U.S. patent application number 11/030567 was filed with the patent office on 2005-12-29 for platinum complexes and methods for inhibiting tumor cell proliferation.
Invention is credited to Kay, Heidi, Palmer, Jay W., Sebti, Said M., Stanko, Joseph A..
Application Number | 20050288365 11/030567 |
Document ID | / |
Family ID | 35506822 |
Filed Date | 2005-12-29 |
United States Patent
Application |
20050288365 |
Kind Code |
A1 |
Kay, Heidi ; et al. |
December 29, 2005 |
Platinum complexes and methods for inhibiting tumor cell
proliferation
Abstract
The subject invention concerns platinum complexes that exhibit
antitumor cell and/or antiparasitic activity. The subject invention
also concerns the use of platinum complexes of the invention to
treat oncological and inflammatory disorders. The platinum
complexes of the invention can also be used to treat or prevent
infection by a virus or a bacterial or parasitic organism in vivo
or in vitro.
Inventors: |
Kay, Heidi; (Wesley Chapel,
FL) ; Palmer, Jay W.; (Sun City Center, FL) ;
Stanko, Joseph A.; (Temple Terrace, FL) ; Sebti, Said
M.; (Tampa, FL) |
Correspondence
Address: |
SALIWANCHIK LLOYD & SALIWANCHIK
A PROFESSIONAL ASSOCIATION
PO BOX 142950
GAINESVILLE
FL
32614-2950
US
|
Family ID: |
35506822 |
Appl. No.: |
11/030567 |
Filed: |
January 6, 2005 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
|
60534575 |
Jan 6, 2004 |
|
|
|
Current U.S.
Class: |
514/492 ;
556/136 |
Current CPC
Class: |
A61P 35/00 20180101;
C07F 15/0093 20130101 |
Class at
Publication: |
514/492 ;
556/136 |
International
Class: |
A61K 031/28; C07F
015/00 |
Claims
We claim:
1. A platinum complex having the structure of a complex selected
from the group consisting of the complex designated herein as
CPA-2, CPA-4, CPA-5, CPA-6, CPA-8, CPA-9, CPA-10, CPA-11, CPA-12,
CPA-13, CPA-14, CPA-15, CPA-16, CPA-17, CPA-18, CPA-19, CPA-20,
CPA-21, CPA-22, CPA-23, CPA-24, CPA-25, CPA-26, CPA-27, CPA-28,
CPA-29, CPA-30, CPA-31, CPA-32, CPA-33, CPA-34, CPA-35, CPA-36,
CPA-37, CPA-38, CPA-39, CPA-40, CPA-41, CPA-42, CPA-43, CPA-44,
CPA-45, CPA-46, CPA-47, CPA-48, CPA-49, CPA-50, CPA-51, CPA-52,
CPA-53, CPA-54, CPA-55, CPA-56, CPA-57, CPA-58, JP3, JP4, JP5, JP6,
GD6, GD2, GD3, GD4, JP13A, JP14C, JP14D, JP15, or a
pharmaceutically acceptable salt thereof.
2. A method for treating or preventing an oncological disorder, an
inflammatory disorder, or a viral, bacterial, or parasitic
infection in a patient, said method comprising administering an
effective amount of a platinum complex of claim 1 to the patient.
Description
CROSS-REFERENCE TO RELATED APPLICATION
[0001] This application claims the benefit of U.S. Provisional
Application Ser. No. 60/534,575, filed Jan. 6, 2004, the disclosure
of which is incorporated herein by reference in its entirety.
BACKGROUND OF THE INVENTION
[0002] Platinum complexes, the prototype of cisplatin, have been
widely used as active anticancer agents (Ardizzoni et al., 1999;
Nitiss, 2002) in a variety of human tumors, including testicular,
ovarian, bladder carcinoma, head and neck, and non-small cell lung
cancers. The outcome of treatments with cisplatin and other
platinum-containing compounds is strongly linked to their
alkylating effects on DNA. However, the potential impact of
platinum-complex-based therapy on cellular signaling and the
therapeutic importance of such interactions have yet to be
explored. Reports show that cisplatin induces activation of members
of the mitogen-activated protein kinase (MAPK) pathways (Persons et
al., 1999; Sanchez-Perez et al., 1998), which may influence
drug-induced apoptosis.
BRIEF SUMMARY OF THE INVENTION
[0003] The subject invention concerns platinum complexes and
methods for treating disease conditions, such as cancer and tumors,
using platinum complexes of the invention.
DETAILED DISCLOSURE OF THE INVENTION
[0004] The subject invention concerns platinum complexes and
methods for inducing apoptosis and inhibiting tumor cell growth and
for treating animals having tumors, cancers, and oncological
disease conditions using the subject platinum complexes. Platinum
complexes of the subject invention are shown in the Table
below:
1 Designation Structure IC50 CisPt 1 A549: 9.9 .+-. 4.0 Calu-1: 8.6
.+-. 45 Panc-1: 4.5 .+-. 1.9 T-24: 1.3 .+-. 0.4 CPA-1 2 A549: 50
.+-. 23 Calu-1: 26 .+-. 11 Panc-1: 19 .+-. 8.1 T-24: 6.2 .+-. 1.1
CPA-2 3 A549: >1000 Calu-1: 497 Panc-1: 890 T-24: 560 CPA-3 4
A549: >250 Calu-1: >250 Panc-1: 94 .+-. 65 T-24:
23/152/>250 CPA-4 ("dirty mix") 5 A549: 50 .+-. 23 Calu-1: 26
.+-. 11 Panc-1: 19 .+-. 8.1 T-24: 6.2 .+-. 1.1 CPA-4 ("clean") same
as above A549: >250 Calu-1: >250 Panc-1: >250 T-24:
>250 CPA-5 6 A549: 213 Calu-1: 181 Panc-1: >250 T-24: 111
CPA-6 7 CPA-7 8 A549: 20 Calu-1: --Panc-1: 6.0 T-24: -- CPA-8 9
A549: >250 Calu-1: --Panc-1: >250 T-24: -- CPA-9 10 A549:
>250 Calu-1: --Panc-1: 6.0 T-24: -- CPA-10 11 A549: >250
Calu-1: --Panc-1: 6.0 T-24: -- CPA-11 12 A549: 138 Calu-1:
--Panc-1: 48 T-24: -- CPA-12 JP1076B
cis-[diaminodichloronitro-C.sub.6H.sub.4O.sub.2--B--O--Pt.sup.IV]-
FW 479.98 rec'd May 13, 1999 13 A549: >250 .mu.M 70 .mu.M
Calu-1: --Panc-1: >250 .mu.M 28 .mu.M T-24: --Panc-1: <1.953
.mu.M 2.3 .mu.M 4.4 .mu.M 2.3 .mu.M CPA-13 JP1078B
Pt.sup.IV(en)HOHN--CO--NH.sub.2)(NO.sub.2)5-FU 64 .mu.M FW 508.39
rec'd May 3, 1999 14 A549: >250 .mu.M >250 .mu.M Calu-1: 149
.mu.M 112 .mu.M Panc-1: <1.953 .mu.M 1.9 .mu.M 2.4 .mu.M 10
.mu.M CPA-14 JP1079
cis-[Pt.sup.IV(NH.sub.3).sub.3Cl.sub.2NO.sub.2-5FU FW 476.2 rec'd
Aug. 13, 1999 15 A549: 18 .mu.M 4.8 .mu.M 38 .mu.M Calu-1: 13 .mu.M
CPA-15 JP1080 cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.-
2NO.sub.2-acetate FW 482.2 rec'd Aug. 13, 1999 16 Panc-1: 3.3 .mu.M
17 .mu.M A549: 13.5 .mu.M 42 .mu.M CPA-16 JP1082
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2--AsO.sub.2 FW 453.0
rec'd Aug. 13, 1999 17 Panc-1: 4.8 .mu.M 20 .mu.M A549: 15 .mu.M 39
.mu.M CPA-17 JP1085A
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2--- SeOOH FW 474.1
rec'd Aug. 13, 1999 18 Panc-1: 5.8 .mu.M 20 .mu.M A549: 14 .mu.M 40
.mu.M CPA-18 JP1087A cis-[Pt.sup.IV(NH.sub.3).su-
b.2Cl.sub.2NO.sub.2-hydroxyurea FW 422.1 rec'd Aug. 13, 1999 19
Panc-1: 5.6 .mu.M 3.0 .mu.M 4.7 .mu.M A549: 13 .mu.M 12 .mu.M
Calu-1: 4.2 .mu.M
[0005]
2 20 Designation A B IC50 CPA-19 JP1081
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO- .sub.2salicylate]FW 483
rec'd Aug. 30, 1999 --NO.sub.2 21 Panc-1: 20 .mu.M 20 .mu.M A549:
37 .mu.M CPA-20 JP1092A --NO.sub.2 --ONO Panc-1: 15 .mu.M
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2(O- NO)] A549: 28
.mu.M FW 392 rec'd Aug. 30, 1999 CPA-21 JP1093B --NO.sub.2 --Cl
Panc-1: 9.2 .mu.M cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2Cl
A549: 4.2 .mu.M FW 382 rec'd Aug. 30, 1999 CPA-22 JP1091A
--NO.sub.2 --ONO.sub.2 Panc-1: 5.4 .mu.M
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.su- b.2NO.sub.2(ONO.sub.2) A549:
4.6 .mu.M FW 408 rec'd Aug. 30, 1999 CPA-23 JP1089A --NO.sub.2
--OPO.sub.3H.sub.2 Panc-1: 3.3 .mu.M
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2(H.sub.2PO.su-
b.4.sup.-) 5.7 .mu.M FW 443 1.9 .mu.M rec'd Aug. 30, 1999 A549: 4.2
.mu.M CPA-24 JP1090A --NO.sub.2 --OSO.sub.3H Panc-1: 5.2 .mu.M
cis-[Pt.sup.IV(NH.sub.3).sub.2Cl.sub.2NO.sub.2(- HSO.sub.4.sup.-)]
9.2 .mu.M FW 443 5.9 .mu.M rec'd Aug. 30, 1999 A549: 3.8 .mu.M
[0006]
3 Designation Structure IC50 CPA-25 JP0097B
cis-[Pt.sup.IV(NH.sub.3).sub.2BrCl.sub.2NO.sub.2 FW 428 rec'd Aug.
30, 1999 22 Panc-1: 5.6 .mu.M 13 .mu.M A549: 8.0 .mu.M CPA-26 1094C
23 Panc-1: 228 .mu.M CPA-27 1084A 24 Panc-1: 31 .mu.M 35 .mu.M
CPA-28 1083A 25 Panc-1: 80 .mu.M CPA-29 1094B 26 Panc-1: 5.7 .mu.M
2.1 .mu.M CPA-30 Eosin 27 Panc-1: 5.7 .mu.M 4.5 .mu.M CPA-31
Citrate 28 Panc-1: 31 .mu.M CPA-32 Dabco 29 Panc-1: 15 .mu.M CPA-33
Tris 30 Panc-1: 10 .mu.M CPA-34 1084C 31 CPA-35 Palmitic 32 Panc-1:
7.7 .mu.M 21 .mu.M CPA-35 1,3-diaminopropane 33 Panc-1: 20 .mu.M
CPA-37 Hydroxydiaminopropane 34 Panc-1: 27 .mu.M CPA-38 Histidine
35 Panc-1: 5.6 .mu.M 36 CPA-39 Fluorescein 37 Panc-1: 171 .mu.M 12
.mu.M CPA-40 2-aminobutyric acid 38 Panc-1: 3.1 .mu.M CPA-41 IBF 39
Panc-1: 1.0 .mu.M CPA-42 CZ 40 Panc-1: 20 .mu.M CPA-43 DNP 41
Panc-1: 3.3 .mu.M 4.5 .mu.M CPA-44 Succinamide 42 Panc-1: 6.0 .mu.M
CPA-45 Acridine 43 Panc-1: 3.0 .mu.M CPA-46 Rhodamine 44 Panc-1:
3.6 .mu.M 4.8 .mu.M CPA-47 1094A 45 Panc-1: 3.3 .mu.M CPA-48 Methyl
thymol blue Panc-1: 8.6 .mu.M
[0007]
4 46 Designation A B IC50 CPA-49 D-+-maltose --NO.sub.2 47 Panc-1:
49 .mu.M CPA-50 Morpholine --NO.sub.2 48 CPA-51
3-aminophthalhydrazide --NO.sub.2 49 CPA-52 Dimethylphthalate
--NO.sub.2 50 CPA-53 2,7-dichlorofluorescein --NO.sub.2 51 CPA-54
8-hydroxyquinoline --NO.sub.2 52 CPA-55 Hydroorotic acid --NO.sub.2
53 CPA-56 Proline --NO.sub.2 54 CPA-57 Stearic acid --NO.sub.2
CH.sub.3(CH.sub.2).sub.16 COOH CPA-58 succinimide --NO.sub.2 55
[0008]
5 Designation Structure IC50 JP3 [Cis-Pt
IV(NH.sub.3).sub.2(Cl).sub.2(NO.sub.2)(5-FU)] 56 Panc-1: 2.6 .mu.M
JP4 [Cis-Pt IV(NH.sub.3).sub.2(Cl).sub.- 2(NO.sub.2)(n-octanol)] 57
Panc-1: 18.5 .mu.M JP5 [Cis-Pt
IV(NH.sub.3).sub.2(Cl).sub.2(NO.sub.2)(salicylate)] 58 Panc-1: 6.2
.mu.M JP6 [Cis-Pt IV(NH.sub.3).sub.2(Cl).sub.2(NO.sub.2)(OP(-
salicylate(OH)O)] 59 Panc-1: 5.5 .mu.M GD6 [Pt IV (1,3
diaminopropane)(Cl).sub.3(NO.sub.2)] 60 Panc-1: 9.5 .mu.M GD2 [cis-
diaminonitrosalicylatohydroselenito-Pt IV] 61 Panc-1: 60 .mu.M GD3
[cis- diaminonitrocaticholatohydroselenito- Pt IV] 62 Panc-1: 49
.mu.M GD4 [cis-diaminonitrocaticholatoarsenito- - Pt IV] 63 Panc-1:
73 .mu.M JP13A [cis-diaminonitrosalicy- latoarsenito- Pt Iv] 64
JP14B [cis-diaminosallcylato-Pt II] 65 JP14C
[cis-diaminonitrosalicylato-(5-FU) Pt IV] 66 JP14D
[Cis-diaminonitrosalycilatoarenito- Pt IV] 67 JP15
[Cis-diaminonitro-bis(ibuprofen)- Pt IV] 68 5-EU Panc-1: 5.0 .mu.M
Calu-1: >250 .mu.M hydroxyurea Panc-1: 1.31 .mu.M Calu-1:
>250 .mu.M Cisplatin 69 Panc-1: 1.3 .mu.M Calu-1: 2.0 .mu.M
[0009] Those platinum complexes designated as CPA-19 through CPA-24
and CPA-49 through CPA-58 have the structure: 70
[0010] wherein the A and B substituents are as identified in the
Table for the particular complex. The substituent can attach to the
platinum atom through any suitable atom therein, e.g., a nitrogen,
sulfur, or oxygen atom. As used herein, "5FU" means
5-fluorouracil.
[0011] The Table shows the results of the various platinum
complexes in MTT assays using cell lines A549, Calu-1, Panc-1, and
T-24. The IC50 for a particular platinum complex is shown in the
far right column of the Table.
[0012] Compounds of the subject invention also include
pharmaceutically-acceptable salts of the subject platinum
complexes. The term pharmaceutically-acceptable salts means salts
of the platinum complexes of the invention which are prepared with
acids or bases, depending on the particular substituents found on
the subject complexes described herein. Examples of a
pharmaceutically-acceptable base addition salts include sodium,
potassium, calcium, ammonium, or magnesium salt. Examples of
pharmaceutically-acceptable acid addition salts include
hydrochloric, hydrobromic, nitric, phosphoric, carbonic, sulphuric,
and organic acids like acetic, propionic, benzoic, succinic,
fumaric, mandelic, oxalic, citric, tartaric, maleic, and the like.
Pharmaceutically-acceptable salts of platinum complexes of the
invention can be prepared using conventional techniques.
[0013] It will be appreciated by those skilled in the art that
certain of the platinum complexes of the invention may contain one
or more asymmetrically substituted carbon atoms which can give rise
to stereoisomers. It is understood that the invention extends to
all such stereoisomers, including enantiomers, and diastereoisomers
and mixtures, including racemic mixtures thereof.
[0014] In one embodiment, methods of the invention comprise
inhibiting proliferation of cancerous or tumorigenic cells using
the platinum complexes of the present invention.
[0015] Platinum complexes of the invention can be delivered to a
cell either through direct contact with the cell or via a carrier
means. Carrier means for delivering compositions to cells are known
in the art and include encapsulating the composition in a liposome
moiety, and attaching the platinum complexes to a protein or
nucleic acid that is targeted for delivery to the target cell.
Published U.S. Patent Application Nos. 20030032594 and 20020120100
disclose amino acid sequences that can be coupled to another
composition and that allows the composition to be translocated
across biological membranes. Published U.S. Patent Application No.
20020035243 also describes compositions for transporting biological
moieties across cell membranes for intracellular delivery.
[0016] The subject invention also concerns methods for treating
oncological or inflammatory disorders in a patient. In one
embodiment, an effective amount of a platinum complex of the
present invention is administered to a patient having an
oncological or inflammatory disorder and who is in need of
treatment thereof. Methods of the invention can optionally include
identifying a patient who is or may be in need of treatment of an
oncological or inflammatory disorder. The patient can be a human or
other mammal, such as a primate (monkey, chimpanzee, ape, etc.),
dog, cat, cow, pig, or horse, or other animals having an
oncological disorder. Means for administering and formulating
platinum complexes for administration to a patient are known in the
art, examples of which are described herein. Oncological disorders
include cancer and/or tumors of the bone, breast, kidney, mouth,
larynx, esophagus, stomach, testis, cervix, head, neck, colon,
ovary, lung, bladder, skin, liver, muscle, pancreas, prostate,
blood cells (including lymphocytes), and brain. Inflammatory
disorders include arthritis, multiple sclerosis, lupus, Crohn's
disease, and related neurological and inflammatory connective
tissue diseases (e.g., Sjogren's syndrome).
[0017] For the treatment of oncological disorders, the platinum
complexes of this invention can be administered to a patient in
need of treatment in combination with other antitumor or anticancer
substances or with radiation therapy or with surgical treatment to
remove a tumor. These other substances or radiation treatments may
be given at the same as or at different times from the platinum
complexes of this invention. For example, the platinum complexes of
the present invention can be used in combination with mitotic
inhibitors such as taxol or vinblastine, alkylating agents such as
cyclophosamide or ifosfamide, antimetabolites such as
5-fluorouracil or hydroxyurea, DNA intercalators such as adriamycin
or bleomycin, topoisomerase inhibitors such as etoposide or
camptothecin, antiangiogenic agents such as angiostatin,
antiestrogens such as tamoxifen, and/or other anti-cancer drugs or
antibodies, such as, for example, GLEEVEC (Novartis Pharmaceuticals
Corporation) and HERCEPTIN (Genentech, Inc.), respectively.
[0018] Many tumors and cancers have viral genome present in the
tumor or cancer cells. For example, Epstein-Barr Virus (EBV) is
associated with a number of mammalian malignancies. The platinum
complexes of the subject invention can be used alone or in
combination with anticancer or antiviral agents, such as
ganciclovir, azidothymidine (AZT), lamivudine (3TC), etc., to treat
patients infected with a virus that can cause cellular
transformation and/or to treat patients having a tumor or cancer
that is associated with the presence of viral genome in the cells.
The platinum complexes of the subject invention can also be used in
combination with viral based treatments of oncologic disease. For
example, platinum complexes of the invention can be used with
mutant herpes simplex virus in the treatment of non-small cell lung
cancer (Toyoizumi et al., 1999).
[0019] The subject invention also concerns methods for treating
bacterial and viral infections of a patient using a platinum
complex of the invention. In one embodiment, an effective amount of
a platinum complex of the invention is administered to a patient
having a bacterial or viral infection. Methods of the invention can
optionally include identifying a patient who is or may be in need
of treatment of a bacterial or viral infection. The patient can be
a human or other mammal, such as a primate (monkey, chimpanzee,
ape, etc.), dog, cat, cow, pig, or horse, or other animal infected
with a bacteria or virus. Bacterial infections that can be treated
according to the present invention include those from
Staphylococcus, Streptococcus, Salmonella, Bacillus, Clostridium,
Pseudomonas, Neisseria, Mycobacterium, and Yersinia. Viral
infections that can be treated according to the present invention
include, but are not limited to, those associated with human
immunodeficiency virus (HIV), human T cell leukemia virus (HTLV),
Papillomavirus (e.g, human papilloma virus), Polyomavirus (e.g.,
SV40, BK virus, DAR virus), orthopoxvirus (e.g., variola major
virus (smallpox virus)), EBV, herpes simplex virus (HSV), hepatitis
virus, Rhabdovirus (e.g., Ebola virus) and cytomegalovirus (CMV).
Platinum compositions of the present invention can also be used to
treat viral diseases in the presence of photodynamic therapy (Cuny
et al., 1999).
[0020] Platinum complexes of the subject invention can also be used
to treat patients infected with a parasitic organism. In one
embodiment, the patient is administered a therapeutically effective
amount of a platinum complex of the present invention. Methods of
the invention can optionally include identifying a patient who is
or may be in need of treatment of a parasitic infection. The
patient can be a human or other mammal, such as a primate (monkey,
chimpanzee, ape, etc.), dog, cat, cow, pig, or horse, or other
animal infected with a parasitic organism. Disease conditions that
can be treated according to the present invention include, but are
not limited to, leishmania, toxoplasmosis, schistosomiasis,
trypanosomiasis, pneumocystis, malaria, and trichinosis. Parasitic
organisms that can cause disease conditions treatable according to
the present invention include, but are not limited to, Leishmania,
Toxoplasma, Schistosoma, Plasmodium, and Trypanosoma. The subject
invention can also be used to treat gastro-intestinal disorders
caused by parasitic organisms such as, Entamoeba, Giardia,
Trichomonas, and nematodes such as Ascaris, Trichuris, Enterobius,
Necator, Ancylostoma, Strongyloides, and Trichinella. In another
embodiment, a platinum complex of the present invention can be
administered to patients prophylactically, wherein an uninfected
patient is traveling to or will be present in an area where
parasitic disease is prevalent or poses a risk to the patient.
Accordingly, the patient can be treated with a composition of the
present invention prior to the patient's exposure to or presence in
the area where parasitic disease is prevalent or poses a risk
and/or prior to infection with the parasitic organism.
[0021] Platinum complexes of the present invention can also be used
to treat biological products in vitro that are contaminated with or
suspected of being contaminated with a virus on a bacterial or
parasitic organism. Biological products which can be treated with a
platinum complexes of the present invention include, but are not
limited to, whole blood, fractionated blood, plasma, serum, whole
organs, or parts of organs, and cells, including blood cells,
muscle cells, skin cells, and neural cells, and products derived
from cells. Products derived from cells which can be treated with a
platinum complex of the present invention include, but are not
limited to, interferons, interleukins, blood clotting factors such
as factor VIII, IX, X, and the like, insulin, polyclonal and
monoclonal antibodies, growth factors, cytokines, and other
products. Treatment of biological products comprises contacting the
product for an effective amount of time and with an effective
amount of a platinum complex of the present invention. If
necessary, the biological product can be subsequently washed,
preferably with a suitable sterile wash solution such as phosphate
buffered saline, to remove the platinum complex that was used to
treat the product.
[0022] Therapeutic application of the subject platinum complexes,
and compositions containing them, can be accomplished by any
suitable therapeutic method and technique presently or
prospectively known to those skilled in the art. The subject
platinum complexes can be administered by any suitable route known
in the art including, for example, oral, nasal, rectal, and
parenteral routes of administration. As used herein, the term
parenteral includes subcutaneous, intravenous, intramuscular, and
intrasternal administration, such as by injection. Administration
of the subject platinum complexes of the invention can be
continuous or at distinct intervals as can be readily determined by
a person skilled in the art.
[0023] Compounds useful in the methods of the subject invention can
be formulated according to known methods for preparing
pharmaceutically useful compositions. Formulations are described in
detail in a number of sources which are well known and readily
available to those skilled in the art. For example, Remington's
Pharmaceutical Science by E. W. Martin describes formulations which
can be used in connection with the subject invention. In general,
the compositions of the subject invention will be formulated such
that an effective amount of the bioactive platinum complex is
combined with a suitable carrier in order to facilitate effective
administration of the composition. The compositions used in the
present methods can also be in a variety of forms. These include,
for example, solid, semi-solid, and liquid dosage forms, such as
tablets, pills, powders, liquid solutions or suspension,
suppositories, injectable and infusible solutions, and sprays. The
preferred form depends on the intended mode of administration and
therapeutic application. The compositions also preferably include
conventional pharmaceutically acceptable carriers and diluents
which are known to those skilled in the art. Examples of carriers
or diluents for use with the subject platinum complexes include
ethanol, dimethyl sulfoxide, glycerol, alumina, starch, and
equivalent carriers and diluents. To provide for the administration
of such dosages for the desired therapeutic treatment, new
pharmaceutical compositions of the invention will advantageously
comprise between about 0.1% and 99%, and especially, 1 and 15% by
weight of the total of one or more of the subject platinum
complexes based on the weight of the total composition including
carrier or diluent.
[0024] The compounds of the subject invention can also be
administered utilizing liposome technology, slow release capsules,
implantable pumps, and biodegradable containers. These delivery
methods can, advantageously, provide a uniform dosage over an
extended period of time. The platinum complexes of the present
invention can also be administered in their salt derivative forms
or crystalline forms known to those of ordinary skill in the
art.
[0025] The subject invention also concerns a packaged dosage
formulation comprising in one or more containers at least one
platinum compound of the subject invention formulated in a
pharmaceutically acceptable dosage.
[0026] All patents, patent applications, provisional applications,
and publications referred to or cited herein are incorporated by
reference in their entirety, including all figures and tables, to
the extent they are not inconsistent with the explicit teachings of
this specification.
Materials and Methods
[0027] Synthesis of NitroPlatinum (Iv) Complexes.
[0028] Cis-diammineoplatinum(II) dichloride (cisplatin) can be
purchased at 99.9% purity from Sigma-Aldrich (#P4394). Using 0.300
grams of Cisplatin (0.00100 moles, FW=300.1), 150 mL of ultra
deionized water and 50 mL of dichloroethane are added to a 250-mL
Erlenmeyer flask. However, hexane or any organic solvents can be
substituted in place of the dichlorethane used here. The choice of
a sixth ligand includes the availability of a nitrogen, sulfur or
oxygen atom in the chemical structure providing a Lewis base for
bonding to the oxidized Pt. Other bondings are possible with
metals, halides (such as HCl) or through chelation or interaction
with pi molecular orbitals. One mole of the chosen ligand per mole
of cisplatin should be weighed and added to the mixture. Organic
solvents, such as dichloroethane, provide solubility for organic
ligands of hydrophobic nature. A magnetic stir bar is placed in the
mixture and the flask placed on a magnetic stir plate in a chemical
fume hood. A lecture bottle of dinitrogen tetroxide is fitted with
a regulator and Teflon hose, with a glass pipet attached to the
hose outlet. The pipet tip is inserted into the lower solvent
(e.g., dichloroethane) and the lecture bottle warmed slightly with
a warm water bath. Nitrogen dioxide gas is released at a rate of
approximately one bubble per second into the stirring mixture. The
gas should be added until all the yellow cisplatin is consumed; the
disappearance of yellow solids and yellow solution will indicate
consumption of the available cisplatin. A blue color is noted to
indicate formation of the nitrosyl intermediate; variations in hue
and duration of this color have been observed. Gas addition is then
terminated (remove the pipet to prevent vacuum suction into the
lecture bottle) and the flask covered in aluminum foil to prevent
light exposure. The flask should be left to stir overnight,
uncovered.
[0029] Additional nitrogen dioxide may be added the next day to
check for completeness of reaction. A blue color would indicate
incomplete oxidation of platinum (II). Normally, this blue fades
within ten minutes. For a colorless ligand, the solution has become
yellow overnight. If blue color remains, allow it to continue
stirring. The mixture requires air for complete oxidation, so
should not be tightly covered. Continued oxidation with air can be
accelerated using air blown through a trap into the Erlenmeyer,
over the liquids. The solvents will evaporate in about two days,
leaving a yellow precipitate, which is the product.
[0030] The precipitate can be purified via recrystallization in
methanol, DMSO, or other suitable solvent. Alternatively, the
product can be purified on silica columns or using HPLC.
[0031] MTT Assay.
[0032] Inhibition of the growth of human tumor cells was carried
out in 96-well plates using the MTT assay Cells were plated and
treated with various concentrations of the platinum complex for 4
days. Cell viability then was determined by adding to the cells 1
mg/ml media of 3-(4-5-dimethyl-thiazol-2-yl)-2,5-diphenyl
tetrazolium bromide (MTT), and incubating the cells for 3 hrs at
37.degree. C. The dye-containing media was replaced by DMSO. After
a 5 min incubation with DMSO, absorbance of the control compared to
inhibitor treated columns of the 96-well plate was then read at 540
nM with a microtiter plate reader and IC50s determined.
[0033] XTT Assay.
[0034] A 96-well plate was used for the assays. Approximately
2.5.times.10.sup.4 cells in log phase were added to each well. A
platinum complex of the invention was dispensed into each well
(dissolved in 20% DMSO and 80% media), with additional media added
as needed to account for uniform volumes. Control wells contained
only cells and media. Each concentration assay was performed in
triplicate. Plates were incubated for 48 hours at 37.degree. C.
with 7.5% CO.sub.2. XTT from MD Biosciences, Quebec, was then added
according to the provided protocol concentrations and allowed to
react for 3 hours. Plates were agitated 5 minutes before reading
absorbance at 475 nm on a Varian Cary 50 spectrophotometer with a
fibre-optic probe. Percent survival as compared to control wells
was plotted against platinum complex concentration.
[0035] It should be understood that the examples and embodiments
described herein are for illustrative purposes only and that
various modifications or changes in light thereof will be suggested
to persons skilled in the art and are to be included within the
spirit and purview of this application and the scope of the
appended claims.
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