COMMUTATION CHIMIQUE DE TAXO-DITERPENOIDES ENTRE DES FORMES ACTIVES A FAIBLE SOLUBILITE ET DES FORMES INACTIVES A HAUTE SOLUBILITE

CHEMICAL SWITCHING OF TAXO-DITERPENOIDS BETWEEN LOW SOLUBILITY ACTIVE FORMS AND HIGH SOLUBILITY INACTIVE FORMS

Abrégé français

On emploie un procédé de commutation chimique cyclique pour rendre solubles et pour rendre insolubles des taxo-diterpénoïdes par rapport à des solvants aqueux. Dans ce procédé on utilise des sels onium 2-halogénés d'aza-arènes pour transformer en dérivés des taxo-diterpénoïdes de manière à modifier leur solubilité dans des solvants aqueux. Le sel onium d'aza-arène comprend une charge délocalisée qui confère des propriétés de polarité et de solubilité dans l'eau aux dérivés de taxo-diterpéroïde. On procède à la solubilisation avec une transformation en dérivé en une seule étape avec le sel onium d'aza-arènes halogénés en position 2. On procède à la désolubilisation en mettant en contact les sels onium de taxo-diterpénoïde-C<n>,2-O-aza-arènes avec une protéine sérique pour déplacer le 2-O-aza-arène et former un intermédiaire de protéine soluble:taxo-diterpénoïde. Cet intermédiaire de protéine soluble:taxo-diterpénoïde se dissocie ensuite avec le temps pour produire un taxo-diterpénoïde bioactif. Les taxo-diterpénoïdes préférés comprennent le taxol, le taxol à substitution C-2 et le Taxotère<TM>. Ces mêmes sels onium de taxo-diterpénoïde-C<n>,2-O-aza-arènes sont utilisés comme promédicaments solubles dans l'eau. Par exemple le taxol-2'-méthylpyridinium tosylate (MPT) se caractérise par une forte solubilité dans l'eau, une activation rapide par la protéine sérique, une bonne stabilité dans la plupart des autres solutions aqueuses, la formation d'un intermédiaire de protéine:taxol et une bonne rétention dans le système circulatoire. La toxicité de la forme activée est comparable ou supérieure à celle du taxol non transformé dérivé. En outre on peut synthétiser du taxol-2'-MPT à l'aide d'une seule et simple étape de réaction entre du taxol et du 2-fluoro-1-MPT. Cette invention peut s'appliquer au taxol et aux substances imitant le taxol comprenant des groupes hydroxyle qui réagissent aux sels onium d'aza-arènes 2-halogénés. Par exemple, le taxol et le Taxotère comprennent chacun des groupes hydroxyle réactifs aux positions 2' et 7. Cette invention peut également s'appliquer à une gamme étendue d'aza-arènes halogénés en position 2.

Abrégé anglais

Cyclic chemical switching method is employed for
solubilizing and desolubilizing taxo-diterpenoids with
respect to aqueous solvents. 2-Halogenated onium salts of
aza-arenes (I and II)
(see figure I and II)
wherein: R0 is a halogen selected from the group consisting
of Cl, Br, F, and I; 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, arenxyl, propargyl, and
fused aryl; R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl; if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6 O-alkyl,
OH, halogen, and fused aryl; if Z2 is C, then R5 is selected
from the group consisting of H, C1-C6 alkyl, allyl, arenxyl,
propargyl, C1-C6 O-alkyl, OH, halogen, and fused aryl; if Z2
is N, then R5 is absent, and S- is a counter ion;
are employed to derivatize taxo-diterpenoids so as to alter
their solubility in aqueous solvents. The onium salt of

aza-arene includes a delocalized charge which imparts
polarity and aqueous solubility to taxo-diterpenoid
derivatives. Solubilization is achieved in a one step
derivatization with the onium salt of 2-halogenated
aza-arenes. Desolubilization is achieved by contacting onium
salts of taxo-diterpenoid-C n,2-O-aza-arenes with serum
protein to displace the 2-O-aza-arene and form a soluble
protein:taxo-diterpenoid intermediate. This
protein:taxo-diterpenoid intermediate then dissociates over time to
provide a bioactive taxo-diterpenoid. Preferred
taxo-diterpenoids include taxol, C-2 substituted taxol, and
Taxotere .TM.. These same onium salts of taxo-diterpenoid-C n,
2-O-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 retention 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 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

-80-
CLAIMS:
1. A cyclic method employing chemical switching for
solubilizing and desolubilizing a taxo-diterpenoid with
respect to an aqueous solvent, an underivatized form of the
taxo-diterpenoid having a low solubility and including a
reactive C n-hydroxyl, the method comprising the following
steps:
Step A: converting the underivatized form of the
taxo-diterpenoid from low solubility to high solubility by
derivatizing the reactive C n-hydroxyl with an onium salt of a
2-halogenated aza-arene to form an onium salt of a taxo-
diterpenoid-C n,2-O-aza-arene derivative having high
solubility, wherein the onium salt of the 2-halogenated aza-
arene is represented by the following structures I and II:
<IMG>
wherein:
R O is a halogen selected from the group consisting
of Cl, Br, F, and I;
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, arenxyl, propargyl, and fused aryl;

-81-
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent, and
S- is a counter ion; and then
Step B: converting the onium salt of the taxo-
diterpenoid-C n,2-O-aza-arene derivative produced in said
Step A from high solubility by contacting the taxo-
diterpenoid-C n,2-O-aza-arene derivative with a serum protein
for displacing the 2-O-aza-arene and forming a protein:taxo-
diterpenoid intermediate, the protein:taxo-diterpenoid
intermediate then dissociating to produce the underivatized
form of the taxo-diterpenoid employed in said Step A.
2. A cyclic method employing chemical switching for
solubilizing and desolubilizing a taxo-diterpenoid with
respect to an aqueous solvent, an underivatized form of the
taxo-diterpenoid having a low solubility and including a
reactive C n-hydroxyl, the method comprising the following
steps:

-82-
Step A: converting the underivatized form of the
taxo-diterpenoid from low solubility to high solubility by
derivatizing the reactive C n-hydroxyl with an onium salt of a
2-halogenated aza-arene to form an onium salt of a taxo-
diterpenoid-C n,2-O-aza-arene derivate having high solubility,
the underivatized form of the taxo-diterpenoid being
represented by formula I as follows:
<IMG>
wherein:
C n is selected from the group consisting of C7
and C2';
R x is selected from the group consisting of Ph
and tBuO;
R10 is selected from the group consisting of OAc
and OHM
R y is selected from the group consisting of benzyl
and the following structures:

-83-
<IMG>
R2' and R7 are each OH;
the onium salt of the taxo-diterpenoid-C n,2-0-aza-arene
derivate produced in said Step A being represented by the
above formula I wherein:
R2' and R7 are each selected from the group
consisting of OH and an onium salt of a 2-O-aza-arene, with
the proviso that at least one of R2' and R7 is said onium
salt of the 2-O-aza-arene, said onium salt of the 2-O-aza-
arene being selected from the group consisting of onium
salts represented by the following formulas II and III:
<IMG>

-84-
wherein:
R0 is oxygen and is bonded to C n;
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, arenxyl, propargyl, and fused aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent; and
S e is a counter ion; and
the onium salt of the 2-halogenated aza-arene employed in
said Step A being selected from the group consisting of
onium salts represented by the above indicated formulas II
and III wherein:

-85-
R0 is a halogen selected from the group consisting
of Cl, Br, F, and I; and then
Step B: converting the opium salt of the taxo-
diterpenoid-C n,2-O-aza-arene derivate produced in said Step A
from high solubility to low solubility by contacting the
taxo-diterpenoid-C n,2-O-aza-arene derivative with a serum
protein for displacing the 2-O-aza-arene and forming a
protein:taxo-diterpenoid intermediate, the protein:taxo-
diterpenoid intermediate then dissociating to produce the
underivatized form of the taxo-diterpenoid employed in said
Step A.
3. ~A method for solubilizing a taxo-diterpenoid with
respect to an aqueous solvent, the taxo-diterpenoid
including a reactive C n-hydroxyl, the method comprising the
following step:
Step A: converting the taxo-diterpenoid from low
solubility to high solubility by derivatizing the C n-hydroxyl
with an opium salt of a 2-halogenated aza-arene to form an
opium salt of a taxo-diterpenoid-C n,2-O-aza-arene derivative
having high solubility, wherein the opium salt of the
2-halogenated aza-arene is represented by the following
structures I and II:
<IMG>
wherein:

-86-
R0 is a halogen selected from the group consisting
of Cl, Br, F, and I;
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, arenxyl, propargyl, and fused aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent; and
S- is a counter ion.
4. ~A method for solubilizing an underivatized form of
a taxo-diterpenoid having a low solubility with respect to
an aqueous solvent, the underivated form of the taxo-
diterpenoid including a reactive C n-hydroxyl, the method
comprising the following steps:

-87-
Step A: converting the underivatized form of the
taxo-diterpenoid from low solubility to high solubility by
derivatizing the reactive C n-hydroxyl with an onium salt of a
2-halogenated aza-arene to form an onium salt of a taxo-
diterpenoid-C n,2-O-aza-arene derivative having high
solubility, the underivatized form of the taxo-diterpenoid
being represented by formula I as follows:
<IMG>
wherein:
C n is selected from the group of C1 and C2';
R x is selected from the group consisting of Ph
and tBuO;
R10 is selected from the group consisting of OAc
and OH;
R y is selected from the group consisting of benzyl
and the following structures:

-88-
<IMG>
R2' and R7 are each OH;
the onium salt of the taxo-diterpenoid-C n,2-O-aza-arene
derivative produced in said Step A being represented by the
above formula I wherein:
R2' and R7 are each selected from the group
consisting of OH and an onium salt of a 2-O-aza-arene, with
the proviso that at least one of R2' and R7 is said onium
salt of the 2-O-aza-arene, said onium salt of the 2-O-aza-
arene being selected from the group consisting of onium
salts represented by the following formulas II and III:
<IMG>
wherein:
R0 is oxygen and is bonded to C n;

-89-
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, arenxyl, propargyl, and fused aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent: and
S~ is a counter ion; and
the onium salt of the 2-halogenated aza-arene employed in
said Step A being selected from the group consisting of
onium salts represented by the above indicated formulas II
and III wherein:
R0 is a halogen selected from the group consisting
of Cl, Br, F, and I.

-90-
5. The method for solubilizing a taxo-diterpenoid as
described in claim 3, comprising the following additional
step:
after said Step A
Step B: converting the onium salt of the taxo-
diterpenoid-C n,2-O-aza-arene derivative produced in said
Step A to a taxo-diterpenoid:protein conjugate by
displacement of 2-O-aza-arene and conjugation with a serum
protein, the taxo-diterpenoid:protein conjugate having high
solubility.
6. A method for converting an onium salt of a taxo-
diterpenoid-C n,2-O-aza-arene derivative into a taxo-
diterpenoid:protein conjugate, the method employing the
following step:
contacting the onium salt of the taxo-diterpenoid-
C n,2-O-aza-arene derivative with a serum protein for
displacing 2-O-aza-arene and conjugating the taxo-
diterpenoid with the serum protein to produce the taxo-
diterpenoid:protein conjugate, the 2-O-aza-arene being
selected from the group consisting of onium salt I and onium
salt II represented by the following formulas:
<IMG>

-91-
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, arenxyl, propargyl, and fused aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent: and
S- is a counter ion.
7. A method for converting an onium salt of a taxo-
diterpenoid-C n,2-O-aza-arene derivative into a taxo-
diterpenoid:protein conjugate, the method employing the
following step:

-92-
contacting the onium salt of the taxo-diterpenoid-
C n,2-O-aza-arene derivative with a serum protein for
displacing 2-O-aza-arene and conjugating the taxo-
diterpenoid with the serum protein to produce the taxo-
diterpenoid:protein conjugate, wherein the onium salt of the
taxo-diterpenoid-C n,2-O-aza-arene derivative produced is
represented by the following formula:
<IMG>
wherein:
C n is selected from the group consisting of C7
and C2';
R x is selected from the group consisting of Ph
and tBuO;
R10 is selected from the group consisting of OAc
and OH;
R y is a C-2 substituent selected from the following
structures:

-93-
<IMG>
R2' and R7 are each selected from the group
consisting of OH and an onium salt of a 2-O-aza-arene, with
the proviso that at least one of R2' and R7 is said onium
salt of the 2-O-aza-arene,
said onium salt of the 2-O-aza-arene being selected from the~
group consisting of onium salt I and onium salt II
represented by the following formulas:
<IMG>
wherein:
R0 is oxygen and is bonded to C n;
Z1 and Z2 are each selected from the group
consisting of C and N;

-94-
Z3 is selected from the group consisting of S
and O;
R1 is selected from the group consisting of C1-C6
alkyl, allyl, arenxyl, propargyl, and fused aryl;
R2 and R6 are each selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, and
fused aryl;
if Z1 is C, then R3 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused 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, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is C, then R5 is selected from the group
consisting of H, C1-C6 alkyl, allyl, arenxyl, propargyl, C1-C6
O-alkyl, OH, halogen, and fused aryl;
if Z2 is N, then R5 is absent; and
S- is a counter ion.

Description

CA 02180445 2004-07-21
-1-
CHEMICAL SWITCHING OF TARO-DITERPENOIDS BETWEEN LOW SOLUBILITY ACTIVE FORMS
AND HIGH SOLUBILITY INACTIVE FORMS
Technical Field:
The invention relates to taxol prodrugs. More particularly, the
invention relates to a method employing derivatization with opium
salts of 2-halogenated aza-arenes for chemically switehiitg ~bctween
low solubility active forms and high solubility inactive forms of ~taxol
0 and taxol memetics.
BackQro~und.:
Taxol;~ an antineoplastic agent originally isolated from Taxus
brevifolia, is approved for usage in the treatment of ovarian cancer
5 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 molar, it has been necessary to formulate
Taxol in a mixture of Cremaphor''"', a polyoxyethylated castor oils and
ethanol in order to achieve a therapeutic concentration. This
0 formulation can induce a variety 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.
5 Although taxol's dramatic efficacy has driven clinical 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
0 introduction of solubilizing functionality that normal metabolic
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 prodrug is rapidly
5 cleared from circulation by the kidneys.
Taxol is only one of a class of taro-diterpenoids having
bioactivity. Another preferred taxo-ditcrpenoid having clinically
*Trade-mark

WO95I18798 ~, 1 PCfIU595100481
-2-
significant activity is Taxotere''T". Unfortunately, all known bioactive
taxo-diterpenoids have a low aqueous solubility.
What is needed is a method for chemically switching taxol and
other taxo-diterpenoids between..a high solubility and low solubility
form in a manner which regulates its rate of clearance from
circulation so that the prodrug is retained for a clinically significant
period after administration.
Taxol itself is known to serve as a chemical switch with
respect to tubulin. Binding of taxol to tubulin prevents its
polymerization and the formation of microtubules. While
unpolymerized tubulin is soluble in aqueous media, polymerization
of tubulin leads to the formation of insoluble microtubules.
Accordingly, the addition or removal of taxol drives the
depolymerization or polymerization of tubulin and, in this manner,
serves as a chemical switch for regulating the solubility of tubuIin.
Summarv:
The invention is a cyclic method employing chemical switching
for solubilizing and desolubilizing taxo-diterpenoids with respect to
2 0 aqueous solvents. The invention employs 2-halogenated opium salts
of aza-arenes to derivatize taxo-diterpenoids so as to alter their
solubility in aqueous solvents. The opium salt of aza-arene includes
a delocalized charge which imparts polarity and aqueous solubility
to taxo-diterpenoid derivatives. Solubilization includes a one step
2 5 derivatization with the opium salt of 2-halogenated aza-arenes.
Contacting opium salts of taxo-diterpenoid-Cn,2-O-aza-arenes with
the serum protein, causes the displacement of 2-O-aza-arene and
the formation of a soluble proteinaaxo-diterpenoid intermediate.
This proteinaaxo-diterpenoid intermediate then dissociates over
3 0 time to provide a bioactive taxo-diterpenoid. Preferred taxo-
diterpenoids 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
3 S hydroxyls on taxol and Taxotere''M are located at C2~ and C~. A
preferred opium salt of 2-halogenated aza-arene is
2-fluoro-1-methylpyridinium tosylate. Other employable opium
SUBSTITUTE SHEET (RULE 281

WO 95/18798 ~ PC1'/US95100481
-3-
salts of 2-halogenated aza-arenes are disclosed by T.Mukaiyama,
Angewandte Chg»iie f979, 18(18), 707-808, incorporated herein by
reference.
More particularly, a first embodiment of the invention is
directed a cyclic method employing chemical switching for
solubilizing and desolubilizing taxo-diterpenoids with respect to
aqueous solvents. Underivatized forms of the taxo-diterpenoid have
low aqueous solubility and include a reactive Cn-hydroxyl, i.e., a
reactive hydroxyl at the Cn position. Preferred reactive Cn
hydroxyls for taxol and TaxotereT"s are located at positions C2~ and
C 7 . The method includes two steps. In the first step, the
underivatized form of the taxo-diterpenoid is converted from low
solubility to high solubility by derivatizing the reactive Cn-hydroxyl
with the opium salt of the 2-halogenated aza-arene to form the
opium salt of a taxo-diterpenoid-Cn,2-O-aza-arene derivative having
high solubility. In the second step, the opium salt of the taxo-
diterpenoid-Cn,2-O-aza-arene derivative is converted from high
solubility to low solubility by contacting the taro-diterpenoid-Cn,2-
O-aza-arene derivative with serum protein for displacing the 2-0-
aza-arene and forming a proteinaaxo-diterpenoid intermediate. The
proteinaaxo-diterpenoid intermediate has a high solubility but then
dissociates over time to produce the underviatized form of the taxo-
diterpenoid employed in the first step, i.e., the taxo-diterpenoid is
released from the proteinaaxo-diterpenoid intermediate. The
2 5 precise nature of the bonding between serum protein and the taxo-
diterpenoid within the proteinaaxo-diterpenoid intermediate has
not been characterized, but can be stable over a period ranging from
minutes to hours. A first alternative embodiment of the invention
are directed to the derivatization of taro-diterpenoids with opium
3 0 salts of 2-halogenated aza-arenes. A second alternative
embodiment is directed to conversion of opium salts of taxo-
diterpenoid-Cn,2-O-aza-arene derivatives to protein:taxo-
diterpenoid intermediates using serum protein. In this second
alternative embodiment, the taxo-diterpenoid-Cn,2-O-aza-arene
3 5 derivative is contacted with serum protein for displacing the 2-O-
aza-arene and forming the proteinaaxo-diterpenoid intermediate.
SUBSTfTUTE SHEET (RULE 2~

wo 9sns~9s ~ ~ ~ d ~ ~ ~ rcT~s9s~ooast
-4-
In a preferred embodiment, the taxo-diterpenoid-2-O-aza-
arenes are represented by the following formula:
t°R O
Rx~ NH to t t° 9 is R~
t2 - s s
ph~0~>.... A is B 8 C 5
t3 ~t7 .
:z H ø D
HO ~Ry OAc z°
wherein Rx is Ph or tBuO; RIO 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~Zt~Zz,RS ~~d~0
~ Qi N
zR~N~O
Rt
S~ Rt S
I $ opium salt ~ opium salt II
wherein Zi and Z2 are each either C or N; Z3 is S or O; RI is selected
from the group consisting of CI-C6 alkyl, allyl, arenxyl, propargyl,
2 0 and fused aryl; R2 and R6 are each selected from the group
consisting of H, CI-C6 alkyl, allyl, arenxyl> propargyl, and fused aryl;
if Z1 is C, then R3 is selected from the group consisting of H, C1-C6
alkyl, allyl, arenxyl, propargyl, CI-C6 O-alkyl, OH, halogen, and fused
aryl; if Z 1 is N, then R3 is absent; R4 and R8 are each selected from
25 the group consisting of H, CI-C6 alkyl, allyl, arenxyl, propargyl, Cl-
C6 O-alkyl, OH, halogen, and fused aryl; and if Z2 is C, then RS is
SUBSTITUTE SHEET (RULE 281

WO 95!18798 ~ PCTlUS95/00481
-5-
selected from the group consisting of H, C1-C6 alkyl, ally!, arenxyl,
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 a C-2 substituent. Preferred C-2 substituents are
represented by the following structures:
I ~ N
Me2N ~ ~ ~ I / / ~~SPh
Brief De crin~inn of the Drawil~:
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
(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
fibrils further magnified to illustrate the helical nature of the
2 5 structure.
Figure 2 B 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
SUBSTITUTE SHEET (RULE 28)

CA 02180445 2004-07-21
-6-
critical aggregation concentration (CAC) of this compound using a
negative uranyl acetate stain and a magnificatioa of x45000.
Figure 3 illustrates a tubulin polymerization-depolymerization
measurements with negative control (triangles), positive taxol
control (diamonds), and taxoI-2'-MPT, i.e., compound Z (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 model: 59b dextrose (triangles), taxol
(diamonds), and taxol-2'-MPT (dots).
etaile~ Descri lion:
The synthesis, physical properties, and pharmaceutical profiles
of water soluble opium salts of taro-diterpenoid-Cn,2-O-aza-arenes
are described.
Synthesis of Taxol-2'-MPT:
2 0 Taxol-2'-MPT (methylpyridinium tosylate), compound 2, was
synthesized according to the method of T.Mukaiyama, Angewandte
Chemie 1979, 18(18), 707-808 .
Taxol (10 mg, 0.012 mM), from NaPm Biochemicals, Boulder CO, USA,
was dried by azeotropic distillation with toluene (2x 1.0 mL) and
2 5 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
3 0 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) (fi. Merck
RP-18 silica, 65 tetrahydrofuran . 35 water, UV/phosphomolybdic
acid) and after thirty minutes of stirring at ambient temperature,
3 5 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) (Clg column, 1mM NH40Ac

w0 95!18798 ~ ~ PCT/US95/00481
_7.
pH 6.5 buffer / methanol gradient, 1.5 mL / min. UV) to give, after
removal of solvent in vaccuo, pure taxol-2'-MPT (2) (12 mg, 93'0
yield) as a white amorphous solid. All spectroscopic data (IH NMR
and HRMS) were in accord with the structure assigned to 2. I H
NMR (CDCI3, 125 MHz) : 1.055 (s, 3 H, C17-H), 1.083 (s, 3 H, CI9-H),
1.724 (s, 3 H, C19-H), 1.858 (m, 1 H, C6- H), 1.913 (s, 3 H, CH3-Ph),
2.193 (s, 3 H, CIO-OC(O)CH3), 2.514 (m, I H, C6-aH), 3.663 (d, 1 H, J =
7.0 Hz, C3-H), 4.110 (d, I H, J = 8.5, C20- H, A of AB), 4.133 (s, 3 H,
N-CH3), 4.230 (d, 1 H, J = 8.5, C20-aH, B of AB), 4.315 (dd, 1 H, J =
1 0 8.7, 10.7, C7-H), 4.901 (dd, 1 H, J = 1.0, 7.7, CS-H), 5.501 (d, 1 H,
C2-H, J = 7.0), 5.702 (bt, I H, C2'-H, J = 8.0), 5.951 (dd, 1 H, C13-H, J
= 1.0, 8.0), 6.120 (bd, 1 H, C3'-H, J = 10.0), 6.181 (s, 1 H, CIO-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 - 7.62 (m, 4 H, Ar-H ), 7.68 - 7.75 (m, 4 H, Ar-H ) ,
8.00 - 8.05 (m, 1 H Ar-H ), 8.23 - 8.28 (m, 1 H, Pyr-H ), 8.41 (m, I H,
Pyr-H). IR (neat, KCI plate) cm-1: 3640 - 3120 (bm), 3030-2870
(bm), 2320 (m), 1720 (s), 1630 (m), 1560 (m), 1500 (m), 1360 (s),
1160 (m), 1070 (m), 700 (m). UV/Vis (CHC13) nm: 254, 280. FAB
HRMS: calc for Cg3H57O14N2~ 945.3810; found: 945.3810
The molecular structures of taxol, compound 1, and of taxol-2'-
MPT, compound 2, are illustated in Scheme IA. The synthesis of
taxol-2'-MPT is illustrated in Scheme 1B.
a
0
--- ~' rosy
1: Taxol = Taxoi oti 2: taxol-2'-methylpyridinium tosylate
2 5 (taxol-2'-MPT)
SUBSTITUTE SHEET (RULE 2~

wo 9snar9a ~ ~ ~ (~~L~~ rcrros9srooaat I
.g.
rosy
b ~ ~~ rod
F
Taxol OH
(CHIC Ha3N, CHzCh
o- Taxol
sox
2
Schemes lA and 1B
synthesis of Taxol-7-MPT:
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 microL,
0.48 mM, 40 equivalents) and 2-fluoro-I- 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
I S 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 (1 H
NMR and HRMS) were as expected.
2 0 synthesis 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
25 with triethylamine (67 microL, 0.48 mM, 40 equivalents)
and
2-fluoro-1- methylpyridinium tosyIate (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 p ure taxol-2'-MPT
(2)
3 0 (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
SUBSTITUTE SHEET (RtiLE 281

WO 95!18798 ~ . PCTIUS95I00481
-9-
,.
yield was 13 mg or 85%. Spectroscopic data NMR and HRMS',
(1H
were as expected .
Alternative syntheticschemes based upon the method of
T
.
Mukaiyama (Angewandte Chemie 1979, 18(18) 707-808) usin
, g a
variety of opium salts 2-halogenated aza-arenes for derivatizing
of
either the 2' or the 7 illustrated in
positions the
of taxol
are
following scheme:
0
t) TESOTf, 2,61utidine, CHzCiz
haloonium salt, 2,6 Iutialne, i ~; Ted \
CH2CI2 ~(/j \ \ CH aloonium salt, 2,6 lutidine,
2C 2
Ph"NH O Ac0 O OH OO
~~On,...
OR s O
HO H
OBz OAc
R~ / I N~-~ ~ ~~ Ij~f / I
XD 0* Alkyl ~Oj~~ ~ C'~ >~ + ,
Alkyl
1 0 X = BF,; TsO; halides
Stabilitv mea ~rements and Kin r;~~
f Taxol ref r .
I5 Due to a difference in retention time using our standard HPLC
conditions (see Fig. 1) and differing ultraviolet absorption maxima
(12801254 = 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
20 results from hydrolysis of pyridinium salts (Fig. lc.). Taxol-2'-MPT
SUBSTITUFE SHEET (RULE 281

WO 95118798 ~ ~ ~ ~ PCTlUS95100481
-lU-
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°lo dextrose, artd 1.596 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 over 21 days.
Taxol-2'-MPT (2) is, however, unstable in 5% HCl (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~Yo 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
2 5 methylene chloride to quench the reaction. Samples were then
analyzed using a Waters Maxima HPLC instrument equipped with an
autoinjector (3.9x300 mm Cl g column equipped with a precolumn.
The flow rate was 1.5 mLlminute. The eluent gradient A-B
extended over 30 minutes. "A" was 80% 80mM ammonium acetate,
3 0 pH 6Ø "B" was 100% methanol. An ultraviolet diode alzay 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.
SUBSTITUTE SHEET (RULE 281

R'O 95118798 ~ PCT/US95/00481
-lI-
't '
~olubilitv Meacnrr~r.,Ants:
The solubility and partition coefficient data for 2 and taxol
were determined using an HPLC method.
TaxoI ~ 2'-MPT-taxol
Solubility in Water < 1.5 X ID 1.7 X 10 -3
> 10000 50
Partition Coefficient
foctanoU [water]
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 for 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 Iimit.
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
2 0 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.
These data clearly show that 2 is significantly more soluble in water
2 5 than the parent taxol. The solubility demonstrated in a range of
aqueous systems is higher than the clinically relevant dosages (3 to
30 mM).
SUBSTITUTE SHEET (RULE 2B~

wo 9sns~9s ~ ~ $ ~ ~ ~ 5 rcTrtrs9srooast
-12-
Self-Assembling- Structures:
,.
While the water solutions of 2 were optically clear at all
concentrations examined,' buffered solutions of concentrations
greater than I x IO-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
10-4 M, a result characteristic of macromolecular structure in
solution. Transmission electron microscopy (TEM) confirmed the
presence of supramolecuIar structures in these solutions. Uniform
aggregates of fibrillar structure (Fig. 2a) with helical conformations
were observed. These structures exhibited varying (up to 800 ~)
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
~.. 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 ~olvmerization-de~ymerization 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 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 2 does act as a
3 0 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
3 5 that calcium chloride promoted depolymerization is suppressed by
taxol but not by taxol-2'-MPT.
SUBSTITUTE SHEET (RULE 28)

WO 95/18'798 21 ~ 0 4 9: 5 P~~595100481
-13-
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 addition
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 (I.0 mg/mL) alone, CaCl2 (0,25 mM) added after
20 minutes. Positive taxol control employed tubulin (1.0 mg/mL)
with taxol (10-6M) and CaCl2 (0.25mM) added after 20 minutes.
The experimental taxol-2'-MPT employed tubulin (1.0 mg/mL) with
taxol-2'-MPT (10-6) and CaCl2 (0.25 mM) added after 20 minutes.
Turbidity was measured as optical density at 340 nm using a
microplate reader (Molecular Devices Thermomax).
Toxicity M ac»rP.nen c;
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 taxol were
similar, although some differences were noted. Both compounds
exhibited IC50 values ranging from 10-5 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
3 0 method from the stock. Plates were incubated at 37 C, S~o C02 in
sterile air in an humidified incubator for 72 hours. An aliquot of 50
L of a solution of 2,3-bis(methoxy-4-nitro-5-sulfophenyl)-5-
[(phenylamino)carbonyl]-1H-tetrazolium hydroxide (XTT), 1 mg
mL-1, in phosphate buffered saline (PBS, 100mM) was added to the
3 5 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
SUBSTfTUTE SHEET (RULE 2~

wo vsnsws ~ ~ g ~ ~ ~ ~' rc'rrtrs9sroo4st
-14-
calculated using the formula: %C = 1 - (OD toxin)(OD growth
control)-1(100).
Efficacy of taxol-2'-MPT in lung tumor xenoeraft:
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 for this study were formulated in sterile PBS without
Cremaphor''n', 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 taxoI-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)2/2. The test
compounds (1.0 microM) were injected LP. on day 1,3, and 7 using
the following media: control. 596 dextrose in water (DSW), triangles;
taxol, suspended in Cremaphor/D5W (5/95, 18.0 mg/kg of animal
3 0 weight), diamonds; and taxol-2'-MPT, dissolved in DSW (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
3 5 Reports, 3 (1972).
SUBSTITUTE ShIEET (RULE 2B)

WO 95118798 ~ ~ PCTIUS95100481
-]5-
Mechanisms of " ayW lPtnaCP~
The mechanism of acid catalyzed taxol (1) release from taxol-
2'-MPT is illustrated in Scheme 2.
C
_ To ~ To
i -" ~~ Ho- Taxol
0
H O ~ ~ Taxol H~ O Taxol
1-methylpyridinone
2
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 proteinaaxol
intermediate. Dialysis of the proteinaaxol 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
proteinaaxol intermediates. The precise nature of the bonding
2 0 between the protein and taxol has not been characterized. Scheme 2
illustrates alternative pathways for MPT release.
SUBSTITUTE SHEET (RULE 2!11

R'O 95118798 2 ~ PCT'1US95/00481
-16-
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 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 memetics 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
CremaphorT"s or ethanol.
SUBSTITUTE SHEET (RULE 28)

WO 95118798 (~ ~ PCl'/US95/00481
-17-
0
&cheme 3
Nu
O'I
Ph' _NH OI' A O Nu
Ph~pn-...
OTES
HO H ~ O
Ogz OAc
I HN~~ HN~OH~ H ASH
Nu=
O
H~p~sn
z
1) TESOTf, 2,frlutidina, CHiCIZ
MFPT, 261utideia, Cl-LlClz I; T~ 2) MFPT, 2,61utidme, CHzCIz
Ph"NH OII ~ O I
Ph' v 'O".... C
OH
___ HO - H
Ogz OAc
Nu
N
SUBSTITUTE SHEET (RULE 28~

wo 95rts~9s 2 ~ $ ~ ~ ~ ~J rcTrus9siooast
. ,t~'.y~~h~-
Synthetic Methods
Preparation of 7-TES deacetylbaccatin III (4)
HO O OH _ H
HO~
~H~'O ~H
HO Ogz pAc HO Ogz OAc
10-deacetylbaccatin III (3)
7-TES deacetylbaccatin III (4). To a solution of 10-
deacetylbaccatin III (3, 3.0 g, 5.5I 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 X50% ethylacetate in petroleum ether) to
1 5 give alcohol 4 (3.39 g> 9190) as a white solid.
Physical Data for 7-TES deacetylbaccatin III (4). R f =
0.32 (silica, 50% ethylacetate in hexanes); IR (thin film) vmax 3464,
2954, 2282, 1710, 1453, 1362, 1271, 1242, 1105, 994 cm-1; ~ H NMR
(500 MHz, CDC13) & 8.06 (dd, J = 8.0, 0.9 Hz, 2 H, Bz), 7.57 (t, J = 7.9
2 0 Hz, 1 H, Bz), 7.44 (t, J = 7.9 Hz, 2 H, Bz), 5.56 (d, J = 7.0 Hz, 1 H, 2-
H),
5.14 (d, J = I .9 Hz, I H, 10-H), 4.92 (d, J = 9.5 Hz, 1 H, 5-H), 4.84-4.78
(m, 1 H, 13-H), 4.37 (dd, J = 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,
20-H), 3.91 (d, J = 7.0 Hz, 1 H, 3-H), 2.48-2.40 (m, 1 H, 6-H), 2.25 (s,
2 5 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, CDCl3) 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,
3 0 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 C35H5pO1oSi 791.2228.
SUBSTITUTE SHEET (RtiLf 2~

WO 95118798 ~ ~ PCTIU895I00481
r. ,
-19-
Preparation of enone 5
S ~ HO O OTES
Ho"..( K I i .-._
HO v H WO HO Y H WO
OBz OAc 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, 2.05
mmol) in CHZC12 (5 mL) was added 4 ~ molecular sieves (200 mg)
and the suspension was stirred at 25 °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 mixture was stirred at 25 °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. 95~'o by TLC. The
reaction mixture was filtered through silica gel, eluted with CHZC12
(100 mL), and concentrated to give enone 5 (1.44 g, 96%) as a white
solid.
Physical Data for Enone 5. R f = 0.5 (silica, 50%
ethylacetate in hexanes); IR (thin film) v",ax 3446, 2957, 2882, 1726,
1672, 1456, 1367, 1243, 1106 cm-1; 1H NMR (500 MHz, CDCI3) 8 8.05
(dd, J = 8.0, 1.0 Hz, 2 H, Bz), 7.61 (t, J = 7.5 Hz, 1 H, Bz), 7.45 (t, J =
7.5
Hz, 2 H, Bz), 5.63 (d, J = 7.5 Hz, 1 H, 2-H), 5.30 (d, J = 2.0 Hz, 1 H, 10-
H), 4.90 (d, J = 8.0 Hz, 1 H, 5-H), 4.36 (dd, J = 10.5, 7.0 Hz, 1 H, 7-H),
4.31 (d, J = 8.5 Hz, 1 H, 20-H), 4.30 (d, J = 2.0 Hz, 1 H, 10-OH), 4.11
2 5 (d, J = 8.5 Hz, I 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, I
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-OH), 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(CH2Cj~)3), 0.60-0.42 (band, 6 H,
3 0 Si(C~CH3)3); 13C NMR (125 MHz, CDC13) 8 208.2, 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,
SUBSTITUTE SHEET (RULE 2E3)

wo 9sns~9s rcrrus9srooast
:-ZO-
5.1; FAB HRMS (NBA / NaI) m / a 657.3070, M + Na+ calcd for
C35H4gO1pSi 657.3095.
Preparation of triol6
C)BZ vmc ....
5 6
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 NH4CI (150 mL) and
the resulting mixture was extracted with CH2C12 (2 x 200 mL). The
organic layer was dried (Na2S04), concentrated, and purified by flash
chromatography (silica, 35 ~ 50% ethylacetate in petroleum ether)
to give enone 5 (270 mg, 199'0) and triol 6 (912 mg, 93% based on
81% conversion).
Physical Data for Triol 6. R p = 0.24 (silica, 509'0
ethylacetate in hexanes); IR (thin film) vm8x 3414, 2957, 2881, 1727,
1664, 1370 cm-1; IH NMR (500 MHz, CDC13) E 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, 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 = 6.5, 4.0 Hz, 1 H, 2-H),
3.57 (d, J = 6.5 Hz, I H, 3-H), 2.78 (d, J = 19.5 Hz, 1 H, 14-H), 2.58 (d,
2 5 4.0 Hz, 1 H, 2-OH), 2.52 (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 H, Me), 0.90 (t, J = 8.0 Hz, 9 H,
Si(CH2C~3)3), 0.60-0.40 (band, 6 H, Si(C~2CH3)3); 13C NMR (125 MHz,
CDC13) b 208.9, 198.5, 170.1, 156.7, 138.8, 83.8, 81.2, 77.6, 75.7, 72.8,
3 0 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 / NaI) m / a 575.2648, M + Na+ calcd for
C28H4409Si 575.2652.
5U8STITUTE SHEET (RULE 28)

2~ 8~~9:5
WO 95118798 PCTIUS95100481
-21-
Preparation of Carbonate 7
HO O OTES
~H
OH OAc
6 O
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. 1N 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 (MgS04), and concentrated to give carbonate 7 (58 mg, 93°!0) as
a white foam.
Physical Data for Carbonate 7. R p = 0.50 (silica, 359b
ethylacetate in hexanes); IR (thin film) v,nax 3438, 2957, 2882, 1820,
1731, 1685, 1370, 1236 cm-i; iH NMR (500 MHz, CDC13) b 5.27 (d, J=
2.5 Hz, 1 H, 10-H), 4.89 (d, J = 9.0 Hz, I H, 5-H), 4.60 (d, J = 9.0 Hz, 1
2 0 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(CHZC$3)3), 0.55
- 0.45 (band, 6 H, Si(C~CH3)3); 13C NMR (125 MHz, CDCI3) 8 208.4,
195.5, I70.5, 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 C29H4201oS i
3 0 579.2626.
SUBSTITUTE SHEET (RULE 28)

w0 95118798 PGTIITS95/00481
. a;.,' , _22_
Preparation of n Butyl-C-2 ester derivative (Alcohol 8)
H
nBuLi
= O THF
li
OAc
O
O g
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 stirred for 1.0 hour The
reaction mixture was poured into a mixture of diethylether (10 mL)
and aqueous NH4C1 (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 a saturated solution of
brine (5 mL), dried (MgS04), concentrated, and purified by flash
chromatography (silica, 35 -.~ 50°lo ethylacetate in hexanes) to give 8
(7.9 mg, 72%) as an amorphous solid.
Physical Data for Alcohol 8. R f = 0.36 (silica, 35%
ethylacetate in petroleum ether); IR (film) vmax 3437, 2962, 2865,
1726, 1671, 1367, 1239, 1105 cm-1; 1H NMR (500 MHz, CDCI3) E 5.36
(d, J = 6.5 Hz, IH, 2-H), 5.26 (d, J = 2.5 Hz, IH, 10-H), 4.89 (br d, J =
2 0 8.0 Hz, 1H, 5-H), 4.47 (d, J = 8.0 Hz, 1H, 20-H), 4.32 (dd, J = 10.5, 6.5
Hz, I H, 7-H), 4.26 (d, J = 2.5 Hz, 1 H, 10-OH), 4.15 (d, J = 8.0 Hz, IH,
20-H), 3.81 (d, J = 6.5 Hz, 1 H, 3-H), 2.73 (d, J = 20.0 Hz, 1 H, 14-H),
2.57 (d, J = 20.0 Hz, 1 H, 14-H), 2.49-2.41 (m, 1 H, 6-H), 2.38-2.23 (m,
2H, OCCH2(CH2)2CH3), 2.06 (s, 3H, Me), 2.04 (s, 3H, Me), 1.90-1.82 (m,
IH, 6-H), 1.67 (s, IH, OH), 1.64 (s, 3H, Me), 1.68-I.52 (m, 2H,
OCCH2CH~CH2CH3), I.41-1.30 (m, 2H, OC(CH2)2C$,2CH3), I.19 (s, 3H,
Me), I.07 (s, 3 H, Me), 0.94-0.86 (band, 12H, CH3 of Bu,
OSi(CH2Cj~)3), 0.58-0.45 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA)
m / a 637.3421, M + H+ calcd for C33Hg2OlOSi 637.3408.
SUBSTITUTE SHEET (RULE 28)

W095l18798 ~~ PCT7US95/00481
-23-
Preparation of vinyl-C-2 ester derivative (Alcohols 9 and
10)
HO O OTES ~ HO O OTES
'~ O
. O or
~O H OAc ~ HO ~ H pAc O
O MgBr
O
HO O OTES
O 10
Alcohots 9 and 10. A solution of carbonate 7 (111.3
mg,
0.192 mmol) in tetrahydrofuran (2 mL) at -78 C was treated with
vinyllithium (3.7 mL of a 0.52 M solution i n 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 CHzCl2 (20 mL) and
aqueous NH4C1 (10
mL), the organic layer was separated, and the aqueous layer was
extracted with CH2Cl2 (3 x 10 mL). The combined
organic layer was
washed with brine (15 mL), dried (MgS04), concentrated, and
purified by flash chromatography (silica, 30 ~ 50% ethylacetate
in
petroleum ether) to give 9 (60.0 mg, 52!0), and 10 (25.7 mg, 24%
)
as white foams.
2 0 Physical Data for Alcohol 9. R f = 0.52 (silica, 50%
ethylacetate in hexanes); IR (film) vn,aa 3442, 2956, 2882, 1727,
1672, 1407, 1368, 1243, 1182, 1110, 1050, 986, 826, 736 cm-I;
IH
NMR (500 MHz, CDC13) b 6.51 (dd, J = 17.0, 1.0
Hz, IH, vinyl H), 6.13
(dd, J = 17.0, 10.5 Hz, IH, vinyl H), 6.00 (dd, = 10.5, 1.0 Hz,
J 1H,
2 S vinyl H), 5.45 (br d, J = 6.5 Hz, 1H, 2-H), J = 2.5 Hz, 1H,
5.30 (d, IO-
SUBSTITUTE SHEET (RULE 281

WO 95/18798 ~ PCTIUS9Sl00481
's
-24-
H), 4.91 (br d, J = 9.5 Hz, 1 H, 5-H), 4.44 (d, J = 8.5 Hz, 1 H, 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, IH, 20-H), 3.88 (d, J = 6.5 Hz, 1H, 3-H), 2.79 (d, J = 20.0 Hz,
1H, 14-H), 2.61 (d, J = 20.0 Hz, 1H, 14-H), 2.48 (ddd, J = 14.5, 9.5, 6.5
Hz, IH, 6-H), 2.09 (s, 3H, Me), 2.08 (s, 3H, Me), 1.89 (ddd, J = 14.5,
10.5, 2.0 Hz, 1H, 6-H), 1.72 (s, 1H, OH), 1.68 (s, 3H, Me), 1.22 (s, 3H,
Me), 1.12 (s, 3H, Me), 0.92 (t, J = 8.0 Hz, 9H, OSi(CHzCj~)3), 0.62-0.46
(band, 6H, OSi(C~CH3)3); FAB HRMS (NBA I CsI) m l a 739.1925, M
+ Cs+calcd for C31H460toSi 739.1915.
Physical Data for Alcohol lU. Rp = 0.24 (silica, 5090
ethylacetate in hexanes); IR (film) v~,aX 3439, 2955, 2881, 1711,
1671, I409, 1365, 1188, ills, 980, 833, 735 cm-1;1H NMR (500
MHz, CDC13) 8 6.48 (br d, J = 17.0 Hz, 1H, vinyl H), 6.10 (dd, J = 17.0,
10.5 Hz, IH, vinyl H), 5.97 (br d, J = 10.5 Hz, IH, vinyl H), 5.47 (br d,
1 5 J = 6.0 Hz, 1H, 2-H), 5.25 (d, J = 2.5 Hz, 1H, 10-H), 4.75 (dd, J = 9.5,
3.5 Hz, 1H, 5-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, 1H, 14-H), 3.24 (d, J = 6.0 Hz, IH, 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, Me), 1.13 (s, 3H,
Me), 0.90 (t, J = 8.0 Hz, 9H, OSi(CHZC~)3), 0.59-0.45 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NBA / CsI) m / a 697.1802, M + Cs+ calcd
for CZ9H4409Si 697.1809.
Preparation of 2-Furyt-C-2 ester derivative (Alcohol 11)
O Li HO O OTES
--~ ~, /
HO o H p c
~O~ O
11
SUBSTfTUTE SHEET (RULE 28j

W095118798 ~ PCT/US95/00481
-25-
Alcohol 11. A solution of carbonate 7 (46 mg, 0.0795 mmol)
in tetrahydrofuran (3 mL) at -78 °C was treated with 2-furyliithium
(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
CH2CI2 (15 mL) and aqueous NH4Cl (20 mL). The organic layer was
separated and the aqueous layer was extracted with CHZCIZ (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.
Physical Data for Alcohol 11. Rp = 0.38 (silica, 20%
ethylacetate in petroleum ether); IR (film) vmax 3442, 2956, 2882,
1727, 1672, 1468, 1300, 1240, 1110, 1007, 733 cm-i;iH NMR (S00
MHz, CDC13) & 7.66-7.64 (m, IH, furan), 7.24 (br d, J = 3.5 Hz, 1H,
furan), 6.58 (dd, J = 3.5, 1.5 Hz, 1H, furan), 5.55 (d, J = 6.5 Hz, 1H, 2-
H), 5.31 (d, J = 2.0 Hz, IH, 10-H), 4.92 (br d, J = 9.0 Hz, IH, 5-H), 4.43
(d, J = 8.5 Hz, 1 H, 20-H), 4.37 (dd, J = 10.5, 6.5 Hz, 1 H, 7-H), 4.32 (d, J
2 0 = 2.0 Hz, 1 H, 10-OH), 4.18 (d, J = 8.5 Hz, 1 H, 20-H), 3.93 (d, J = 6.5
Hz,
IH, 3-H), 2.88 (d, J = 20.0 Hz, 1H, 14-H), 2.63 (d, J = 20.0 Hz, IH, 14-
H), 2.55-2.37 (m, IH, 6-H), 2.15 (s, 3H, Me), 2.09 (s, 3H, Me), 1.93-
1.87 (m, IH, 6-H), 1.81 (s, IH, 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(CHZC~)3), 0.62-0.42
2 5 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA / NaI) m / a 669.2717, M
+ Na+calcd for C33H46O1iSi 669.2707.
Preparation of 2-thiophenyl-C-2 ester derivative (Alcohol
12)
H~ES S ~ HO O OTES
---~ ., /
j - H _~~ HO H
~O OAc
II S O OAc
O ~ ~ O
12
SUBSTITUTE SHEET (RULE 28)

WO 9SI18798 ~ ~ g ~ PCTIUS95100481
-26-
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) and stirred for 0.5
hour The reaction mixture was poured into a mixture of
diethylether (10 mL) and aqueous NH4CI (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 (MgS04), concentrated, and purified by
flash chromatography (silica, 10 -> 3596 ethylacetate in hexanes) to
give 7 (16.5 mg, 33%), 12 (36.8 mg, 96% based on 6796 conversion)
as an amorphous solid.
Physical Data for Alcohol 12. Rp = 0.56 (silica, 50°!0
ethylacetate in hexanes); IR (film) vmax 3403, 2945, 2881, 1717,
1669, 1520, 1413, 1360, 1248, 1078; IH NMR (500 MHz, CDCI3) 8
7.84 (dd, J = 3.5, L0 Hz, 1H, thiophene), 7.64 (d, J = 1.0, 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, IH> 20-H), 4.35 (dd, J = 10.5 Hz, 6.5 Hz,
1 H, 7-H), 4.29 (d, J = 2.5 Hz, 1 H, 10-OH), 4.19 (d, J = 8.5 Hz, 1 H, 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), 1.7I (s, 3H,
2 5 Me), 1.21 (s, 3H, Me), 1.13 (s, 3H, Me), 0.91 (t, J = 8.0 Hz, 9H,
OSi(CH2C~)3), 0.56-0.49 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA)
m l a 663.2655, M t H+ calcd for C33H46OIOSSi 663.2659.
SUBSTITUTE SHEET (RIiLE 2Ei~

wo 9s1t8798 ~ ~ ~ ~ /~ ~ ~ PCTlUS95r00481
-27-
Preparation of 3-thiophenyl-C-2 ester derivatives (Alcohol
13 and 14)
HO O
OTES
H =~"
i QAC
OAC
[S /~O
7 13
ES
HO H
.O fl
'S /~(O
14
Alcohols 13 and 14. A solution of carbonate 7 (107.9 mg,
0.186 mmoI) in tetrahydrofuran (6.2 mL) at -78 °C was treated with
3-thienyllithium (2.76 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. 1986, 27, 5155) and stirred
for 1.5 hour The reaction mixture was poured into a mixture of
CHZCIZ (15 mL) and aqueous NH4Cl (20 mL), the organic layer was
separated, and the aqueous layer was extracted with CHzCIz (2 x 10
mL). The combined organic layer was washed with brine (10 mL),
dried (Mg50q), concentrated, and purified by flash chromatography
(silica, 20 ~ 30% ethylacetate in hexanes) to give 7 (16.9 mg, 1696),
13 (87.0 mg, 8396 based on 84% conversion), and hydrolyzed C4
2 0 acetate 14 (C4-hydrolyzed side product, 9.7 mg, 10% based on 84%
conversion) as amorphous solids.
Physical Data for Alcohol 13. R f = 0.74 (silica, 50°l0
ethylacetate in hexanes), 0.4I (silica, 10% ethylacetate in benzene, 3
SUBSTITUTE SHEET (RULE 2B)

WO 95118798 PCT/US95100481
_2g_
i
elutions); IR (thin film) vmax 3442, 3110, 2956, 2882, 1725, 1672,
1410, 1368, 1244, 1198, 1101, 988, 825, 744 cm-I;1H NMR (500
MHz, CDC13) 8 8.I8 (dd, J = 3.0, 1.2 Hz, IH, thiophene), 7.54 (dd, J =
5.0, 1.2 Hz, 1H, thiophene), 7.37 (dd, J = 5.0, 3.0 Hz, IH, thiophene),
5.56 (dd, J = 6.5, I.0 Hz, IH, 2-H), 5.31 (d, J = 2.5 Hz, IH, 10-H), 4.92
(dd, J = 7.5, 2.0 Hz, IH, 5-H), 4.40-4.34 (m, 2H, 20-H, 7-H), 4.31 (d, J
= 2.5 Hz, IH, 10-OH), 4.15 (d, J = 8.5 Hz, 1H, 20-H), 3.93 (d, J = 6.5 Hz,
1 H, 3-H), 2.88 (d, J = 20 Hz, 1 H, 14-H), 2.63 (dd, J = 20.0, 1.0 Hz, 1 H,
14-H), 2.47 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, 6-H), 2.18 (s, 3H, Me), 2.10
(s, 3H, Me), 1.89 (ddd, J = 14.5, 10.5, 2.0 Hz, 1H, 6-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(CH2Cj33)3), 0.62-0.48 (band, 6H, Si(Cj~CH3)3); FAB HRMS
(NBA / CsI) m / a 795.1640, M + Cs+ calcd for C33H46OlOSSi
795.1635.
Physical Data for Alcohol 14: Rg = 0.54 (silica, 50°10
ethylacetate in hexanes); IR (thin film) vmax 3437, 3108, 2955, 2880,
1709, 1674, 1605, 1520, 1410, 1360, 1258, 1194, 1103, 1004, 829,
744 cm-I; 1H NMR (500 MHz, CDC13) 8 8.15 (dd, J = 3.0, I.0 Hz, IH,
thiophene), 7.49 (dd, J = 5.0, I.0 Hz, 1H, thiophene), 7.35 (dd, J = 5.0,
2 0 3.0 Hz, 1 H, 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,
1 H, 20-H), 4.32 (d, J = 2.5 Hz, 1 H, 10-OH), 4.15 (d, J = 8.5 Hz, 1 H, 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 = 6.0 Hz, 1 H, 3-H), 2.91 (br s, 1 H, OH), 2.61 (d, J = 19.5 Hz,
IH, 14-H), 2.38 (ddd, J = 14.5, 9.5, 6.0 Hz, 1H, 6-H), 2.09 (s, 3H, Me),
L99 (ddd, J = 14.5, 11.5, 3.5 Hz, 1H, 6-H), 1.81 (br s, 1H, OH), 1.65 (s,
3H, Me), I.24 (s, 3H, Me), 1.16 (s, 3H, Me), 0.91 (t, J = 8.0 Hz, 9H,
OSi(CH2CIj3)3), 0.60-0.46 (band, 6H, OSi(Cj~CH3)3); FAB HRMS (NBA
CsI) m l a 753.1530, M + Cs+ calcd for C31H44O9SSi 753.1530.
SUBSTITUTE SHEET (RULE 28j

WO 95118798 PCT/US95100451
-29-
Preparation of 2-pyridinyl-C-2 ester derivatives (Alcohol
15, 16 and trio! 6)
E S I Nw a
H _w
OAc O vwc
I N\
O
5
HO O OTES
HO O OTES
O
HO ~HH pA
16 6
10 Alcohol 15 and 16, and trio! 6. A solution of
carbonate 7 (62.6 mg, 0.108 mmol) in tetrahydrofuran (5.4 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, prepared from
2-bromopyridine and t-Butyllithium; methodology from Malmberg,
15 H.; Nilsson, M. Tetrahedron, 1986, 42, 3981) and stirred for 1.3 hour
The reaction mixture was poured into a mixture of ethylacetate ( 10
mL) and aqueous NH4C1 (5 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 ~ 100% ethylacetate in petroleum ether) to give 6 (16.3 mg,
27°Jo), 15 (28.0 mg, 399'0), and 16 (8.4 mg, 1386) as amorphous solids.
SUBSTITUTE SHEET (RULE 28)

WO 95118798 2 ~ ~ PCl'IU595100481
-30-
Physical Data for Alcohol 15. Rp - 0.60 (silica,
ethylacetate); 1 H NMR (500 MHz, CDCI3) b 8.77 (ddd, J = 4.5, 1.7, 1.0
Hz, IH, pyridine), 8.05 (br d, J = 7.5 Hz, IH, pyridine), 7.89 (ddd, J =
7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J = 7.5, 4.5, 1.0 Hz, IH,
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, 1 H, 20-H), 3.96 (d, J = 6.5 Hz, 1 H, 3-H), 2.98
(d, J = 20.0 Hz, IH, 14-H), 2.71 (dd, J = 20.0, 1.0 Hz, 1H, 14-H), 2.50
1 0 (s, IH, 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, IH, 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,
OSi(CH2C~3)3), 0.63-0.47 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA /
CsI) m / a 790.2060, M + Cst calcd for C34H47O1oNSi 790.2024.
Physical Data for Alcohol 16. Rg - 0.45 (silica,
ethylacetate); IR (film) vmaX 3435, 2954, 2879, 1732, 1674, 1589,
1362, 1305, 1241, 1116, 998, 829, 741 cm-t; IH NMR (500 MHz,
CDC13) 8 8.73 (br d, J = 4.5 Hz, IH, pyridine), 8.15 (br d, J = 7.5 Hz, IH,
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, IH, pyridine), 5.53 (dd, J = 7.5, I.O, IH, 2-H), 5.30 (d,
J = 2.5 Hz, 1 H, 10-H), 4.84 (dd, J = 9.5, 3.0 Hz, I H, 5-H), 4.81 (br s, 1 H,
OH), 4.31 (d, J = 2.5 Hz, IH, 10-OH), 4.25 (s, 2H, 20-CHZ), 3.97 (dd, J =
1 I .5, 6.5 Hz, 1 H, 7-H), 3.31 (d, J = 19.5 Hz, I H, 14-H), 3.23 (d, J = 7.5
Hz, IH, 3-H), 2.57 (br d, J = 19.5 Hz, IH, 14-H), 2.43 (ddd, J = 14.5,
2 5 9.5, 6.5 Hz, IH, 6-H), 2.11 (s, 3H, Me), 1.95 (ddd, J = 14.5, 11.5, 3.0
Hz, IH, 6-H), 1.92 (br s, IH, OH), 1.70 (s, 3H, Me), 1.24 (s, 3H, Me),
1.I7 (s, 3H, Me), 0.91 (t, J = 8.0 Hz, 9H, OSi(CHZC~)3), 0.60-0.46
(band, 6H, OSi(C~CH3)3).
Physical Data for Triol 6. Rp = 0.24 (silica, 50% ethylacetate
3 0 in hexanes); IR (thin film) v",aX 3414, 2957, 2881, 1727, 1664, 1370
cm-1; iH 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, 5-H), 4.63 (d, J = 9.5 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 = 6.5, 4.0 Hz, 1 H, 2-H), 3.57 (d, J = 6.5 Hz, 1
3 5 H, 3-H), 2.78 (d, J = 19.5 Hz, 1 H, 14-H), 2.58 (d, 4.0 Hz, I H, 2-OH),
2.52 (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, I H, 6-H), 1.68 (s, 3 H, Me), 1.21 (s, 3 H, Me), 1.04
SUBSTITUTE SHEET (RtiLE 28j

WO 95118798 ~ ~ PCTIUS95100481
-31-
(s, 3 H, Me), 0.90 (t, J= 8.0 Hz, 9 H, Si(CHZC -~I )3), 0.60-0.40 (band, 6
H, Si(C~CH3)3); 13C NMR (125 MHz, CDC13) & 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 / NaI) m /
a 575.2648, M + Na+ calcd for CZgH440gSi 575.2652.
Preparation of 3-pyridinyl-C-2 ester derivative (Alcohol
17)
HO O OTES
a
o ---.,
O OAc
O
1 0 7 O 17
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 (methodology from Parham, W.E.;
Piccirilli,
R. M. J. Org. Chem. 1977, 42, 257), was added a solution of
carbonate 7 (133.1 mg, 0.230 mmol) in tetrahydrofuran (2 mI,) via
cannula. The resulting solution was stirred for 1 h, allowed to warm
2 0 to -78 °C, stirred for 1 h, and poured into a mixture of
ethylacetate
(10 mL) and aqueous NH4Cl (10 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 (10
mL), dried (MgS04), concentrated, and purified by flash
2 5 chromatography (silica, 70 -> 95% ethylacetate in petroleum ether)
to give 7 (64.8 mg, 49%) and 17 (43.9 mg, 57~o based on 51 %
conversion) as an amorphous solid.
Physical Data for Alcohol 17. R f - 0.56 (silica,
ethylacetate); IR (film) vmax 3435, 2956, 2882, 1731, 1671, 1592,
30 1366, 1280, 1240, 1109, 991, 824, 739 cm-~; 1H NMR (500 MHz,
SUBSTITUTE SHEET (RIiLE 2B)

R'O 95118798 2 PCT/US95/00481
-32-
CDC13) b 9.24 (br s, 1H, pyrsdine), 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), 5.66 (d, J = 6.5 Hz, 1H, 2-H), 5.32 (s, 1H,
10-H), 4.92 (dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.38 (dd, J = 10.5, 6.5 Hz,
IH, 7-H), 4.32 (br s, IH, OH), 4.30 (d, J = 8.5 Hz, 1H, 20-H), 4.13 (d, J
= 8.5 Hz, IH, 20-H), 3.96 (d, J = 6.5 Hz, 1H, 3-H), 2.92 (d, J = 19.5 Hz,
1H, 14-H), 2.66 (d, J = 19.5 Hz, 1H, 14-H), 2.48 (ddd, J = 15.5, 9.5, 6.5
Hz, 1H, 6-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,
1 0 Me), 1.16 (s, 3H, Me), 0.92 (t, J = 8.0 Hz, 9H, OSi(CHZCj~)3), 0.62-0.48
(band, 6H, OSi(CHZCH3)3); FAB HRMS (NBA / CSI) m / a 790.2030, M
+ Cs+calcd for C34H4701pNSi 790.2024.
Preparation of 4-N, N-dimethylaniline-C-2 ester derivative
(Alcohol 18)
Me2N ~ ~ ~ ~~ ~ ES
H =~
OAC
18
Alcohol 18. A solution of carbonate 7 (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
2 5 minutesutes The reaction mixture was poured into a mixture of
CHzCl2 (35 mL) and aqueous NH4C1 (20 mL), the organic layer was
separated, and the aqueous layer was extracted with CHZC12 (2 x 20
mL). The combined organic layer was washed with brine (20 mL),
dried (MgS04), concentrated, and purified by flash chromatography
3 0 (silica, 10 -> 35% ethylacetate in petroleum ether) to give 18 (55.0
mg, 30~Yo) as an amorphous solid.
SUBSTITUTE SHEET (RULE 261

WO 95118798 PC17US95/00481
-33-
Physical Data for Alcohol 18. R f = 0.26 (silica, 35~Yo
ethylacetate in hexanes); IR (film) vmax 3414, 2924, 1706, 1669,
1605, 1530, I094; I H NMR (500 MHz, CDC13 ) 8 7.90 (d, J = 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
S (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, 1 H, 20-H), 4.36 (dd, J = 10.5, 6.5 Hz, 1 H, 7-H), 4.31 (d, J = 2.5
Hz, IH, 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, NMeg), 2.93 (s, IH, 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(CH2Cjj3)3), 0.56-0.49 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA /
NaI) m / a 722.3354, M + Na+ calcd for C37Hg3OlONSi 722.3336.
Preparation of 1-naphthalene-C-2 ester derivative (Alcohol
19)
HO O OTES
i
/ HO v H
I O
O
19
2 0 Alcohol 19. A solution of carbonate 7 (47 rng, 0.0812 mmol)
in tetrahydrofuran (2 mL) at -78 °C was treated with I-
lithionaphthalene (6.3 mL of a 0.32 M solution in diethylether, 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 into a mixture of
CHZC12 (15 mL) and aqueous NH4CI (20 mL), the organic Iayer was
separated, and the aqueous layer was extracted with CHZC12 (2 x 10
mL). The combined organic layer was washed with brine (10 mL),
SUBSTITUTE SHEET (RULE 28)

w0 95!18798 2 i g PCT/US95100481
-34-
dried (MgS04) 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 (20% ethylacetate
in petroleum ether); IR (film) vn,ax 3442, 2954, 2882, 1724, 1671,
1461, 1362, 1279, 1228, 1195, 1092, 987, 826, 736 cm-I; 1H NMR
(500 MHz, CDCl3) 8 8.66 (s, IH, naphthalene), 8.07 {dd, J = 9.0, 2.0 Hz,
IH, naphthalene), 7.97-7.89 (m, 3H, naphthalene), 7.68-7.57 (m, 2H,
naphthalene), 5.71 (br d, J = 6.5 Hz, IH, 2-H), 5.35 (d, J = 2.5 Hz, IH,
I O-H), 4.94 (br d, J = 8.0 Hz, I H, 5-H), 4.41 (dd, J = I 1.0, 7.0 Hz, 1 H,
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, 1 H, 20-H), 4.00 (d, J = 6.5 Hz, 1 H, 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, IH,
6-H), 2.27 (s, 3H, Me), 2.13 (s, 3H, Me), 1.94-1.87 (m, 1H, 6-H), 1.86
(s, IH, OH), 1.75 (s, 3H, Me), 1.25 (s, 3H, Me), 1.20 (s, 3H, Me), 0.94 (t,
1 5 J = 8.0 Hz, 9H, OSi(CH2Cj~)3), 0.63-0.49 (band, 6H, OSi(C~CH3)3); FAB
HRMS (NBA) m / a 707.3270, M + H+ calcd for C39H5oO loci 707.3252.
Preparation of phenyl acetylide-C-2 ester derivative
(Alcohol 20)
HO O OTES
O
0 0
\'p H OAc
O
v
7 20
Alcohol 20. A solution of carbonate 7 (5.0 mg, 0.00864
mmol) in tetrahydrofuran (0.5 mL) at -78 °C was treated with
2 5 lithium phenylacetylide from Aldrich Chemical Company Inc. (0.13
mL of a 1.0 M solution in tetrahydrofuran, 0.13 mmol) and stirred
for 0.5 hour The reaction mixture was treated with aqueous NH4C 1
(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
SUBSTITUTE SHEET (RULE 28)

w0 95118798 ~ ~ PCTIUS95100481
-35-
concentrated to give a 9 , : . ] mixture of carbonate 7 and alcohol 2 0
(5.0 mg, 95%) as a film.
Physical Data for Alcohol 20. R f = 0.59 (50% ethylacetate
in hexanes); 1H NMR (300 MHz, CDCI3) b 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, IH,
10-H), 4.90 (br d, J = 7.5 Hz, IH, 5-H), 4.67 (d, J = 8.5 Hz, 1H, 20-H),
4.44 (d, J = 8.5 Hz, 1 H, 20-H), 4.37-4.30 (m, 1 H, 7-H), 4.28 (d, J = 2.5
Hz, IH, 10-OH), 3.88 (d, J = 6.5 Hz, IH, 3-H), 2.85 (d, J = 20.2 Hz, IH,
I4-H), 2.63 (d, J = 20.2 Hz, IH, 14-H), 2.55-2.47 (m, 1 H, 6-H), 2.I1
1 0 (s, 3 H, OAc), 2.08 (s, 3 H, 18-Me), 1.94-1.85 (m, I H, 6-H), 1.67 (s, 3
H, Me), 1.41 (s, 3 H, Me), 1.21 (s, 3 H, Me), 0.91 (t, J = 8.0 Hz, 9H,
OSi(CHZC~)3), 0.59-0.42 (band, 6H, OSi(C~CH3)3).
Preparation of Hydroxycarbamate-C-2 ester derivative
(Alcohol 21)
H~ES H~~ES
~ NH \~~V~~Z
./ H ~~O HBO '~H
O O=Arc
N
H O
7
21
Alcohol 21. A solution of carbonate 7 (5 .0 mg, 0.00864
mmol) in MeOH (0.5 mL) at 25 °C was treated with n-Butyl-NH2 from
Aldrich Chemical Company dnc. (0.05 mL, 0.506 mmol) and stirred
for 10 minutes. The reaction mixture was concentrated and purified
by flash chromatography (silica, 30 -~ 50% ethylacetate in petroleum
ether) to give 21 (5.2 mg, 92%) as an amorphous solid.
Physical Data for Alcohol 21. R f = 0.13 (silica, 30%
ethylacetate in petroleum ether); IR (film) vmax 3434, 2957, 2881,
1711, 1671, 1368, 1243, 1108, 987, 829 cm-1; IH NMR (500 MHz,
CD C 13 ) 8 5.27 (d, J = 2.0 Hz, 1 H, 10-H), 5.23 (d, J = 6.5 Hz, 1 H, 2-H),
4.91 (br d, J = 8.0 Hz, IH, 5-H), 4.79 (t, J = 6.0 Hz, 1H, NH), 4.47 (d, J =
3 0 8.5 Hz, 1H, 20-H), 4.34 (dd, J = 11.0, 7.0 Hz, 1H, 7-H), 4.30 (d, J = 2.5
Hz, 1H, 10-OH), 4.28 (d, J = 8.5 Hz, 1H, 20-H), 3.78 (d, J = 6.5 Hz, 1H,
SUBSTITUTE SHEET (RULE 2B~

R'O 95II879S ~, PCTlU595100481
-36-
3-H), 3.29-3.I2 (m, 2H, N~ICj~), 2.70 (d, J = 20.0 Hz, IH, 14-H), 2.60
(d, J = 20.0 Hz, IH, 14-H), 2.51-2.42 (m, IH, 6-H), 2.24 (s> iH, OH),
2.06 (s> 3H, Me), 2.05 (s, 3H, Me), 1.94-1.86 (m, IH, 6-H), 1.69 (s, 3H,
Me), 1.55-1.46 (m, 2H, NHCHZCj~), 1.40-1.30 (m, 2H, NHCHZCHZCj~),
1.21 (s, 3H, Me), 1.09 (s, 3H, Me), 0.95-0.80 (m, 12H, CH3 of Bu,
OSi(CHgC~3)3), 0.61-0.47 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA /
NaI) m / a 674.3336, M + Na+ calcd for C33H5301oNSi 674.3336.
Preparation of NN-methyl-phenyl-hydroxycarbamate-C-Z
ester derivative (Alcohol 22)
HO O OTES
LiM
Y
_ HO ~ H~
~ N--
M a O 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 (5 mL) and aqueous NH4CI (5 mL), the
2 0 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 (MgS04), concentrated, and purified
by flash chromatography (silica, 15 -~ 3596 ethylacetate in hexanes)
to give 7 (2.5 mg, 50%) and 22 (2.8 mg, 93% based on 50%
2 5 conversion) as an amorphous solid.
Physical Data for Alcohol 22. R f = 0.22 (silica 35%
ethylacetate in petroleum ether); iH NMR (500 MHz, CDCl3) 8 7.45-
7.18 (band, SH), 5.25 (br d, J = 6.5 Hz, 1H, 2-H), 5.20 (d, J = 2.5 Hz,
IH, 10-H), 4.70 (br d, J = 8.0 Hz, IH, 5-H), 4.26 (d, J = 2.5 Hz, IH, 10-
3 0 OH), 4.22 (dd, J = 10.5, 6.5 Hz, IH, 7-H), 4.19 (d, J = 8.5 Hz, IH, 20-H),
4.16 (d, J = 8.5 Hz, IH, 20-H), 3.58 (d, J = 7.0 Hz, 1H, 3-H), 3.27 (s, 3H,
SUBSTITUTE SHEET (RULE 28)

w0 95118798 PCT/US95I00481
-37-
MeN), 2.52 (d, J = 20.0 Hz, IH; 14=H), (d, J = Hz, IH, 14-H),
2.35 20.0
2.40-2.31 (m, 1H, 6-H), 2.03 (s, IH, OH), 1.97 (s, Me),1.85-L76
3H,
(m, 1H, 6-H), I.66 (s, 3H, Me), 1.57 (s, Me), 1.18 3H, Me),
3H, (s, 1.08
(s, 3H, Me), 0.87 (t, J = 8.0 Hz, 9H, OSi(CH2C~)3), (band,
0.55-0.43 6H,
OSi(Cjj2CH3)3); FAB HRMS (NBA) m/ a 686.3358, + calcd
M H+ for
C36H51010NSi 686.3361.
Preparation of Thioether-C-2 ester derivative (Alcohol 23)
H
~O vac HO p H OA
SPh
1 0 23
Alcohol 23. A solution of vinyl ester 9 (55.6 mg, 0.0916
mmol) and 4-dimethylaminutesopyridine from Aldrich Chemical
Company Inc. (DMAP, 1.8 mg, 0.0147 mmol) in CHZC12 (4.3 mL) at 25
°C was treated with PhSH from Aldrich Chemical Company Inc.
(0.030 mL, 0.292 nl) and stirred for I.5 hour The reaction
mixture was concentrated and purified by flash chromatography
(silica, 30% ethylacetate in petroleum ether) to give 23 (58.1, 88%)
as a white solid.
2 0 Physical Data for Alcohol 23. R p = 0.37 (silica, 30%
ethylacetate in hexanes), 0.34 (10% ethylacetate in PhH, 2 elutions);
IR (film) v,nax 3441, 3057, 2956, 2883, 1732, 1672, 1600, 1367,
1238, 1111, 988, 825, 739 cm -1; IH NMR (500 MHz, CDCl3) 8 7.39-
7.24 (band, 5H), 5.44 (d, J = 6.5 Hz, IH, 2-H), 5.28 (d, J = 2.5 Hz, 1H,
10-H), 4.90 (dd, J = 9.5, 2.0 Hz, IH, 5-H), 4.38 (d, J = 8.0 Hz, IH, 20-
H), 4.33 (dd, J = 10.5, 6.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), 2.76 (d, J = 19.5 Hz, IH, 14-H), 2.72-2.58 (m, 3H,
14-H, CH2C$,2SPh), 2.47 (ddd, J = 14.5, 9.5, 6.5 Hz, IH, 6-H), 2.39 (s,
3 0 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), L23 (s, 3H, Me), 1.12 (s, 3H, Me),
SUBSTITUTE SHEET (RULE 2B1

R'O 95/18798 ~ PCTIUS95J00481
-38-
0.92 (t, J = 8.0 Hz, 9H, OSi(CH2C -~I3)3), 0.61-0.47 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NB~i / CsI) m / a 849.2085, M + Cs+ calcd
for C3~H5201oSSi 849.2105. '
S Preparation of intermediates 25-27 and 2-furanyl-C-2-
taxoid (28)
OTES
a
p O H O~O O O vwc
TESO~ Ph
O y ~ ~ ~ O 25
N
O 24 BZ ~b
Ac0 O OR wcu a OTES
BzNH Q
Ph~II 0.... ~ HO....
OR = . = . O
OHO ~ H OAc O o O O H OAc
27 : R = TES d ~ . ~ O 26
28: R=H ~ v
Acetate 25. A solution of alcohol 11 and 4-
dimethylaminopyridine (DMAP, 100 mg, 0.819 mmoI) in CH2CI2 (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
CH2CIg (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 CHzCl2 (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgS04),
concentrated, and purified by preparative TLC (silica, 1090
ethylacetate in benzene, 3 eIutions) to give 25 (36 mg, 66% from
carbonate 7) as a white foam.
Physical Data for Acetate 25. R f = 0.38 (20% ethylacetate
in petroleum ether); IR (film) vmax 3509, 2956, 2881, 1727, 1674,
SUBSTITUTE SHEET (RULE 28~

218445
w0 95118798 PC1'/US95100481
-39-
1469, 1371, 1299, 1227, 1108, 746 cm-1; IH NMR (500 MHz, CDC13) &
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), 5.59 (d, J = 6.5 Hz, 1H, 2-H), 4.92 (br d, J = 7.5
Hz, 1H, 5-H), 4.46 (dd, J = 10.5, 6.5 Hz, IH, 7-H), 4.42 (d, J = 8.5 Hz,
1H, 20-H), 4.16 (d, J = 8.5 Hz, 1 H, 20-H), 3.87 (d, J = 6.5 Hz, 1 H, 3-H),
2.89 (d, J = 20.0 Hz, IH, 14-H), 2.63 (d, J = 20.0 Hz, IH, 14-H), 2.59-
2.48 (m, IH, 6-H), 2.22 (s, 3H, Me), 2.17 (s, 3H, Me), 2.14 (s, 3H, Me),
1.90-1.83 (m, IH, 6-H), I.65 (s, 3H, Me), 1.25 (s, 3H, Me), 1.18 (s, 3H,
Me), 0.91 (t, J = 8.0 Hz, 9H, OSi(CH2Cjj~)3), 0.64-0.52 (band, 6H,
1 0 OSi(C$,2CH3)3); FAB HRMS (NBA / CsI) m / a 821.1966, M + Cs+ calcd
for C3gH4gO12Si 821.1969.
Alcohol 26. A solution of enone 25 (36 mg, 0.0523 mmol) in
MeOH (3 mL) containing two drops of CH3COOH at 0 °C was treated
1 S with NaBH4 (200 mg, 5.29 mmol, added by 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
CH2C12 (2 x 10 mL). The combined organic layer was washed with
20 brine (5 mL), dried (MgS04), concentrated, and purified by
preparative TLC (silica, 50% ethylacetate in petroleum ether) to give
26 (30 mg, 83°k ) as an amorphous solid.
Physical Data for Alcohol 26. R f = 0.42 (silica, 50%
ethylacetate in petroleum ether); 1H NMR (300 MHz, CDC13) b 7.62
25 (br s, IH, furan), 7.25 (d, J = 3.5 Hz, IH, furan), 6.58 (d, J = 3.5 Hz,
IH,
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, I H, 13-H), 4.48 (dd, J = 10.5, 7.5 Hz,
1H, 7-H), 4.38 (d, J = 8.0 Hz, IH, 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
3 0 (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(CH2C j~3 )3 ), 0.65-0.50 (band, 6H,
OSi(C~CH3)3)~
35 DiTES taxoid 27. To a solution of alcohol 26 (30.0 mg,
0.0434 mmol, previously azeotroped twice with benzene) and [3-
lactam 24 (28.0 mg, 0.0734 mmol, previously azeotroped twice with
SUBSTITUTE SHEET (RULE 281

w0 95!18798 PCl'/US95I00481
-40-
benzene) in THF (2 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, L; Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.;
Brigaud, T. Tetrahedron 1992, 48, 6985-7012), at 0 °C was added
N aN ( S i M a 3 )g (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 CHZCIZ (10 mL) and aqueous NH4CI (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 (MgS04), concentrated, and purified by
preparative TLC (silica, 60% ethylacetate in petroleum ether) to give
27 (12 mg, 26%) 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 (I 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
2 0 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
(MgS04), concentrated, and purified by preparative TLC (silica, 60%
ethylacetate in petroleum ether) to give 28 (3 mg, 640) as a
2 5 colorless film.
Physical Data for Taxoid 28. Rp = 0.1 (50% ethylacetate
in petroleum ether); IR (film) vmax 3383, 2933, 2898, 1729, 1649,
1519, 1242, 1110, 1071 cm-I; 1H NMR (500 MHz, CDC13) b 7.77-7.73
(m, 2H), 7.68-7.66 (m, IH, furan), 7.55-7.33 (band, 9H), 6.98 (d, J =
30 9.0 Hz, 1H, NH), 6.58 (dd, J = 3.5, 1.5 Hz, IH, 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, 1 H, 6-H), 2.47 (d, J = 4.0
35 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, IH, 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).
SUBSTITUFE SHEET (RULE 28)

wo vsns~9s 2 ~ ~ ~ ~ ~ ~ 8crnrs9srooast
-41-
Preparation of 2-thiophenyl-C-2 taxol (32)
-OTES
a
S vEac
____ ' ~ O 29
TESO~ Ph
12 N~
O 24 BZ ,~ b
s~cv a OTES
BzNH O
Ph~~ . HO~~
OR ' O
SHO H OAc
O 30
Acetate 29. A solution of alcohol 12 (36.0 mg, 0.0543 mmol)
and 4-dimethylaminopyridine (DMAP, 33.0 mg, 0.270 mmol) in
CHZCI2 (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 CHZC12 (10 mL), treated with aqueous NaHC03 (7 mL), and
stirred vigorously for 0.5 h. 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 (5 mL), dried
(MgSOø), concentrated, and purified by flash chromatography (silica,
10 -~ 35R6 ethylacetate in hexanes) to give 29 (29.5 mg, 77%) as an
amorphous solid.
Physical Data for Acetate 29. Rf = 0.56 (silica, 50%
ethylacetate in petroleum ether); IR (film) vmax 3457, 2956, 1712,
2 0 1669, 1525, 1413, 1376, 1264, 1227, 1073; 1H NMR (500 MHz,
CDC13) 8 7.84 (dd, J = 4.0, 1.5 Hz, 1H, thiophene), 7.63 (dd, J = 5.0, 1.5
Hz, 1H, thiophene), 7.13 (dd, J = 5.0, 4.0 Hz, 1H, thiophene), 6.56 (s,
SUBSTITUTE SHEET (RULE 28)

w0 95118798 ~ ~ PCTIU595/00481
-42-
IH, 10-H), 5.58 (br d, J = 6.5 Hz, 1H, 2-H), 4.90 (br d, J = 8.0 Hz, 1H,
5-H), 4.44 (dd, J = 10.5, 7.0 Hz, 1H, 7-H), 4.42 (d, J = 8.5 Hz, IH, 20-
H), 4.18 (d, J = 8.5 Hz, IH, 20-H), 3.85 (d, J = 6.5 Hz, IH, 3-H), 2.91 (d,
J = 19.5 Hz, IH, 14-H), 2.64 (dd, J = 19.5, 1.0 Hz, IH, 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, IH, 6-H), 1.65 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me),
0.88 (t, J = 8.0 Hz, 9H, OSi(CHZC~3)3), 0.59-0.53 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NBA / Csi) m l a 837.1736, M + Cs+ calcd
for C35H4gOlISSi 837.1741.
Alcohol 30. A solution of enone 29 (29.0 mg, 0.0411 mmol)
in MeOH (5 mL) at 0 °C was treated with NaBH4 (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
1 5 N H 4CI (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 (5 mL),
dried (MgS04), concentrated, and purified by flash chromatography
(silica, 25 --> 50% ethylacetate in petroleum ether) to give 29 (4.0
mg, 14%) and 30 (14.7 mg, 599'o based on 86°6 conversion) as an
amorphous solid.
Physical Data for Alcohol 30. R f = 0.34 (silica, SORE
ethylacetate in petroleum ether); IR (film) vmax 3478, 2946, 2892,
1717, 1520, 1365, 1238, 1083; IH NMR (500 MHz, CDCI3) & 7.85 (dd,
2 5 J = 3.5, 1.5 Hz, 1 H, thiophene), 7.61 (dd, J = 5.0, 1.5 Hz, I H,
thiophene), 7.12 (dd, J = 5.0, 3.5 Hz, IH, ttniophene), 6.43 (s, IH, 10-
H), 5.51 (d, J = 7.0 Hz, 1H, 2-H}, 4.94 (br d, J = 7.5 Hz, IH, 5-H), 4.83-
4.77 (m, 1H, 13-H), 4.45 (dd, J = 10.5, 7.5 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,
3 0 IH, 3-H), 2.55-2.48 (m, IH, 6-H), 2.24 (s, 3H, Me), 2.26-2.21 (m, 2H,
14-CHZ), 2.16 (d, J = 1.0 Hz, 3H, 18-Me), 2.15 (s> 3H, Me), 2.00 (d, J =
S.0 Hz, 1H, OH), 1.90-1.82 (m, 1H, 6-H), 1.66 (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,
OSiCH2C~)3), 0.59-0.55 (band, 6H, OSi(CHZCH3)3); FAB HRMS (NBA
3 S CsI) m / a 839.1893, M + Cs+ calcd for C35HSpO 11 SSi 839.1897.
SUBSTITUTE SHEET (RULE 28<

WO 95118798 ~ ~ PCT/US95/00481
-43-
DiTES taxoid . 31. To a solution of alcohol 30 (14.5 mg,
0.0205 mmol, previously azeotroped twice with benzene) and (3-
lactam 24 (16.0 mg, 0.0420 mmol, previously azeotroped twice with
benzene) in THF (L0 mL), prepared from the Ojima-Holton protocol
S (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, L; Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.;
Brigaud, T. Tetrahedron 1992,48, 6985-7012), at -78 °C was added
NaN(SiMe3)2 (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 NH4Cl (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 (5 mL), dried (MgS04), concentrated, and
purified by flash chromatography (silica, 10 -~ 35°lo ethylacetate in
hexanes) followed by preparative TLC (silica, IS% ethylacetate in
benzene) to give 30 (3.0 mg, 21%) and 31 (7.6 mg, 43% based on
79% conversion) as a white solid.
Physical Data for DiTES taxoid 31. R g = 0.48 (silica, 50%
ethylacetate in hexanes); IR (film) vm~ 3382, 2913, 2850, 1722,
1653,1461,1243,1083, 1014; 1H NMR (500 MHz, CDC13) b 7.90 (br d, J
= 4.0 Hz, IH, thiophene), 7.74 (d, J = 8.0 Hz, 2H, NBz), 7.62 (br d, J
=5.0 Hz, 1H, thiophene), 7.48 (t, J = 7.0 Hz, IH, Ar), 7.42-7.28 (band,
7H, Ar), 7.14 (dd, J = 5.0, 4.0 Hz, IH, thiophene), 7.10 (d, J = 9.0 Hz,
2 5 1 H, NH), 6.42 (s, 1 H, 10-H), 6.20 (br t, J = 9.0 Hz, 1 H, 13-H), 5.65
(br
d, J = 9.0 Hz, IH, 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 = 1.5 Hz, IH, 2'-H), 4.44 (dd, J = 11.0, 6.5 Hz,
IH, 7-H), 4.43 (d, J = 8.5 Hz, 1H, 20-H), 4.26 (d, J = 8.5 Hz, IH, 20-H),
3.77 (d, J = 7.0 Hz, 1H, 3-H), 2.51 (s, 3H, Me), 2.54-2.47 (m, 1H, 6-H),
3 0 2.34 (dd, J = 15.0, 9.5 Hz, IH, 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,
IH, 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(CHzC~j3)3), 0.79 (t, J = 8.0 Hz, 9H, Si(CH2C~3)3), 0.57-
0.55 (band, 6H, Si(C -j~I CH3)3), 0.45-0.40 (band, 6H, Si(C,~CH3)3); FAB
3 S HRMS (NBA / CsI) m / a 1220.3685 M + Cs+ calcd for CS~H~~014NSSi2
1220.3658.
SUBSTITUTE SHEET (RULE 281

WO 95118798 ~ ~ ~ ~ PCTIUS95100481
-44-
Taxoid 32. A solution 3 of ; silyl ether 31 (7.5 mg, 0.00689
mmol) in THF (0.8 mL) at 25 °~ was treated with HF~pyridine (0.150
mL) and stirred for 1 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 IO mL). The combined organic layer was washed, with brine (5
mL), dried (MgS04), concentrated, and purified by flash
chromatography (silica, 50 -> 100% ethylacetate in petroleum ether)
1 0 to give 32 (4.2 mg, 7lgb) as a colorless film.
Physical Data for Taxoid 32. R f = 0.44 (silica, 7590
ethylacetate in petroleum ether); IR (film) vmaX 3417, 2929, 1716,
1649, 1521, 1460, 1417, 1368, I247, 1076; IH 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, IH, thiophene), 7.51-7.32 (band,
8H, Ar), 7.14 (dd, J = 5.0, 4.0 Hz, IH, thiophene), 6.96 (d, J = 9.0 Hz,
1H, NH), 6.24 (s> 1H, 10-H), 6.19 (br t, J = 9.0 Hz, 1H, 13-H), 5.75 (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), 4.76 (dd, J = 5.0, 2.5 Hz, 1H, 2'-H), 4.41 (d, J = 8.5
2 0 Hz, 1 H, 20-H), 4.40-4.33 (m, 1 H, 7-H), 4.24 (d, J = 8.5 Hz, 1 H, 20-H),
3.73 (d, J = 7.0 Hz, 1H, 3-H), 3.52 (d, J = 5.0 Hz, IH, 2'-OH), 2.58-2.49
(m, 1H, 6-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-CHZ), 2.22 (s, 3H, Me), 1.91-1.83 (m, 1H, 6-H), 1.76
(s, 3H, Me), 1.66 (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 C45H~9N014S
992.1928.
SUBSTITUTE SHEET (RULE 2B)

w0 95118798 PCT/US95100481
-45-
Preparation of 3-thiophenyl-C-2 taxol (36)
a
TESO~ ~Ph ~O v.
S ~ O 33
13 O 24 Z ~ b
wcu a pR
BzNH O
c
ES
O R H~H~'O
O OAc ~ p OAc
i ~
35 : R = TES~ S~ 34
O 36:R=H ~d O
Acetate 33. A solution of alcohol 13 (68.4 mg, 0.103 mmol)
and 4-dimethylaminopyridine (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 CH2CI2 (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 CHgCIZ (2 x 10 mL). The
combined organic layer was washed with brine (5 mL), dried
(MgS04), concentrated, and purified by flash chromatography (silica,
30% ethylacetate in hexanes) to give 33 (66.0 mg, 91 °k) as an
amorphous solid.
Physical Data for Acetate 33. R f = 0.48 (silica, 10%
ethylacetate in benzene, 3 elutions); IR (film) va,ax 3518, 2956,
2881, 1727, 1676, 1520, 1460, 1371, 1236, 1098, 985, 824, 744 cm-
1. 1H NMR (500 MHz, CDC13) b 8.19 (dd, J = 3.0, 1.1 Hz, 1H, thiophene),
2 0 7.55 (dd, J = 5.0, 1.1 Hz, 1H, thiophene), 7.38 (dd, J = 5.0, 3.0 Hz, 1H,
thiophene), 6.58 (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, IH, 5-H), 4.47 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.38
SUBSTITUTE SHEET (RULE 28)

~~.8p~.4~
w0 95f18798 PCTIUS9S100481
-46-
(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, IH, 3-H), 2.89 (d, J = 20 Iiz,~. IFI, 14-H), 2.64 (br d, J = 20 Hz, 1H,
14-H), 2.54 (ddd, J = 14.5, 9:5, 6.5 Hz, IH, 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, IH, 6-H),
1.85 (s> 1H, (~I), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.19 (s, 3H, Me),
0.92 (t, J = 8.0 Hz, 9H, OSi(CHzC~3)3), 0.65-0.54 (band, 6H,
OSi(C~CH3)3); FAIT HRMS (NBA I CsI) m l a 837.1760, M+Cs+ calcd
for C35H4gOlISSi 837.1741.
Alcohol 34. A solution of enone 33 (57.3 mg, 0.0813 mmol)
in MeOH-THF (5 : 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 CHZC12 (10 mL), treated with
aqueous NH4CI (5 mL), and stirred for 10 min. The organic layer
was separated and the aqueous layer was extracted with CHZCIZ (2 x
10 mL). The combined organic layer was washed with brine (5 mL),
dried (MgS04), concentrated, and purified by flash chromatography
(silica, 30% ethylacetate in hexanes) to give 33 (6.8 mg, 12%) and 3 4
(45.2 mg, 89% based on 88~Yo conversion) as an amorphous solid.
Physical Data for Atcohot 34. R f = 0.48 (silica, 50%
ethylacetate in hexanes); IR (film) vmax 3520, 2953, 2881, 1719,
1520, 1370, 1238, 1100, 979, 823, 746 cm-1; iH NMR (500 MHz,
CDCI3) 8 8.20 (dd, J = 3.0, 1.0 Hz, IH, thiophene), 7.57 (dd, J = 5.0, 1.0
Hz, IH, thiophene), 7.35 (dd, J = 5.0, 3.0 Hz, IH, thiophene), 6.45 (s,
IH, 10-H), 5.54 (d, J = 7.0 Hz, IH, 2-H), 4.96 (br d, J = 8.5 Hz, IH, 5
H), 4.82 (br dd, J = 12.0, 8.0 Hz, IH, 13-H), 4.48 (dd, J = I0.5, 6.5 Hz,
1H, 7-H), 4.36 (d, J = 8.5 Hz, IH, 20-H), 4.15 (d, J = 8.5 Hz, IH, 20-H),
3.85 (d, J = 7.0 Hz, 1H, 3-H), 2.53 (ddd, J = 14.5, 9.5, 6.5> IH, 6-H),
2.27 (s, 3H, Me), 2.28-2.21 (m, 2H, 14-CHZ), 2.18 (s, 6H, Me, Me),
2.03 (s, I H, OH), 1.87 (ddd, J = 14.5, 10.5, 2.0 Hz, I H, 6-H), 1.67 (s,
3 0 3H, Me), 1.65 (s, 1H, OH), 1.18 (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J
= 8.0 Hz, 9H, OSi(CHgC~)3), 0.64-0.50 (band, 6H, OSi(C~CHg)3); FAB
HRMS ( NBA / CsI) m / a 839.1908 M + Cs+ calcd for C3gH5oO ISSi
839.1897.
DiTES taxoid 35. To a solution of alcohol 34 (19.5 mg,
0.0276 mmol, previously azeotroped twice with benzene) and (3-
lactam 24 (27.5 mg, 0.0721 mmol, previously azeotroped twice with
SUBSTITUTE SHEET (RULE 2~

WO 95118798 ~ PC1YUS95100481
-47-
benzene) in THF (1.4 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, L; Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.;
Brigand, T. Tetrahedron 1992, 48, 6985-7012), at 0 °C was added
NaN(SiMe3)2 (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 CH2CIg (10 mL) and aqueous NH4C1 (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 (MgS04), concentrated, and purified by
flash chromatography (silica, 20 -> 30 % ethylacetate in hexanes) to
give 34 (I.1 mg, 6%) and 35 (17.3 mg, 61% based on 94%
conversion) as a white solid.
Physical Data for DiTES taxoid 35. Rf = 0.86 (silica, 50%
ethylacetate in hexanes); IR (film) vmax 3519, 3437, 2953, 2879,
1726, 1666, 1515, 1483, 1369, 1240, 1100, 979, 825, 746 cm-1;1H
NMR (500 MHz, CDC13) b 8.32 (dd, J = 3.0, 1.2 Hz, 1H, thiophene),
7.76-7.73 (m, 2H), 7.60 (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), 6.26 (br t,
J = 9.0 Hz, 1H, 13-H), 5.72 (dd, J = 9.0, 2.0 Hz, IH, 3'-H), 5.61 (d, J =
7.0 Hz, 1H, 2-H), 4.95 (dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.70 (d, J = 2.0 Hz,
IH, 2'-H), 4.48 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.37 (d, J = 8.5 Hz, IH,
20-H), 4.23 (d, J = 8.5 Hz, IH, 20-H), 3.81 (d, J = 7.0 Hz, IH, 3-H),
2.56-2.49 (m, IH, 6-H), 2.54 (s, 3H, Me), 2.35 (dd, J = 15.5, 9.0 Hz,
1H, 14-H), 2.17 (s, 3H, Me), 2.07 (dd, J = 15.5, 9.0 Hz, IH, 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, IH, OH), 1.20 (s, 3H, Me), 1.18 (s, 3H, Me), 0.93 (t, J = 8.0 Hz,
9H, OSi(CH2Cj~3)3), 0.81 (t, J = 8.0 Hz, 9H, OSi(CH2C~)3), 0.63-0.53
3 0 (band, 6H, OSi(CJ~CH3)3), 0.52-0.36 (band, 6H, OSi(C~CH3)3); FAB
HRMS (NBA / CsI) m / a 1220.3675, M + Cs+ calcd for
C57H7~014SSi2N 1220.3658.
Taxoid 36. A solution of silyl ether 35 (17.3 mg, 0.0159
3 5 mmol) in THF (0.6 mL) at 25 °C was treated with HF~pyridine (0.150
mL) and stirred for 2 h. The reaction mixture was poured into a
mixture of eihylacetate (10 mL) and aqueous NaHC03 (5 mL) and the
SUBSTITUTE SHEET (RULE 2~

5
w0 95/18798 ,~, PCT/US95l00481
-4g-
resulting mixture was starred 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 (MgS04), concentrated, and purified by preparative TLC
(silica, 2596 ethylacetate in petroleum ether) to give 36 (7.7 mg,
563fo) as a colorless film.
Preparation of Taxoid 36. Rf _ 0.11 (silica, SO~Yo
ethylacetate in hexanes); IR (film) va,ax 3496, 3434, 2940, 1723,
1648, 1519, 1370, 1243, 1071, 975 cm-t; IH NMR (500 MHz, CDC13) 8
8.32 (dd, J = 3.0, I.0 Hz, 1H, thiophene), 7.75-7.72 (m, 2H), 7.60 (dd,
J = 5.0, 1.0 Hz, IH, thiophene), 7.53-7.33 (band, 9H), 6.95 (d, J = 9.0
Hz, 1H, NH), 6.28-6.23 (m, 2H, 10-H, 13-H), 5.81 (dd, J = 9.0, 2.0 Hz,
1H, 3'-H), 5.58 (d, J = 7.0 Hz, 1H, 2-H), 4.95 (dd, J = 9.5, 2.0 Hz, IH, 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),
1 5 4.36 (d, J = 8.5 Hz, 1H, 20-H), 4.22 (d, J = 8.5 Hz, IH, 20-H), 3.78 (d, J
= 7.0 Hz, 1 H, 3-H), 3.49 (d, J = 4.5 Hz> 1 H, 2'-OH), 2.55 (ddd, J = 14.5,
9.5, 6.5 Hz, IH, 6-H), 2.45 (br s, 1H, OH), 2.40 (s, 3H, Me), 2.34 (dd, J
= 15.5, 9.0 Hz, IH, 14-H), 2.25 (dd, J = 15.5, 9.0 Hz, IH, 14-H), 2.24 (s,
3H, Me), 1.89 (ddd, J = 14.5, 11.0, 2.0 Hz, IH, 6-H), 1.81 (d, J = 2.0 Hz,
3H, 18-Me), 1.74 (br s, 1H, OH), 1.67 (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 C49H49014NS 992.1928.
SUBSTITUTE SHEET (RtiLE 2(~

1 w0 95118798 PCT'/US95/00481
-49-
Preparation of 2-pyridinyl-C-2 taxol (40)
Ac
a
' H =~.
p vwc ego, , Ph N O O OAc
/ O 37
/ O 15 ~N.
O 24 Bz
b
Ac0 O OR Aco v OTES
BzNH O
Ph~O~-~' ~ HO~~
OR ' , O . ' D
NHO p H OAc HO O H OAc
39 : R - TES'1 d ~ / O 38
40 : R = ~IH
Acetate 37. A solution of alcohol 15 (23.2 mg, 0.0353 mmol)
and 4-dimethylaminopyridine (DMAP, 12.9 mg, 0.106 mmol) in
CH2CI2 (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 (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 (MgS04), concentrated, and purified by flash
chromatography (silica, 70 ~ 100% ethylacetate in petroleum ether)
to give 37 (19.0 mg, 77%) as an amorphous solid.
Preparation of Acetate 37. Rf = 0.58 (silica, ethylacetate);
IR (film) v",aX 3482, 2954, 2881, 1730, 1675, 1370, 1304, 1231,
1118, 987, 823, 739 cm-1; ~H NMR (500 MHz, CDC13) & 8.77 (ddd, J=
4.5, 1.7, 1.0 Hz, 1H, pyridine), 8.05 (ddd, J = 8.0, 1.0, L0 Hz, IH,
pyridine), 7.89 (ddd, J = 8.0, 8.0, 1.7 Hz, 1H, pyridine), 7.53 (ddd, J =
SUBSTITUTE SHEET (RULE 2~

WO 95/18798 PC'T/US9510048t
-$~-
8.0, 4.5, 1.0 Hz, IH, pyridine), 6.59 (s, 1H, 10-H), 5.65 (dd, J = 6.6, 1.0
Hz, 1H, 2-H), 4.92 (dd, J = 9.5, 2.0 Hz, IH, 5-H), 4.48 (dd, J = 10.5, 7.0
Hz, 1H, 7-H), 4.35 (d, J = 8 ~5 Hz; IH, 20-H), 4.26 (dd, J = 8.5, 1.0 Hz,
IH, 20-H), 3.91 (d, J = 6.S Hz, IH, 3-H), 3.00 (d, J = 20.0 Hz, IH, 14-
H), 2.7I (dd, J = 20.0, 1.0 Hz, IH, 14-H), 2.54 (ddd, J = 14.5, 9.5, 7.0
Hz, IH, 6-H), 2.53 (s, 1H, OH), 2.23 (s, 3H, Me), 2.18 (s, 3H, Me), 2.14
(s, 3H, Me), 1.88 (ddd, J = 14.5, 10.5, 2.0 Hz, IH, 6-H), 1.70 (s, 3H,
Me), 1.27 (s, 3H, Me), 1.20 (s, 3H, Me), 0.92 (t, 9H, J = 8.0 Hz,
OSi(CHZC~3)3), 0.64-0.52 (band, 6H, OSi(CFj2CH3)3); FAB HRMS (NBA /
CsI) m le 832.2139, M + Cs+catcd for C36H49O11NSi 832.2129.
Alcohol 38. A solution of enone 37 (47.6 mg, 0.0680 mmol)
in MeOH-THF (5 : 1, 3.8 mL) at 0 °C was treated with NaBH4 (46.0
mg, 1.22 mmol, added by portions) and stirred for 1.5 h. The
I S reaction mixture was diluted with ethylacetate (10 mL), treated
with aqueous NH4CI (5 mL), and stirred for 10 min. The organic
layer was separated and the aqueous layer was extracted with
ethylacetate (2 x 10 mL). The combined organic layer was washed
with brine (5 mL), dried (MgS04), concentrated, and purified by
2 0 flash chromatography (basic alumina, ethylacetate -~ 10°k MeOH in
ethylacetate) to give 27f (28.0 mg, 59%) as an amorphous solid.
Physical Data for Alcohol 38. R p _ 0.36 (silica,
ethylacetate); IR (film) v",$X 3487, 2951, 2880, 1736, 1583, 1369,
1307, 1236, 1132, 983, 824, 739 cm-t; tH NMR (500 MHz, CDCI3) b
25 8.79 (dm, J = 4.5 Hz, 1H, pyridine), 8.13 (br d, J = 7.5 Hz, IH,
pyridine), 7.88 (ddd, J = 7.5, 7.5, 1.7 Hz, 1H, pyridine), 7.SI (ddd, J =
7.5, 4.5, 1.0 Hz, IH, pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J = 7.0 Hz,
IH, 2-H), 4.96 (dd, J = 9.5, 2.0 Hz, IH, 5-H), 4.85 (br t, J = 8.0 Hz, IH,
I3-H), 4.49 (dd, J = 10.5, 6.5 Iiz, 1H, 7-H), 4.31 (d, J = 8.0 Hz, 1H, 20-
3 0 H), 4.25 (d, J = 8.0 Hz, IH, 20-H), 3.89 (d, J = 7.0 Hz, 1H, 3-H), 2.53
(ddd, J = 14.5, 9.5, 6.5 Hz, IH, 6-H), 2.36-2.11 (m, 2H, 14-CHZ), 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, IH, 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(CHZC~3)3), 0.65-0.51
3 5 (band, 6H, OSi(C~CH3)q); FAB HRMS (NBA / CsI) m / a 834.2311, M
+ Cs+calcd for C36Hg1O11NSi 834.2286.
SUBSTITUTE SHEET (RIiLE 281

t WO 95118798 PCT/US95100481
-51-
DiTES taxoid 39. To a solution of alcohol 38 (10.3 mg,
0.0147 mmol, previously azeotroped twice with benzene) and [3-
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,
56, I681-1683; Ojima, L; Habus, L; Zhao, M.; Zucco, M.; Park, Y.H.;
Sun, C. M.; Brigaud, 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 NH4Cl (5
mI,). 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 (MgS04), concentrated, and
purified by preparative TLC (silica, 60% ethylacetate in petroleum
ether) to give 39 (2.7 mg, 17°!0) as a film.
Physical Data for DiTES taxoid 39. Rf = 0.28 (silica, 50%
ethylacetate in hexane); IR (film) vmax 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 = 4.5 Hz, IH, pyridine), 8.21 (d, J =
8.0 Hz, IH, 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, IH, 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, IH, 7-H), 4.32 (br s, 2H, 20-CHZ), 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, 9.5 Hz, IH, 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, IH, 6-
H), 1.72 (s, 3H, Me), 1.22 (s, 3H, Me), 1.19 (s, 3H, Me), 0.93 (t, J = 8.0
3 0 Hz, 9H, OSi(CH2C~)3), 0.81 (t, J = 8.0 Hz, 9H, OSi(CHZC~j3)3), 0.64-0.34
(band, 12H, OSi(C~CH3)3); FAB HRMS (NBA / CsI) m / a 1215.4065,
M + Cs+calcd for CSgH7g014N2Si2 1215.4046.
Taxoid 40. A solution of silyl ether 39 (2.7 mg, 0.00249
3 5 mmol) in THF (0.4 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
SUBSTITUTE SHEET (RtiLE 28)

w0 95118798 PCl'/US95I00481
-$Z-
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 oiganic layer was washed with brine (5
mL), dried (MgS04), concentrated, and purified by preparative TLC
(silica, ethylacetate) to give 40 (0.8 mg, 38%) as a colorless film.
Physical Data for Taxoid 40. R f = 0.54 (silica,
ethylacetate); iH 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), 6.98 (d, J
1 0 = 9.0 Hz, 1H, NH), 6.30-6.24 (m, 2H, 10-H, 13-H), 5.82 (dd, J = 9.0, 2.5
Hz, 1H, 3'-H), 5.67 (d, J = 7.0 Hz, 1H, 2-H), 4.95 (dd, J = 10.0, 2.0 Hz,
1H, 5-H), 4.81 (dd, J = 4.5, 2.5 Hz, 1H, 2'-H), 4.41 (ddd, J = 11.0, 7.0,
4.5 Hz, 1 H, 7-H), 4.31 (s, 2H, 20-CH2), 3.81 (d, J = 7.0 Hz, 1 H, 3-H),
3.52 (br s, 1H, OH), 3.50 (d, J = 4.5 Hz, 1H, 2'-OH), 2.56 (ddd, J = 14.5,
1 5 9.5, 7.0 Hz, 1H, 6-H), 2.46 (d> J = 4.0 Hz, 1H, 7-OH), 2.43-2.30 (m, 2H,
14-CH2), 2.38 (s, 3H, OAc), 2.25 (s, 3H, OAc), 1.90 (ddd, J = 14.5, 11.0,
2.0 Hz, 1H, 6-H), 1.81 (s, 3H, Me), 1.71 (s, 3H, Me), 1.26 (s, 3H, Me),
1.15 (s, 3H, Me).
SUBSTITUTE SHEET (RULE 28j

WO 95118798 PCT/US95100481
-53-
Prepartion of 3-pyridinyl-C-2-taxol (44)
17 ~~24 ~Bz ~
~b
wcu a OR Ac0 O OTES
BzNH O
Ph~O~,.. c
~ HO~~~-
OR : O D
/ 10 p H OAC / O 3 H
O OAc
N ~ ~ 43 : R a TES'~ ~ 1 42
O 44 : R = H s--J d O
Acetate 41. A solution of alcohol 17 (42.9 mg, 0.0652 mmol)
and 4-dimethylaminopyridine (DMAP, 23.9 mg, 0.196 mmol) in
CHZC12 (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 (MgS04), concentrated, and purified by flash
chromatography (silica, ethylacetate) to give 41 (43.5 mg, 95%) as a
white solid.
Physical Data for Acetate 41. R f - 0.61 (silica,
ethylacetate); IR (film) va,ax 3470, 3327, 2955, 2881, 1731, 1675,
1592, 1370, 1279, 1229, 1108, 822, 738 cm-1; IH NMR (500 MHz,
CDCI3) 8 9.23 (br s, 1H, pyridine), 8.79 (br s, 1H, pyridine), 8.30 (ddd,
J = 8.0, 2.0, 2.0 Hz, 1H, pyridine), 7.43 (dd, J = 8.0, 5.0 Hz, 1H,
SUBSTITUTE SHEET (RULE 2~

W0 95/18798 PC1YU895I0048t
-54-
pyridine), 6.58 (s, IH, 10-H), 5.70 (dd, J = 6.5, 1.0 Hz, IH, 2-H), 4.91
(dd, J = 9.5, 2.0 Hz, 1H, 5-H), 4.47 (dd, J = I0.5, 7.0 Hz, IH, 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 = 6.5
Hz, 1H, 3-H), 2.93 (d, J = 20.0 Hz, 1H, I4-H), 2.68 (dd, J = 20.0, 1.0 Hz,
IH, 14-H), 2.53 (ddd, J = 14.5, 9.5, 7.0 Hz, IH, 6-H), 2.24 (br s, IH,
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, 6-H), 1.66 (s, 3H, Me), 1.26 (s, 3H, Me), 1.18 (s,
3H, Me), 0.90 (t, J = 8.0 Hz, 9H, OSi(CHZC~I3)3), 0.63-0.51 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NBA / CsI) m /e 832.2145, M + Cs+ calcd
for C36H4gOIINSi 832.2129.
Alcohol 42. A solution of enone 41 (39.8 mg, 0.0569 mmol)
in MeOH-THF (5 : 1, 3.1 mL) at 0 C was treated with NaBH4 (65.0
mg, 1.72 mmol, added by portions) and stirred for 1.5 h. The
1 5 reaction mixture was diluted with ethylacetate( 10 mL), treated
with aqueous NH4Cl (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 (MgS04), concentrated,
and purified by
flash chromatography (silica, ethylacetate) 41 (3.7 mg, 9qb)
to give
and 42 (24.3 mg, 67% based on 91~ conversion) as an amorphous
solid.
Physical Data for Alcohol 42. R p - 0.42 (silica,
ethylacetate); IR (film) vmax 3490, 2953, 2881, 1727, 1592, 1369,
1235, 1110, 822, 740 cm-1; 1H NMR (500 MHz, CDC13) 8 9.30 (d, J =
2.0 Hz, 1H, pyridine), 8.81 (dd, J = 5.0, 2.0 Hz, IH, pyridine), 8.35
(ddd, J = 8.0, 2.0, 2.0 Hz, IH, pyridine), 7.44 (dd, J = 8.0, 5.0 Hz, IH,
pyridine), 6.46 (s, 1H, 10-H), 5.64 (d, J = 7.0 Hz, 1H, 2-H), 4.96 (dd, J
= 9.5, 1.5 Hz, I H, 5-H), 4.83 (br dd, J = 12.5, 7.5 Hz, 1 H, 13-H), 4.49
3 0 (dd, J = 10.5, 6.5 Hz, 1H, 7-H), 4.28 (d, J = 8.0 Hz, IH, 20-H), 4.15 (d,
J
= 8.0 Hz, 1H, 20-H), 3.89 (d, J = 7.0 Hz, IH, 3-H), 2.53 (ddd, J = 14.5,
9.5, 6.5 Hz, 1H, 6-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.I9
3 S (s, 3H, Me), 1.04 (s, 3H, Me), 0.92 (t, J = 8.0 Hz, 9H, OSi(CH2C~)3),
0.64-0.51 (band, 6H, OSi(C~CH3)3); FAB HRMS (NBA / CsI) m / a
834.2270, M + Cs+ calcd for C36Hg1O l 1 NSi 834.2286.
SUBSTITUFE SHEET (RULE 2~

w0 95118798 PCT/US95100481
-55-
DiTES taxoid 43. To a solution of alcohol 42 (12.6 mg, 0.018
mmol, previously azeotroped twice with benzene) and (3-lactam 2 4
(17.0 mg, 0.0446 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, 164568q; Ojima, L; Habus, L;
Zhao, M.; Georg, G. L; Jayasinghe, L. R. J. Org. Chem. 1991, 56, 1681-
1683; Ojima, L; Habus, L; Zhao, M.; Zucco, M.; Park, Y.H.; Sun, C. M.;
Brigand, T. Tetrahedron 1992, 48, 6985-7012), was added
1 0 N aN ( S i M a 3 )2 (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 (IO mL) and aqueous NH4C1 (5 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 (MgS04), concentrated, and
purified by flash chromatography (silica, 50 ~ 95% ethylacetate in
hexanes) to give 42 (1.0 mg, 89b) and 43 (8.6 mg, 48% based on 92%
conversion) as a white solid.
Physical Data for DiTES taxoid 43. Rf = 0.40 (silica , 50%
ethylacetate in hexanes); IR (film) vmax 3433, 2955, 2880, 1730,
1662, 1370, 1238, 1112, 1018, 985, 824, 740 cm-i;lH NMR (500
MHz, CDC13) 8 9.34 (d, J = 2.0 Hz, 1H, pyridine), 8.82 (dd, J = 5.0, 2.0
Hz, IH, 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), 6.22 (br t, J = 9.0 Hz, 1H, 13-H), 5.74-5.66 (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 = I0.5,6.5 Hz, IH, 7-H), 4.30 (d, J = 8.0 Hz, 1H, 20-H), 4.21
(d, J = 8.0 Hz, IH, 20-H), 3.86 (d, J = 7.0 Hz, 1H, 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
3 0 (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), 1.22 (s, 3H, Me), 1.18 (s,
3H, Me), 0.93 (t, J = 8.0 Hz, 9H, OSi(CHZC -~I )3), 0.82 (t, J = 8.0 Hz, 9H,
OSi(CHZC~3)3), 0.65-0.37 (band, 12H, OSi(C~CH3)3, OSi(Cj~CH3)3);
FAB HRMS (NBA / Csi) m / a 1215.4066, M + Cs+ calcd for
3 S C9gH7g014NySi2 1215.4046.
SUBSTITUTE SHEET (RULE 2~

wo 9sns~9s ~, ~ $ ~ ~ ~ ~ PCTIUS9510048t
-56-
Taxoid 44. A solution of silyi ether 43 (6.4 mg, 0.0059
mmol) in THF (0.4 mL) at 25 °C wad ueated with HF~pyridine (0.160
mL) and stirred for 1.25 h. The reaction mixture was poured into a
mixture of ethylacetate (10 mI.)- 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 (MgS04), concentrated, and purified by preparative TLC
(silica, ethylacetate) to give 44 (3.8 mg, 75g'o) as a colorless film.
Physical Data for Taxoid 44. R f _ 0.59 (silica,
ethylacetate); IR (film) vm$X 3396, 2928, 1728, 1644, 1371, 1273,
1241, 1111, 1071 cm-~; ~H NMR (500 MHz, CDCl3) 8 9.34 (br s, IH,
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 =
1 5 9.0 Hz, iH, NH), 6.27 (s, IH, 10-H), 6.23 (br t, J = 9.0 Hz, IH, 13-H),
5.78 (dd, J = 9.0, 2.5 Hz, 1H, 3'-H), 5.69 (d, J = 7.0 Hz, IH, 2-H), 4.95
(dd, J = 9.5, 2.0 Hz, IH, 5-H), 4.79 (dd, J = 5.5, 2.5 Hz, IH, 2'-H), 4.41
(ddd, J = 11.0, 6.5, 4.0 Hz, IH, 7-H), 4.29 (d, J = 8.5 Hz, 1H, 20-H),
4.20 (d, J = 8.5 Hz, IH, 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, 6.5 Hz, 1H, 6-H), 2.49 (d, J
= 4.0 Hz, IH, 7-OH), 2.43-2.26 (m, 2H, 14-CH2), 2.38 (s, 3H, Me), 2.24
(s, 3H, Me), 1.89 (ddd, J = 14.5, 11.0, 2.0 Hz, IH, 6-H), 1.83 (s, iH,
OH), 1.82 (s, 3H, Me), 1.69 (s, 3H, Me), 1.25 (s, 3H, Me), 1.14 (s, 3H,
Me); FAB HRMS (NBA / CsI) m / a 987.2325, M + Cs+ calcd for
2 5 C46H50014N2 987.2316.
SUBSTITUTE SHEET (RULE 28~

w0 95118798 218 fl 4 9 ~ PCTIUS95100481
-57-
Preparation of 4-N, N-dimethylaniline-C-2 taxol (48)
HO ~O OTES _ Ac0 O OTES
O°C ~ 1 I -a a O
J H ~O ~ H W
j OAc ~O n OAc
Me2N
O 18 ~-=~ p 45
TESO, Ph
~ b
BzNH 1 1 1O ~ ACO ~O' OR O' 24N'Bz \ Ac ~. OTES
Ph~O"..l ~ T 1 ~-c-.- HO~~~.
O R H~/H~O
O OAc p O'A~c
Me2N ~ I 47 : R - TES MezN ~ I 46
O 48:R=H ~d O
Acetate 45. A solution of alcohol 18 (50.0 mg, 0.0714 mmol)
and 4-dimethylaminopyridine (DMAP, 26.0 mg, 0.213 mmol) in
CHZC12 (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 CH2C12 (10 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
CH2C12 (2 x 10 mL). The
combined organic layer was washed with brine (5 mL,), dried
(MgS04), concentrated, and purified by flash chromatography (silica,
1030 ethylacetate in benzene) to give 45 (41.0 mg, 77%) as an
amorphous solid.
Physical Data for Acetate 45. R f = 0.27 (silica, 35!0
ethylacetate in hexanes); IR (film) vma x 3425, 2945, 1722, 1674,
1605, 1365, 1275, 1232, 1179, 1094; tH NMR (500 MHz, CDCI3) b
7.89 (d, J = 9.0 Hz, 2H, Ar), 6.64 (d, 9.0 Hz, 2H, Ar), 6.56
J = (s, 1H,
10-H), 5.64 (d, J = 6.5 Hz, IH, 2-H), 4.90(br d, J = 8.0 Hz, 1H,
5-H),
4.45 (dd, J = 10.5, 7.0 Hz, 1 H, 7-H),
4.36 (d, J = 9.0 Hz, 1 H, 20-H), 4.11
SUBSTITUTE SHEET (RULE 28)

w0 95/18798 PCTlUS95100481
-$8-
(d, J = 9.0 Hz, IH, 20-H), 3.85 (d, J = 6.5 Hz, 1H, 3-H), 3.05 (s, 6H,
NMe2), 2.90 (d, J = 20.0 Hz, 1H, I4-H), 2.62 (d, J = 20.0 Hz, 1H, 14-H),
2.51 (ddd, J = 14.0, 8.0, 7.0, IH, 6-H), 2.20 (s, 3H, Me), 2.16 (s, 3H,
Me), 2.15 (s, 3H, Me), 2.04 (s, LH, OH), L84 (ddd, J = 14.0, 10.5, 2.0
Hz, IH, 6-H), 1.63 (s, 3H, Me), 1.23 (s, 3H, Me), 1.16 (s, 3H, Me), 0.89
(t, J = 8.0 Hz, 9H, OSi(CH~Cjj3)3), 0.58-0.53 (band, 6H, OSi(Cj~CH3)3);
FAB HRMS (NBA / CsI) m / a 874.8589, M + Cs+ calcd for
C39H55O11NSi 874.8594.
Alcohol 46. A solution of enone 4$ (40.0 mg, 0.0539 mmol)
in MeOH-THF (5.8 : 1, 4.1 mL) at 0 °G was treated with NaBH4 (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
CHZCl2 (15 mL), treated with aqueous NH4Cl (5 mL), and stirred for
10 min. The organic layer was separated and the aqueous layer was
extracted with CH2CI2 (2 x 10 mL). The combined organic layer was
washed with brine (5 mL), dried (MgS04), concentrated, and purified
by flash chromatography (silica, 25 -~ 50% ethylacetate in petroleum
ether) to give 4$ (6.0 mg, I5~'o) and 46 (30.0 mg, 889'o based on 859b
2 0 conversion) as an amorphous solid.
Physical Data for Alcohol 46. Rf = 0.30 (silica, SOg'o
ethylacetate in petroleum ether); 1H NMR (500 MHz, CDC13) & 7.93 (d,
J = 9.0 Hz, 2H, Ar), 6.64 (d, J = 9.0 Hz, 2H, Ar), 6.42 (s, IH, IO-H), 5.57
(d, J = 7.0 Hz, 1H, 2-H), 4.94 (br d, J = 8.0 Hz, IH, 5-H), 4.83-4.75 (m,
1H, 13-H), 4.46 (dd, J = 10.5, 6.5 Hz, IH, 7-H), 4.34 (d, J = 8.5 Hz, IH,
20-H), 4.13 (d, J = 8.5 Hz, IH, 20-H), 3.82 (d, J = 7.0 Hz, 1H, 3-H), 3.04
(s, 6H, Me2N), 2.54-2.44 (m, IH, 6-H), 2.26 (s, 3H, Me), 2.23 (d, J =
7.5 Hz, 2H, 14-CHZ), 2.16 (s, 6H, Me, Me), 2.08 (d, J = 4.5 Hz, 1H, OH),
1.89-L80 (m, 2H, 6-H, OH), 1.64 (s, 3H, Me), 1.16 (s, 3H, Me), 1.01 (s,
3 0 3H, Me), 0.89 (t, J = 8.5 Hz, 9H, OSi(CHZCj~)3), 0.62-0.48 (band, 6H,
OSi(C~CH3)3).
DiTES taxoid 47. To a solution of alcohol 46 (14.0 mg,
0.0188 mmol, previously azeotroped twice with benzene) and [3-
3 5 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, L;
SUBSTITUTE SHEET (RLiLE 28J

WO 95/18798 PCTIUS95/00481
-59-
Habus, L; Zhao, M.; Georg, G. I:; 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), was
added NaN(SiMe3)2 (0.056 mL of a L0 M solution in THF, 0.056
mmol) dropwise. The resulting solution was stirred for 20 min and
poured into a mixture of CHZCIZ (10 mL) and aqueous NH4CI (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 (MgS04), concentrated, and purified
by flash chromatography (silica, 10 ~ 1596 ethylacetate in benzene,
then 50°Io ethylacetate in petroleum ether) to give 47 (12.0 mg,
57°.h)
as a white solid.
Physical Data for DiTES taxoid 47. Rp = 0.26 (silica, 15%
ethylacetate in PhH); IR (film) vmBX 3425, 2946, 2882, 1722, 1669,
1600, 1365, 1275, 1238, 1179, 1094 cm-1; IH NMR (500 MHz, CDCl3)
8 7.96 (d, J = 9.0 Hz, 2H, Ar), 7.77-7.72 (m, 2H), 7.54-7.26 (band, 8H),
7.I2 (d, J = 8.5 Hz, 1 H, NH), 6.69 (d, J = 9.0 Hz, 2H), 6.43 (s, 1 H, 10-H),
6.23 (br t, J = 9.0 Hz, 1 H, 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), 3.04 (s, 6H, MegN), 2.55-2.46
(m, 1H, 6-H), 2.53 (s, 3H, OAc), 2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H),
2.15 (s, 3H, Me), 2.09 (dd, J = 15.5> 9.0 Hz, 1H, 14-H), 2.00 (d, J = 1.0
Hz, 3H, Me), 1.92-1.84 (m, 2H, 6-H, OH), 1.67 (s, 3H, Me), 1.20 (s, 3H,
2 5 Me), 1.16 (s> 3H, Me), 0.90 (t, J = 8.0 Hz, 9H, OSi(CHzC~)3), 0.79 (t, J =
8.0 Hz, 9H, OSi(CHZCj33)3), 0.63-0.35 (band, 12 H, OSi(C~CH3)3); FAB
HRMS (NBA / CsI) m / a 1257.4503, M + Cs+ calcd for
C61 Hs4014N2S i2 1257.4515.
3 0 Taxoid 48. A solution of silyl ether 47 (12.0 mg, 0.0107
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 min. The organic layer was
3 5 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"
SUBSTITUTE SHEET {RULE 281

2~8~4~
WO 95118798 PCTIU895/00481
-60-
chromatography (silica, 50 ~ ; 75~ ethylacetate in petroleum ether)
to give 48 (8.0 mg, 84°x) as a colorless film.
Physical Data for Taxoid 48. R f = 0.44 (silica, 7596
ethylacetate in petroleum ether); IR (film) vmax 3414, 2914, 2850,
1722, 1664, 1660, 1371, 1275, 1243, 1179 cm-I; IH NMR (500 MHz,
CDC13) 8 7.95 (d, J = 9.0 Hz, 2H), 7.77-7.72 {m, 2H), 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, IH,
10-H), 6.20 (br t, J = 9.0 Hz, IH, 13-H), 5.76 (dd, J = 9.0, 2.5 Hz, IH,
3'-H), 5.62 (d, J = 7.0 Hz, IH, 2-H), 4.93 (br d, J = 7.5 Hz, 1H, 5-H),
1 0 4.76 (dd, J = 5.0, 2.5 Hz, 1 H, 2'-H), 4.37 (ddd, J = 11.5, 6.5, 4.0 Hz, I
H,
7-H), 4.34 (d, J = 8.5 Hz, IH, 20-H), 4.18 (d, J = 8.5 Hz, IH, 20-H), 3.73
(d, J = 7.0 Hz, IH, 3-H), 3.57 (d, J = 5.0 Hz, IH, 2'-OH), 3.04 (s, 6H,
Me2N), 2.58-2.48 (m, IH, 6-H), 2.44 (d, J = 4.0 Hz, 1H, 7-OH), 2.37 (s,
3H, Me), 2.30-2.25 (m, 2H, 14-CH2), 2.22 (s, 3H, Me), 1.95 (s, IH, OH),
1 5 1.88-1.81 (m, IH, 6-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 / Csd) m / a
1029.2760, M + Cs+ calcd for C49H56N2014 1029.2786.
SUBSTITUTE SHEET (RULE 2~

PCT/US95J00481
WO 95/18798
-61-
Preparation of 1-naphthalene-C-2-taxol (52)
HO O OTES _ ACO O OTES
a
' HO ~ H ~A '~ / ' HQ O H ~A
49
TESO~ Ph
~b
Ac0 O
Ac0 O OR O~~ez OTES
BzN
Ph 0.... ~ Hp.,..
OR / ' HO ~ H pAc O / ' HO ~ H pAc C
52-R-__HE~d W ~ O 50
Acetate 49. A solution of previous alcohol 1 9 and 4-
dimethylaminopyridine (DMAP, 100 mg, 0.819 mmol) in CHgCl2 (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
CHZC12 (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 CH2C12 (2 x 10 mL). The combined
organic layer was washed with brine (5 mL), dried (MgS04),
concentrated, and purified by preparative TLC (silica, 10%
1 5 ethylacetate in benzene) to give 49 (54.1 mg, ~ 89R6 from carbonate
7) as an amorphous solid.
Physical Data for Acetate 49. R f = 0.27 (20% ethylacetate
in petroleum ether); IR (film) vmax 3416, 2953, 2879, 1726, 1676,
1370, 1224, 1089 cm-l; IH NMR (500 MHz, CDCI3) 8 8.66 (s, 1H,
2 0 naphthalene), 8.06 (dd, 1H, J = 9.0, 2.0 Hz, naphthalene), 7.98-7.89
(m, 3H, naphthalene), 7.68-7.55 (m, 2H, naphthalene), 6.61 (s, 1H,..
10-H), 5.75 (d, J = 7.0 Hz, 1H, 2-H), 4.95 (br d, J = 8.0 Hz, 1H, 5-H),
SUBSTITUTE SHEET (RULE 26~

w0 95118798 PCTIUS95/00481
2~~fl~4~
-62-
4.50 (dd, J = 10.5, 7.0 Hz, IH, 7-H), 4.35 (d, J = 8.5 Hz, 1H, 20-H), 4.16
(d, J = 8.5 Hz, 1 H, 20-H), 3.96 (d, J = 8.5 Hz, 1 H, 20-H), 3.96 (d, J = 7.0
Hz, 1H, 3-H), 3.03 (d, J = 20.0 Hz, 1~I, 14-H), 2.70 (d, J= 20.0 Hz, 1H,
14-H), 2.61-2.50 (m, 2H, 6-H, OH), 2.27 (s, 3H, Me), 2.24 (s, 3H, Me),
2.21 (s, 3H, Me), 1.91-1.83 (m, IH, 6-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(CHgC~j3)3), 0.66-0.57
(band, 6H, OSi(C~CH3)3); FAB HRMS (NBA / CsI) mle 881.2326, M
+ Cs+ calcd for C41Hg2011Si 881.2333.
Alcohol 50. A solution of enone 49 (54.1 mg, 0.0722 mmol)
in MeOH (10 mL) at 25 °C was treated with NaBHø (54.5 mg, 1.44
mmol, added by portions) and stirred for 2.0 h. The reaction
mixture was diluted with CHZC12 (10 mL), treated with aqueous
N H 4Cl (5 mL), and stirred for 10 min. The organic layer was
1 5 separated and the aqueous layer was extracted with CH2C12 (2 x 10
mL). The combined organic layer was washed with brine (5 mL),
dried (MgS04), concentrated, and purified by preparative TLC (silica,
20°Jo ethylacetate in petroleum ether) to give 50 (26 mg, 48°Jo
) as an
amorphous solid.
2 0 Physical Data for Alcohol 50. R f = 0.12 (2096 ethylacetate
in petroleum ether); IR (film) v,oaX 3524, 2953, 1719, 1369, 1231,
1093, 829 cm-1; 1H NMR (500 MHz, CDCI3) b 8.70 (s, 1H,
naphthalene), 8.11 (dd, J = 8.5, 1.5 Hz, 1H, naphthalene), 7.96-7.86
(m, 3H, naphthalene), 7.65-7.54 (m, 2H, naphthalene), 6.45 (s, 1H,
25 10-H), 5.68 (d, J = 7.0 Hz, IH, 2-H), 4.98 (br d, J = 8.0 Hz, IH, 5-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, IH, 20-H), 4.19 (d, J = 8.5 Hz, IH, 20-H), 3.93 (d, J = 7.0 Hz,
IH, 3-H), 2.58-2.50 (m, 1H, 6-H), 2.41-2.14 (m, 3H, 14-CHz, 13-OH),
2.37 (s, 3H, Me), 2.21 (br s, 3H, Me), 2.19 (s, 3H, Me), 1.92-1.84 (m,
3 0 1H, 6-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(CHZC~)3), 0.65-0.51 (band, 6H,
OSi(Cjj2CH3)3); FAB HRMS (NBA / CsI) m / a 883.2484, M + Cs~- calcd
for C41H54011Si 883.2490.
3 5 DiTES taxoid 51. To a solution of alcohol 50 (20.0 mg,
0.0266 mmoI, previously azeotroped twice with benzene) and p-
lactam 24 (20.0 mg, 0.0525 mmol, previously azeotroped twice with
SUBSTITUTE SHEET (RLiLE 281

w0 95118798 21 g ~ 4 4 ~ PCTIUS95100481
-63-
benzene) in THF (1.1 mL) at -78 °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,
56, 1681-1683; Ojima, L; Habus, L; Zhao, M.; Zucco, M.; Park, Y.H.;
Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was
added NaN(SiMe3)Z (0.065 mL of a 1.0 M solution in THF,
0.065mmo1) dropwise. The resulting solution was stirred for 10 min
and poured into a mixture of CH2C12 (10 mL) and aqueous NH4CI (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 (MgS04), concentrated, and purified
by preparative TLC (silica, 20% ethylacetate in petroleum ether) to
give 51 (18.7 mg, 62°!0) as a white solid.
Taxoid 52. 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 (5 mL) and the
resulting mixture was stirred for 10 min. The organic layer was
2 0 separated and the aqueous layer was extracted with ethylacetate (2
x 10 mL). The combined organic layer was washed with brine (S
mL), dried {MgS04), concentrated, and purified by preparative TLC
(silica, 50% ethylacetate in petroleum ether) to give 52 (12.8 mg,
86%) as a colorless film.
2 5 Physical Data for Taxoid 52. R f = 0.16 (silica, 50%
ethylacetate in petroleum ether); IR (film) vn,ax 3420, 2967, 2896,
1721, 1652, 1519, 1370, 1233, 1073, 776 cm-i; 1H NMR (500 MHz,
CDC13) b 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 (bs d, J =
3 0 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), 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 =
3 5 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, Me),
2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H), 2.25 (dd, J = 15.5, 9.0 Hz, lf-I,
SUBSTITUTE SHEET (RULE 281

wo 9sns~9s 2 ~ ~ ~ 4 ~ ~ rc~rius9srooasi 1
-64-
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 CS 1 H 53 N O 14
1036.2520
Preparation of thioether-C-2 taxol (56)
OTES , Ac0 ,O OTES
- ~-.a O
HO H OA v HO v H pA
SPh---"' SPh--"'
O O 53
23 TESO, Ph
~b
BzNH O \ Ac 1 I/O. Or R ~ 24 Bz
~O~~~ ~ H
OR = , O
HO = H
O OAc
SPhJ " 55 : R = TES~ SPh--"' 54
O 56:R=H ~d O
1 0 Acetate 53. A solution of alcohol 23 (25.2 mg, 0.0351 mmol)
and 4-dimethylaminopyridine (DMAP, 12.2 mg, 0.0999 mmol) in
CHZCIZ (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 CHzCI~ (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 CHZC12 (2 x 10 mL). The
combined organic layer was washed with brine (5 mL), dried
(MgS04), concentrated, and purified by flash chromatography (silica,
30°Jo ethylacetate in petroleum ether) to give 53 (25.3 mg, 9586) as a
2 0 colorless oil.
Physical Data for Acetate 53. R f = 0.41 (silica, 10%
ethylacetate in benzene, 2 elutions); IR (film) vmax 3471, 2954,
SUBSTiTUFE SHEET (RULE 2~

wo 95/t8798 ~ ~ $ ~ ~ ~ ~ - PCTlUS9510048t
-G5-
2881, 1729, 1675, 1370, 1226, 986, 824, 738 cm-i;iH NMR (500
MHz, CDC13) 8 7.38-7.25 (band; SH, SPh), 6.54 (s, 1H, 10-H), 5.49 (br
d, J = 6.5 Hz, IH, 2-H), 4.90 (dd, J = 9.5, 2.0 Hz, 1H, 5-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~SPh), 2.78 (d, J = 20.0 Hz, 1H, 14-H), 2.72-2.58 (m, 3H,
C$,ZCH2SPh, 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), 1.23 (s, 3H, Me), 1.19 (s,
1 0 3H, Me), 0.91 (t, J = 8.0 Hz, 9H, OSi(CHZC~3)3), 0.64-0.52 (band, 6H,
OSi(C~CH3)3); FAB HRMS (NBA / CsI) m / a 891.2225, M + Cs+ calcd
for C39H54~11SSi 891.2210.
Alcohol 54. A solution of enone 53 (24.4 mg, 0.032 mmol) in
1 5 MeOH-THF (5 : 1, 1.9 mL) at 0 °C was treated with NaBHq (18.1 mg,
0.48 mmol, added by portions) and stirred for 1.25 h. The reaction
mixture was diluted with CHZC12 (5 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 5 mL). The
20 combined organic layer was washed with brine (5 mL), dried
(MgS04), concentrated, and purified by flash chromatography (silica,
30% ethylacetate in hexanes) to give 54 (14.6 mg, 60%) as an
amorphous solid.
Physical Data for Alcohol 54. R p = 0.11 (silica, 30°k
25 ethylacetate in hexanes); IR (film) vmaX 3487, 2938, 2880, 1729,
1586, 1369, 1234, 977, 738 cm-i; iH NMR (500 MHz, CDCl3) b 7.40
7.23 (band, SH, SPh), 6.42 (s, 1H, 10-H), 5.43 (d, J = 7.0 Hz, 1H, 2-H),
4.94 (dd, J = 9.5, 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
3 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~CH2SPh), 2.51 (ddd, J = 14.5, 9.5, 6.5 _
Hz, 1H, 6-H), 2.25 (dd, J = 15.5, 9.5 Hz, 1H, I4-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, IH, OH), 1.99-1.96 (m, 1H, OH), 1.86
3 5 (ddd, J = 14.5, 10.5, 2.0 Hz, 1H, 6-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(CHZC~)3), 0.64-0.50
SUBSTITUTE SHEET (RULE 28)

w0 95118798 2 ~ PC1YU595100481
-66-
(band, 6H, Si(C~CH3)3); FAB HRMS (NBA / CsI) m l a 893.2350, M +
Cs+calcd for C39H56O11SSi 893.2367.
DiTES taxoid 55. To a. solution of alcohol 54 (21.8 mg,
0.0286 mmol, previously azeotroped twice with benzene) and (3-
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 Abstr. 1990, 114, 164568q; Ojima, L;
Habus, L; Zhao, M.; Georg, G. L; Jayasinghe, L. R. J. Org. Chem. 1991,
IO 56, 1681-1683; Ojima, L; Habus, L; Zhao, M.; Zucco, M.; Park, Y.H.;
Sun, C. M.; Brigaud, T. Tetrahedron 1992, 48, 6985-7012), was
added NaN(SiMe3)2 (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 into a mixture of CHZCIZ (10 mL) and aqueous NH4C1 (5 mL).
The organic layer was separated and the aqueous layer was
extracted with CHZCIz (2 x 5 mL). The combined organic layer was
washed with brine (5 mL), dried (MgS04), concentrated, and purified
by flash chromatography (silica, 15 -a 30 ~ 50% ethylacetate in
petroleum ether) to give 35 (13.8 mg, 4296) as an amorphous solid.
Physical Data for DiTES taxoid 55. Rp = 0.40 (silica, 3086
eehylacetate in hexanes); IR (film) vm8x 3437, 2952, 2879, 1735,
1662, 1482, 1369, 1236, 1128, 981, 740 cm-l IH NMR (500 MHz,
CDC13) b 7.82-7.76 (m, 2H), 7.54-7.I6 (band, 13H), 7.11 (d, J = 9.0 Hz,
IH, NH), 6.41 (s, 1H, 10-H), 6.18 ( br t, J = 9.0 Hz, IH, 13-H), 5.62 (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, IH, 5-H), 4.64 (d, J = 2.0 Hz, 1H, 2'-H), 4.42 (dd, J = 10.5,
6.5 Hz, IH, 7-H), 4.40 (d, J = 8.0 Hz, IH, 20-H), 4.21 (d, J = 8.0 Hz, IH,
20-H), 3.70 (d, J = 7.0 Hz, 1H, 3-H), 3.23-3.17 (m, 2H, Cj~SPh), 2.78-
3 0 2.69 (m, IH, HC~CH2SPh), 2.67-2.57 (m, IH, ~,CHCHZSPh), 2.55-2.46
(m, 2H, 6-H, OH), 2.38 (s, 3H, Me), 2.27-2.10 (m, 2H, 14-CHZ), 2.16 (s,
3H, Me), 1.98 (d, J = 1.0 Hz, 3H, Me), 1.89 (ddd, J = 14.0, I L0, 2.0 Hz,
IH, 6-H), 1.64 (s, 3H, Me), 1.18 (s, 3H, Me), 1.17 (s, 3H., Me), 0.91 (t, J
= 8.0 Hz, 9H, OSi(CHZC$3)3), 0.81 (t, J = 8.0 Hz, 9H, OSi(CH2C~)3),
3 5 0.64-0.36 (band, 12 H, OSi(Cj~CH3)3); FAB HRMS (NBA / CsI) m / a
M + Cs+ 1274.4125 calcd for C61Hg3014SSiz 1274.4127.
SUBSTITUTE SHEET (RULE 281

w0 95118798 ~ PCTlUS95/00481
-67-
Taxoid 56. A solution of silyl ether 55 (8.1 mg, 0.0071
mmol) in THF (0.5 mL) at 25 °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 (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 (MgS04), concentrated, and purified by preparative TLC
(silica, 60°!o ethylacetate in petroleum ether) to give 56 (3.2 mg,
499bj as a colorless film.
Physical Data for Taxoid 56. R E = 0.39 (silica, 6096
ethylacetate in petroleum ether); IR (film) v,nax 3426, 2928, 1731,
1642, 1371, 1238, 1070, 739, 709 cm-1; 1H NMR (500 MHz, CDC13) b
7.80-7.75 (m, 2H), 7.55-7.18 (band, 13 H), 6.94 (d, J = 9.0 Hz, 1H,
1 5 NH), 6.23 (s, iH, 10-H), 6.19 (br t, J = 9.0 Hz, 1H, 13-H), 5.74 (dd, J =
9.0, 2.5 Hz, iH, 3'-H), 5.47 (d, J = 7.0 Hz, iH, 2-H), 4.93 (dd, J = 9.5,
2.0 Hz, 1H, 5-H), 4.74 (dd, J = 5.0, 2.5 Hz, iH, 2'-H), 4.38 (d, J = 8.0 Hz,
1H, 20-H), 4.35 (ddd, J = 11.0, 6.5 Hz, 4.5 Hz, iH, 7-H), 4.21 (d, J = 8.0
Hz, 1H, 20-H), 3.67 (d, J = 7.0 Hz, iH, 3-H), 3.51 (d, J = 5.0 Hz, IH, 2'-
2 0 OH), 3.28-3.14 (m, 2H, Cj~SPh), 2.77-2.68 (m, 1H, HCj~CH2SPh), 2.67-
2.59 (m, iH, gCHCHgSPh), 2.54 (ddd, J = 14.5, 9.5, 6.5 Hz, 1H, 6-H),
2.44 (d, J = 4.5 Hz, iH, 7-OH), 2.36 (dd, J = 15.5, 9.0 Hz, 1H, 14-H),
2.26 (br s, iH, OH), 2.23 (s, 3H, Me), 2.21 (s, 3H, Me), 2.18 (dd, J =
15.5, 9.0 Hz, iH, 14-H), 1.88 (ddd, J = 14.5, 11.0, 2.0 Hz, 1H, 6-H),
2 5 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
C49H55014NS 1046.2398.
3 0 Preparation of MPA taxoid 57
H
HO H
N~ i i0
S /
CH~COO' O
36 57
SUBSTITUTE SHEET (RULE 261

CA 02180445 2004-07-21
-68-
MPA taxoid 57. A solution of taxoid 3 6 (4.3 mg, 0.005
mmol) and triethylamine (0.0033 mL, 0.0237 mmol) in CH2Clz (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 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 was directly purified by HPLC (Vydak'~ RP-18, 22.5
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 36 (0.8 mg, 1996) and 57 (4.1
mg, 100 based on 8196 conversion) as a colorless film.
Physical Data for taxoid 57 1 H NMR (500 MHz, CDC13 ) 8
10.5 (d. J = 7.5 Hz, I H), 8.44 (ddd, J = 9.0, 7.5, 2.0 Hz, 1 H), 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, 1 H, thiophene), 7.48-7.34 (band, 7H), 7.16-7.12 (m, 1 H),
2 0 6.53-6.43 (m, 1H, 2'-H), 6.21 (s, 1H, 10-H), 6.03 (dd, J = 10.5, 8.0 Hz,
1 H, 3'-H), 5.82 (br t, J = 9.0 Hz, 1 H, 13-H), 5.44 (d, J = 7.0 Hz, 1 H, 2-
H), 4.90 (dd, J = 9.5, 2.0 Hz, 1 H, 5-H), 4.33 (dd, J = 11.0, 6.5 Hz, 1 H, 7-
H), 4.30 (d, J = 8.0 Hz, 1H, ZO-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 5 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, J = 14.5, 11.5, 2.0 Hz, IH, 6-H), 1.78 (br s, 3H, 18-
Me), 1.64 (s, 3H, Me), 1.61 (dd, J = 16.0, 7.0 Hz, 1H, 14-H), I.18 (dd, J
= 16.0, 9.0 Hz, 1H, 14-H), 1.I3 (s, 3 H, Me), 1.08 (s, 3 H, Me).
*Trade-mark

PCT/US95100481
WO 95118798
-69-
Preparation of MPA taxoid 58
Ac0 O OH
BzNH -O BzNH Q
C ph 0....
OH ~ O , ' O
HO p H OAc
~N~ S
CHsC00'
32 58
MPA taxoid 58. A solution of taxoid 32 (1.0 equiv.) and
triethylamine (4.7 equiv.) in CH2C12 (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: 20% MeOH in 20 mM NH40Ac, B:
1 5 100% MeOH, 9 mL / min, RT = 26.12) to give 58 as a colorless film.
Preparation of MPA taxoid 59
cH,coo-
2 0 32 ss
MPA taxoid 59. The synthesis of the taxoid-7-MPA 5 9
differs only slightly from the synthesis of taxoid-2'-MPA 58. The C-
2 taxoid 32 is dissolved in methylene chloride (.006 M) and treated
25 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
SUBSTITUTE SHEET (RULE 28)

wo 9sns~9s ~ ~ 8 ~ ~ ~ ~ rcrnJS9s~ooasi
.70.
mixture is then directly purified by HPLC (Vydak RP-18, 22.5 x 3
mm, A -~ B 0.5 h linear, A: 2096 MeOH in 20 mM NH40Ac, B: 100°k
MeOH, 9 mL / min, RT = 26.12) to give 59 . as a colorless film.
Preparation of MPA taxoid 60
BzNH O O OH CH~COO'
~I ~
R,~O,~..
OH ' O
~O HOAc
's_ ~~o
36 gp
MPA taxoid 60. The synthesis of the taxoid-7-MPA 6 0
differs only slightly from the synthesis of taxoid-2'-MPA 57. The C
2 taxoid 3b 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, 22.5 x 3
I 5 mm, A -> B 0.5 h linear, A: 2096 MeOH in 20 mM NH40Ac, B: 100%
MeOH, 9 mL / min, RT = 26.12) to give 60 as a colorless film.
SUBSTITUTE SHEET (RULE 2~

W095I18798 ~ PCTIUS95100481
-71-
Preparation of C-2-taxoid-2'-methyl-pyridinium salts
0
~w ,
o =
x. ~
I. X . CH9C00; TaO; BFI; halides II.
_ ~' O ~'' S S ~ N N
62 58 57 62 63
Ms2N ~ / ~ ~ / / ~~SPh
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 CH2C12
(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
1 5 HPLC (Vydak RP-18, 22.5 x 3 mm, A -> B 0.5 h linear, A: 20% MeOH
in 20 mM NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give
(62-66) II. as a colorless film.
SUBSTITUTE SHEET (RULE 28)

W095/18798
PCl'IUS95100481
-72-
Preparation of C-2-taxoid-7-methyl-pyridinium salts
x
X a CHsC00; TaO; BFI; ha8des O
I.
II.
O
,~ ~ S S
N~ ~ N~
s
67 59 60 68 ~ 69
MozN ~ / ~ ~ \ ~ ~~SPh
70 71 72
C-2-taxoid-7-opium salts 67-72. The synthesis of the
taxoid-7-methyl-pyridinium salts (67-72) II, differs only slightly
from the synthesis of taxoid-2'-methyl-pyridinium salts (62-66) II.
The C-2 taxoid (67-72) I is dissolved in methylene chloride (.006
M) and treated sequentially with triethylamine (40 equivalents) and
2-fluoro-I-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, 22.5 x 3 mm, A ~ B 0.5 h linear, A: 20°!o MeOH in 20 mM
NH40Ac, B: I00°k MeOH, 9 mL / min, RT = 26.12) to give (67-72) II
as a colorless film.
SUBSTITUTE SHEET (RULE 2B~

R'O 95118798 PCTlUS95100481
-73-
Preparation of C-2-taxoid-bis-2',7-methyl-pyridinium salts
x
X=CH:COv; isv;ur~;naaaes
I. II.
0
R= '~ 1 / ~ 1S/ ~1 / ~ I ~ ~ I ~
73 74 75 76 77
..»
~ ,~~SPh
78 79 80
C-2-taxoid-bis-2',7-onium salts 73-80. The synthesis of
C-2-taxoid-bis-2',7-methyl-pyridinium salts II (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 equivalents) Aldrich Chemicals, and allowed
to stir at ambient temperature for 18 hours. 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% MeOH in 20 mM NH40Ac, B: 100% MeOH, 9 mL /
1 5 min, RT = 26.12) to give (73-80) II as a colorless film.
SUBSTfTUTE SHEET (RULE 28)

w0 95118798 PCTIUS95I00481
-74-
Preparation of C-2-taxoid-2'-benzothiazolium salts
BzNH
~N~ O
X ~ * A~kf'~
O
H
I. X . CPi3C00'; TsO; BFI; halides
II.
R=_ t'' lo/ ~ ls/ ''y.ls/ ~ I N. I''l,
81 82 83 84 ~ 85
..",»
MaZN ~ ~ ~ I ~ ~ ~~SPh
86 87 88
C-2-taxoid-2'-benzothiazoliu m salts 81-88. A solution
of taxoid 81-88 (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 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: 20~Yo MeOH
in 20 mM NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give
81-88 as a colorless film.
SUBSTITUTE SHEET (RULE 28j

WO 95f 18798 2 ~ $ ~ 4 ~ ~ PCTlIJS95100481
-75-
Preparation of C-2-taxoid-7-benzothiazolium salts
BzNH O BzNH O "~'~ N + X'
Alkyl
Ph~O~,.. -~ Ph~O,
OH OH = .,
R
X = CHyC00; TsO; BFI; he8das O
II.
1S/ ~'~. / ~ I / 5'n. I
89 90 91 92 93
.,..»
Mash ~ / ~ I / / ~~SPh
94 95 96
C-2-taxoid-7-benzothiazolium salts (89-96). The
synthesis of the taxoid-7-benzothiazolium salts (89-96) II, differs
only slightly from the synthesis of taxoid-2'-benzothiazolium salts
(81-88) II. The C-2 taxoid (89-96) 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:
20% MeOH in 20 mM NH40Ac> B: 100% MeOH, 9 mL / min, RT =
1 5 26.12) to give (89-96) II as a colorless film.
SUBSTITUTE SHEET (RtiLE 2~

WO 95/18798
PC1'/U595100481
-76-
Preparation of C-2-taxoid-2'-benzoxazolium salts
Ac0 O OH Ac0 O OH
Bz~~ BzN
1 _ . _
Ph 0....
ph 0....
O ' _
OH ° O O
HO - H - O ~ I ~Y HO = H '
p OAc N p OAc
X- + Alkyl
O O
I, X s CH9C00'; TsO; BFI halides
II.
O
1S/ ~ S/ I N' I N
97 98 99 100 ~ 101
MezN ~ I ~ I / / ~~SPh
102 103 104
C-2-taxoid-2'-benzoxazolium salts 97-104. A solution of
taxoid (97-104) I. (1.0 equiv.) and triethylamine (4.7 equiv.) in
C H 2 C 12 (0.025 M) at 25 °C is treated with 2-chloro-3-
ethylbenzoxazolium tetrafluoroborate from AIdrich 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, 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: 20°lo MeOH
in 20 mM NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give
(97-104) II. as a colorless film.
SUBSTITUTE SHEET (RtiLE 28)

21~~~~5
w0 95118798 PCT/U895100481
-77-
Preparation of C-2-taxoid'-7-benzoxazolium salts
BzN~ Ac0 O OH
Ph
OH - O
HO - H a
p OAc
R1 R
O X = CHaC00; TsO; BFI; hdidw O
I.
II.
R= ~ ~O/ ~ ~S/ y4. S/ ~ ~ N tS. ~ Nw
105 108 107 108 109
MeyN ~ ~ ~ I ~ ~ ~~SPh
110 111 112
C-2-taxoid--7-benzoxazolium salts (105-112). The
synthesis of the taxoid-7-benzoxazolium salts (105-112) II, differs
only slightly from the synthesis of taxoid-2'-benzoxazolium salts
(97-104) II. The C-2 taxoid (105-112) 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 mixture is then directly purified by HPLC (Vydak RP-
18, 22.5 x 3 mm, A ~ B 0.5 h linear, A: 20% MeOH in 20 mM
1 5 NH~OAc, B: 1009'o MeOH, 9 mL / min, RT = 26.12) to give (105-112)
II as a colorless film.
SUBSTITUTE SHEET (RULE 28~

w0 95/18798 ~ ~ ~ PCT'lUS95100481
-78-
Preparation of C-2-taxoid-2'-pyrimidinium salts
-- X a CF13C00; TsO; BFI; halides II.
O
113 114 115 116 ~ 117
MezN ~ / ~ ~ ~, / ~~SPh
118 119 120
C-2-taxoid-2'-pyrimidinium salts 113-120. A solution
of taxoid (113-120) I. (1.0 equiv.) and triethyiamine (4.7 equiv.) in
C H 2 C12 (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, 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: 20% MeOH in 20 mM
NH40Ac, B: 100% MeOH, 9 mL / min, RT = 26.12) to give (113-120)
II. as a colorless film.
SUBSTITUTE SHEET (RULE 28~

WO 95118798 PCTIUS95100481
-79-
.°.
Preparation of C-2-taxoiil-7-pyrimidinium salts
N+ X'
Alkyl
OH ~ , H~ :
HO\
X = CH~COO; TsO; BFI; hslidea
II.
R- ~ 10/ ~ 's/ '~. / I I i '~ I .,~
121 122 123 124 125
Me2N ~ / ~ I / / ~~SPh
126 127 128
C-2-taxoid-7-pyrimidinium salts (121-128). The
synthesis of the taxoid-7-pyrimidinium salts (121-128) II, differs
only slightly from the synthesis of taxoid-2'-pyrimidinium salts
( 1 13 - 12 0 ) I I . 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 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: 20% MeOH in 20 mM NH40Ac, B: 100°k MeOH, 9 mL / nun,
1 5 RT = 26.12) to give (121-128) II as a colorless film.
SUBSTITUTE SHEET (RULE 2B)

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