mg, G09'o) as an amorphous solid.
Physical Data for Alcohol 54. R~ = 0.11 (silica, 300
ethylacetate in hexanes); IR (film) vma,; 3487, 2938, 2880,
1729, 1586, 1369, 1234, 977, 738 cm-1; 1H NMR (500 MHz,
CDC13) 8 7.40-7.23 (band, 5H, SPh), 6.42 (s, 1H, 10-H), 5.43 (d, J
= 7.0 Hz, 1H, 2-H), 4.94 (dd, J= 9.~, 2.0 Hz, 1H, 5-H), 4.85-4.78
(m, 1H, 13-H), 4.43 (dd, J= 10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J=
8.0 Hz, 1H,20-H),4.18(d,J=8.0 Hz, 1H,20-H),3.74(d,J=7.0
Hz, 1H, 3-H), 3.25-3.15 (m, 2H, C~2SPh), 2.71-2.57 (m, 2H,
C~?CH?SPh), 2.51 (ddd, J= 14.5, 9.5, G.5 Hz, 1H, G-H), 2.25 (dd, J
= 15.5, 9.5 Hz, 1H, 14-H), 2.16 (s, 3H, Me), 2.15 (d, J = 1.0 Hz,
3H, 18-Me), 2.15 (s, 3H; Me), 2.09 (dd, J= 15.5, 7.0 Hz, 1H, 14-
H), 2.05 (br s, 1H, OH), 1.99-1.96 (m, 1H, OH), 1.86 (ddd, J =
14.5, 10.5, 2.0 Hz, 1H, G-H), 1.63 (s, 3H, Me), 1.15 (s, 3H, Me),
1.04 (s, 3H, Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH~C~i_3)3), O.G4-0.50
(band, GH, Si(C~?CH3)3); FAB HRMS (NBA / CsI) m / a 893.2350,
M + Cs+ calcd for C39H>~Ol lSSi 893.2367.
DiTES taxoid 55. To a solution of alcohol 54 (21.8 mg,

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0.0286 mmol, previously azeotroped twice with benzene) and
~i-lactam 24 (33.0 mg, 0.0866 mmol, previously azeotroped
twice with benzene) in THF ( 1.1 mL) at 0 °C, prepared from the
Ojima-Holton protocol (Holton, R.A: Chem Absrr. '1990, 114,
164S68q; Ojima, L; Habus, L; Zhao, M.; Georg, G. L; Jayasinghe,
L. R. J. Org. Chem. 1991, SG, 1681-1683; Ojima, L; Habus, L;
Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.; Brigaud, T.
Tetrahedron 1992, 48, G98S-7012), was added NaN(SiMe3)?
(0.086 mL of a 1.0 M solution in THF, 0.086 mmol) dropwise.
The resulting solution was stirred for 20 min and poured ilzto a
mixture of CHZCI? ( 10 mL) and aqueous NH~CI (S mL). The
organic layer was separated and the aqueous layer was
extracted with CH?C1? (2 x S mL). The combined organic layer
was washed with brine (5 mL), dried .(MgSO~), concentrated,
1 S and purified by flash chromatography ( silica, 1 S -~ 3 0 -~ S 090
ethylacetate in petroleum ether) to give 55 (I3.8 mg, 420) as
an amorphous solid.
Physical Data for DiTES taxoid S S . Rf = 0.40 ( silica,
309'o ethylacetate in hexanes); IR (film) v",~,,; 3437, 2952, 2879,
1735, 1662, 1482, 1369; 1236, 1128; 981, 740 cm-1; 1H NMR
(500 MHz, CDCl3) 8 7.82-7.76 (m, 2H), 7.54-7.16 (band, 13H),
7.11 (d, J= 9.0 Hz, 1H, NH), 6.41 (s, 1H, 10-H), 6.18 ( br t, J= 9.0
Hz, 1H, 13-H), S.G2 (dd, J= 9.0, 2.0 Hz; 1H, 3'-H), 5.49 (d, J= 7.0
Hz, 1H, 2-H), 4.93 (dd, J = 9.5, 2.0 Hz, 1H, S-H), 4.64 (d, J = 2.0
2S Hz, IH, 2'-H), 4.42 (dd, J= 10.5, G.S Hz, 1H, 7-H), 4.40 (d, J= 8.0
Hz, IH, 20-H), 4.21 (d, J= 8.0 Hz, 1H, 20-H), 3.70 (d, J= 7.0 Hz,
1H, 3-H), 3.23-3.17 (m, 2H, C,~?SPh), 2.78-2.69 (m, 1H,

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HC$CH2SPh), 2.67-2.57 (m, 1H, ~CHCH2SPh), 2.SS-2.4G (m, 2H,
6-H, OH), 2.38 (s, 3H, Me), 2.27-2.10 (m, 2H, 14-CH?), 2.1G (s,
3H, Me), 1.98 ( d, J = 1.0 Hz, 3H, Me), 1.89 (ddd, J = 14.0, 11.0,
2.0 Hz, 1H, 6-H), 1.64 (s, 3H, Me), 1.18 (s, 3H, Me), 1.17 (s, 3H.,
S Me), 0.91 (t, J= 8.0 Hz, 9H, OSi(CH2C~3)3), 0.81 (t, J= 8.0 Hz, 9H,
OSi(CH2C,~3)3), 0.64-0.36 (band, 12 H, OSi(C~?CH3)3); FAB HRMS
(NBA / CsI) m / a M + Cs+ 1274.4125 calcd for Ct,IH~~O1.~SSi?
1274.4127.
Taxoid 5 6. A solution of ,silyl ether S S ( 8.1 mg, 0.0071
mmol) in THF (O.S mL) at 2S °C was treated with HF~pyridine
(0.150 mL) and stirred for 3.75 h. The reaction mixture was
poured into a mixture of ethylacetate ( 10 mL) and aqueous
NaHC03 (S mL) and the resulting mixture was stirred for 10
1S min. The organic layer was separated and the aqueous layer
was extracted with ethylacetate (2 x 10 mL). The combined
organic layer was washed with brine ( S mL), dried ( MgSO~),
concentrated, and purified by preparative TLC ( silica, 60°0
ethylacetate in petroleum ether) to give S 6 ( 3.2 mg, 4990) as a
colorless film.
Physical Data for Taxoid 5 6. R f = 0.3 9 ( silica, 6090
ethylacetate in petroleum ether); IR (film) vn,~~ 3426, 2928,
1731, 1642, 1371, 1238, 1070, 739, 709 cm-1; 1H NMR (500
MHz, CDC13) 8 7.80-7.7S (m,, 2H), 7.SS-7.18 (band, 13 H), 6.94
2S (d, J= 9.0 Hz, 1H, NH), 6.23 (s, 1H, 10-H), G.19 (br t, J= 9.0 Hz,
1H, 13-H), 5.74 (dd, J= 9.0, 2.S Hz, 1H, 3'-H), 5.47 (d, J= 7.0 Hz,
1H, 2-H), 4.93 (dd, J= 9.5, 2.0 Hz, 1H, S-H), 4.74 (dd, J= 5.0, Z.S

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Hz, 1H, 2'-H), 4.38 (d, J= 8.0 Hz, 1H, 20-H), 4.35 (ddd, J= 11.0,
6.5 Hz, 4.5 Hz, 1H, 7-H), 4.21 (d, J= 8..0 Hz, 1H, 20-H), 3.67 (d, J
= 7.0 Hz, 1H, 3-H), 3.51 (d, J= 5.0 Hz, 1H, 2'-OH), 3.28-3.14 (m,
2H, C~2SPh), 2.77-2.68 (m, 1H, HC~CH~SPh), 2.67-2.59 (m, 1H,
$CHCH2SPh), 2.54 (ddd, J= 14.5, 9.5, 6.S Hz, 1H, 6-H), 2.44 (d, J
= 4.5 Hz, 1H, 7-OH), 2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H), 2.26
(br s, 1H, OH), 2.23 (s, 3H, Me), 2.21 (s, 3H, Me), 2.18 (dd, J =
15.5, 9.0 Hz, 1H, 14-H), 1.88 (ddd; J= 14.5, 11.0, 2.0 Hz, 1H, 6-
H), 1.75 (d, J = 1.0 Hz, 3H, Me), 1.63 (s, 3H, Me), 1.24 (s, 3H,
Me), 1.10 (s, 3H, Me); FAB HRMS (NBA / CsI) m / a 1046.2410,
M + Cs+ calcd for C.~9H~~Oz4NS 1046.2398.
Preparation of MPA taxoid S 7
Ac0 O OH ACO O OH
BzNH 0 BzNH O
Ph.~II ~0.... ph~0 ~...
w
OH = . O \ O : H _ O
O p H OAc I ~ N\ ~ O ~p OAc
S ~ ~ CH3C00' S O
36 5~
MPA taxoid 57. A solution of taxoid 36 (4.3 mg, 0.005
mmol) and triethylamine (0.0033 mL, 0.0237 mmol) in CH?Ch
(0.2 mL) at 25 °C was treated with 2-fluoro-1-
methylpyridinium p-toluenesulfonate (2.1 mg, 0.0075 mmol)
and stirred for 35 min. The clear colorless solution rapidly
turned to a clear pale yellow. The course of the reaction was

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monitored through thin layer chromatography ( TLC ) ( E. Merck
RP- 18 silica, 65 tetrahydrofuran: 3 5 water, UV/phospho-
molybidic acid) and after thirty minutes of stirring at ambient
temperature, judged complete as no taxol remained and only
one compound was apparent by TLC. The reaction mixture was
directly purified by HPLC (Vydak RP-18, 22.5 x 3 mm, A -> B
0.5 h linear, A: 20~o MeOH in 20 mM NH~OAc, B: 100% MeOH, 9
mL / min, RT = 2 6.12 ) to give 3 6 ( 0.8 mg, 199'0) and 5 7 ( 4.1
mg, 1009'o based on 819% conversion) as a colorless film.
Physical Data for taxoid 5 7 1 H NMR ( 500 MHz, CDC13 )
b 10.5 (d, J = 7.5 Hz, 1H), 8.44 (ddd, J = 9.0, 7.5, 2.0 Hz, 1H),
8.33-8.29 (m, 2H), 8.15 (dd, J= 3.0, 1.0 Hz, 1H, thiophene), 8.12
(br d, J= 6.0 Hz, 1H), 7.84 (br d, J= 8.5 Hz, 1H), 7.74-7.69 (m,
2H), 7.53 (dd, J= 5.0, 1.0 Hz, 1H, thiophene), 7.48-7.34 (band,
7H), 7.16-7.12 (m, 1H), 6.53-6.43 (m, 1H, 2'-H), 6.21 (s, 1H, 10-
H), 6.03 (dd, J= 10.5, 8.0 Hz, 1H, 3'-H), 5.82 (br t, J= 9.0 Hz, 1H,
13-H), 5.44 (d, J= 7.0 Hz, 1H, 2-H), 4.90 (dd, J= 9.5, 2.0 Hz, 1H,
5-H), 4.33 (dd, J= 11.0, 6.5 Hz, 1H, 7-H), 4.30 (d, J= 8.0 Hz, 1H,
20-H), 4.15 (d, J= 8.0 Hz, 1H, 20-H), 4.08 (s, 3H, N+Me), 3.68 (d,
J= 7.0 Hz, 1H, 3-H), 2.58-2.49 (m, 1H, 6-H), 2.52 (s, 3H, OAc),
2.21 (s, 3H; OAc), 2.04 (s, 3H, OAc), 2.02 (br s, 2H, OH, OH), 1.88
(ddd, - 14.5, 11.5, 2.0 Hz, 1H, 6-H), 1.78 (br s, 3H, 18-Me),
1.64 ( 3H, Me), 1.61 (dd, J= 16.0, 7.0 Hz, 1H, 1.4-H), 1.18 (dd, J
= 16.G .0 Hz, 1H, 14-H), 1.13 (s, 3 H, Me), 1.08 (s, 3 H, Me).

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Preparation of MPA taxoid S 8
Ac0 O OH Ac0 O OH
BzNH -O BzNH
~r,~~o".. c~r, o"..
OH H ~ O ~ O HO = H = O
HO ~ OAc I ~ N+ S p OAc
S
O CH3C00' ~ ~ O
32 58
S
MPA taxoid S 8. A solution of taxoid 3 2 ( 1.0 equiv.)
and triethylamine (4.7 equiv.) in CH?Cl? (0.025 M) at 25 °C is
treated with 2-fluoro-1-methylpyridinium p-toluenesulfonate
from Aldrich Chemical company inc. ( 1.5 equiv.) and stirred for
35 minutes. The course of the reaction was monitored through
thin layer chromatography (TLC)(E. Merck RP- 18 silica, 65
tetrahydrofuran: 35 water, UV/phospho-molybidic acid) and
after thirty minutes of stirring at ambient temperature, judged
complete as no taxol remained and only one compound was
apparent by TLC. The reaction mixture is then directly purified
by HPLC (Vydak RP-18, 22.5 x 3 mm, A -~ B 0.5 h linear, A:
209'o MeOH in 20 mM NH.~OAc, B: 100% MeOH, 9 mL / min, RT =
26.12) to give 58 as a colorless film.

CA 02180444 1996-07-03
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~ 89
Preparation of MPA taxoid 5 9
~I~
Ago 0 o~N+ c H,c o a
BzNH O
----.. Ph' Y 'O~,..
OH
HO ~ H pAC
o
32 59
MPA taxoid 5 9. The synthesis of the taxoid-7-MPA 5 9
differs only slightly fromw the synthesis of taxoid-2'-MPA S 8.
The C-2 taxoid 3 2 is dissolved in methylene chloride ( .006 M)
and treated sequentially with triethylamine (40 equivalents)
and 2-fluoro-1-methyl-pyridinium tosylate ( 10 equivalents)
Aldrich Chemicals, and allowed to stir at ambient temperature
for 5 minutes. The reaction mixture is then directly purified
by HPLC ( Vydak RP-18, 2 2.5 x 3 mm, A -~ B 0.5 h linear, A:
209'o MeOH in 20 mM NH~OAc, B: 100~Yo MeOH, 9 mL / min, RT
26.12 ) to give 5 9 as a colorless film.
Preparation of MPA taxoid fi0
A O OH CH~COO'
BzNH O
r _
Ph y,.. -~. P~
OH HO ~ H pAc O
S ~
O
3s so

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MPA taxoid 60. The synthesis of the taxoid-7-MPA 60
differs only slightly from the synthesis of taxoid-2'-MPA 5 7.
The C-2 taxoid 36 is dissolved in methylene chloride (.006 M)
and treated sequentially with triethylamine (40 equivalents)
S and 2-fluoro-1-methyl-pyridinium tosylate ( ZO equivalents)
Aldrich Chemicals, and allowed to stir at ambient temperature
for S minutes. The reaction mixture is then directly purified
by HPLC (Vydak RP-18, 22.5 x 3 mm, A ~ B 0.5 h linear, A:
20'6 MeOH in 20 mM NH40Ac, B: 100~o MeOH, 9 mL / min, RT =
26.12) to give 60 as a colorless film.
Preparation of C-2-taxoid-2'-methyl-pyridinium salts
ACOO OH Ac00 OH
Bz[JH O BzNH O
Ph' v 'O".. Pn~a,..
off ~ , o ~' ~ a = H : o
HO ~ H pAc ~ N\ HO p OAc
R 1, X. R O
O
X a CH~COO; TtO; BFa; halides II.
% MaiN ~ ~ ~ I ~ ~ ~~gPh
62 58 57 62 63 64 65 66
C-2-taxoid-2'-opium salts 62-66. A solution of
taxoid (62-66) I. ( 1.0 equiv.) and triethylamine (4.7 equiv.) in
CHZC12 (0.025 M) at 25 °C is treated with 2-fluoro-1-
methylpyridinium p-toluenesulfonate from Aldrich Chemical

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WO 95/I8804 PCTlUS95/00478
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company inc. (1.5 equiv.) and stirred for 35 minutes. The
course of the reaction was monitored through thin layer
chromatography (TLC)(E. Merck RP- 18 silica, 65
tetrahydrofuran: 35 water, W/phospho-molybidic acid) and
S after thirty minutes of stirring at ambient temperature, judged
complete as no taxol remained and only one compound was
apparent by TLC. The reaction mixture is then directly purified
by HPLC (Vydak RP-18, 22.5 x 3 mm, A -> B 0.5 h linear, A:
209~b MeOH in 20 mM hTH40Ac, B: 1009~o MeOH, 9 mL / min, RT =
26.12) to give (62-66) II. as a colorless film.
Preparation of C-2-taxoid-7-methyl-pyridinium salts
O OH . Ac0 O ~ X'
BZNH O 8zNH .
a,... -----... Pn
OH HO : H f O OH H~ : H O
p OAc
R-l R1
O X = CH~C00': TaO: BF,,; halides O
I. 11.
Ro ~ ,O~ ~ 1S~ 1S~ ~ I ~ 1 I ~ MaZH ~ I ~ I / / /~SPh
67 59 80 68 69 70
C-2-taxoid-7-onium salts .67-72. The synthesis of
the taxoid-7-methyl-pyridinium salts ( 6 7-7 2 ) I I, differs only
slightly from the synthesis of taxoid-2'-methyl-pyridinium
salts ( 6 2 -6 6 ) II. The C-2 taxoid ( 6 7 - 7 2 ) I is dissolved in

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methylene chloride (.006 M) and treated sequentially with
triethylamine (40 equivalents) and 2-fluoro-1-methyl-
pyridinium tosylate ( 10 equivalents) Aldrich Chemicals, and
allowed to stir at ambient temperature for S minutes. The
S reaction mixture is then directly purified by HPLC (Vydak RP-
18, 22.5 x 3 mm, A --~ B O.S h linear, A: 209'o MeOH in 20 mM
NH~OAc, B: 1009'o MeOH, 9 mL / min, RT = 26.12) to give (67-
7 2 ) II as a colorless film.
Preparation of C-2-taxoid-bis-2',7-methyl-pyridinium
salts
ACO O OH ACO O ~ X'
BzNH BzNH O
Pr, o".. Pf,~~o....
OH = . O ~ O . H : O
HO ~~ H OAc I ~N~ HO p OAc
R~ X. R~l
O X = CH3C00'; TsO; BFI; halides O
I. II.
/ / SPh
\ / \ / \ / I I \ / ~ I ,
~a ~a ~ ~s ~s ~ ~~ ~e ~s eo
C-2-taxoid-bis-2',7-onium salts 73-80. The
1S synthesis of C-2-taxoid-bis-2',7-methyl-pyridinium salts I I
(73-80), differs from the synthesis of taxoid-7-methyl-
pyridinium salts (67-72) II only with respect to reaction time.
The C-2 taxoid (73-80) I is dissolved in methylene chloride
(.006 M) and treated sequentially with triethylamine (40
equivalents) and 2-fluoro-1-methyl-pyridinium tosylate ( 10

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equivalents) Aldrich Chemicals, and ~.llowed to stir at ambient
temperature for 18 hours. The reaction mixture is then
direc tly purified by HPLC ( Vydak RP-18, 2 2.5 x 3 mm, A -~ B
0.5 h linear, A: 2096 MeOH in 20 mM NH40Ac, B: 10096 MeOH, 9
mL / min, RT = 26.12) to give (73-80) II ~as a colorless film.
Preparation of C-2-taxoid-2'-benzothiazolium salts
Ac00 OH
8i H O BzNH O
OH \ I N~,.O HO o H O~ O
X. r Alkyl R 1~/
O
X a CH~COO'; TsO; BF4; hvlidw
'8/ ,S/ ~ I. ~~_SAvtN ~ ~ ~ I ~ / ,~~8Ph
Bt 82 83 84 85 86 g7 g8
IO
C-2-taxoid-2'-benzothiazolium salts 81-88. A
solution of taxoid ( 81-88 ) I ( 1.0 equiv.) and triethylamine (4.7
equiv.) in CH?Cl2 (0:025 M) at 25 °C is treated with 2-fluoro-I-
methylpyridinium p-toluenesulfonate from Aldrich Chemical
15 company inc. (1.5 equiv.) and stirred for 35 minutes. The course
of the reaction was monitored through thin layer chromatography
(TLC)(E. Merck RP- 18 silica, 65 tetrahydrofuran: 35 water,
IN/phospho-molybidic acid) and after thirty minutes of stirring
at ambient temperature, judged complete as no taxol remained
20 and only one compound was apparent by TLC. The reaction
mixture is directly purified by HPLC (Vydal: RP-18, 22.5 x 3 mm,

CA 02180444 1996-07-03
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-~ B 0.5 h linear, A: 209~o MeOH in 20 mM NH.~OAc, B: 100%
MeOH, 9 mL / min, RT = 2 6.12 ) to give ( 81- 8 8 ) II. as a
colorless film.
Preparation of C-2-taxoid-7-benzothiazolium salts
v i
Ac0 O pH Ac0 O O N + X '
BzNH O BzNH O '
Alkyl
Pr,~o~.. ~- Pr,~~o~...
OH ' OH . ~ p
O - H
HO /p H OAc HO O OAc
R 1~ R_1
O ~ X s CN~COO; TsO; BF,; hltldvc O
I. I I.
~ MszN ~ ~ ~ I ~ ~ ~~gPh
89 90 9~ 92 93 94 95 96
C-2-taxoid-7-benzothiazolium salts (89-96). The
synthesis of the taxoid-7-benzothiazolium salts ( 8 9 - 9 6 ) II,
differs only slightly from the synthesis of taxoid-2'-
benzothiazolium salts ( 81-8 8 ) II. The C-2 taxoid ( 8 9-9 6 ) I is
dissolved ; in methylene chloride (.006 M) and treated
sequentially with triethylamine (40 equivalents) and 2-fluoro-
3-ethylbenzothiazolium tetrafluoroborate ( 10 equivalents)
Aldrich Chemicals, and allowed to stir at ambient temperature
for 5 minutes. The reaction mixture is then directly purified
by HPLC (Vydak RP-18, 22.5 x 3 mm, A -~ B 0.5 h linear, A:
209'o MeOH in 20 mM NH.~OAc, B: 100% MeOH, 9 mL / min, RT =
26.12) to give (89-96) II as a colorless film.

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Preparation of C-2-taxoid-2'-benzoxazolium salts
Ac00 AC 0 OH
BZNH O OH BZNH O
Pn."~'a,..
~o b
OH ~ p ~ /~ Hp = H O
HO ~ H pAt ''~ N p,, OAt
X ~ " Alkyl
O O
I. X . CIi~C00'; TAO: BF,~; halides
II.
R= ~ \0/ r \S/ \S/ ~ ~ ~ ~ ~ Me:N ~ ~ ~ ~ ~ ~ ~~SPh
97 98 gg 100 101 102 103 104
C-2-taxoid-2'-benzoxazolium salts 97-104. A solution
of taxoid 97-104 L( 1.0 equiv.) and triethylamine (4.7 equiv.) in
CH2C1? (0.025 M) at 2~ °C is treated with 2-fluoro-1-
methylpyridinium p-toluenesulfonate from Aldrich Chemical
company inc. ( 1.5 equiv. ) and stirred for 3 S minutes. The course
of the reaction was monitored through. thin layer chromatography
(TLC)(E. Merck RP- 18 silica, 6~ tetrahydrofuran: 3~ water,
UV/phospho-rnolybidic acid) and after thirty minutes of stirring
at ambient temperature, judged complete as no taxol remained
and only one compound was apparent by TLC. The reaction
1~ mixture is then directly purified b~~ HPLC (Vyda~ RP-18, 22.~ x 3
mm, A --> B 0.5 h linear, A: 20~Yo MeOH in 20 mM I~TH.~OAc, B: 100%
MeOH, 9 mL / min, RT = 26.12) to give 97-104 II. as a colorless
film.

CA 02180444 1996-07-03
PCT/US95/00478
wo 9s/issoa
- 96 -
Preparation of C-2-taxoid-7-benzoxazolium salts
s~ \ /
Ae00 OH At 0 O~p w X.
BzNH O B~H ., Alkyl
~,%~o~... Ph o ..
off
OH HO : H : 0 HO ~ H i
p OAC 0 OAt
R~ R
X s CIi,,C00; T10; BFI; hllidel
1. ~I.
~/ ~ \S/ \5/ ~ I ~ I ~ M~~N \ / ~ I ~ ~ ,~i~,~SPh
105 106 107 106 109 »0 171 X12
S C-2-taxoid-7-benzoxazolium salts (105-112). The
synthesis of the taxoid-7-benzoxazolium salts ( 10 S -112 ) II,
differs only slightly from the synthesis of taxoid-2'-
benzoxazolium salts ( 9 7-104 ) II. The C-2 taxoid ( 10 S -1 I 2 ) I is
dissolved in methylene chloride (.006 M) and treated sequentially
with triethylamine (40 equivalents) and 2-chloro-3-
ethylbenzoxazolium tetrafluoroborate from Aldrich Company ( 10
equivalents) Aldrich Chemicals, and allowed to stir at ambient
temperature for 5 minutes. The reaction miaaure is then directly
purified by HPLC (~'yda~ RP-1$, 22.~ x 3 mm, A -~ B 0.~ h linear,
A: 209'o MeOH in 20 mM NH.~OAc, B: 1009~o MeOH, 9 mL / min, RT =
26.12 ) to give ( 105-112 ) II as a colorless film.

CA 02180444 1996-07-03
WO 95/18804 ~ PCT/U895/00478
- 97 -
Preparation of C-2-taxoid-2'-pyrimidinium salts
A O OH
Ac00 OH Bz~NH O
BzNH
Pn o~ ~ ~ Pn~ ;~'a...
OH ' O : H ~ O
HO ~ H pAc + HO O OAc
R , X.~~YI R Oi
I ~ -.~O X s CH~COO'; TsO; BF,~; htlides I I.
R= ~ 10~ ~ ~S~ 1ST ~ I ~ ~ i MesN ~ ~ ~ ~ i % ~~SPh
113 114 115 116 117 11$ 119 120
S C-2-taxoid-2'-pyrimidinium salts 113-120. A
solution of taxoid ( 113-120) I. ( 1.0 equiv.) and triethylamine
(4.7 equiv.) in CH2C12 (0.025 M) at 25 °C is treated with 2-
chloro-methyl-pyrimidinium fluoride from Aldrich Company
( 1.5 equiv.) and stirred for 35 minutes. The course of the
reaction was monitored through thin layer chromatography
(TLC)(E. Merck RP- 18 silica, ~5 tetrahydrofuran: 35 water,
W/phospho-molybidic acid) and after thirty minutes of
stirring at ambient temperature, judged complete as no taxol
remained and only one compound was apparent by TLC. The
reaction mixture is then directly purified by HPLC (Vydak RP-
18, 22.5 x 3 mm, A -~ B 0.5 h linear, A: 20~'o MeOH in 20 mM
NH40Ac, B: 100°Yo MeOH, 9 mL / min, RT ~ 2 6.12 ) to give ( 113 -
120) II. as a colorless film.

CA 02180444 1996-07-03
WO 95/18804 ~ PCT/US95/00478
_ 98 _
Preparation of C-2-taxoid-7-pyrimidinium salts
Ac0 O OH X
BzNH _. BzNH 0
P,,~p .. Pn~_'~jj~_
OH s OH
HO = O
O H OAc
X s C11~C00'; TsO; BF,; halidecR O
I. II.
% Mesh ~ ~ ~ I ~ ~ ~~gPh
t21 122 123 124 125 126 127 128
C-2-taxoid-7-pyrimidinium salts ( 121-128 ) . The
synthesis of the taxoid-7-pyrimicimium salts ( 121-12 8 ) II,
differs only slightly from the synthesis of taxoid-2'-
pyrimidinium salts ( 113 -12 0 ) II. The C-2 taxoid ( 121-12 8 )
I is dissolved in methylene chloride (.006 M) and treated
sequentially with triethylamine (40 equivalents) and 2-chloro-
methyl-pyrimidinium fluoride from Aldrich Company ( 10
equivalents), and allowed to stir at ambient temperature for ~
minutes. The reaction mixture is then directly purified by
HPLC (Vydak RP-18, 22.~ x 3 mm, A -~ B 0.5 h linear, A: 20%
MeOH in 20 mM NH.~OAc, B: 100% MeOH, 9 mL / min, RT =
2 G.12 ) to give ( 121-12 8 ) a as a colorless film.

Dessin représentatif