U.S. patent application number 11/931080 was filed with the patent office on 2008-02-28 for vinorelbine derivatives.
This patent application is currently assigned to AMR TECHNOLOGY, INC.. Invention is credited to Jeffrey M. RALPH, Ian L. SCOTT, Matthew E. VOSS.
Application Number | 20080051425 11/931080 |
Document ID | / |
Family ID | 34676709 |
Filed Date | 2008-02-28 |
United States Patent
Application |
20080051425 |
Kind Code |
A1 |
SCOTT; Ian L. ; et
al. |
February 28, 2008 |
VINORELBINE DERIVATIVES
Abstract
The present invention relates to novel vinorelbine derivatives.
Pharmaceutical compositions containing these compounds as well as
processes of preparation and processes of use for treatment of
various conditions are also disclosed.
Inventors: |
SCOTT; Ian L.; (Woodinville,
WA) ; RALPH; Jeffrey M.; (Niskayuna, NY) ;
VOSS; Matthew E.; (Nassau, NY) |
Correspondence
Address: |
NIXON PEABODY LLP - PATENT GROUP
CLINTON SQUARE
P.O. BOX 31051
ROCHESTER
NY
14603-1051
US
|
Assignee: |
AMR TECHNOLOGY, INC.
5429 Main Street P.O. Box 2587
Manchester Center
VT
05255-2587
|
Family ID: |
34676709 |
Appl. No.: |
11/931080 |
Filed: |
October 31, 2007 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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11697370 |
Apr 6, 2007 |
|
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|
11931080 |
Oct 31, 2007 |
|
|
|
11003583 |
Dec 3, 2004 |
7235564 |
|
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11697370 |
Apr 6, 2007 |
|
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60527194 |
Dec 4, 2003 |
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Current U.S.
Class: |
514/283 ;
540/478 |
Current CPC
Class: |
C07D 519/04 20130101;
C07D 519/00 20130101; A61P 35/00 20180101 |
Class at
Publication: |
514/283 ;
540/478 |
International
Class: |
A61K 31/4745 20060101
A61K031/4745; C07D 491/14 20060101 C07D491/14 |
Claims
1. A compound of the following chemical formula: ##STR45## where:
R.sub.1 is alkyl; alkenyl; alkynyl; CN; SR.sub.5; or CF.sub.3;
R.sub.2=alkyl or CH(O); R.sub.5=hydrogen, alkyl, alkenyl, alkynyl,
aryl, or heterocyclyl; or a pharmaceutically acceptable salt
thereof, wherein the alkyl and alkenyl groups may be branched,
straight, unsubstituted, and/or substituted and wherein the aryl,
alkynyl, and heterocyclyl groups are substituted or
unsubstituted.
2. A compound of the following chemical formula: ##STR46## where:
R.sub.1=alkyl or SR.sub.5; R.sub.5=hydrogen, alkyl, alkenyl,
alkynyl, aryl, or heterocyclyl; or a pharmaceutically acceptable
salt thereof, wherein the alkyl and alkenyl groups may be branched,
straight, unsubstituted, and/or substituted and wherein the aryl,
alkynyl, and heterocyclyl groups are substituted or
unsubstituted.
3. A compound of the following chemical formula: ##STR47## where:
R.sub.1=alkyl; or a pharmaceutically acceptable salt thereof,
wherein the alkyl group may be branched, straight, unsubstituted,
and/or substituted.
4. A compound of the following chemical formula: ##STR48## where:
R.sub.5=alkyl; or a pharmaceutically acceptable salt thereof,
wherein the alkyl group may be branched, straight, unsubstituted,
and/or substituted.
Description
[0001] This application is a continuation of U.S. patent
application Ser. No. 11/697,370, filed Apr. 6, 2007, which is a
continuation of U.S. patent application Ser. No. 11/003,583, filed
Dec. 3, 2004, now U.S. Pat. No. 7,235,564, which claims benefit of
U.S. Provisional Patent Application Ser. No. 60/527,194, filed Dec.
4, 2003, which are hereby incorporated by reference in their
entirety.
FIELD OF THE INVENTION
[0002] The present invention relates to vinorelbine derivatives
which are potent inhibitors of cellular mitosis and proliferation,
as well as pharmaceutical compositions, preparation processes, and
methods of use for treatment of various conditions.
BACKGROUND OF THE INVENTION
Cellular Proliferation and Cancer
[0003] The disruption of external or internal regulation of
cellular growth can lead to uncontrolled cellular proliferation and
in cancer, tumor formation. This loss of cellular growth control
can occur at many levels and, indeed, does occur at multiple levels
in most tumors. Under these circumstances, although tumor cells can
no longer control their own proliferation, such cells still must
use the same basic cellular machinery employed by normal cells to
drive their growth and replication.
Mitosis and Spindle Formation
[0004] In a process known as mitosis, cancer cells, like all
mammalian cells, multiply through replication and segregation of
the original chromosomes. Following DNA replication in the S phase,
the cells progress in the G2 phase. During the G2 phase, cells
continue to increase in mass and prepare for mitosis. If chromosome
damage is present in the G2 phase, the affected cell responds by
activating the G2 phase checkpoint, which prevents progression into
mitosis. In the absence of DNA damage or following repair of
damage, the G2 phase cells then enter the M phase in which the
identical pairs of chromosomes are separated and transported to
opposite ends of the cell. The cell then undergoes division into
two identical daughter cells.
[0005] In a process known as spindle formation, the cell utilizes
the mitotic spindle apparatus to separate and pull apart the
chromosomes. This apparatus, in part, consists of a network of
microtubules that form during the first stage of mitosis.
Microtubules are hollow tubes that are formed by the assembly of
tubulin heterodimers from alpha- and beta-tubulin. The assembly of
tubulin into microtubules is a dynamic process with tubulin
molecules being constantly added and subtracted from each end.
Vinca Compounds as Inhibitors of Mitosis and Cellular
Proliferation
[0006] In general, vinca compounds are known to be inhibitors of
mitosis and cellular proliferation. In particular, the
antiproliferative activity of the vinca alkaloid class of drugs has
been shown to be due to their ability to bind tubulin. Assembly of
tubulin into microtubules is essential for mitosis and the binding
of the vincas to tubulin leads to cell cycle arrest in M phase and
subsequently to apoptosis. For example, at low concentrations,
these compounds interfere with the dynamics of microtubule
formation. At higher concentrations, they cause microtubule
disassembly, and at still higher concentrations, the formation of
tubulin paracrystals.
[0007] Moreover, the anti-cancer activity of vinca alkaloids is
generally believed to result from a disruption of microtubules
resulting in mitotic arrest. However, cytotoxicity of vinca
alkaloids also has been demonstrated in non-mitotic cells.
Considering the role of microtubules in many cellular processes,
the cytotoxic action of vinca alkaloids may involve contributions
from inhibition of non-mitotic microtubule-dependent processes.
[0008] Cytotoxicity may also be a consequence of changes in
membrane structure resulting from the partitioning of vinca
alkaloids into the lipid bilayer. Studies with another tubulin
binding compound, taxol, have shown that cell cycle arrest was not
a precondition for apoptosis by agents of this type. Therefore, the
anti-cancer activity of vinca alkaloids may be the result from
disruption of a number of distinct microtubule-dependent and
possibly microtubule-independent processes.
[0009] The assembly of tubulin into microtubules is a complex
process involving dynamic instability (i.e. the switching between
periods of slow growth and rapid shortening at both ends of the
microtubule), and treadmilling (i.e. the addition of tubulin to one
end of the microtubule occurring at the same rate as loss of
tubulin from the other). Low concentrations of vinca alkaloids have
been shown to bind to the ends of the microtubules and suppress
both microtubule instability and treadmilling during the metaphase
stage of mitosis. For example, vinca alkaloids have been shown to
stabilize microtubule plus ends and destabilize microtubule minus
ends. Although the spindle is retained under these conditions,
there is frequently abnormal alignment of condensed chromosomes. At
higher concentrations of vinca alkaloids, the spindle is not
present and the chromosome distribution resembles that of
prometaphase cells. At both low and high concentrations of vincas,
mitotic arrest results from activation of metaphase-anaphase
checkpoint. The molecular basis of this checkpoint is a negative
signal sent from the kinetochore of chromosomes that are not
attached to microtubules. This signal prevents the activation of
pathways that result in the initiation of anaphase events.
[0010] Although there is a common binding site for the vinca
alkaloids on tubulin, the members of this class do behave
differently. The relative overall affinities for .beta.-tubulin
binding are
vincristine>vinblastine>vinorelbine>vinflunine, but there
is no significant difference in the affinity of all four drugs for
tubulin heterodimers. The discrepancy has primarily been explained
by differences in the affinities of vinca-bound heterodimers for
spiral polymers and the binding of drug to unliganded polymers. For
example, tubulin spirals induced by vinflunine are significantly
smaller than those induced by vinorelbine.
[0011] In addition, vinca alkaloids also differ in their effects on
microtubule dynamics. Vinflunine and vinorelbine suppress dynamic
instability through: slowing the microtubule growth rate,
increasing the mean duration of a growth event and reducing the
duration of shortening. In contrast, vinblastine reduces the rate
of shortening and increases the percentage of time the microtubules
spend in the attenuated state. Vinblastine, vinorelbine, and
vinflunine all suppress treadmilling, with vinblastine displaying
the greatest potency.
In Vivo Properties
[0012] The vinca derivatives fall into the general class of
cytotoxic anti-cancer agents and, as such, suffer from the same
problem as all cytotoxics--i.e., toxicity. Vincristine and
vinblastine are neurotoxic. Vinorelbine, which is structurally very
similar to vinblastine and vincristine and is only slightly less
potent, is less neurotoxic. This change in toxicity cannot be
explained by examination of the binding affinity of these compounds
for tubulin alone. It has been postulated to arise from an increase
in sensitivity to changes in microtubule dynamics in tumor cells
and, as described above, these compounds have been shown to have
subtly different effects. It could also arise from changes in
cellular uptake of the drug. Vinflunine is not very potent in vitro
yet is active in vivo, and this has been attributed to its superior
cellular uptake. There are also quite significant differences in
the profile of efficacy of vinca alkaloids. Vincristine has found
wide use in the treatment of hematologic malignancies including
leukemias and lymphomas. It is also widely used in pediatric solid
tumors and, in the past, in small cell lung cancer. Vinblastine is
an important component of the combination regimen that is curative
for testicular cancer. Vinorelbine is quite different and has found
use mainly in breast cancer and non-small cell lung cancer.
[0013] There remains a need for novel vinca derivatives with
improved pharmacological and therapeutic properties, improved
processes for the preparations of such vinca derivative compounds,
corresponding pharmaceutical compositions, and methods of use.
[0014] The present invention is directed to achieving these
objectives.
SUMMARY OF THE INVENTION
[0015] The present invention relates to a compound of Formula (I)
as follows: ##STR1## where:
[0016] R.sub.1=alkyl; [0017] alkenyl; [0018] alkynyl; [0019] aryl;
[0020] heterocyclyl; [0021] halogen; [0022] CN; [0023] CH(O);
[0024] COR.sub.5; [0025] SO.sub.2NHNH.sub.2; [0026]
SO.sub.2NR.sub.5NH.sub.2; [0027] SO.sub.2NR.sub.5NHR.sub.6; [0028]
SO.sub.2NR.sub.5NR.sub.6R.sub.7; [0029] SO.sub.2NHNHR.sub.5; [0030]
SO.sub.2NHNR.sub.5R.sub.6; [0031] CO.sub.2R.sub.5; [0032] SR.sub.5;
[0033] SSR.sub.5; [0034] SOR.sub.5; [0035] SO.sub.2R.sub.5; [0036]
SO.sub.2NHR.sub.5; [0037] SO.sub.2NR.sub.5R.sub.6; [0038]
B(OR.sub.5).sub.2; [0039] CF.sub.3; [0040] SH; [0041]
SO.sub.2NH.sub.2; [0042] NH.sub.2; [0043] NHR.sub.5; [0044]
NHCOR.sub.5; [0045] NHSO.sub.2R.sub.5; [0046] NR.sub.5R.sub.6;
[0047] NR.sub.5COR.sub.6; or [0048] NR.sub.5SO.sub.2R.sub.6; [0049]
R.sub.5 and R.sub.6 can form a ring
[0050] R.sub.2=alkyl or CH(O);
[0051] R.sub.3=hydrogen, alkyl, or C(O)R.sub.5;
[0052] R.sub.4=hydrogen or C(O)R.sub.5;
[0053] R.sub.5 and R.sub.6 each are independently alkyl, alkenyl,
alkynyl, aryl, or heterocyclyl;
[0054] X.dbd.OR.sub.5, NR.sub.5R.sub.6, NHNH.sub.2,
NHNHC(O)R.sub.5, OH, NHR.sub.5, NH.sub.2, or NHNHC(O)H;
[0055] R.sub.4 and X may be linked together with intervening atoms
to form a ring; or a pharmaceutically acceptable salt thereof,
wherein the alkyl and alkenyl groups may be branched, straight,
unsubstituted, and/or substituted and wherein the aryl, alkynyl,
and heterocyclyl groups are substituted or unsubstituted.
[0056] Most preferably:
[0057] R.sub.1=alkyl; [0058] alkenyl; [0059] alkynyl; [0060] aryl;
[0061] heterocyclyl; [0062] halogen; [0063] CN; [0064] CH(O);
[0065] COR.sub.5; [0066] CO.sub.2R.sub.5; [0067] SR.sub.5; [0068]
SSR.sub.5; [0069] SH; [0070] NH.sub.2; [0071] NHR.sub.5; or [0072]
NR.sub.5R.sub.6;
[0073] More preferably:
[0074] R.sub.1=alkyl; [0075] alkenyl; [0076] alkynyl; [0077]
halogen; [0078] CN; [0079] SR.sub.5; [0080] SSR.sub.5; [0081] SH;
[0082] NH.sub.2; [0083] NHR.sub.5; [0084] NR.sub.5R.sub.6; where
R.sub.5 and R.sub.6 form a ring
[0085] Another aspect of the present invention relates to a process
for preparation of a derivative product compound of Formula (I) as
follows: ##STR2## where:
[0086] R.sub.1 is: [0087] alkyl; [0088] alkenyl; [0089] alkynyl;
[0090] aryl; [0091] heterocyclyl; [0092] CN; [0093] CH(O); [0094]
COR.sub.5; [0095] SO.sub.2NHNH.sub.2; [0096]
SO.sub.2NR.sub.5NH.sub.2; [0097] SO.sub.2NR.sub.5NHR.sub.6; [0098]
SO.sub.2NR.sub.5NR.sub.6R.sub.7; [0099] SO.sub.2NHNHR.sub.5; [0100]
SO.sub.2NHNR.sub.5R.sub.6; [0101] CO.sub.2R.sub.5; [0102] SR.sub.5;
[0103] SSR.sub.5; [0104] SOR.sub.5; [0105] SO.sub.2R.sub.5; [0106]
SO.sub.2NHR.sub.5; [0107] SO.sub.2NR.sub.5R.sub.6; [0108]
B(OR.sub.5).sub.2; [0109] CF.sub.3; [0110] SH; [0111]
SO.sub.2NH.sub.2; [0112] NH.sub.2; [0113] NHR.sub.5; [0114]
NHCOR.sub.5; [0115] NHSO.sub.2R.sub.5; [0116] NR.sub.5R.sub.6;
[0117] NR.sub.5COR.sub.6; or [0118] NR.sub.5SO.sub.2R.sub.6; [0119]
R.sub.5 and R.sub.6 can form a ring;
[0120] R.sub.2=alkyl or CH(O);
[0121] R.sub.3=hydrogen, alkyl, or C(O)R.sub.5;
[0122] R.sub.4=hydrogen or C(O)R.sub.5;
[0123] R.sub.5 and R.sub.6 each are independently alkyl, alkenyl,
alkynyl, aryl, or heterocyclyl;
[0124] X.dbd.OR.sub.5, NR.sub.5R.sub.6, NHNH.sub.2,
NHNHC(O)R.sub.5, OH, NHR.sub.5, NH.sub.2, or NHNHC(O)H;
[0125] R.sub.4 and X may be linked together with intervening atoms
to form a ring; or a pharmaceutically acceptable salt thereof,
wherein the alkyl and alkenyl groups may be branched, straight,
unsubstituted, and/or substituted and wherein the aryl, alkynyl,
and heterocyclyl groups are substituted or unsubstituted. The
process involves converting an intermediate compound of formula:
##STR3##
[0126] wherein Y is a halogen, under conditions effective to
produce the product compound of Formula (I).
[0127] Another aspect of the present invention relates to a process
for preparation of a derivative product compound of Formula (I) as
follows: ##STR4## where:
[0128] R.sub.1 is: [0129] halogen;
[0130] R.sub.2=alkyl or CH(O);
[0131] R.sub.3=hydrogen, alkyl, or C(O)R.sub.5;
[0132] R.sub.4=hydrogen or C(O)R.sub.5;
[0133] R.sub.5 and R.sub.6 each are independently alkyl, alkenyl,
alkynyl, aryl, or heterocyclyl;
[0134] X.dbd.OR.sub.5, NR.sub.5R.sub.6, NHNH.sub.2,
NHNHC(O)R.sub.5, OH, NHR.sub.5, NH.sub.2, or NHNHC(O)H;
[0135] R.sub.4 and X may be linked together with intervening atoms
to form a ring; or a pharmaceutically acceptable salt thereof,
wherein the alkyl and alkenyl groups may be branched, straight,
unsubstituted, and/or substituted and wherein the aryl, alkynyl,
and heterocyclyl groups are substituted or unsubstituted. The
process involves halogenating a starting material of the formula:
##STR5## under conditions effective to form the derivative product
compound
[0136] The present invention also relates to a method for
inhibiting cell proliferation in mammals, which comprises
administering a therapeutically effective amount of the compound of
Formula (I) to the mammal.
[0137] The present invention also relates to a method for treating
a condition in mammals, which comprises administering a
therapeutically effective amount of the compound of Formula (I) to
the mammal. The condition can be bacterial infection, allergy,
heart disease, AIDS, Human T-lymphotropic virus I infection, Human
herpesvirus 3, Human herpesvirus 4, Human papillomavirus, diabetes
mellitus, rheumatoid arthritis, Alzheimer's Disease, inflammation,
arthritis, asthma, malaria, autoimmune disease, eczema, Lupus
erythematosus, psoriasis, rheumatic diseases, Sjogren's syndrome,
and viral infection.
[0138] The present invention also relates to a pharmaceutical
composition of matter, which comprises the compound of Formula (I)
and one or more pharmaceutical excipients.
DETAILED DESCRIPTION OF THE INVENTION
[0139] The present invention relates to novel vinorelbine
derivatives, corresponding pharmaceutical compositions, preparation
processes, and methods of use for treatment of various
conditions.
[0140] In general, the novel compounds of the vinca family of
compounds of the present invention, include derivatives of
vinorelbine. In accordance with the present invention, such
derivative compounds are represented by the chemical structures of
Formula (I) as shown herein.
[0141] In particular, the present invention relates to a compound
of Formula (I) as follows: ##STR6## where:
[0142] R.sub.1 is: [0143] alkyl; [0144] alkenyl; [0145] alkynyl;
[0146] aryl; [0147] heterocyclyl; [0148] halogen; [0149] CN; [0150]
CH(O); [0151] COR.sub.5; [0152] SO.sub.2NHNH.sub.2; [0153]
SO.sub.2NR.sub.5NH.sub.2; [0154] SO.sub.2NR.sub.5NHR.sub.6; [0155]
SO.sub.2NR.sub.5NR.sub.6R.sub.7; [0156] SO.sub.2NHNHR.sub.5; [0157]
SO.sub.2NHNR.sub.5R.sub.6; [0158] CO.sub.2R.sub.5; [0159] SR.sub.5;
[0160] SSR.sub.5; [0161] SOR.sub.5; [0162] SO.sub.2R.sub.5; [0163]
SO.sub.2NHR.sub.5; [0164] SO.sub.2NR.sub.5R.sub.6; [0165]
B(OR.sub.5).sub.2; [0166] CF.sub.3; [0167] SH; [0168]
SO.sub.2NH.sub.2; [0169] NH.sub.2; [0170] NHR.sub.5; [0171]
NHCOR.sub.5; [0172] NHSO.sub.2R.sub.5; [0173] NR.sub.5R.sub.6;
[0174] NR.sub.5COR.sub.6; or [0175] NR.sub.5SO.sub.2R.sub.6; [0176]
R.sub.5 and R.sub.6 can form a ring;
[0177] R.sub.2=alkyl or CH(O);
[0178] R.sub.3=hydrogen, alkyl, or C(O)R.sub.5;
[0179] R.sub.4=hydrogen or C(O)R.sub.5;
[0180] R.sub.5 and R.sub.6 each are independently alkyl, alkenyl,
alkynyl, aryl, or heterocyclyl;
[0181] X.dbd.OR.sub.5, NR.sub.5R.sub.6, NHNH.sub.2,
NHNHC(O)R.sub.5, OH, NHR.sub.5, NH.sub.2, or NHNHC(O)H;
[0182] R.sub.4 and X may be linked together with intervening atoms
to form a ring; or a pharmaceutically acceptable salt thereof,
wherein the alkyl and alkenyl groups may be branched, straight,
unsubstituted, and/or substituted and wherein the aryl, alkynyl,
and heterocyclyl groups are substituted or unsubstituted.
[0183] In one embodiment, the present invention relates to a
compound where R.sub.3=acetyl.
[0184] In another embodiment, the present invention relates to a
compound where R.sub.4=hydrogen.
[0185] In another embodiment, the present invention relates to a
compound where X.dbd.OMe.
[0186] In another embodiment, the present invention relates to a
compound where R.sub.3=acetyl, R.sub.4 hydrogen, and X.dbd.OMe.
[0187] In another embodiment, the present invention relates to a
compound where R.sub.2=CH(O).
[0188] In another embodiment, the present invention relates to a
compound where R.sub.2=alkyl.
[0189] Representative examples of the compounds of Formula (I) are
set forth in Table 1 below: TABLE-US-00001 TABLE 1 Compounds of
Formula (I) Example NAME OF VINCA Number COMPOUND OF FORMULA (I)
COMPOUND 1 ##STR7## 11'-bromovinorelbine 2 ##STR8##
11'-iodovinorelbine 3 ##STR9## 11'-vinylvinorelbine 4 ##STR10##
11'-(3-oxohex-1-enyl) vinorelbine 5 ##STR11## 11'-(2-tert-butoxy
carbonylvinyl) vinorelbine 6 ##STR12## 11'-(carboxyvinyl)
vinorelbine 7 ##STR13## 11'-(methoxycarbonyl ethylsulfanyl)
vinorelbine 8 ##STR14## 11'-thiovinorelbine 9 ##STR15##
11'-(methoxycarbonyl methylsulfanyl) vinorelbine 10 ##STR16##
11'-(methylsulfanyl) vinorelbine 11 ##STR17## 11'-(ethylsulfanyl)
vinorelbine 12 ##STR18## 11'-(4-hydroxybutyl sulfanyl)vinorelbine
13 ##STR19## 11'-(3-hydroxypropyl sulfanyl)vinorelbine 14 ##STR20##
11'-(2-hydroxyethyl sulfanyl)vinorelbine 15 ##STR21##
11'-(2-fluorobenzyl sulfanyl)vinorelbine 16 ##STR22##
11'-(2-chlorobenzyl sulfanyl)vinorelbine 17 ##STR23##
11'-(phenysulfanyl) vinorelbine 18 ##STR24## 11'-(3-hydroxypheny
sulfanyl)vinorelbine 19 ##STR25## 11'-(3-hydroxyethyl
sulfinyl)vinorelbine 20 ##STR26## 11'-(3-hydroxypropyl
sulfinyl)vinorelbine 21 ##STR27## 11'-ethynylvinorelbine 22
##STR28## 11'-hexynylvinorelbine 23 ##STR29##
11'-(4-methylpentynyl) vinorelbine 24 ##STR30## 11'-(3-methoxy
propynyl)vinorelbine 25 ##STR31## 11'-cyanovinorelbine 26 ##STR32##
11'-acetylvinorelbine 27 ##STR33## 11'-(methoxycarbonyl)
vinorelbine 28 ##STR34## 11'-(2,2,2-trichloro ethoxycarbonyl)
vinorelbine 29 ##STR35## 11'-(2,2-dichloroethoxy
carbonyl)vinorelbine 30 ##STR36## 11'-phenylvinorelbine 31
##STR37## 11'-(3-hydroxyphenyl) vinorelbine 32 ##STR38##
11'-(3,5-dimethyl isoxazol-4yl)vinorelbine 33 ##STR39##
3,11'-dimethyl vinorelbine 34 ##STR40## 3-methyl-11'-iodo
vinorelbine 35 ##STR41## 11'-aminovinorelbine 36 ##STR42##
11'-(4-methoxyphenyl amino)vinorelbine
[0190] In yet another embodiment of the present invention, a
complex can be formed which includes 2 structures of Formula (I)
joined together at their R.sub.1 groups, wherein each R.sub.1 is
--S--.
[0191] The synthetic reaction schemes for the preparation of
compounds of Formula (I) are depicted below.
[0192] A synthetic scheme for preparing compounds of Formula (I) is
shown in Scheme 1 below. A vinca alkaloid is treated with either
N-iodosuccinimide to introduce an iodine in the 11'-position or
subjected to enzymatic bromination to introduce a bromine in the
11'-position. Pd-mediated coupling is then used to introduce other
functionality at this position. This methodology can be used to
introduce alkyl, alkenyl, alkynyl, aryl, heterocyclyl, acyl and
formyl groups and to form sulphides. Each of these groups can then
be subjected to further derivitization following stand methods of
organic synthesis. ##STR43##
[0193] In practicing either of the above processes, a variety of
catalysts may be utilized, such as palladium chloride, palladium
acetate, tetrakis(triphenylphosphine)palladium(0),
tris(dibenzylideneacetone)dipalladium(0),
dichlorobis(triphenylphosphine)palladium(II),
benzylchlorobis(triphenylphosphine)palladium(II),
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II),
tetrakis(triphenylphosphine)palladium, or
bis(triphenylphosphine)palladium(II)dichloride. Additionally, the
catalyst reactivity can be modified by addition of appropriate
ligands or additives. Representative ligands or additives include:
2-(dicyclohexylphosphino)-2',4',6'-tri-i-propyl-1,1'-biphenyl,
2-(dicyclohexylphosphino)-2',6'-dimethoxy-1,1'-biphenyl, PPh.sub.3,
t-Bu.sub.3P, CuI, or CuBr.
[0194] Based on the results obtained in the standard
pharmacological test procedures described below, the compounds of
the present invention are useful in inhibiting cellular
proliferation in a mammal by administering to such mammal an
effective amount of compound(s) of the present invention.
[0195] In particular, such vinca derivatives are useful as
antineoplastic agents. More particularly, the compounds of the
present invention are useful for inhibiting the growth of
neoplastic cells, causing cell death of neoplastic cells, and
eradicating neoplastic cells. The compounds of the present
invention are, therefore, useful for treating solid tumors, (e.g.,
sarcomas), carcinomas, (e.g., astrocytomas), lymphomas, (e.g.,
adult T-cell lymphoma), different cancer disease types, (e.g.,
prostate cancer, breast cancer, small cell lung cancer, ovarian
cancer, (Hodgkin's Disease), and other neoplastic disease states
(e.g., leukemias, particularly adult T-cell leukemias).
[0196] Since vinca compounds are known to be tubulin inhibitors,
the compounds of the present invention would also be expected to be
useful in treating the following conditions: bacterial infection;
allergy; heart disease; AIDS; Human T-lymphotropic virus 1
infection; Human herpesvirus 3; Human herpesvirus 4; Human
papillomavirus; diabetes mellitus; rheumatoid arthritis;
Alzheimer's Disease; inflammation; arthritis; asthma; malaria;
autoimmune disease; eczema; Lupus erythematosus; psoriasis;
rheumatic diseases; Sjogren's syndrome; and viral infection.
[0197] The vinca derivatives of the present invention can be
administered alone as indicated above, or utilized as biologically
active components in pharmaceutical compositions with suitable
pharmaceutically acceptable carriers, adjuvants and/or
excipients.
[0198] In accordance with the present invention, the compounds
and/or corresponding compositions can be introduced via different
administration routes, which include orally, parenterally,
intravenously, intraperitoneally, by intranasal instillation, or by
application to mucous membranes, such as, that of the nose, throat,
and bronchial tubes.
[0199] The active compounds of the present invention may be orally
administered, for example, with an inert diluent, or with an
assimilable edible carrier, or they may be enclosed in hard or soft
shell capsules, or they may be compressed into tablets.
[0200] The quantity of the compound administered will vary
depending on the patient and the mode of administration and can be
any effective amount. The quantity of the compound administered may
vary over a wide range to provide in a unit dosage an effective
amount of from about 0.01 to 20 mg/kg of body weight of the patient
per day to achieve the desired effect. The amount of active
compound in such therapeutically useful compositions is such that a
suitable dosage will be obtained. Preferred compositions according
to the present invention are prepared so that an oral dosage unit
contains between about 1 and 250 mg of active compound.
[0201] For example, with oral therapeutic administration, these
active compounds may be incorporated with excipients and used in
the form of tablets, capsules, elixirs, suspensions, syrups, and
the like. Such compositions and preparations should contain at
least 0.1% of active compound. The percentage of the compound in
these compositions may, of course, be varied and may conveniently
be between about 2% to about 60% of the weight of the unit.
[0202] The tablets, capsules, and the like may also contain a
binder such as gum tragacanth, acacia, corn starch, or gelatin;
excipients such as dicalcium phosphate; a disintegrating agent such
as corn starch, potato starch, alginic acid; a lubricant such as
magnesium stearate; and a sweetening agent such as sucrose,
lactose, or saccharin. When the dosage unit form is a capsule, it
may contain, in addition to materials of the above type, a liquid
carrier, such as a fatty oil.
[0203] Various other materials may be present as coatings or to
modify the physical form of the dosage unit. For instance, tablets
may be coated with shellac, sugar, or both.
[0204] These active compounds and/or pharmaceutical compositions
may also be administered parenterally. Solutions of these active
compounds and/or compositions can be prepared in water. Dispersions
can also be prepared in glycerol, liquid polyethylene glycols, and
mixtures thereof in oils.
[0205] Illustrative oils are those of animal, vegetable, or
synthetic origin, for example, peanut oil, soybean oil, or mineral
oil. In general, water, saline, aqueous dextrose and related sugar
solution, and glycols such as, propylene glycol or polyethylene
glycol, are preferred liquid carriers, particularly for injectable
solutions. Under ordinary conditions of storage and use, these
preparations contain a preservative to prevent the growth of
microorganisms.
[0206] The pharmaceutical forms suitable for injectable use include
sterile aqueous solutions or dispersions and sterile powders for
the extemporaneous preparation of sterile injectable solutions or
dispersions. In all cases, the pharmaceutical form of the present
invention must be sterile and must be fluid to the extent that easy
syringability exists. It must be stable under the conditions of
manufacture and storage and must be preserved against the
contaminating action of microorganisms, such as bacteria and fungi.
The carrier can be a solvent or dispersion medium containing, for
example, water, ethanol, polyol (e.g., glycerol, propylene glycol,
and liquid polyethylene glycol), suitable mixtures thereof, and
vegetable oils.
[0207] The compounds and/or pharmaceutical compositions of the
present invention may also be administered directly to the airways
in the form of an aerosol. For use as aerosols, the compounds of
the present invention in solution or suspension may be packaged in
a pressurized aerosol container together with suitable propellants,
for example, hydrocarbon propellants like propane, butane, or
isobutane with conventional adjuvants. The materials of the present
invention also may be administered in a non-pressurized form such
as in a nebulizer or atomizer.
[0208] Some of the compounds of the present invention can be in the
form of pharmaceutically acceptable acid-addition and/or base
salts. All of these forms of salts are within the scope of the
present invention.
[0209] Pharmaceutically acceptable acid addition salts of the
compounds of the present invention include salts derived from
nontoxic inorganic acids, such as hydrochloric acid, nitric acid,
phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid,
hydrofluoric acid, phosphorous acid, and the like, as well as the
salts derived from nontoxic organic acids, such as aliphatic mono-
and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy
alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and
aromatic sulfonic acids, etc. Such salts thus include sulfates,
pyrosulfates, bisulfates, sulfites, bisulfites, nitrates,
phosphates, monohydrogenphosphates, dihydrogenphosphates,
metaphosphates, pyrophosphates, chlorides, bromides, iodides,
acetates, trifluoroacetates, propionates, caprylates, isobutyrates,
oxalates, malonates, succinate suberates, sebacates, fumarates,
maleates, mandelates, benzoates, chlorobenzoates, methylbenzoates,
dinitrobenzoates, phthalates, benzenesulfonates, toluenesulfonates,
phenylacetates, citrates, lactates, malates, tartrates,
methanesulfonates, and the like. Also contemplated are salts of
amino acids, such as arginates, gluconates, and galacturonates
(see, for example, Berge S M. et al., "Pharmaceutical Salts,"
Journal of Pharmaceutical Science, 66:1-19 (1997), which is hereby
incorporated by reference in its entirety).
[0210] The acid addition salts of said basic compounds are prepared
by contacting the free base forms with a sufficient amount of the
desired acid to produce the salt in the conventional manner.
[0211] Pharmaceutically acceptable base addition salts are formed
with metals or amines, such as alkali and alkaline earth metals or
organic amines. Examples of metals used as cations are sodium,
potassium, magnesium, calcium, and the like. Examples of suitable
amines are N,N-dibenzylethylenediamine, chloroprocaine, choline,
diethanolamine, dicyclohexylamine, ethylenedianline,
N-methylglucamine, and procaine (see, for example, Berge S. M. et
al., "Pharmaceutical Salts," Journal of Pharmaceutical Science,
66:1-19 (1997), which is hereby incorporated by reference in its
entirety).
[0212] The base addition salts of the acidic compounds are prepared
by contacting the free acid form with a sufficient amount of the
desired base to produce the salt in the conventional manner.
[0213] Certain of the compounds of the present invention can exist
in unsolvated forms as well as solvated forms, including hydrated
forms. In general, the solvated forms, including hydrated forms,
are equivalent to unsolvated forms and are intended to be
encompassed within the scope of the present invention.
[0214] The present invention can be used in conjunction with other
known cancer treatments, including other chemotherapeutic agents
and radiation.
EXAMPLES
[0215] The following examples are provided to illustrate
embodiments of the present invention but are no means intended to
limit its scope.
[0216] Spectroscopic analysis of products described in the
experimental procedures below were performed with conventional or
standard scientific instrumentation known in the art. Proton NMR
spectra were obtained on a Bruker AC 300 spectrometer at 300 MHz or
a Bruker 500 MHz spectrometer at 500 MHz and were referenced to
tetramethylsilane as an internal standard. Mass spectra were
obtained on either a Shimadzu QP-5000 or a PE Sciex API 150 Mass
Spectrometer.
Example 1
Preparation of 11'-Bromovinorelbine
[0217] A solution of N-iodosuccinimide (288 mg, 1.28 mmol) in
trifluoroacetic acid/methylene chloride (1:1, 40 mL) was cooled to
approximately 0.degree. C. in an ice water jacketed addition funnel
then added dropwise to vinorelbine tartrate (1.35 g, 1.28 mmol) in
trifluoroacetic acid/methylene chloride (1:1, 60 mL) at -15.degree.
C. The temperature was monitored by an internal thermometer and
maintained at -15.+-.3.degree. C. during the course of the addition
(45 min). After stirring for 0.5 h, additional N-iodosuccinamide
(15 mg, 0.067 mmol) in trifluoroacetic acid/methylene chloride
(1:1, 5 mL at 0.degree. C. was added dropwise to drive the reaction
to completion. The reaction mixture was then poured carefully into
a rapidly stirring mixture of 10% sodium
sulfite/chloroform/saturated sodium hydrogencarbonate (1:1:2, 400
mL). Solid sodium hydrogencarbonate was then added in small
portions until gas evolution stopped. The solution was then
extracted with choroform (3.times.100 mL) and the combined extracts
were washed with 10% sodium sulfite (50 mL) and brine (50 mL) then
dried over magnesium sulfate. The solvent was remove in vacuo to
provide 11'-iodovinorelbine (1.21 g, quantitative) as a tan foam
which was carried forward without further purification: .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 8.14 (d, J=1.3 Hz, 1H), 7.45 (dd,
J=8.6, 1.5 Hz, 1H), 7.21 (d, J=8.6 Hz, 1H), 6.34 (s, 1H), 6.20 (bs,
1H), 5.79-5.87 (m, 2H), 5.28 (d, J=10.4 Hz, 1H), 4.68 (d, J=14.3
Hz, 1H), 4.56 (d, J=14.2 Hz, 1H), 3.94 (d, J=17.0 Hz, 1H), 3.86 (s,
3H), 3.76 (s, 3H), 3.74 (s, 3H), 3.51-3.64 (m, 3H) 3.33 (m, 1H),
3.22 (m, 1H), 3.11 (m, 1H), 2.72 (s, 3H), 2.54-2.72 (m, 5H), 2.31
(m, 1H), 2.04-2.16 (m, 3H), 2.02 (s, 3H), 1.73-1.88 (m, 2H), 1.66
(m, 1H), 1.34 (m, 1H), 1.12 (t, J=7.5 Hz, 3H), 0.67 (t, J=7.3 Hz,
3H); ESI MS m/z 905 [M+H].sup.+.
Example 2
Preparation of 11'-Iodovinorelbine
[0218] To an ice cold solution of vinorelbine ditartrate (0.084 g,
0.286 mmol) in trifluoroacetic acid (6 mL) under nitrogen was added
a solution of N-iodosuccinimide (0.084 g, 0.365 mmol) in
trifluoroacetic acid (3 mL) dropwise and the mixture was stirred at
0.degree. C. for 1.25 hours. The reaction was quenched by the
dropwise addition of saturated NaHCO.sub.3 and the mixture
extracted with dichloromethane. The organic layer was washed with
saturated NaHCO.sub.3, dried with Na.sub.2SO.sub.4, filtered and
concentrated under reduced pressure. Purification by flash
chromatography (silica gel, chloroform to 25% methanol in acetone)
gave 11'-iodovinorelbine (220 mg, 85%). .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 8.06 (d, J=1 Hz, 1H), 7.40 (dd, J=9, 1 Hz, 1H),
7.16 (dd, J=9 Hz, 1H), 6.31 (s, 1H), 6.25 (s, 1H), 5.84 (dd, J=10,
4 Hz, 1H), 5.74 (br, 1H), 5.45 (s, 1H), 5.30 (s, 1H), 5.26 (d, J=10
Hz, 1H), 4.31 (d, J=13 Hz, 1H), 4.23 (d, J=13 Hz, 1H), 3.85 (s,
3H), 3.76 (s, 3H), 3.73 (s, 3H), 3.65 (d, J=16 Hz, 1H), 3.58 (s,
1H), 3.35 (s, 1H), 3.37-3.15 (m), 3.00 (q, J=7 Hz, 1H), 2.72 (s,
3H), 2.75-2.50 (m, 3H), 2.58 (s, 1H), 2.31 (m, 2H), 2.15-1.98 (m,
3H), 2.02 (s, 3H), 1.78 (m, 1H), 1.70-1.45 (m, 2H), 1.40-1.20 (m,
1H), 1.09 (t, J=7 Hz, 3H), 0.68 (t, J=7 Hz, 3H); ESI MS m/z 905
[M+H].sup.+.
Example 3
Preparation of 11'-Vinylvinorelbine
[0219] A solution of 11'-iodovinorelbine (45 mg, 0.050 mmol) in DME
(0.5 mL) and water (0.2 mL) was deoxygenated with argon for 3
minutes. The reaction vessel was charged with
2,4,6-trivinylcyclotriboroxane pyridine complex (13 mg, 0.055
mmol), Pd(PPh.sub.3).sub.4 (7.5 mg, 0.070 mmol), and
K.sub.2CO.sub.3 (7.6 mg, 0.055 mmol) and the mixture was heated to
80-90.degree. C. After 2 h, the reaction appeared complete by ESI
mass spectral analysis. The reaction mixture was diluted with
saturated NaHCO.sub.3 (8 mL) and extracted with EtOAc (2.times.2
mL). The combined extracts were dried (Na.sub.2SO.sub.4) and
concentrated to a brown solid which was purified by flash
chromatography (silica gel, [CHCl.sub.3/MeOH/NH.sub.4OH
(40:18:2)]/CH.sub.2Cl.sub.2, 1:99 to 10:90) to yield
11'-vinylvinorelbine (15 mg, 31%) as awhite solid: .sup.1HNMR (500
MHz, CD.sub.3OD) .delta. 10.23 (br s, 1H), 7.73 (s, 1H), 7.34 (s,
1H), 6.82 (dd, J=17.5, 11 Hz, 1H), 6.61 (s, 1H), 6.41 (s, 1H),
5.94-5.91 (m, 1H), 5.87-5.86 (m, 1H), 5.74-5.70 (m, 1H), 5.63 (d,
J=10.5 Hz, 1H), 5.31 (s, 1H), 5.13-5.11 (m, 1H), 4.93-4.90 (m, 1H),
4.70-4.67 (m, 1H), 4.07 (d, J=17.0 Hz, 1H), 3.94-3.87 (m, 5H),
3.81-3.71 (m, 8H), 3.41 (d, J=16.0 Hz, 1H), 3.19-3.09 (m, 2H),
3.43-3.40 (m, 1H), 3.19-3.09 (m, 2H), 2.85-2.78 (m, 4H), 2.64-2.59
(m, 1H), 2.35-2.29 (m, 1H), 2.18-2.06 (m, 6H), 1.96-1.92 (m, 1H),
1.73-1.69 (m, 1H), 1.49-1.43 (m, 1H), 1.14 (t, J=7.5 Hz, 3H), 0.72
(t, J=7.5 Hz, 3H); ESI MS m/z 805 [M+H].sup.+.
Example 4
Preparation of 11'-(3-Oxohex-1-enyl)vinorelbine
[0220] A solution of 11'-iodovinorelbine (0.61 g, 0.674 mmol) in
toluene (6 mL) was deoxygenated with argon, Pd(OAc).sub.2 (0.01 g,
0.045 mmol), PPh.sub.3 (0.02 g, 0.76 mmol) and triethylamine (0.15
mL, 1.08 mmol) were added and the mixture purged with argon again.
The mixture was heated to 70.degree. C., 1-hexen-3-one (0.2 mL, 1.7
mmol) was added and the heating continued for 5 h. The mixture was
cooled, deoxygenated again, Pd(OAc).sub.2 (0.01 mg, 0.045 mmol) and
1-hexen-3-one (0.78 mL, 0.67 mmol) were added and the mixture was
heated at 75.degree. C. overnight. After cooling to room
temperature, the mixture was diluted with EtOAc, filtered, and
concentrated under reduced pressure. Purification by reverse phase
chromatography (C18; MeCN/water containing 0.05% of NH.sub.4OH)
gave 11'-(3-oxohex-1-enyl)vinorelbine (18 mg, 31%). .sup.1H NMR
(300 MHz, CD.sub.3OD) .delta. 8.00 (s, 1H), 7.82 (d, J=16 Hz, 1H),
7.48 (d, J=14 Hz, 1H), 7.35 (d, J=15 Hz, 1H), 7.84 (d, J=16 Hz,
1H), 6.32 (s, 1H), 6.30 (s, 1H), 5.82 (dd, J=10, 4 Hz, 1H), 5.75
(d, J=4 Hz, 1H), 5.30 (s, 1H), 5.27 (d, J=10 Hz, 1H), 4.34 (s, 1H),
3.85 (s, 3H), 3.75 (s, 3H), 3.76-3.70 (m, 1H), 3.73 (s, 3H), 3.64
(m, 1H), 3.57 (s, 1H), 3.45-3.15 (m), 3.00 (m, 2H), 2.72 (s, 3H),
2.75-50 (m, 3H), 2.30 (m, 2H), 2.12-2.02 (m, 3H), 2.02 (s, 3H),
1.93 (s, 1H), 1.80-1.40 (m, 5H), 1.27 (m), 1.10 (t, J=7 Hz, 3H),
1.02 (t, J=7 Hz, 3H), 0.90 (m, 4H), 0.67 (t, J=7 Hz, 3H); ESI MS
m/z 875 [M+H].sup.+.
Example 5
Preparation of 11'-(2-tert-Butoxycarbonylvinyl)vinorelbine
[0221] Palladium(II) acetate (3.0 mg, 0.012 mmol),
triphenylphosphine (6.0 mg, 0.024 mmol), and triethylamine (35
.mu.L, 0.24 mmol) were added to a solution of 11'-iodovinorelbine
(107 mg, 0.118 mmol) in toluene (2 mL). The reaction mixture was
deoxygenated with an argon purge and tert-butyl acrylate (35 .mu.L,
0.236 mmol) was then added. The reaction mixture was heated to
70.degree. C. overnight. After cooling to room temperature, the
reaction mixture was diluted with CH.sub.2Cl.sub.2 (20 mL),
filtered through Celite, and concentrated to dryness. Purification
by column chromatography (silica gel, CH.sub.3OH/CH.sub.2Cl.sub.2,
7:93) gave a brown solid (48 mg, 45%) which was further purified by
reverse phase chromatography (C18, Waters Symmetry; isocratic 80%
acetonitrile/water, 0.05% NH.sub.4OH) to give
11'-(2-tert-butoxycarbonylvinyl)vinorelbine as a yellow solid (25
mg, 23%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.85 (s, 1H),
7.68 (d, J=15.9 Hz, 1H), 7.35 (dd, J=10.5, 1.2 Hz, 1H), 7.26 (d,
J=8.5, Hz, 1H), 6.33 (d, J=16.9 Hz, 1H), 6.23 (d, J=6.5 Hz, 2H),
5.75 (dd, J=10.1, 3.6 Hz, 1H), 5.66 (d, J=4.9 Hz, 1H), 5.23 (s,
1H), 5.19 (d, J=10.1 Hz, 1H), 4.26-4.17 (m, 2H), 3.77 (s, 3H), 3.68
(s, 3H), 3.65 (s, 3H), 3.58-3.50 (m, 3H), 3.30-3.08 (m, 3H), 2.93
(dd, J=15.4, 7.5 Hz, 1H), 2.64 (s, 3H), 2.64-2.41 (m, 3H),
2.24-2.14 (m, 2H), 2.09-1.86 (m, 3H), 1.94 (s, 3H), 1.76-1.68 (m,
1H), 1.62-1.37 (m, 2H), 1.47 (s, 9H), 1.32-1.21 (m, 1H), 1.02 (t,
J=7.4 Hz, 3H), 0.62 (t, J=7.3 Hz, 3H); ESI MS m/z 905
[M+H].sup.+.
Example 6
Preparation of 11'-(Carboxyvinyl)vinorelbine Trifluoroacetate
[0222] A solution of 11'-(2-tert-butoxycarbonyl-vinyl)vinorelbine
(25 mg, 0.03 mmol) in CH.sub.2Cl.sub.2 (1.0 mL) was treated with
TFA (0.3 mL) at 0.degree. C. and then stirred at room temperature
for 1 h. The reaction mixture was poured slowly into saturated
NaHCO.sub.3 (30 mL) and the mixture was extracted with
CH.sub.2Cl.sub.2 (60 mL). The organic layer was washed with brine,
dried over Na.sub.2SO.sub.4, and concentrated to give a yellow
solid (19 mg, 83%). The solid was treated with CH.sub.2Cl.sub.2 (1
mL) and a drop of trifluoroacetic acid. The solution was evaporated
to give 11'-(carboxyvinyl)vinorelbine trifluoroacetate as a yellow
solid (23.6 mg, 98%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
10.38 (s, 1H), 7.86 (s, 1H), 7.72 (d, J=15.9 Hz, 1H), 7.38-7.29 (m,
2H), 6.57 (s, 1H), 6.35 (d, J=15.9 Hz, 1H), 6.31 (s, 1H), 5.82 (dd,
J=9.9, 4.3 Hz, 1H), 5.81-5.77 (m, 1H), 5.54 (d, J=9.8 Hz, 1H), 5.20
(s, 1H), 4.84 (d, J=14.5 Hz, 1H), 4.58 (d, J=14.5 Hz, 1H),
4.02-3.83 (m, 3H), 3.79 (s, 3H), 3.71 (s, 3H), 3.66 (s, 3H),
3.71-3.62 (m, 4H), 3.32 (d, J=12.5 Hz, 1H), 3.14-2.97 (m, 2H),
2.76-2.69 (m, 1H), 2.69 (s, 3H), 2.57-248 (m, 1H), 2.25-2.16 (m,
1H), 2.10-2.02 (m, 4H), 1.96 (s, 3H), 1.63-1.54 (m, 1H), 1.41-1.34
(m, 1H), 1.04 (t, J=7.4 Hz, 3H), 0.61 (t, J=7.2 Hz, 3H); ESI MS m/z
849 [M+H].sup.+.
Example 7
Preparation of 11'-(Methoxycarbonylethylsulfanyl)vinorelbine
vinorelbine Tartrate
[0223] Step 1: A flask containing 11'-iodovinorelbine (113 mg, 0.13
mmol), 1,1'-bis(diphenylphosphino)ferrocene (28 mg, 0.05 mmol), and
tris(dibenzylideneacetone)dipalladium(0) (11.4 mg, 0.012 mmol) was
deoxygenated with nitrogen then triethylamine (35 .mu.L, 0.25 mmol)
and NMP (1.1 mL) were added via syringe. The resulting solution was
purged again with nitrogen before the addition of methyl
3-mercaptopropionate (28 .mu.L, 0.25 mmol). The mixture was heated
at 60.degree. C. for 4 h. After cooling, the reaction mixture was
diluted with ethyl acetate, washed with water and brine, dried over
Na.sub.2SO.sub.4, and concentrated under vacuum. Purification by
flash chromatography (silica gel,
CHCl.sub.3/MeOH/Et.sub.3N=99:1:0.5), followed by reverse phase
chromatography (C18; MeCN/water with 0.1% of TFA) gave
11'-(methoxycarbonylethylsulfanyl)vinorelbine trifluoroacetate(22
mg, 21%). .sup.1H NMR (500 MHz, MeOD) .delta. 7.85 (d, J=Hz, 1H),
7.31 (d, J=8 Hz, 1H), 7.25 (dd, J=8, 2 Hz, 1H), 6.32 (s, 1H), 6.28
(s, 1H), 5.83 (dd, J=10, 4 Hz, 1H),5.76 (d, J=4 Hz, 1H), 5.31 (s,
1H), 5.27 (d, J=10 Hz, 1H), 4.38 (s, 2H), 3.85 (s, 3H), 3.76 (s,
3H), 3.76-3.70 (m, 1H), 3.73 (s, 3H), 3.64 (s, 3H), 3.58 (s, 1H),
3.41 (d, J=14 Hz, 1H), 3.37-3.32 (m), 3.21 (td, J=9, 5 Hz, 1H),
3.11 (t, J=7 Hz, 2H), 3.03 (dd, J=16, 8 Hz, 1H), 2.72 (s, 3H), 2.65
(br d, J=16 Hz, 1H), 2.62-2.55 (m, 4H), 2.40 (br dd, J=13, 5 Hz,
1H), 2.27 (td, J=10, 6 Hz, 1H), 2.12-2.02 (m, 3H), 2.02 (s, 3H),
1.79 (ddd, J=13, 11, 5 Hz, 1H), 1.63 (dd, J=14, 7 Hz, 1H),
1.62-1.58 (m, 1H), 1.32 (dd, J=14, 7 Hz, 1H), 1.10 (t, J=7 Hz, 3H),
0.69 (t, J=7 Hz, 3H); ESI MS m/z 898 [M+H].sup.+.
[0224] Step 2: To a stirred solution of 11'-[3-(methyl
3-mercaptopropionoate)]vinorelbine (16 mg, 0.018 mmol) in
ether/MeOH (2.0 mL/0.2 mL) at room temperature was added a solution
of L-tartaric acid (5.9 mg, 0.039 mmol) ether/MeOH (2.0 mL/0.2 mL)
and the resulting slurry was stirred at room temperature for 10
min, and refluxed for 12 min. Another 2 mL of ether was added and
the mixture cooled to 0.degree. C. The solid was collected by
filtration, washed with ether, and dried under vacuum to give the
tartrate salt (9.2 mg, 43%); mp: 140-170.degree. C. (dec.); .sup.1H
NMR (500 MHz, MeOD) 37.92 (s, 1H), 7.39 (d, J=8 Hz, 1H), 7.31 (dd,
J=8, 1 Hz, 1H), 6.36 (s, 1H), 6.30 (s, 1H), 5.90-5.84 (m, 2H), 5.33
(d, J=10 Hz, 1H), 5.29 (s, 1H), 4.93 (d, J=15 Hz, 1H), 4.68 (d,
J=15 Hz, 1H), 4.43 (s, 3.4 H), 4.12 (d, J=17 Hz, 1H), 3.87 (s, 3H),
3.77 (s, 3H), 3.75 (s, 3H), 3.78-3.74 (m, 1H), 3.68 (br d, J=14 Hz,
1H), 3.64 (s, 3H), 3.61 (s, 1H), 3.40 (dd, J=16, 5 Hz, 1H), 3.15
(t, J=7 Hz, 2H), 3.13 (m, 1H), 2.88 (dd, J=13, 4 Hz, 1H), 2.80-2.73
(m, 2H), 2.74 (s, 3H), 2.65-2.54 (m, 1H), 2.61 (t, J=7 Hz, 2H),
2.48-2.40 (m, 1H), 2.19-2.09 (m, 3H), 2.03 (s, 3H), 1.99-1.91 (m,
1H), 1.86-1.78 (m, 1H), 1.64 (m, 1H), 1.36 (m, 1H), 1.14 (t, J=7
Hz, 3H), and 0.69 (t, J=7 Hz, 3H); ESI MS m/z 898 [M+H].sup.+.
Example 8
Preparation of 11'-Thiovinorelbine Trifluoroacetate
[0225] 11'-Iodovinorelbine (44 mg, 0.049 mmol),
thiotriisopropysilyl potassium salt (33 mg, 0.156 mmol), and
tetrakis(triphenylphosphine)palladium(0) (11 mg, 0.010 mmol) were
combined in benzene/tetrahydrofuran (2.5 mL, 4:1) and the reaction
was deoxygenated by bubbling argon through the solution for 30 min.
The mixture was heated to 65.degree. C. for 1 h then diluted with
ethyl acetate (15 mL). The organic solution was washed with
saturated NaHCO.sub.3 (2.times.5 mL) and brine (5 mL), dried over
MgSO.sub.4, and evaporated to dryness in vacuo. The residue was
purified by reverse phase chromatography (C18, acetonitrile/water,
0.05% TFA) to provide 11'-thiovinorelbine trifluoroacetate (22.7
mg, 45% yield) as a white powder after lyophilization: .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 10.24 (bs, 1H), 7.70 (s, 1H), 7.28
(d, J=8.4 Hz, 1H), 7.13 (dd, J=8.5, 1.7 Hz, 1H), 6.61 (s, 1H), 6.41
(s, 1H), 5.92 (m, 1H), 5.86 (m, 1H), 5.62 (d, J=10.5 Hz, 1H), 5.30
(s, 1H), 4.84 (m, 1H), 4.64 (d, J=14.5, 1H), 4.06 (d, J=16.9 Hz,
1H), 3.94-3.68 (m, 6H), 3.88 (s, 3H), 3.81 (s, 3H), 3.75 (s, 3H),
3.40 (d, J=15.4 Hz, 1H), 3.14 (m, 3H), 2.81 (m, 1H), 2.78 (s, 3H),
2.59 (m, 1H), 2.30 (m, 1H), 2.15 (q, J=7.4 Hz, 2H), 2.06 (s, 3H),
1.93 (m, 1H), 1.68 (m, 1H), 1.46 (m, 1H), 1.13(t, J=7.5 Hz, 3H),
0.70 (t, J=7.3 Hz, 3H); ESI MS m/z 811 [M+H].sup.+.
Example 9
Preparation of 11'-(Methoxycarbonylmethylsulfanyl)vinorelbine
Trifluoroacetate
[0226] Methyl thioglycolate (63 mg, 0.596 mmol),
11'-iodovinorelbine (54 mg, 0.060 mmol), triethylamine (145 mg,
1.43 mmol) and tris(dibenzylideneacetone)dipalladium(0) (5.4 mg,
0.006 mmol) were combined in N-methyl-2-pyrrolidinone (1.5 mL) and
the reaction mixture was deoxygenated by bubbling argon through the
solution for 30 min. The mixture was heated at 60.degree. C. for 8
h then diluted with ethyl acetate (20 mL). The organic solution was
washed with saturated NaHCO.sub.3 (2.times.5 mL) and brine (5 mL),
dried over MgSO.sub.4, and evaporated to dryness in vacuo. The
residue was purified by reverse phase chromatography (C18,
acetonitrile/water, 0.05% TFA) to provide
11'-(methoxycarbonylmethylsulfanyl)vinorelbine trifluoroacetate
(5.8 mg, 9% yield) as a white powder after lyophilization: .sup.1H
NMR (500 MHz, CD.sub.3OD) .delta. 10.38 (bs, 1H), 7.90 (s, 1H),
7.36 (d, J=8.5 Hz, 1H), 7.30 (dd, J=8.5, 1.6 Hz, 1H), 6.63 (s, 1H),
6.41 (s, 1H), 5.93 (dd, J=10.4, 5.4 Hz, 1H), 5.87 (d, J=4.1 Hz,
1H), 5.62 (d, J=10.4 Hz, 1H), 5.31 (s, 1H), 4.88 (d, J=14.7 Hz,
1H), 4.67 (d, J=14.5, 1H), 4.08 (d, J=16.8 Hz, 1H), 3.95-3.55 (m,
8H), 3.88 (s, 3H), 3.81 (s, 3H), 3.75 (s, 3H), 3.65 (s, 3H), 3.39
(m, 1H), 3.14 (m, 2H), 2.83 (dd, J=13.7, 4.5 Hz, 1H), 2.78 (s, 3H),
2.63 (m, 1H), 2.31 (m, 1H), 2.16 (q, J=7.5 Hz, 2H), 2.08 (m, 1H),
2.06 (s, 3H), 1.94 (m, 1H), 1.70 (m, 1H), 1.47 (m, 1H), 1.14 (t,
J=7.5 Hz, 3H), 0.70 (t, J=7.3 Hz, 3H); ESI MS m/z 883
[M+H].sup.+.
Example 10
Preparation of 11'-(Methylsulfanyl)vinorelbine Trifluoroacetate
[0227] 11'-Iodovinorelbine (70 mg, 0.077 mmol),
tris(dibenzylideneacetone)dipalladium(0) (11 mg, 0.011 mmol),
1,1'-bis(diphenylphosphino)ferrocene (26 mg, 0.046 mmol),
1-methyl-2-pyrrolidinone (0.5 mL), and triethylamine (22 .mu.L)
were combined in a resealable glass test tube. Argon was bubbled
through the solution for 10 min. Methanethiol (0.29 mL of a 4 N
solution in NMP, 1.2 mmol) was added, the test tube sealed, and the
mixture was heated to 65.degree. C. After 5 h, additional
methanethiol (0.39 mL of a 4 N solution in NMP, 1.5 mmol) was
added, and the mixture was heated to 65.degree. C. overnight. After
cooling, the mixture was diluted with ethyl acetate (75 mL), washed
with saturated NH.sub.4Cl (3.times.15 mL), water and brine, then
dried (Na.sub.2SO.sub.4) and evaporated to dryness under vacuum.
The residue was purified by reverse phase chromatography (C18,
acetonitrile/water, 0.05% TFA) to provide
11'-(methylsulfanyl)vinorelbine trifluoroacetate (10 mg, 12% yield)
as a white powder after lyophilization: .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 10.25 (br s, 1H), 7.71(d, J=1.0 Hz, 1H), 7.33
(d, J=8.5 Hz, 1H), 7.18 (dd, J=8.5, 1.5 Hz, 1H), 6.63 (s, 1H), 6.41
(s, 1H), 5.93 (dd, J=10.5, 4.5 Hz, 1H), 5.87 (d, J=4.5 Hz, 1H),
5.63 (d, J=10.5 Hz, 1H), 5.31 (s, 1H), 4.90 (d, J=14.5 Hz, 1H),
4.67 (d, J=14.5 Hz, 1H), 4.07 (d, J=16.5 Hz, 1H), 3.95-3.88 (m,
4H), 3.83 (s, 3H), 3.81-3.71 (m, 7H), 3.44-3.40 (m, 1H), 3.19-3.08
(m, 2H), 2.83 (dd, J=13.5, 4.5 Hz, 1H), 2.74 (s, 3H), 2.64-2.58 (m,
1H), 2.49 (s, 3H), 2.35-2.29 (m, 1H), 2.18-2.06 (m, 6H), 1.98-1.91
(m, 1H), 1.73-1.68 (m, 1H), 1.48-1.43 (m, 1H), 1.14 (t, J=7.5 Hz,
3H), 0.71 (t, J=7.3 Hz, 3H); ESI MS m/z 825 [M+H].sup.+.
Example 11
Preparation of 11'-(Ethylsulfanyl)vinorelbine Trifluoroacetate
[0228] A solution of 11'-iodovinorelbine (70 mg, 0.07 mmol) in NMP
(1.5 mL) was deoxygenated with argon for 10 minutes. The reaction
vessel was charged with 1,1'-bis(diphenylphosphino)ferrocene (20
mg, 0.035 mmol), tris(dibenzylideneacetone)dipalladium(0) (10 mg,
0.011 mmol) and Et.sub.3N (0.10 mL, 0.72 mmol). The mixture was
stirred for 20 min at room temperature, and then ethanethiol (0.10
mL, 1.3 mmol) was added and then stirred at 60.degree. C.
overnight. The mixture was cooled to room temperature, diluted with
EtOAc (100 ml) and washed with saturated NH.sub.4Cl (3.times.10 mL)
and brine (3.times.10 mL), dried (Na.sub.2SO.sub.4) and
concentrated. The residue was filtered through a pad of silica gel,
eluted with CH.sub.2Cl.sub.2/MeOH (4:1, 2.times.100 mL). The
filtration was concentrated, purified by reverse phase
chromatography (C-18, acetonitrile/water, 0.05% TFA) to give
11'-(ethylsulfanyl)vinorelbine trifluoroacetate (10 mg, 15%) as a
light yellow solid: .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 10.29
(s, 1H), 7.81 (s, 1H), 7.34 (d, J=8.5 Hz, 1H), 7.23 (dd, J=8.5, 1.5
Hz, 1H), 6.67 (s, 1H), 6.41 (s, 1H), 5.93 (dd, J=10.5, 5.0 Hz, 1H),
5.86 (d, J=4.5 Hz, 1H), 5.63 (d, J=10.5 Hz, 1H), 5.31 (s, 1H), 4.90
(d, J=14.5 Hz, 1H), 4.65 (d, J=14.5 Hz, 1H), 4.06 (d, J=17 Hz, 1H),
3.93 (dd, J=15.0, 4.5 Hz, 2H), 3.89 (s, 3H), 3.81 (s, 3H), 3.78 (s,
2H), 3.75 (s, 3H), 3.72 (s, 1H), 3.43 (d, J=15.5 Hz, 1H), 3.24-3.16
(m, 1H), 3.10 (dd, J=15.5, 7.5 Hz, 1H), 2.90 (q, J=7.0 Hz, 2H),
2.82 (dd, J=13.5, 4.5 Hz, 1H), 2.79 (s, 3H), 2.61 (dd, J=15.5, 12.5
Hz, 1H), 2.36-2.28 (m, 1H), 2.16 (q, J=7.5 Hz, 2H), 2.07 (s, 3H),
1.97-1.89 (m, 1H), 1.70 (dd, J=14.5, 7.5 Hz, 1H), 1.47 (dd, J=14.5,
7.5 Hz, 1H), 1.90-1.85 (m, 1H), 1.70 (dd, J=14.5, 7.5 Hz, 1H), 1.47
(dd, J=14.5, 7.5 Hz, 1H), 1.24 (t, J=7.0 Hz, 3H), 1.13 (t, J=7.5
Hz, 3H), 0.71 (t, J=7.0 Hz, 3H); ESI MS m/z 839 [M+H].sup.+.
Example 12
Preparation of 11'-(4-Hydroxybutylsulfanyl)vinorelbine
[0229] 1,1'-Bis(diphenylphosphino)ferrocene (8 mg, 0.014 mmol),
tris(dibenzylideneacetone)dipalladium(0) (3.0 mg, 0.004 mmol), and
triethylamine (10 .mu.L, 0.07 mmol) were added o a solution of
11'-iodovinorelbine (31.6 mg, 0.035 mmol) in NMP (1 mL). The
reaction mixture was deoxygenated with an argon purge and
4-mercapto-1-butanol (8 .mu.L, 0.07 mmol) was then added. The
reaction mixture was heated to 60.degree. C. overnight. After
cooling to room temperature, the reaction mixture was diluted with
CH.sub.2Cl.sub.2 (20 mL), washed with water and brine, then dried
over Na.sub.2SO.sub.4 and concentrated to an oily residue.
Purification by column chromatography (silica gel,
CH.sub.3OH/CH.sub.2Cl.sub.2, 5:95) gave a brown solid (13 mg, 42%)
which was further purified by reverse phase chromatography (C18,
Waters Symmetry; isocratic 70% acetonitrile/water, 0.05%
NH.sub.4OH) to give 11'-(4-hydroxybutylsulfanyl)vinorelbine as a
brown solid (9 mg, 30%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.70 (s, 1H), 7.18-7.10 (m, 2H), 6.21 (d, J=5.2 Hz, 2H), 5.73 (dd,
J=10.1, 3.7 Hz, 1H), 5.64 (d, J=4.4 Hz, 1H), 5.21 (s, 1H), 5.16 (d,
J=10.1 Hz, 1H). 4.21-4.15 (m, 2H), 3.75 (s, 3H), 3.65 (s, 3H), 3.62
(s, 3H), 3.58-3.41 (m, 4H), 3.30-3.08 (m, 2H), 2.90 (dd, J=15.6,
6.6 Hz, 1H), 2.84-2.80 (m, 2H), 2.61 (s, 3H), 2.57-2.43 (m, 3H),
2.23-2.11 (m, 2H), 2.02-1.91 (m, 3H), 1.91 (s, 3H), 1.75-1.50 (m,
6H), 1.41-1.18 (m, 5H), 0.99 (t, J=7.4 Hz, 3H), 0.59 (t, J=7.2 Hz,
3H); ESI MS m/z 883 [M+H].sup.+.
Example 13
Preparation of 11'-(3-Hydroxypropylsulfanyl)vinorelbine
[0230] 1,1'-Bis(diphenylphosphino)ferrocene (12 mg, 0.022 mmol),
tris(dibenzylideneacetone)dipalladium(0) (5.0 mg, 0.006 mmol), and
triethylamine (15 .mu.L, 0.11 mmol) were added to a solution of
11'-iodovinorelbine (50 mg, 0.055 mmol) in NMP (1 mL). The reaction
mixture was deoxygenated with an argon purge and
3-mercapto-1-propanol (10 .mu.L, 0.11 mmol) was then added. The
reaction mixture was heated to 60.degree. C. overnight. After
cooling to room temperature, the reaction mixture was diluted with
CH.sub.2Cl.sub.2 (20 mL), washed with water and brine, then dried
over Na.sub.2SO.sub.4 and concentrated to an oily residue.
Purification by column chromatography (silica gel,
CH.sub.3OH/CH.sub.2Cl.sub.2, 7:93) gave a brown solid (20 mg, 42%)
which was further purified by reverse phase chromatography (C18,
Waters Symmetry; isocratic 70% acetonitrile/water, 0.05%
NH.sub.4OH) to give 11'-(3-hydroxypropylsulfanyl)vinorelbine as a
brown solid (10 mg, 19%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta.
7.81 (s, 1H), 7.29-7.21 (m, 2H), 6.33 (s, 2H), 5.84 (dd, J=10.4,
4.0 Hz, 1H), 5.75 (d, J=4.0 Hz, 1H), 5.32 (s, 1H), 5.28 (d, J=10.1
Hz, 1H). 4.32-4.21 (m, 2H), 3.86 (s, 3H), 3.77 (s, 3H), 3.73 (s,
3H), 3.69-3.58 (m, 3H), 3.39-3.19 (m, 2H), 2.98 (dd, J=7.1, 7.3 Hz,
3H), 2.72 (s, 3H), 2.69-2.54 (m, 3H), 2.33-2.23 (m, 2H), 2.11-2.03
(m, 3H), 2.03 (s, 3H), 1.81 (dd, J=7.3, 6.8 Hz, 2H), 1.86-1.76 (m,
1H), 1.71 (dd, J=7.9, 7.1 Hz, 1H), 1.63 (dd, J=6.9, 7.4 Hz, 1H),
1.53-1.43 (m, 1H), 1.36-1.30 (m, 3H), 1.10 (t, J=7.5 Hz, 3H), 0.71
(t, J=7.2 Hz, 3H); ESI MS m/z 869 [M+H].sup.+.
Example 14
Preparation of 11'-(2-Hydroxyethylsulfanyl)vinorelbine
[0231] 1,1'-Bis(diphenylphosphino)ferrocene (12 mg, 0.022 mmol),
tris(dibenzylideneacetone)dipalladium(0) (5.0 mg, 0.006 mmol), and
triethylamine (15 .mu.L, 0.11 mmol) were added to a solution of
11'-iodovinorelbine (50.5 mg, 0.056 mmol) in NMP (1 mL). The
reaction mixture was deoxygenated with an argon purge and
2-mercaptoethanol (8 .mu.L, 0.11 mmol) was then added. The reaction
mixture was heated to 60.degree. C. for 4 h. After cooling to room
temperature, the reaction mixture was diluted with CH.sub.2Cl.sub.2
(20 mL), washed with water and brine, then dried over
Na.sub.2SO.sub.4 and concentrated to an oily residue. Purification
by column chromatography (silica gel, CH.sub.3OH/CH.sub.2Cl.sub.2,
5:95) gave a brown solid (20 mg, 42%) which was further purified by
reverse phase chromatography (C18, isocratic 70%
acetonitrile/water, 0.05% NH.sub.4OH) to give
11'-(2-hydroxyethylsulfanyl)vinorelbine as a brown solid (7 mg,
15%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 7.76 (s, 1H),
7.23-7.15 (m, 2H), 6.23 (d, J=2.1 Hz, 2H), 5.76 (dd, J=10.1, 3.5
Hz, 1H), 5.66 (d, J=4.4 Hz, 1H), 5.23 (s, 1H), 5.19 (d, J=10.2 Hz,
1H). 4.25-4.14 (m, 2H), 3.77 (s, 3H), 3.68 (s, 3H), 3.64 (s, 3H),
3.61-3.50 (m, 4H), 3.30-3.09 (m, 2H), 2.93 (dd, J=7.0, 6.8 Hz, 3H),
2.64 (s, 3H), 2.67-2.46 (m, 3H), 2.24-2.14 (m, 2H), 2.09-1.94 (m,
3H), 1.94 (s, 3H), 1.54 (dd, J=7.0, 7.5 Hz, 1H), 1.39 (dd, J=7.8,
7.6 Hz, 1H), 1.81-1.21 (m, 5H), 1.01 (t, J=7.4 Hz, 3H), 0.62 (t,
J=7.2 Hz, 3H); ESI MS m/z 855 [M+H].sup.+.
Example 15
Preparation of 11'-(2-Fluorophenylmethyl)vinorelbine
Trifluoroacetate
[0232] A solution of 11'-iodovinorelbine (70 mg, 0.07 mmol) in NMP
(1.5 mL) was deoxygenated with argon for 10 minutes. The reaction
vessel was charged with 1,1'-bis(diphenylphosphino)ferrocene (20
mg, 0.035 mmol), tris(dibenzylideneacetone)dipalladium(0) (10 mg,
0.011 mmol) and Et.sub.3N (0.10 mL, 0.13 mmol). The mixture was
stirred for 20 min at room temperature, and then
2-fluorophenylmethane-thiol (20 mg, 0.141 mmol) was added and then
stirred at 60.degree. C. overnight. The mixture was cooled to room
temperature, diluted with EtOAc (100 ml) and washed with saturated
NH.sub.4Cl (3.times.10 mL) and brine (3.times.10 mL). The organic
layer was dried (Na.sub.2SO.sub.4), filtered, and concentrated. The
residue was filtered through a pad of silica gel, eluted with
CH.sub.2Cl.sub.2/MeOH (4:1, 2.times.100 mL). Purification by
reverse phase chromatography (C18, acetonitrile/water, 0.05% TFA)
gave 11'-(2-fluorobenzylsulfanyl)vinorelbine trifluoroacetate as an
off-white solid (3 mg, 4%): .sup.1H NMR (500 MHz, CD.sub.3OD)
.delta. 10.38 (s, 1H), 7.82 (s, 1H), 7.38 (dt, J=8.0, 1.5 Hz, 1H),
7.32 (d, J=8.5 Hz, 1H), 7.26-7.03 (m, 4H), 6.16 (s, 1H), 6.41 (s,
1H), 5.94 (dd, J=10.5, 4.0 Hz, 1H), 5.86 (d, J=4.5 Hz, 1H), 5.64
(d, J=11 Hz, 1H), 5.31 (s, 1H), 4.72 (dd, J=8.5, 4.0 Hz, 1H), 4.66
(d, J=14.5 Hz, 1H), 4.15-4.05 (m, 3H), 3.95-3.84 (m, 3H), 3.81 (s,
3H), 3.76 (s, 3H), 3.75-3.72 (m, 3H), 3.46-3.42 (m, 1H), 3.22-3.18
(m, 1H), 3.08 (dd, J=16.8, 8.0 Hz, 1H), 2.84-2.81 (m, 1H), 2.78 (s,
3H), 2.74 (s, 3H), 2.64-2.44 (m, 2H), 2.38-2.31 (m, 2H), 2.21-2.13
(m, 2H), 2.06 (s, 3H), 1.70 (dd, J=14.5, 7.2 Hz, 1H), 1.46 (dd,
J=15.0, 7.5 Hz, 1H), 1.13 (t, J=7.5 Hz, 3H), 0.71 (t, J=7.5 Hz,
3H); ESI MS m/z 919 [M+H].sup.+.
Example 16
Preparation of 11'-(2-Chlorophenylmethyl)vinorelbine
Trifluoroacetate
[0233] A solution of 11'-iodovinorelbine (70 mg, 0.07 mmol) in NMP
(1.5 mL) was deoxygenated with argon for 10 minutes. The reaction
vessel was charged with 1,1'-bis(diphenylphosphino)ferrocene (20
mg, 0.036 mmol), tris(dibenzylideneacetone)dipalladium(0) (10 mg,
0.01 mmol) and Et.sub.3N (0.10 mL, 0.13 mmol). The mixture was
stirred for 20 min at room temperature, and then
2-chlorobenzenmethanethiol (0.10 mL, 0.75 mmol) was added and then
stirred at 60.degree. C. overnight. The mixture was cooled to room
temperature, diluted with methylene chloride (100 mL) and washed
with saturated NH.sub.4Cl (3.times.10 mL) and brine (3.times.10
mL), dried over Na.sub.2SO.sub.4, and evaporated to dryness in
vacuo. The residue was purified by flash chromatography (silica
gel, CH.sub.2Cl.sub.2/MeOH 10:1) and then by reverse phase
chromatography (C18, acetonitrile/water, 0.05% TFA) to give
11'-(2-chlorobenzylsulfanyl)vinorelbine trifluoroacetate (9 mg,
yield 12%) as a light yellow solid: .sup.1H NMR (500 MHz,
CD.sub.3OD) 310.35 (s, 1H), 7.80 (s, 1H), 7.36 (dd, J=8.5, 1.0 Hz,
1H), 7.31 (d, J=8.5 Hz, 1H), 7.20-7.17 (m, 2H), 7.12-7.01 (m, 2H),
6.63 (s, 1H), 6.41 (s, 1H), 5.94 (dd, J=10.5, 5.0 Hz, 1H), 5.86 (d,
J=4.5 Hz, 1H), 5.64 (d, J=10.5 Hz, 1H), 5.30 (s, 1H), 4.98 (d,
J=14.5 Hz, 1H), 4.65 (d, J=14.5 Hz, 1H), 4.22 (d, J=13.0 Hz, 1H),
4.15 (d, J=12.5 Hz, 1H), 4.05 (d, J=16.5 Hz, 1H), 3.93 (dd, J=15.5,
5.5 Hz, 2H), 3.88 (s, 3H), 3.81 (s, 3H), 3.76 (s, 3H), 3.75-3.72
(m, 2H), 3.71 (s, 2H), 3.42 (d, J=16.0 Hz, 1H), 3.20-3.14 (m, 1H),
3.09 (dd, J=16.0, 7.5 Hz, 1H), 2.83-2.79 (m, 1H), 2.78 (s, 3H),
2.64-2.57 (m, 1H), 2.34-2.27 (m, 1H), 2.15 (q, J=7.0 Hz, 2H), 2.06
(s, 3H), 1.98-1.88 (m, 1H), 1.70 (dd, J=14.5, 7.5 Hz, 1H), 1.47
(dd, J=14.5, 7.5 Hz, 1H), 1.29 (s, 1H), 1.13 (t, J=7.5 Hz, 3H),
0.92-0.83 (m, 1H), 0.70 (t, J=7.5 Hz, 3H); ESI MS m/z 935
[M+H].sup.+.
Example 17
Preparation of 11'-(Phenysulfanyl)vinorelbine Ditartrate
[0234] Step 1: To a deoxygenated solution of 11'-iodovinorelbine
(105 mg, 0.116 mmol) in NMP (2.5 mL) was added
1,1'-bis(diphenylphosphino)ferrocene (36 mg, 0.065 mmol),
tris(dibenzylideneacetone)dipalladium(0) (16 mg, 0.017 mmol) and
Et.sub.3N (33 .mu.L, 0.24 mmol). The mixture was stirred for 10 min
at room temperature, benzenethiol (26 mg, 0.232 mmol) was added and
the mixture was stirred at 60.degree. C. overnight (16 h), then
cooled to room temperature, diluted with methylene chloride (15 ml)
and washed with saturated NH.sub.4Cl (5 mL). The organic layer was
dried (Na.sub.2SO.sub.4), filtered, and concentrated. Purification
by plug chromatography (silica gel, MeOH/CH.sub.2Cl.sub.2, 3:97),
followed by reverse phase chromatography (C18 column, CH.sub.3CN,
H.sub.2O, 0.05% NH.sub.4OH) and then flash chromatography (silica
gel, MeOH/CHCl.sub.3, 1:99) afforded 11'-(phenysulfanyl)vinorelbine
(23 mg, 22%) as a pale yellow solid: mp 202-206.degree. C.; .sup.1H
NMR (500 MHz, CD.sub.3OD) 37.89 (d, J=2 Hz, 1H), 7.35 (d, J=6 Hz,
1H), 7.25 (d, J=6 Hz, 1H), 7.22-7.16 (m, 2H), 7.16-7.07 (m, 3H),
6.32 (s, 1H), 6.31 (s, 1H), 5.84 (dd, J=6, 5 Hz, 1H), 5.73 (s, 1H),
5.32 (s, 1H), 5.27 (d, J=10 Hz, 1H), 4.31 (d, J=13 Hz, 1H), 4.23
(d, J=12 Hz, 1H), 3.85 (s, 3H), 3.76 (s, 3H), 3.74 (s, 3H), 3.61
(br s, 1H), 3.58 (s, 1H), 3.37 (d, J=15 Hz, 1H), 3.25-3.16 (m, 2H),
3.00 (dd, J=15, 7 Hz, 1H), 2.72 (s, 3H), 2.68-2.55 (m, 3H),
2.32-2.33 (m, 2H), 2.12-2.04 (m, 3H), 2.02 (s, 3H), 1.85-1.77 (m,
1H), 1.68-1.59 (m, 1H), 1.53 (br s, 1H), 1.39-1.25 (m, 2H),
1.04-1.10 (m, 3H), 0.70 (t, J=7 Hz, 3H); ESI MS m/z 887
[M+H].sup.+.
[0235] Step 2: To a solution of 11'-(phenysulfanyl)vinorelbine (9.9
mg, 0.0112 mmol) in MeOH (100 .mu.L) and Et.sub.2O (2 mL) was added
a solution of L-tartaric acid (4.5 mg, 0.030 mmol) in MeOH (120
.mu.L) and Et.sub.2O (2 mL). The solid was collected by filtration
to yield 11'-(phenysulfanyl)vinorelbine ditartrate (10.5 mg, 79%)
as a white solid: mp 239-242.degree. C. (dec); .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 8.04 (br s, 1H), 7.45 (d, J=8 Hz, 1H),
7.32 (d, J=8 Hz, 1H), 7.23 (t, J=7 Hz, 2H), 7.17 (d, J=7 Hz, 2 H),
7.14 (t, J=7 Hz, 1H), 6.37 (s, 1H), 6.33 (br s, 1H), 5.88 (s, 2H),
5.34 (d, J=10 Hz, 1H), 5.30 (s, 1H), 4.90 (m), 4.69 (d, J=14 Hz,
1H), 4.44 (s, 4H), 4.08 (d, J=17 Hz, 1H), 3.88 (s, 3H) 3.77 (s,
3H), 3.76 (s, 3H), 3.76-3.70 (m, 2H), 3.62 (s, 1H), 3.44 (d, J=16
Hz, 1H), 3.15 (dd, J=15, 7 Hz, 1H), 2.94-2.76 (m, 3H), 2.74 (s,
3H), 2.62 (t, J=12 Hz, 1H), 2.47 (br m, 1H), 2.19-2.09 (m, 3H),
2.03 (s, 3H), 1.96 (br s, 1H), 1.84 (br s, 1H), 1.69-1.60 (m, 1H),
1.43-1.33 (m, 1H), 1.13 (t, J=7 Hz, 3H), 0.70 (t, J=7 Hz, 3H); ESI
MS m/z 887 [M+H].sup.+.
Example 18
Preparation of 11'-(3-Hydroxyphenysulfanyl)vinorelbine
Ditartrate
[0236] Step 1: To a flask containing 11'-iodovinorelbine (100 mg,
0.111 mmol) was added 1,1'-bis(diphenylphosphino)ferrocene (36 mg,
0.065 mmol), tris(dibenzylideneacetone)dipalladium(0) (15 mg, 0.016
mmol) NMP (2.0 mL) and Et.sub.3N (33 .mu.L, 0.24 mmol). The mixture
was stirred for 10 min at room temperature. A solution of
3-hydroxythiophenol (28 mg, 0.22 mmol) in NMP (0.5 mL) was added
and the mixture was stirred at 60.degree. C. overnight (16 h). The
mixture was cooled to room temperature, diluted with methylene
chloride (15 ml) and saturated NH.sub.4Cl (5 mL) was added. The
organic layer was dried (Na.sub.2SO.sub.4), filtered, and
concentrated. Purification by plug chromatography (silica gel,
eluent: MeOH/CH.sub.2Cl.sub.2, 3:97) followed by reverse phase
chromatography (C8 column, CH.sub.3CN, H.sub.2O, 0.05% NH.sub.4OH),
flash chromatography (silica gel, 0.5% to 4% MeOH/CHCl.sub.3), and
then reverse phase chromatography (C18, H.sub.2O, CH.sub.3CN, 0.1%
TFA) gave 11'-(3-hydroxyphenysulfanyl)vinorelbine trifluoroacetate
(10 mg, 10%) as a white solid: mp 236-242.degree. C. (dec); .sup.1H
NMR (500 MHz, CD.sub.3OD) .delta. 7.93 (s, 1H), 7.36 (d, J=8 Hz,
1H), 7.27 (dd, J=8, 1 Hz, 1H), 7.01 (t, J=8 Hz, 1H), 6.62 (d, J=8
Hz, 1H), 6.54-6.49 (m, 2H), 6.31 (d, J=9 Hz, 1H), 5.84 (dd, J=10, 5
Hz, 1H), 5.74 (br s, 1H), 5.32 (s, 1H), 5.27 (d, J=10 Hz, 1H), 4.33
(d, J=13 Hz, 1H), 4.26 (d, J=13 Hz, 1H), 3.86 (s, 3H), 3.76 (s,
3H), 3.74 (s, 3H), 3.65-3.57 (m, 2H), 3.42-3.32 (m, 1H), 3.26-3.18
(m, 2H), 3.02 (dd, J=15, 8 Hz, 1H), 2.72 (s, 3H), 2.70-2.56 (m,
3H), 2.34-2.25 (m, 2H), 2.13-1.98 (m, 7H), 1.85-1.78 (m, 1H),
1.67-1.60 (m, 1H), 1.56 (br s, 1H), 1.45-1.39 (m, 1H), 1.38-1.30
(m, 1H), 1.07 (t, J=7 Hz, 3H), 0.71 (t, J=7 Hz, 3H); ESI MS m/z 903
[M+H].sup.+.
[0237] Step 2: To a solution of 11'-(phenysulfanyl)vinorelbine (10
mg, 0.0113 mmol) in MeOH (0.1 mL) and Et.sub.2O (2 mL) was added a
solution of L-tartaric acid (10 mg, 0.067 mmol) in MeOH (0.1 mL)
and Et.sub.2O (1 mL). Collection of the precipitate by filtration
gave 11'-(3-hydroxyphenysulfanyl)vinorelbine ditartrate (10.5 mg,
79%) as a white solid: mp 204-210.degree. C. (dec); .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 8.03 (s, 1H), 7.44 (d, J=8 Hz, 1H),
7.31 (d, J=8 Hz, 1H), 7.03 (t, J=8 Hz, 1H), 6.65 (d, J=7 Hz, 1H),
6.66-6.30 (m, 2H), 6.41-6.35 (m, 2H), 6.90-6.84 (m, 2H), 5.36 (d,
J=10 Hz, 1H), 5.30 (s, 1H), 4.90 (d, 1H), 4.66 (d, J=14 Hz, 1H),
4.43 (s, 4H), 4.11 (m, 1H), 3.88 (s, 3H), 3.79-3.67 (m, 8H), 3.63
(s, 1H), 3.50-3.42 (m, 1H), 3.14 (dd, J=15, 7 Hz, 1H), 2.95-2.83
(m, 3H), 2.75 (s, 3H), 2.65-2.52 (m, 2H), 2.19-2.05 (m, 3H), 2.03
(s, 3H), 1.96-1.85 (m, 2H), 1.68-1.59 (m, 1H), 1.42-1.33 (m, 1H),
1.12 (t, J=7 Hz, 3H), 0.71 (t, J=7 Hz, 3H); ESI MS m/z 903
[M+H].sup.+.
Example 19
Preparation of 11'-(3-Hydroxyethylsulfinyl)vinorelbine
Trifluoroacetate
[0238] Hydrogen peroxide (30%, 10 .mu.L, 0.35 mmol) was added
dropwise to a solution of 11'-(2-hydroxyethylsulfanyl)vinorelbine
(36.0 mg, 0.04 mmol) in CH.sub.3CO.sub.2H/H.sub.2O (2:1, 1.5 mL) at
0.degree. C. The reaction mixture was stirred at 0.degree. C. for
10 min and then at room temperature for 1.5 h. The reaction mixture
was poured slowly into saturated NaHCO.sub.3 (30 mL) and then
extracted with CH.sub.2Cl.sub.2 (60 mL). The organic layer was
washed with brine, dried over Na.sub.2SO.sub.4, and concentrated.
Purification by preparative TLC (silica gel, MeOH/CH.sub.2Cl.sub.2,
3:17) provided the two diasteromers of
11'-(2-hydroxyethylsulfinyl)vinorelbine as yellow solids
(diasteromer 1:6.8 mg, first to elute and diasteromer 2:5.0 mg,
32%).
[0239] Diasteromer 1 (6.8 mg) was treated with CH.sub.2Cl.sub.2 (1
mL) and a drop of TFA. The solution was evaporated to give
11'-(2-hydroxyethylsulfinyl)vinorelbine trifluoroacetate as a
yellow solid (8.9 mg, 97%): .sup.1H NMR (300 MHz, CD.sub.3OD)
.delta. 10.70 (s, 1H), 8.01 (s, 1H), 7.53 (d, J=8.6 Hz, 1H), 7.43
(dd, J=8.6, 1.3 Hz, 1H), 6.64 (s, 1H), 6.32 (s, 1H), 5.83 (dd,
J=10.7, 3.4 Hz, 1H), 5.80-5.79 (m, 1H), 5.55 (d, J=10.8 Hz, 1H),
5.21 (s, 1H), 4.85 (d, J=14.5 Hz, 1H), 4.63 (d, J=14.6 Hz, 1H),
4.03-3.63 (m, 10H), 3.79 (s, 3H), 3.72 (s, 3H), 3.66 (s, 3H),
3.37-3.32 (m, 1H), 3.00-3.29 (m, 3H), 2.79-2.72 (m, 1H), 2.70 (s,
3H), 2.60-2.51 (m, 1H), 2.24-2.17 (m, 1H), 2.10-2.03 (m, 3H), 1.97
(s, 3H), 1.92-1.89 (m, 1H), 1.61 (dd, J=14.4, 7.3 Hz, 1H), 1.41
(dd, J=14.2, 7.2 Hz, 1H), 1.04 (t, J=7.4 Hz, 3H), 0.60 (t, J=7.2
Hz, 3H); ESI MS m/z 871 [M+H].sup.+.
Example 20
Preparation of 11'-(3-Hydroxypropylsulfinyl)vinorelbine
Trifluoroacetate
[0240] Hydrogen peroxide (30%, 10 .mu.L, 0.35 mmol) was added
dropwise to a solution of 11'-(3-hydroxypropylsulfanyl)vinorelbine
(18.0 mg, 0.04 mmol) in CH.sub.3CO.sub.2H/H.sub.2O (2:1, 1.5 mL) at
0.degree. C. The reaction mixture was stirred at 0.degree. C. for
10 min and then at room temperature for 1 h. The reaction mixture
was poured slowly into saturated NaHCO.sub.3 (30 mL) and the
mixture was extracted with CH.sub.2Cl.sub.2 (60 mL). The organic
layer was washed with brine, dried over Na.sub.2SO.sub.4, and
concentrated. Purification by preparative TLC (silica gel,
MeOH/EtOAc, 6:4) provided 11'-(3-hydroxypropylsulfinyl)vinorelbine
as a yellow solid (12 mg, 67%). The solid was treated with
CH.sub.2Cl.sub.2 (1 mL) and a drop of TFA. The solution was
evaporated to give 11'-(3-hydroxypropylsulfinyl)vinorelbine
trifluoroacetate as a yellow solid (14.7 mg, 98%): .sup.1H NMR (300
MHz, CD.sub.3OD) .delta. 10.81 (s, 1H), 8.09 (d, J=2.9 Hz, 1H),
7.63 (dd, J=8.6, 3.4 Hz, 1H), 7.52 (ddd, J=8.6, 1.6, 1.6 Hz, 1H),
6.75 (d, J=3.0 Hz, 1H), 6.43 (s, 1H), 5.94 (dd, J=10.4, 5.4 Hz,
1H), 5.93-5.88 (m, 1H), 5.67 (d, J=10.5 Hz, 1H), 5.32 (s, 1H), 4.97
(dd, J=14.7, 7.1 Hz, 1H), 4.73 (dd, J=14.6, 2.8 Hz, 1H), 4.14-3.71
(m, 7H), 3.90 (s, 3H), 3.82 (s, 3H), 3.77 (s, 3H), 3.68-3.61 (m,
2H), 3.43 (d, J=15.8 Hz, 1H), 3.25-2.97 (m, 4H), 2.89-2.81 (m, 1H),
2.81 (s, 3H), 2.71-2.62 (m, 1H), 2.36-2.30 (m, 1H), 2.22-214 (m,
3H), 2.08 (s, 3H), 2.03-1.81 (m, 3H), 1.75-1.67 (m, 1H), 1.52 (dd,
J=14.1, 6.7 Hz, 1H), 1.15 (t, J=7.4 Hz, 3H), 0.73-0.66 (m, 3H); ESI
MS m/z 885 [M+H].sup.+.
Example 21
Preparation of 11'-Ethynylvinorelbine Ditartrate
[0241] Step 1: A solution of 11'-iodovinorelbine (550 mg, 0.608
mmol), in toluene (11 mL) and Et.sub.3N (6 mL) was deoxygenated
with argon Copper iodide (6 mg, 0.032 mmol) and
PdCl.sub.2(PPh.sub.3).sub.2 (19 mg) were added and the mixture was
purged again with argon. Trimethylsilylacetylene (0.13 mL, 0.917
mmol) was added and the mixture stirred at 55.degree. C. overnight.
The reaction mixture was cooled to room temperature, diluted with
EtOAc (20 mL) and washed with saturated NH.sub.4Cl (10 mL). The
organic layer was dried (Na.sub.2SO.sub.4), filtered and
concentrated. Purification by column chromatography (silica gel,
MeOH/CH.sub.2Cl.sub.2, 4:96) gave
11'-(trimethylsilylethynyl)vinorelbine (400 mg), which was used
directly in the next step.
[0242] Step 2: To a solution of
11'-(Trimethylsilylethynyl)vinorelbine (400 mg) in methanol (9 mL)
was added potassium carbonate (catalyst) and the mixture was
stirred at room temperature for 12 h. The solvent was removed under
reduced pressure and the residue was taken up in CH.sub.2Cl.sub.2
(20 mL). The mixture was washed with H.sub.2O (2.times.15 mL). The
organic layer was dried (Na.sub.2SO.sub.4), filtered, and
concentrated. Purification by column chromatography (silica gel,
0.5% to 4% MeOH, CH.sub.2Cl.sub.2, 0.1% Et.sub.3N), followed by
reverse phase chromatography (C18 column, H.sub.2O/CH.sub.3CN, 0.1%
TFA) gave 11'-ethynylvinorelbine trifluoroacetate (55 mg, 11%) as a
dark yellow solid: mp 207-211.degree. C.; .sup.1H NMR (500 MHz,
CD.sub.3OD) .delta. 7.90 (s, 1H), 7.29 (d, J=8 Hz, 1H), 7.24 (dd,
J=8, 1 Hz, 1H), 6.32 (s, 1H), 6.28 (s, 1H), 5.84 (dd, J=10, 4 Hz,
1H), 5.75 (d, J=5 Hz, 1H), 5.31 (s, 1H), 5.27 (d, J=10 Hz, 1H),
4.33 (d, J=13 Hz, 1H), 4.26 (d, J=13 Hz, 1H), 3.85 (s, 3H), 3.76
(s, 3H), 3.73 (s, 3H), 3.65 (d, J=17 Hz, 1H), 3.58 (s, 1H),
3.39-3.32 (m, 2H), 3.28-3.17 (m, 2H), 3.01 (dd, J=16, 8 Hz, 1H),
2.72 (s, 3H), 2.69-2.55 (m, 2H), 2.35-2.24 (m, 1H), 2.12-1.98 (m,
6H), 1.84-1.76 (m, 1H), 1.68-1.59 (m, 1H), 1.54 (br s, 1H),
1.37-1.28 (m, 1H), 1.09 (t, J=7 Hz, 3H), 0.69 (t, J=7 Hz, 3H); ESI
MS m/z 803 [M+H].sup.+.
[0243] Step 3: To a solution of 11'-ethynylvinorelbine
trifluoroacetate (83 mg, 0.103 mmol) in MeOH (0.2 mL) and Et.sub.2O
(3 mL) was added a solution of L-tartaric acid (16 mg, 0.106 mmol)
in MeOH (0.2 mL) and Et.sub.2O (1.5 mL). Collection of the
precipitate by filtration gave 11'-ethynylvinorelbine ditartrate
(74.2 mg, 65%) as an orange solid. mp 190.degree. C. (dec). .sup.1H
NMR (500 MHz, CD.sub.3OD) .delta. 7.99 (s, 1H), 7.37 (d, J=8 Hz,
1H), 7.29 (dd, J=8, 1 Hz, 1H), 6.41 (s, 1H), 6.36 (s, 1H),
5.88-5.81 (m, 2H), 5.37 (d, J=10 Hz, 1H), 5.29 (s, 1H), 4.90 (d,
J=15 Hz, 1H), 4.65 (d, J=15 Hz, 1H), 4.39 (s, 4H), 4.15 (d, J=17
Hz, 1H), 3.88 (s, 3H), 3.77 (s, 3H), 3.76 (s, 3H), 3.75-3.68 (m,
2H), 3.62 (s, 1H), 3.47 (dd, J=16, 5 Hz, 1H), 3.36-3.33 (m, 2H),
3.13 (dd, J=16, 8 Hz, 1H), 2.97-2.89 (m, 2H), 2.86 (dd, J=14, 4 Hz,
1H), 2.75 (s, 3H), 2.67-2.55 (m, 2H), 2.18-2.09 (m, 3H), 2.03 (s,
3H), 1.96-1.85 (m, 2H), 1.68-1.59 (m, 1H), 1.42-1.33 (m, 1H), 1.13
(t, J=7 Hz, 3H), 0.71 (t, J=7 Hz, 3H); ESI MS m/z 803
[M+H].sup.+.
Example 22
Preparation of 11'-Hexynylvinorelbine
[0244] To a solution of 11'-iodovinorelbine (140 mg, 0.154 mmol) in
THF (5 mL) was added triethylamine (1 mL), CuI (2.86 mg, 15 mmol)
and (PPh.sub.3).sub.2PdCl.sub.2 (10.5 mg, 15 mmol) and the mixture
deoxygenated with argon. 1-Hexyne (45 mg, 310 mmol) was added and
the mixture was sealed and heated to 60.degree. C. After 4 h, CuI
(1.4 mg, 7 mmol), (PPh.sub.3).sub.2PdCl.sub.2 (5.5 mg, 8 mmol) and
1-hexyne (55 .mu.L) were added and the mixture deoxygenated,
sealed, and heated to 60.degree. C. overnight. The mixture was
diluted with dichloromethane, washed with water and brine, dried
(Na.sub.2SO.sub.4), and concentrated under reduced pressure.
Purification by flash chromatography (silica gel, eluent 2%
methanol in dichloromethane containing 0.5% triethylamine) followed
by reverse phase chromatography (C18 column, methanol/water) gave
11'-hexynylvinorelbine (18.8 mg, 14%). .sup.1H NMR (300 MHz,
CD.sub.3OD) .delta. 7.77 (s, 1H), 7.26 (d, J=8 Hz, 1H), 7.15 (dd,
J=8, 1 Hz, 1H), 6.35 (s, 1H), 6.26 (s, 1H), 5.83 (dd, J=10, 5 Hz,
1H), 5.90-5.78 (m, 1H), 5.30 (s, 1H), 5.22 (d, J=10 Hz, 1H), 4.12
(s, 2 H), 4.42 (d, J=13 Hz, 1H), 4.39 (d, J=13 Hz, 1H), 3.85 (s,
3H), 3.76 (m, 1H), 3.75 (s, 3H), 3.73 (s, 3H), 3.57 (s, 1H), 3.43
(d, J=13 Hz, 1H), 3.39-3.27 (m, 1H), 3.20 (dt, J=9, 5 Hz, 1H), 3.06
(dd, J=16, 7 Hz, 1H), 2.71 (s, 3H), 2.65 (d, J=18 Hz, 1H), 2.58 (s,
1H), 2.50-2.48 (m, 3H), 2.38 (dt, J=9, 5 Hz, 1H), 2.14-2.00 (m,
3H), 2.01 (s, 3H), 1.90 (br s, 1H), 1.80 (m, 1H), 1.70-1.42 (m),
1.40-1.25 (m), 1.10 (t, J=7 Hz, 3H), 0.98 (t, J=7 Hz, 3H), 0.68 (t,
J=7 Hz, 3H); ESI MS m/z 859 [M+H].sup.+.
Example 23
Preparation of 11'-(4-Methylpentynyl)vinorelbine
[0245] 11'-Iodovinorelbine (53 mg, 0.058 mmol), copper(I) iodide
(1.6 mg, 0.0088 mmol), dichlorobis(triphenylphosphine)palladium(II)
(4.1 mg, 0.0059 mmol), toluene (1.2 mL), and triethylamine (0.8 mL)
were combined in a resealable glass test tube. Argon was bubbled
through the solution for 10 min. 4-Methyl-1-pentyne (41.3 mg, 0.351
mmol) was added, the test tube sealed, and the mixture was heated
at 55.degree. C. for 1.5 h. Saturated NaHCO.sub.3 (5 mL) was added
and the mixture was extracted with ethyl acetate (3.times.5 mL).
The combined organic extracts were washed with brine (5 mL), dried
over MgSO.sub.4, and evaporated to dryness in vacuo. The residue
was purified by prep TLC (silica gel, methylene chloride/methanol,
95/5) to provide 11'-(4-methylpentynyl)vinorelbine (9 mg, 18%
yield) as a white powder: .sup.1H NMR (500 MHz, CD.sub.3OD) 37.76
(s, 1H), 7.25 (d, J=8.5 Hz, 1H), 7.15 (dd, J=8.5, 1.5 Hz, 1H), 6.32
(s, 1H), 6.26 (s, 1H), 5.87-5.82 (m, 1H), 5.76 (d, J=4.0 Hz, 1H),
5.30 (s, 1H), 5.26 (d, J=10.0 Hz, 1H), 4.89-4.79 (m, 1H), 4.35 (br
s, 2H), 3.85 (s, 3H), 3.75-3.65 (m, 7H), 3.57 (s, 1H), 3.45-3.28
(m, 2H), 3.23-3.15 (m, 2H), 3.06-2.99 (m, 1H), 2.71 (s, 3H),
2.68-2.54 (m, 3H), 2.40-2.23 (m, 4H), 2.12-2.02 (m, 9H), 1.93-1.77
(m, 2H), 1.66-1.57 (m, 2H), 1.35-1.28 (m, 2H), 1.11-1.06 (m, 9H),
0.69 (t, J=7.5 Hz, 3H); ESI MS m/z 859 [M+H].sup.+.
Example 24
Preparation of 11'-(3-Methoxypropynyl)vinorelbine
Trifluoroacetate
[0246] 11'-Iodovinorelbine (68 mg, 0.075 mmol), copper(I) iodide
(2.1 mg, 0.011 mmol), dichlorobis(triphenylphosphine)palladium(II),
(5.3 mg, 0.008 mmol) toluene (1.2 mL), and triethylamine (0.8 mL)
were combined in a resealable glass test tube. Argon was bubbled
through the solution for 30 min. Methyl propargyl ether (32 mg,
0.451 mmol) was added, the test tube sealed, and the mixture was
heated at 55.degree. C. for 1.5 h. Saturated NaHCO.sub.3 (5 mL) was
added and the mixture was extracted with ethyl acetate (3.times.5
mL). The combined organic extracts were washed with brine (5 mL),
dried over MgSO.sub.4, and evaporated to dryness in vacuo. The
residue was purified by reverse phase chromatography (C18,
acetonitrile/water, 0.05% TFA) to provide
11'-(3-methoxypropynyl)vinorelbine trifluoroacetate (8.3 mg, 10%
yield) as a white powder after lyophilization: .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 10.40 (bs, 1H), 7.86 (s, 1H), 7.35 (d,
J=8.4 Hz, 1H), 7.23 (dd, J=8.4, 1.3 Hz, 1H), 6.64 (s, 1H), 6.41 (s,
1H), 5.93 (dd, J=10.4, 5.4 Hz, 1H), 5.87 (d, J=3.7 Hz, 1H), 5.63
(d, J=10.4 Hz, 1H), 5.30 (s, 1H), 4.87 (m, 1H), 4.60 (d, J=14.5 Hz,
1H), 4.32 (s, 2H), 4.07 (d, J=17.0 Hz, 1H), 3.96-3.71 (m, 4H), 3.88
(s, 3H), 3.81 (s, 3H), 3.76 (s, 3H), 3.71 (s, 1H), 3.43 (s,3 H)
3.42 (m, 1H), 3.11 (m, 2H), 2.81 (m, 1H), 2.79 (s, 3H), 2.61 (m,
1H), 2.31 (m, 1H), 2.15 (q, J=7.4 Hz, 2H), 2.09 (m, 2H), 2.06 (s,
3H), 1.98 (m, 1H), 1.69 (m, 1H), 1.47 (m, 1H), 1.14 (t, J=7.5 Hz,
3H), 0.70 (t, J=7.2 Hz, 3H); ESI MS m/z 847 [M+H].sup.+.
Example 25
Preparation of 11'-Cyanovinorelbine Ditartrate
[0247] Step 1: A mixture of
tris(dibenzylideneacetone)dipalladium(0) (21 mg, 0.0232 mmol),
1,1'-bis(diphenylphosphino)ferrocene (26 mg, 0.0464 mmol),
Zn(CN).sub.2 (54 mg, 0.46 mmol) in DMF (1.0 mL) was deoxygenated
with argon a solution of 11'-iodovinorelbine (210 mg, 0.23 mmol) in
DMF (7 mL) was added and the mixture was purged again with argon.
The reaction mixture was heated at 65.degree. C. for 3.5 h, then
cooled to 25.degree. C. and diluted with EtOAc. The organic layer
was washed with 5% LiCl, brine, dried (Na.sub.2SO.sub.4), and
concentrated. Purification by reverse phase chromatography (C8,
CH.sub.3CN/H.sub.2O, 0.05% NH.sub.4OH) gave 11'-cyanovinorelbine
(15.3 mg, 8%). .sup.1H NMR (500 MHz, MeOD) .delta. 8.18 (s, 1H),
7.49 (d, J=9 Hz, 1H), 7.40 (d, J=9 Hz, 1H), 6.32 (s, 1H), 6.27 (s,
1H), 5.83 (dd, J=10, 4 Hz, 1H), 5.76 (d, J=4 Hz, 1H), 5.29 (s, 1H),
5.27 (s, 1H), 4.37 (s, 2H), 3.85 (s, 3H), 3.75 (s, 3H), 3.74 (s,
3H), 3.58 (s, 3H), 3.20 (dt, J=9, 5 Hz, 1H), 3.00 (dd, J=16, 8 Hz,
1H), 2.72 (s, 3H), 2.70-2.58 (m, 3H), 2.38 (dd, J=14, 8 Hz, 1H),
2.30 (dt, J=11, 6 Hz, 1H), 2.07-2.01 (m, 6H), 1.77 (m, 1H), 1.61
(m, 2H), 1.34 (m, 4H), 1.09 (t, J=7 Hz, 3H), 0.67 (t, J=7 Hz, 3H);
ESI MS m/z 804 [M+H].sup.+.
[0248] Step 2: 11'-Cyanovinorelbine (68 mg, 0.075 mmol) in
methylene chloride (1 mL) was treated with a solution of L-tartaric
acid (22.5 mg, 0.150 mmol) in methanol (0.20 mL). After 4 hours,
reaction mixture was reduced to dryness. The residue was
lyophilized with water and acetonitrile to provide
11'-cyanovinorelbine ditartate (80 mg, 96% yield) as a white
powder: .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 8.26 (br s, 1H),
7.57 (d, J=8.5 Hz, 1H), 7.48 (d, J=8.5 Hz, 1H), 6.35 (s, 1H),
6.27-6.25 (m, 1H), 5.88-5.84 (m, 2H), 5.33 (d, J=10.5 Hz, 1H), 5.28
(s, 1H), 4.77-4.71 (m, 1H), 4.42 (s, 4H), 4.17-4.10 (m, 1H),
3.88-3.82 (m, 4H), 3.78-3.72 (m, 7H), 3.68-3.61 (m, 2H), 3.44-3.43
(m, 1H), 3.39-3.30 (m, 1H), 3.17-3.11 (m, 2H), 2.92-2.88 (m, 1H),
2.76-2.71 (m, 5H), 2.68-2.60 (m, 1H), 2.43-2.40 (m, 1H), 2.18-2.07
(m, 3H), 2.02 (s, 3H), 1.99-1.94 (m, 1H), 1.82-1.76 (m, 1H),
1.64-1.60 (m, 1H), 1.41-1.29 (m, 2H), 1.14 (t, J=7.5 Hz, 3H), 0.66
(t, J=7.5 Hz, 3H); ESI MS m/z 804 [M+H].sup.+.
Example 26
Preparation of 11'-Acetylvinorelbine Ditartrate
[0249] 11'-(Trimethylsilanylethynyl)vinorelbine (320 mg, 0.365
mmol) was added to an ice-water cold solution of CH.sub.2Cl.sub.2
(10 mL) and TFA (10 mL), and the reaction mixture was stirred at
room temperature. After 20 min, the reaction was quenched by the
addition of saturated NaHCO.sub.3, and the pH was adjusted to 8
using 3 N NaOH and saturated NaHCO.sub.3. The resulting suspension
was extracted with EtOAc (250 mL). The organic solution was washed
with water and brine, then dried (Na.sub.2SO.sub.4) and evaporated
to dryness under vacuum. Purification by column chromatography
(silica gel, 95.5:4:0.5 CH.sub.2Cl.sub.2/MeOH/Et.sub.3N) followed
by reverse phase chromatography (C18, acetonitrile/water, 0.05%
TFA) gave the product as the TFA salt. After conversion to the free
base, treatment with 2 equivalents of L-tartaric acid gave
11'-acetylvinorelbine L-tartaric acid salt as a white solid:
.sup.1H NMR (500 MHz, CD.sub.3OD) 8.57 (d, J=1.0 Hz, 1H), 7.88 (dd,
J=8.5, 1.5 Hz, 1H), 7.48 (d, J=8.5 Hz, 1H), 6.41 (s, 1H), 6.37 (s,
1H), 5.88-5.83 (m, 2H), 5.38 (d, J=10.0 Hz, 1H), 5.29 (s, 1H), 5.03
(d, J=15.0 Hz, 1H), 4.71 (d, J=14.5 Hz, 1H), 4.40 (s, 4 H), 4.14
(d, J=17.0 Hz, 1H), 3.88 (s, 3H), 3.78-3.74 (m, 8H), 3.63 (s, 1H),
3.46 (dd, J=15, 4.5 Hz, 1H), 3.27-3.32 (m, 1H), 3.14 (dd, J=16.0,
8.0 Hz, 1H), 2.96 (s, 1H), 2.92-2.86 (m, 2H), 2.75 (s, 3H), 2.70
(s, 3H), 2.64 (dd, J=15.5, 12.0 Hz, 1H), 2.18-2.09 (m, 3H), 2.03
(s, 4H), 1.95-1.86 (m, 2H), 1.66-1.61 (m, 1H), 1.42-1.37 (m, 1H),
1.15 (t, J=7.5 Hz, 3H), 0.69 (t, J=7.5 Hz, 3H); ESI MS m/z 821
[M+H].sup.+.
Example 27
Preparation of 11'-(Methoxycarbonyl)vinorelbine
Trifluoroacetate
[0250] Carbon monoxide was bubbled through a solution of
11'-iodovinorelbine (30 mg, 0.033 mmol), triethylamine (33 mg,
0.331 mmol) and bis(triphenylphosphine)palladium(II) dichloride
(4.6 mg, 0.007 mmol) in a mixture of DMF/methanol (2 mL, 1:1) for 5
min. The reaction mixture was heated at 50.degree. C. for 12 h
under one atmosphere of carbon monoxide (balloon). The solution was
diluted with ethyl acetate (20 mL) then washed with satd
NaHCO.sub.3 (2.times.5 mL) and brine (5 mL), dried over MgSO.sub.4,
and evaporated to dryness in vacuo. The residue was purified by
reverse phase chromatography (C18, acetonitrile/water, 0.05% TFA)
to provide 11'-(methoxycarbonyl)vinorelbine trifluoroacetate (10
mg, 28% yield) as a white powder after lyophilization: .sup.1H NMR
(500 MHz, CD.sub.3OD) .delta. 10.64 (bs, 1H), 8.50 (s, 1H), 7.85
(dd, J=8.6, 1.4 Hz, 1H), 7.44 (d, J=8.6 Hz, 1H), 6.66 (s, 1H), 6.41
(s, 1H), 5.92 (dd, J=10.4, 5.2 Hz, 1H), 5.88 (m, 1H), 5.63 (d,
J=10.6 Hz, 1H), 5.31 (s, 1H), 4.95 (d, J=14.6 Hz, 1H), 4.71 (d,
J=14.5 Hz, 1H), 4.09 (d, J=17.0 Hz, 1H), 3.89 (d, J=14.2 Hz, 1H),
3.91-3.69 (m, 5H), 3.91 (s, 3H), 3.88 (s, 3H), 3.81 (s, 3H), 3.77
(s, 3H), 3.37 (d, J=14.5 Hz, 1H), 3.13 (m, 2H), 2.84 (dd, J=13.3,
4.2 Hz, 1H), 2.79 (s, 3H), 2.65 (m, 1H), 2.29 (m, 1H), 2.17 (q,
J=7.3 Hz, 2H), 2.08 (m, 1H), 2.06 (s, 3H), 1.94 (m, 1H), 1.69 (m,
1H), 1.47 (m, 1H), 1.14 (t, J=7.5 Hz, 3H), 0.69 (t, J=7.3 Hz, 3H);
ESI MS m/z 837 [M+H].sup.+.
Example 28
Preparation of 11'-(2,2,2-Trichloroethoxycarbonyl)vinorelbine
[0251] Carbon monoxide was bubbled through a solution of
11'-iodovinorelbine (151 mg, 0.167 mmol), triethylamine (169 mg,
1.67 mmol) and bis(triphenylphosphine)palladium(II) dichloride (23
mg, 0.033 mmol) in a mixture of DMF/2,2,2-trichloroethanol (4 mL,
1:1) for 5 min. The reaction mixture was heated at 50.degree. C.
for 14 h under one atmosphere of carbon monoxide (balloon). The
solution was diluted with ethyl acetate (35 mL) then washed with
saturated NaHCO.sub.3 (2.times.20 mL) and brine (20 mL), dried over
MgSO.sub.4, and evaporated to dryness in vacuo. The residue was
purified by reverse phase chromatography (C18, acetonitrile/water,
0.05% TFA) to provide
11'-(2,2,2-trichloroethoxycarbonyl)lvinorelbine Trifluoroacetate(78
mg, 40% yield) as a white powder after lyophilization: .sup.1H NMR
(500 MHz, CD.sub.3OD) 38.62 (s, 1H), 7.97 (dd, J=8.7, 1.5 Hz, 1H),
7.52 (d, J=8.7 Hz, 1H), 6.37 (s, 2H), 5.87 (m, 2H), 5.39 (d, J=10.1
Hz, 1H), 5.30 (s, 1H), 5.11 (d, J=12.2 Hz, 1H), 5.07 (d, J=12.1 Hz,
1H), 4.98 (d, J=14.7 Hz, 1H), 4.74 (d, J=14.7 Hz, 1H), 4.09 (m,
1H), 3.92-3.72 (m, 5H), 3.88 (s, 3H), 3.77 (s, 3H), 3.76 (s, 3H),
3.63 (s, 1H), 3.48 (m, 1H), 3.33 (m, 1H), 3.16 (m, 1H), 2.88 (dd,
J=13.5, 4.6 Hz, 1H), 2.75 (s, 3H), 2.64 (dd, J=15.4, 12.5 Hz, 1H),
2.16 (q, J=7.4 Hz, 2H), 2.12 (m, 1H), 2.03 (s, 3H), 1.98 (m, 1H),
1.87 (m, 1H), 1.64 (m, 1H), 1.40 (m, 1H), 1.14 (t, J=7.5 Hz, 3H),
0.69 (t, J=7.3 Hz, 3H); ESI MS m/z 953 [M+H].sup.+.
Example 29
Preparation of 11'-(2,2-dichloroethoxycarbonyl)vinorelbine
Trifluoroacetate
[0252] The second eluting fraction formed during the preparation of
11'-carboxyvinorelbine (see Example 33) was purified by reverse
phase chromatography (C18, acetonitrile/water, 0.05% TFA) to
provide 11'-(2,2-dichloroethoxycarbonyl)vinorelbine
trifluoroacetate (2.4 mg, 3.5% yield) as a white powder after
lyophilization: .sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 10.71
(bs, 1H), 8.55 (s, 1H), 7.90 (dd, J=8.6, 1.4 Hz, 1H), 7.48 (d,
J=8.7 Hz, 1H), 6.63 (s, 1H), 6.41 (s, 1H), 5.91 (dd, J=10.4, 4.7
Hz, 1H), 5.88 (d, J=4.2 Hz, 1H), 5.61 (d, J=10.0 Hz, 1H), 5.31 (s,
1H), 4.97 (d, J=14.7 Hz, 1H), 4.73 (m, 3H), 4.09 (d, J=17.0 Hz,
1H), 3.95 (d, J=13.3 Hz, 1H), 3.89 (s, 3H), 3.86-3.74 (m, 2H), 3.81
(s, 3H), 3.77 (s, 3H), 3.71 (s, 1H), 3.66 (m, 2H), 3.33 (m, 1H),
3.09 (m, 3H), 2.85 (dd, J=13.5, 4.2 Hz, 1H), 2.78 (s, 3H), 2.65 (m,
1H), 2.28 (m, 1H), 2.17 (q, J=7.4 Hz, 2H), 2.10 (m, 1H), 2.06 (s,
3H), 1.96 (m, 1H), 1.69 (m, 1H), 1.47 (m, 1H), 1.14 (t, J=7.5
Hz,3H), 0.69 (t, J=7.3 Hz, 3H); ESI MS m/z 919 [M+H].sup.+.
Example 30
Preparation of 11'-Phenylvinorelbine Trifluoroacetate
[0253] To a solution of 11'-iodovinorelbine (38 mg, 0.04 mmol) in
dioxane (1 mL) was added phenylboronic acid (10 mg, 0.08 mmol) and
Cs.sub.2CO.sub.3 (68 mg, 0.21 mmol). The mixture was deoxygenated
with an argon purge, and
[1,1'-bis(diphenylphospino)ferrocene]dichloropalladium (5 mg, 0.006
mmol) was added. The resulting mixture was deoxygenated again and
then heated to 60.degree. C. for 2 h. The reaction mixture was
cooled to room temperature, diluted with CH.sub.2Cl.sub.2, and
filtered through Celite. The filtrate was washed with water and
brine, and then dried (MgSO.sub.4). Purification by column
chromatography (silica gel, CH.sub.2Cl.sub.2/MeOH, 9:1) followed by
prep-TLC (silica gel, EtOAc/MeOH, 7:3) gave 11'-phenylvinorelbine
(9 mg, 26%). The solid was dissolved in CH.sub.2Cl.sub.2 (1 mL) and
treated with a drop of TFA. The solution was evaporated to give
11'-phenylvinorelbine trifluoroacetate (11.3 mg, quantitative):
.sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 10.1 (s, 1H), 7.89 (s,
1H), 7.58-7.56 (m, 2H), 7.36 (s, 2H), 7.31 (dd, J=7.3, 6.9 Hz, 2H),
7.21-7.16 (m, 1H), 6.55 (s, 1H), 6.33 (s, 1H), 5.85-5.77 (m, 2H),
5.55 (d, J=11.1 Hz, 1H), 5.22 (s, 1H), 4.93 (d, J=14.5 Hz, 1H),
4.59 (d, J=14.4 Hz, 1H), 3.99 (d, J=17.1 Hz, 1H), 3.85-3.63 (m,
6H), 3.80 (s, 3H), 3.71 (s, 3H), 3.68 (s, 3H), 3.35 (d, J=15.3 Hz,
1H), 3.14-2.99 (m, 2H), 2.77-2.69 (m, 1H), 2.69 (s, 3H), 2.54 (dd,
J=14.3, 12.1 Hz, 1H), 2.25-2.18 (m, 1H), 2.10-2.03 (m, 3H), 1.97
(s, 3H), 1.89-1.84 (m, 1H), 1.63 (dd, J=14.6, 7.5 Hz, 1H), 1.38
(dd, J=14.5, 7.2 Hz, 1H), 1.04 (t, J=7.4 Hz, 3H), 0.67 (t, J=7.2
Hz, 3H); ESI MS m/z 855 [M+H].sup.+.
Example 31
Preparation of 11'-(3-Hydroxyphenyl)vinorelbine
Trifluoroacetate
[0254] To a solution of 11'-iodovinorelbine (87 mg, 0.10 mmol) in
dioxane (1 mL) was added 3-hydroxyphenylboronic acid (27 mg, 0.19
mmol) and Cs.sub.2CO.sub.3 (157 mg, 0.480 mmol). The mixture was
deoxygenated with argon, and PdCl.sub.2(dppf).sub.2 (8 mg, 0.01
mmol) was added. The resulting mixture was deoxygenated with argon
again and then heated to 60.degree. C. for 7 h and then to
70.degree. C. overnight. The reaction mixture was cooled to room
temperature, diluted with CH.sub.2Cl.sub.2, and filtered through
Celite. The filtrate was washed with water and brine, dried
(Na.sub.2SO.sub.4), and the solvent evaporated in vacuo.
Purification by column chromatography (silica gel,
CH.sub.2Cl.sub.2/MeOH, 9:1), followed by reverse phase
chromatography (C18, acetonitrile/water, 0.05% TFA) gave
11'-(3-hydroxyphenyl)vinorelbine trifluoroacetate (5 mg, 4.5%) as a
white solid after lyophilization: .sup.1H NMR (500 MHz, CD.sub.3OD)
.delta. 7.96 (s, 1H), 7.49-7.47 (m, 2H), 7.23 (t, J=8.0 Hz, 1H),
7.14 (d, J=7.5 Hz, 1H), 7.09-7.08 (m, 1H), 6.74 (dd, J=8.0, 1.5 Hz,
1H), 6.56-6.50 (m, 1H), 6.44 (s, 1H), 5.94-5.89 (m, 2H), 5.60 (d,
J=11.0 Hz, 1H), 5.34 (s, 1H), 5.03-5.00 (m, 1H), 4.76-4.73 (m, 1H),
4.08 (d, J=16.5 Hz, 1H), 3.90-3.72 (m, 10H), 3.56-3.52 (m, 2H),
3.45-3.38 (m, 2H), 3.18-3.03 (m, 3H), 2.91-2.87 (m, 1H), 2.78 (s,
3H), 4.70-4.63 (m, 1H), 2.39-2.34 (m, 1), 2.19-2.14 (m, 2H),
2.07-1.96 (m, 5H), 1.77-1.72 (m, 1H), 1.48-1.43 (m, 1H), 1.06-1.11
(m, 4H), 0.78-0.75 (m, 3H); ESI MS m/z 871 [M+H].sup.+.
Example 32
Preparation of 11'-(3,5-Dimethylisoxazol-4-yl)vinorelbine
Trifluoroacetate
[0255] To a solution of 11'-iodovinorelbine (72 mg, 0.08 mmol) in
dioxane (1 mL) was added 3,5-dimethylisoxazole-4-boronic acid (22
mg, 0.16 mmol) and Cs.sub.2CO.sub.3 (130 mg, 0.4 mmol). The mixture
was deoxygenated with an argon purge, and
[1,1'-bis(diphenylphospino)ferrocene]dichloropalladium ( (4 mg,
0.005 mmol) was added. The resulting mixture was deoxygenated again
and then heated to 60.degree. C for 7 h. The reaction mixture was
cooled to room temperature, diluted with CH.sub.2Cl.sub.2, and
filtered through Celite. The filtrate was washed with water and
brine, and then dried (MgSO.sub.4). Purification by column
chromatography (silica gel, CH.sub.2Cl.sub.2/MeOH, 9:1) followed by
prep-TLC (silica gel, CH.sub.2Cl.sub.12/MeOH 9:1) gave
11'-(3,5-dimethylisoxazol-4-yl)vinorelbine (14 mg, 20%). The solid
was dissolved in CH.sub.2Cl.sub.2 (1 mL) and treated with a drop of
TFA. The solution was evaporated to give
11'-(3,5-dimethylisoxazol-4-yl)vinorelbine trifluoroacetate (17.6
mg, 90%): .sup.1H NMR (300 MHz, CD.sub.3OD) .delta. 10.2 (s, 1H),
7.56 (s, 1H), 7.38 (d, J=8.4 Hz, 1H), 7.01 (dd, J=8.4, 1.4 Hz, 1H),
6.66 (s, 1H), 6.32 (s, 1H), 5.85-5.77 (m, 2H), 5.56 (d, J=9.8 Hz,
1H), 5.22 (s, 1H), 4.85 (d, J=14.6 Hz, 1H), 4.59 (d, J=14.5 Hz,
1H), 4.00-3.85 (m, 2H), 3.80-3.62 (m, 5H), 3.80 (s, 3H), 3.71 (s,
3H), 3.67 (s, 3H), 3.34 (d, J=15.7 Hz, 1H), 3.15-2.98 (m, 2H),
2.75-2.70 (m, 1H), 2.70 (s, 3H), 2.54 (dd, J=15.5, 12.2 Hz, 1H),
2.30 (s, 3H), 2.25-2.17 (m, 1H), 2.15 (s, 3H), 2.10-2.02 (m, 3H),
1.97 (s, 3H), 1.86-1.84 (m, 1H), 1.59 (dd, J=14.5, 7.4 Hz, 1H),
1.40 (dd, J=14.5, 7.3 Hz, 1H), 1.04 (t, J=7.4 Hz, 3H), 0.64 (t,
J=7.2 Hz, 3H); ESI MS m/z 874 [M+H].sup.+.
Example 33
Preparation of 3,11'-Dimethylvinorelbine Trifluoroacetate
[0256] Dimethylzinc (2.0 M in toluene, 0.16 mL, 0.32 mmol) was
added to 11'-iodovinorelbine (148 mg, 0.16 mmol) and
[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (13 mg,
0.016 mmol) in anhydrous 1,4-dioxane (3 mL) under nitrogen. The
reaction mixture was heated at 45.degree. C. for 10 h then quenched
by the addition of saturated NaHCO.sub.3 (8 mL). After extraction
with chloroform (3.times.10 mL) the combined organic extracts were
washed with brine (5 mL), dried over MgSO.sub.4, and evaporated to
dryness in vacuo. The residue was initially purified by reverse
phase chromatography (C18, acetonitrile/water, 0.05% TFA) to
provide two separate products. Purification of the first fraction
by reverse phase chromatography (C18, acetonitrile/water, 0.05%
TFA) gave 3,11'-dimethylvinorelbine trifluoroacetate (16.0 mg, 9.5%
yield) as a white powder after lyophilization; .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 10.3 (bs, 1H), 7.58 (s, 1H), 7.37 (d,
J=8.4 Hz, 1H), 7.09 (d, J=8.4 Hz, 1H), 6.49 (s, 1H), 6.44 (s, 1H),
5.95 (dd, J=10.5,4.2 Hz, 1H), 5.88 (d, J=4.2 Hz, 1H), 5.63 (d,
J=10.4 Hz, 1H), 5.31 (s, 1H), 5.01 (d, J=14.7 Hz, 1H), 4.75 (d,
J=14.7 Hz, 1H), 4.16 (d, J=16.6 Hz, 1H), 4.08 (d, J=16.6 Hz, 1H),
3.96 (dd, J=15.7, 5.5 Hz, 1H), 3.88-3.75 (m, 3H), 3.88 (s, 3H),
3.82 (s, 3H), 3.75 (s, 3H), 3.66 (s, 1H), 3.61 (d, J=13.0 Hz, 1H),
3.46 (d, J=15.7 Hz, 1H), 3.18 (m, 2H), 3.06 (s, 3H), 2.96 (dd,
J=13.0, 4.5 Hz, 1H), 2.79 (s, 3H), 2.63 (dd, J=15.7, 12.1 Hz, 1H),
2.38 (m, 1H), 2.17 (q, J=7.5 Hz, 2H), 2.08 (m, 1H), 2.07 (s, 3H),
1.98 (m, 1H), 1.74 (m, 1H), 1.46 (m, 1H), 1.16 (t, J=7.4 Hz, 3H),
0.78 (t, J=7.3 Hz, 3H); ESI MS m/z 807 [M+H].sup.+.
Example 34
Preparation of 3-Methyl-11'-iodovinorelbine Trifluoroacetate
[0257] The second eluting fraction formed during the preparation of
3,11'-dimethylvinorelbine (see Example 53) was purified by reverse
phase chromatography (C18, acetonitrile/water, 0.05% TFA) to give
3-methyl-11'-iodovinorelbine trifluoroacetate (8.8 mg, 4.7% yield);
.sup.1H NMR (500 MHz, CD.sub.3OD) .delta. 8.20 (s, 1H), 7.52 (dd,
J=8.6, 1.3 Hz, 1H), 7.31 (d, J=8.6 Hz, 1H), 6.46 (m, 1H), 6.44 (s,
1H), 5.94 (dd, J=10.5, 4.2 Hz, 1H), 5.88 (d, J=4.4 Hz, 1H), 5.63
(d, J=10.7 Hz, 1H), 5.31 (s, 1H), 5.00 (d, J=14.8 Hz, 1H), 4.74 (d,
J=16.5 Hz, 1H), 4.17 (d, J=16.4 Hz, 1H), 4.05 (d, J=16.5 Hz, 1H),
3.95 (dd, J=15.8, 5.4 Hz, 1H), 3.87-3.75 (m, 3H), 3.88 (s, 3H),
3.82 (s, 3H), 3.76 (s, 3H), 3.67 (s, 1H), 3.58 (d, J=13.3 Hz, 1H),
3.47 (d, J=15.9 Hz, 1H), 3.18 (m, 2H), 3.06 (s, 3H), 2.96 (dd,
J=13.1, 8.6 Hz, 1H), 2.79 (s, 3H), 2.64 (m, 1H), 2.38 (m, 1H), 2.17
(q, J=7.3 Hz, 2H), 2.08 (m, 1H), 2.07 (s, 3H), 1.98 (m, 1H), 1.73
(m, 1H), 1.46 (m, 1H), 1.16 (t, J=7.5 Hz, 3H), 0.75 (t, J=7.3 Hz,
3H); ESI MS m/z 919 [M+H].sup.+.
Example 35
Preparation of 11'-Aminovinorelbine Trifluoroacetate
[0258] 11'-Aminovinorelbine was prepared according to the scheme
below. ##STR44##
[0259] Step 1: A solution of 11'-iodovinorelbine (211 mg, 0.233
mmol) in CH.sub.2Cl.sub.2 (5 mL) was charged with
N,N-diisopropylethylamine (0.41 g, 2.33 mmol) and
tert-butyldimethylsilyl trifluoromethanesulfonate (123 mg, 0.466
mmol). After 1 h, the reaction mixture was diluted with ethyl
acetate (30 mL), then washed with saturated NaHCO.sub.3 (2.times.10
mL) and brine (10 mL). The solution was dried (MgSO.sub.4) and
concentrated to a brown solid which was purified by flash
chromatography (silica gel, 20:79:1 to 50:49:1 ethyl
acetate/hexanes/triethylamine) to yield
11'-iodo-3-(tert-butyldimethylsilanyloxy)vinorelbine (180 mg, 76%)
as a white solid: ESI MS m/z 1019 [M+H].sup.+.
[0260] Step 2: 11'-Iodo-3-(tert-butyldimethylsilanyloxy)vinorelbine
(172 mg, 0.169 mmol), benzophenone imine (71 .mu.L, 0.42 mmol), and
NaOt-Bu (48 mg, 0.50 mmol) were dissolved in anhydrous toluene (1.5
mL) while stirring under argon atmosphere in a resealable tube. The
mixture was deoxygenated with argon at room temperature for 3 min
then tris(dibenzylideneacetone)dipalladium(0) (15.5 mg, 16.9
.mu.mol) and
2-(dicyclohexylphosphino)-2',4',6'-tri-i-propyl-1,1'-biphenyl (16.1
mg, 33.9 .mu.mol) were added. The reaction vessel was sealed and
the mixture heated to 80.degree. C. for 4 h. The reaction mixture
was cooled to room temperature, diluted with EtOAc, filtered
through diatomaceous earth, and concentrated to provide crude
12'-benzhydrylidene-amino-3-(tert-butyldimethylsilanyloxy)vinorelbine:
ESI MS m/z 1072 [M+H].sup.+.
[0261] Step 3: A solution of crude
12'-benzhydrylideneamino-3-(tert-butyldimethylsilanyloxy)vinorelbine
(181 mg, 0.17 mmol) in methanol (1.0 mL) was treated with NaOAc
(123 mg, 1.50 mmol) and hydroxylamine hydrochloride (81 mg, 1.1
mmol). After 6 h, the reaction mixture was concentrated to dryness.
The residue was diluted with saturated NaHCO.sub.3 (10 mL) and
extracted with CH.sub.2Cl.sub.2 (2.times.10 mL). The combined
extracts were dried (Na.sub.2SO.sub.4), concentrated, and purified
by reverse phase chromatography (C18, acetonitrile/water, 0.05%
TFA) to provide
11'-amino-3-(tert-butyldimethylsilanyloxy)vinorelbine
trifluoroacetate (25 mg, 16% yield) as a white powder after
lyophilization: ESI MS m/z 908 [M+H].sup.+.
[0262] Step 4: A solution of
11'-amino-3-(tert-butyldimethylsilanyloxy)vinorelbine
trifluoroacetate (20 mg, 0.17 mmol) in THF (1.0 mL) was treated
with Bu.sub.4NF (80 .mu.L of a 1 N solution in THF, 0.080 mmol).
After 3 h, the reaction appeared complete as indicated by ESI mass
spectral analysis. The reaction was diluted with saturated
NaHCO.sub.3 (10 mL) and extracted with CH.sub.2Cl.sub.2 (2.times.10
mL). The combined extracts were dried (Na.sub.2SO.sub.4) and then
concentrated. The residue was purified by reverse phase
chromatography (C18, acetonitrile/water, 0.05% TFA) to provide
11'-aminovinorelbine trifluoroacetate (15 mg, 75% yield) as a white
powder after lyophilization: .sup.1H NMR (500 MHz, CD.sub.3OD)
.delta. 10.73 (br s, 1H), 7.66 (d, J=2.0 Hz, 1H), 7.51 (d, J=9 Hz,
1H), 7.11 (dd, J=8.5, 2.0 Hz, 1H), 6.72 (s, 1H), 6.40 (s, 1H),
5.93-5.88 (m, 2H), 5.65 (d, J=10.5 Hz, 1H), 5.29 (s, 1H), 4.81-4.79
(m, 1H), 4.72-4.70 (m, 1H), 4.13-4.07 (m, 2H), 3.93-3.88 (m, 4H),
3.82-3.80 (m, 4H), 3.78-3.70 (m, 7H), 3.38-3.32 (m, 1H), 3.17-3.08
(m, 2H), 2.86-2.80 (m, 4H), 2.65-2.59 (m, 1H), 2.32-2.26 (m, 1H),
2.19-2.12 (m, 3H), 2.06 (s, 3H), 1.9-1.94 (m, 1H), 1.64-1.65 (m,
1H), 1.52-1.48 (m, 1H), 1.44 (t, J=7.5 Hz, 3H), 0.65 (t, J=7.5 Hz,
3H); ESI MS m/z 794 [M+H].sup.+.
Example 36
Preparation of 11'-(4-Methoxyphenylamino)vinorelbine
Trifluoroacetate
[0263] Step 1: 11'-Iodo-3-(tert-butyldimethylsilanyloxy)vinorelbine
(52.6 mg, 0.0522 mmol), p-anisidine (15 mg, 0.13 mmol), and NaOt-Bu
(16 mg, 0.16 mmol) were dissolved in anhydrous toluene (1.5 mL)
while stirring under argon atmosphere in a resealable tube. The
reaction mixture was deoxygenated with an argon purge at room
temperature for 3 min then tris(dibenzylideneacetone)dipalladium(0)
(4.7 mg, 5.2 .mu.mol) and
2-(dicyclohexylphosphino)-2',4',6'-tri-isopropyl-1,1'-biphenyl (4.9
mg, 10 .mu.mol) were added. The reaction vessel was sealed and the
mixture heated to 80.degree. C. for 4 h. The reaction mixture was
cooled to room temperature, diluted with EtOAc, filtered through
diatomaceous earth, and then concentrated to provide crude
11'-(4-methoxyphenylamine)-3-(tert-butyldimethylsilanyloxy)vinorelbine:
ESI MS m/z 1014 [M+H].sup.+.
[0264] Step 2: A solution of
11'-(4-methoxyphenylamino)-3-(tert-butyl-dimethylsilanyloxy)vinorelbine
(17 mg, 0.016 mmol) in THF (1.0 mL) was treated with Bu.sub.4NF (50
.mu.L of a 1 N solution in THF, 0.050 mmol). After 1.5 h, the
reaction appeared complete as indicated by ESI mass spectral
analysis. The reaction mixture was diluted with saturated
NaHCO.sub.3 (10 mL) and extracted with CH.sub.2Cl.sub.2 (2.times.10
mL). The combined extracts were dried (Na.sub.2SO.sub.4),
concentrated, and purified by reverse phase chromatography (C18,
acetonitrile/water, 0.05% TFA) to provide
11'-(4-methoxyphenylanime)vinorelbine trifluoroacetate (16 mg, 13%
yield) as a white powder after lyophilization: .sup.1H NMR (500
MHz, CD.sub.3OD) .delta. 9.97 (br s, 1H), 7.33-7.27 (m, 2H), 7.03
(d, J=7 Hz, 2H), 6.91 (d, J=8 Hz, 1H), 6.82 (d, J=8.5 Hz, 2H), 6.63
(s, 1H), 6.42 (s, 1H), 5.94 (dd, J=10.5, 4.0 Hz, 1H), 5.86-5.85 (m,
1H), 5.63 (d, J=9.5 Hz, 1H), 5.32 (s, 1H), 5.76 (d, J=14.5 Hz, 1H),
5.68-5.64 (m, 1H), 4.03 (d, J=16.5 Hz, 1H), 3.97-3.81 (m, 8H),
3.79-3.70 (m, 8H), 3.47-3.44 (m, 1H), 3.20-3.07 (m, 3H), 2.84-2.78
(m, 4H), 2.62-2.57 (m, 1H), 2.37-2.32 (m, 1H), 2.16-2.03 (m, 7H),
1.95-1.91 (m, 1H), 1.68-1.64 (m, 1H), 1.47-1.44 (m, 1H), 1.12 (t,
J=7.5 Hz, 3H), 0.76 (t, J=7.5 Hz, 3H); ESI MS m/z 900
[M+H].sup.+.
Example 37
Description of Biological Assays
A. HeLa GI.sub.50 Determinations
[0265] Growth inhibition (GI.sub.50) values were measured on the
human cervical carcinoma cell line, HeLa S-3, which were selected
for growth on plastic. The HeLa cell assay was based on the
description of Skehan et al., J. Natl. Cancer Inst, 82:1107-12
(1990), which is hereby incorporated by reference in its entirety.
HeLa cells were plated at 2.times.10.sup.4 cells/well in 96 well
plates. One day later, a control plate was fixed by the addition of
TCA to 5%. After five rinses with tap water, the plate was
air-dried and stored at 4.degree. C. Test compounds were added to
the remaining plates at 10-fold dilutions. Two days later, all
plates were fixed as described above. Cells were then stained by
the addition of 100 SAL per well of 0.4% sulforhodamine B (SRB) in
1% acetic acid for 30 min at 4.degree. C. Wells were then quickly
rinsed 5.times. with 1% acetic acid and allowed to air dry. The SRB
was then solubilized by the addition of 100 .mu.L per well of
unbuffered 10 mM Tris base. Dye was quantified by measuring
absorbance at 490 nm on a Molecular Devices microplate reader.
Growth inhibition was calculated according to the following
equation: GI=100.times.(T-T.sub.0)/(C-T.sub.0), where the optical
density (OD) of the test well after 2 days of treatment was T, the
OD of the wells in the control plate on day 0 was T.sub.0 and C was
the OD of untreated wells. Plots of percent growth inhibition
versus inhibitor concentration were used to determine the
GI.sub.50.
B. MCF-7 GI.sub.50 Determinations
[0266] Growth inhibition (GI.sub.50) values were measured on the
human breast carcinoma line, MCF-7. MCF-7 cells were plated at
2.times.10.sup.4 cells/well in 96 well plates and grown for 24
hours in drug free media. On day 2, test compounds were added to
the plates at 10-fold dilutions. Four days later, cells were fixed
by the addition of glutaraldehyde to 0.75%. After 30 min, the fixed
cells were extensively rinsed with distilled water and dried at
room temperature for one hour. The cells were then stained with a
0.2% crystal violet solution for one hour at room temperature.
Unbound stain was removed by ten rinses with tap water and plates
were allowed to air dry for 30 min. The crystal violet was then
solubilized by the addition of 10% acetic acid for 15 min and
quantified by measuring absorbance at 570 nm on a Molecular Devices
microplate reader. Growth inhibition was calculated according to
the following equation: GI=100.times.(T/T.sub.0), where the optical
density (OD) of the test well after 4 days of treatment was T, the
OD of the wells in the control plate on day 0 was T.sub.0. Plots of
percent growth inhibition versus inhibitor concentration were used
to determine the GI.sub.50. TABLE-US-00002 TABLE 3 Growth
Inhibition (GI.sub.50) of HeLa Cells for Compounds of the Current
Invention. HeLa Cells MCF-7 Cells Example GI.sub.50 (nM) GI.sub.50
(nM) 1 5 3 2 25 7 3 3 7 4 400 300 5 300 600 6 800 >1000 7 200
400 8 40 60 9 20 30 10 0.3 1 11 5 20 12 20 40 13 20 30 14 30 50 15
200 400 16 300 500 17 20 300 18 30 300 19 300 >1000 20 40 300 21
0.4 5 22 40 50 23 300 300 24 100 300 25 1 6 26 4 20 27 30 50 28 300
>1000 29 70 300 30 70 300 31 300 600 32 300 400 33 30 50 34 30
30 35 300 600 36 300 400
C. NCI Sixty Cell Line Data
[0267] The following data in Table 4 summarize the growth
inhibition properties of several compounds of the present invention
against 60-human transformed cell lines. These data were
cooperatively obtained at the National Cancer Institute in their
60-cell line growth inhibition assay according to published
procedures (Boyd, M. R., "Anticancer Drug Development Guide,"
Preclinical Screening, Clinical Trials, and Approval; Teicher, B.
Ed.; Humana Press; Totowa, N.J., 23-42 (1997), which is hereby
incorporated by reference in its entirety). TABLE-US-00003 TABLE 4
In Vitro Growth Inhibition (GI.sub.50) of NCI Human Transformed
Cell Lines of Several Compounds of the Current Invention. Cancer
Type Cell Line 1 GI.sub.50 (nM) 2 GI.sub.50 (nM) Breast BT-549
<10 -- Breast HS 578T 679 19.2 Breast MCF7 <10 11.3 Breast
MDA-MB- <10 77.2 231/ATCC Breast MDA-MB-435 <10 1370 Breast
NCI/ADR-RES 152 5280 Breast T-47D -- -- CNS SF-268 <10 78.8 CNS
SF-295 <10 27 CNS SF-539 <10 37.9 CNS SNB-19 -- 62.5 CNS
SNB-75 15100 40.4 CNS U251 <10 42.3 Colon COLO 205 -- 17.9 Colon
HCC-2998 -- <10 Colon HCT-116 <10 -- Colon HCT-15 <10 1070
Colon HT29 <10 10.3 Colon KM12 <10 92.7 Colon SW-620 <10
37.8 Leukemia CCRF-CEM <10 37.9 Leukemia HL-60(TB) <10 71
Leukemia K-562 <10 12.4 Leukemia MOLT-4 -- 24.1 Leukemia
RPMI-8226 <10 16.7 Leukemia SR >100000 26.1 Melanoma LOX IMVI
<10 31.3 Melanoma M14 <10 >100000 Melanoma MALME-3M 53.6
65 Melanoma SK-MEL-2 <10 1400 Melanoma SK-MEL-28 -- 91.4
Melanoma SK-MEL-5 -- 18.8 Melanoma UACC-257 5730 3430 Melanoma
UACC-62 <10 25.6 Non-Small Cell A549/ATCC <10 58.6 Lung
Non-Small Cell EKVX -- 72.6 Lung Non-Small Cell HOP-62 <10 37
Lung Non-Small Cell HOP-92 1240 2250 Lung Non-Small Cell NCI-H226
<10 83.3 Lung Non-Small Cell NCI-H23 <10 52.8 Lung Non-Small
Cell NCI-H322M <10 -- Lung Non-Small Cell NCI-H460 <10 43.2
Lung Non-Small Cell NCI-H522 <10 <10 Lung Ovarian IGROV1
<10 32.4 Ovarian OVCAR-3 <10 -- Ovarian OVCAR-4 -- 264
Ovarian OVCAR-5 -- 225 Ovarian OVCAR-8 <10 51.7 Ovarian SK-OV-3
-- 63.3 Prostate DU-145 <10 25.7 Prostate PC-3 <10 57.3 Renal
786-0 <10 -- Renal A498 <10 -- Renal ACHN 3230 1430 Renal
CAKI-1 <10 -- Renal RXF 393 10300 60.5 Renal SN12C <10 70.8
Renal TK-10 <10 298 Renal UO-31 <10 674 Renal RPMI-8226
<10 16.7
[0268] Although the invention has been described in detail for the
purpose of illustration, it is understood that such detail is
solely for that purpose, and variations can be made therein by
those skilled in the art without departing from the spirit and
scope of the invention which is defined by the following
claims.
* * * * *