U.S. patent application number 10/416259 was filed with the patent office on 2004-05-27 for combination preparation of a biological response modifier and an anticancer agent and uses thereof.
Invention is credited to Young, Aiping H.
Application Number | 20040101511 10/416259 |
Document ID | / |
Family ID | 4167585 |
Filed Date | 2004-05-27 |
United States Patent
Application |
20040101511 |
Kind Code |
A1 |
Young, Aiping H |
May 27, 2004 |
Combination preparation of a biological response modifier and an
anticancer agent and uses thereof
Abstract
The present invention provides anticancer biological response
modifier combinations. In accordance with an aspect of the present
invention, there is provided a combination comprising: (i) a
composition comprising small molecular weight components of less
than 3000 daltons, and having the following properties: is
extracted from bile of animals; is capable of stimulating monocytes
and/or macrophages in vitro and/or in vivo; is capable of
modulating tumor necrosis factor production and/or release;
contains no measurable level of IL-1.alpha., IL-1.beta., TNF, IL-6,
IL-8, IL4, GM-CSF or IFN-gamma; is not cytotoxic to human
peripheral blood mononuclear cells; is not an endotoxin; and (ii)
one or more anticancer agent(s), wherein said combination has
therapeutic synergy or improves the therapeutic index in the
treatment of cancer over the composition or the anticancer agent(s)
alone. Another aspect of the present invention provides the use of
this combination in the manufacture of a medicament or a
pharmaceutical kit and in the treatment of cancer.
Inventors: |
Young, Aiping H; (Ontario,
CA) |
Correspondence
Address: |
SALIWANCHIK LLOYD & SALIWANCHIK
A PROFESSIONAL ASSOCIATION
2421 N.W. 41ST STREET
SUITE A-1
GAINESVILLE
FL
326066669
|
Family ID: |
4167585 |
Appl. No.: |
10/416259 |
Filed: |
January 2, 2004 |
PCT Filed: |
November 8, 2001 |
PCT NO: |
PCT/CA01/01558 |
Current U.S.
Class: |
424/85.2 ;
424/537; 424/649; 424/85.4; 514/44A; 514/449; 514/50 |
Current CPC
Class: |
A61K 38/20 20130101;
A61K 35/413 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 38/21 20130101; A61K 35/413 20130101;
A61P 35/00 20180101; A61K 38/21 20130101; A61K 45/06 20130101; A61K
38/20 20130101; A61P 35/02 20180101 |
Class at
Publication: |
424/085.2 ;
424/085.4; 424/537; 424/649; 514/044; 514/050; 514/449 |
International
Class: |
A61K 048/00; A61K
038/21; A61K 038/20; A61K 035/37; A61K 033/24 |
Foreign Application Data
Date |
Code |
Application Number |
Nov 8, 2000 |
CA |
2,325,361 |
Claims
We claim:
1. A combination comprising: (a) a composition comprising small
molecular weight components of less than 3000 daltons, and having
the following properties: (i) is extracted from bile of animals;
(ii) is capable of stimulating monocytes and/or macrophages in
vitro and/or in vivo; (iii) is capable of modulating tumor necrosis
factor production and/or release; (iv) contains no measurable level
of IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or
IFN-gamma; (v) is not cytotoxic to human peripheral blood
mononuclear cells; (vi) is not an endotoxin; and (b) one or more
anticancer agent(s), wherein said combination has therapeutic
synergy or improves the therapeutic index in the treatment of
cancer over the composition or the anticancer agent(s) alone.
2. The combination according to claim 1, wherein said anticancer
agent(s) is selected from the group consisting of a
chemotherapeutic drug, radiation, a gene therapy and an antisense
oligonucleotide.
3. The combination according to claim 2, wherein said anticancer
agent(s) is a chemotherapeutic drug, interleukin or interferon.
4. The combination according to any one of claims 1, 2 or 3,
wherein at least one of said one or more anticancer agent(s) is a
chemotherapeutic drug.
5. The combination of claim 4, wherein the chemotherapeutic drug is
gemcitabine, 5-fluorouracil, dacarbazine, taxol, taxotere,
cisplatin or mitoxantrone.
6. Use of the combination according to any one of claims 1, 2, 3, 4
or 5 in the manufacture of a medicament.
7. Use of the combination according to any one of claims 1, 2, 3, 4
or 5 in the manufacture of a pharmaceutical kit.
8. A pharmaceutical kit comprising: (a) a dosage unit of a
composition and a pharmaceutically acceptable carrier wherein the
composition comprises small molecular weight components of less
than 3000 daltons, and has the following properties: (i) is
extracted from bile of animals; (ii) is capable of stimulating
monocytes and/or macrophages in vitro and/or in vivo; (iii) is
capable of modulating tumor necrosis factor production and/or
release; (iv) contains no measurable level of IL-1.alpha.,
IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma; (v) is not
cytotoxic to human peripheral blood mononuclear cells; (vi) is not
an endotoxin; and (b) a dosage unit of one or more chemotherapeutic
drug(s) and a pharmaceutically acceptable carrier, said (a) and (b)
being provided in amounts that have therapeutic synergy or that
improve the therapeutic index in the treatment of cancer over the
composition or the chemotherapeutic drug(s) alone.
9. The kit according to claim 8, wherein said one or more
chemotherapeutic drug(s) is gemcitabine, 5-fluorouracil,
dacarbazine, taxol, taxotere, cisplatin or mitoxantrone.
10. A pharmaceutical composition comprising: (a) a composition
comprising small molecular weight components of less than 3000
daltons, and having the following properties: (i) is extracted from
bile of animals; (ii) is capable of stimulating monocytes and/or
macrophages in vitro and/or in vivo; (iii) is capable of modulating
tumor necrosis factor production and/or release; (iv) contains no
measurable level of IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4,
GM-CSF or IFN-gamma; (v) is not cytotoxic to human peripheral blood
mononuclear cells; (vi) is not an endotoxin; (b) one or more
chemotherapeutic drug(s); and (c) a pharmaceutically acceptable
carrier; wherein said pharmaceutical composition has therapeutic
synergy or improves the therapeutic index in the treatment of
cancer over the composition or the chemotherapeutic drug(s)
alone.
11. The pharmaceutical composition according to claim 10, wherein
at least one of said one or more chemotherapeutic drug(s) is
gemcitabine, 5-fluorouracil, dacarbazine, taxol, taxotere,
cisplatin or mitoxantrone.
12. The pharmaceutical composition according to claim 10 or 11,
formulated into a sterile solution, a lyophilate, a pill, a tablet,
a cream, a capsule, a suppository, a gelatin capsule, a soft
gelatin capsule, a gel, a membrane or a tubelet.
13. The combination according to any one of claims 1, 2, 3, 4 or 5
for use in the treatment of cancer.
14. The combination according to claim 13, wherein said composition
and said one or more anticancer agent(s) are suitable for separate,
concurrent or simultaneous administration.
15. The combination according to claim 13 or 14, wherein said
cancer is pancreatic cancer, melanoma, breast cancer, prostate
cancer, ovarian cancer, endometrial cancer, lung cancer, Kaposi's
sarcoma, leukemia, lymphoma, gastric cancer, colon cancer,
colorectal cancer, esophageal cancer, renal cancer, head or neck
cancer.
16. The combination according to claim 13, wherein said cancer is
melanoma, said anticancer agent is dacarbazine and said anticancer
agent is suitable for concurrent administration with the
composition.
17. The combination according to claim 13, wherein said cancer is
breast cancer, said anticancer agent is taxol and said anticancer
agent is suitable for concurrent administration with the
composition.
18. The combination according to any one of claims 13, 14, 15, 16
or 17, wherein said composition and/or said anticancer agent(s) are
suitable for administration via oral, topical, rectal, parenteral,
local, inhalant or intracerebral delivery.
19. The combination according to claim 18, wherein said parenteral
delivery is achieved via intramuscular injection.
20. The pharmaceutical composition according to any one of claims
10, 11 or 12 for use in the treatment of cancer.
21. The pharmaceutical composition according to claim 20, wherein
said cancer is pancreatic cancer, melanoma, breast cancer, prostate
cancer, ovarian cancer, endometrial cancer, lung cancer, Kaposi's
sarcoma, leukemia, lymphoma, gastric cancer, colon cancer,
colorectal cancer, esophageal cancer, renal cancer, head or neck
cancer.
22. The pharmaceutical composition according to claim 20, wherein
said cancer is melanoma and said anticancer agent is
dacarbazine.
23. The pharmaceutical composition according to claim 20, wherein
said cancer is breast cancer and said anticancer agent is
taxol.
24. The pharmaceutical composition according to any one of claims
20, 21, 22 or 23, wherein said pharmaceutical composition is
suitable for administration via oral, topical, rectal, parenteral,
local, inhalant or intracerebral delivery.
25. The pharmaceutical composition according to claim 24, wherein
said parenteral delivery is achieved via intramuscular
injection.
26. Use of the pharmaceutical composition according to any one of
claims 10, 11 or 12, for administration to a patient in need
thereof.
27. Use of the combination according to any one of claims 1, 2, 3,
4 or 5, for administration to a patient in the treatment of
cancer.
28. The use according to claim 27, wherein said composition and one
or more anticancer agent(s) are administered separately,
concurrently or simultaneously.
29. A method for treating cancer, comprising the step of
administering a therapeutically effective amount of the combination
of any one of claims 1, 2, 3, 4 or 5 to a patient in need of such
the
30. A method for treating cancer comprising administering a
therapeutically effective amount of a composition and one or more
anticancer agent(s) to a patient in need thereof, wherein said
composition comprises small molecular weight components of less
than 3000 daltons, and has the following properties: (a) is
extracted from bile of animals; (b) is capable of stimulating
monocytes and/or macrophages in vitro and/or in vivo; (c) is
capable of modulating tumor necrosis factor production and/or
release; (d) contains no measurable level of IL-1.alpha.,
IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma; (e) is not
cytotoxic to human peripheral blood mononuclear cells; (f) is not
an endotoxin; and wherein said composition and said anticancer
agent(s) are formulated for administration to a patient in need
thereof.
31. The method according to claim 30, wherein said anticancer
agent(s) is selected from the group consisting of a
chemotherapeutic drug, radiation, a gene therapy and an antisense
olig
32. The method according to claim 31, wherein said anticancer
agent(s) is a chemotherapeutic drug, interleukin or interferon
33. The method according to any one of claims 30, 31 or 32, wherein
at least one of said one or more anticancer agent(s) is a
chemotherapeutic drug.
34. The method according to claim 33, wherein the chemotherapeutic
drug is gemcitabine, 5-fluorouracil, dacarbazine, taxol, taxotere,
cisplatin or mitoxantrone.
35. The method according to any one of claims 29, 30, 31, 32, 33 or
34, wherein said cancer is pancreatic cancer, melanoma, breast
cancer, prostate cancer, ovarian cancer, endometrial cancer, lung
cancer, Kaposi's sarcoma, leukemia, lymphoma, gastric cancer, colon
cancer, colorectal cancer, esophageal cancer, renal cancer, head or
neck cancer.
36. The method according to any one of claims 29, 30, 31, 32, 33,
34 or 35, wherein said combination is formulated into a sterile
solution, a lyophilate, a pill, a tablet, a cream, a capsule, a
suppository, a gelatin capsule, a soft gelatin capsule, a gel, a
membrane or a tubelet.
37. The method according to any one of claims 29, 30, 31, 32, 33,
34, 35 or 36, wherein said administering is achieved by means of
oral, topical, rectal, parenteral, local, inhalant, or
intracerebral delivery.
38. The method of claim 37, wherein said parenteral delivery is
achieved via intramuscular injection.
39. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein said cancer is pancreatic cancer.
40. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein said cancer is melanoma.
41. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein said cancer is breast cancer.
42. The method of claim 39 wherein one of the one or more
anticancer agent(s) is gemcitabine.
43. The method of claim 39 wherein one of the one or more
anticancer agent(s) is 5-fluorouracil.
44. The method of claim 41 wherein one of the one or more
anticancer agent(s) is dacarbazine.
45. The method of claim 41 wherein one of the one or more
anticancer agent(s) is taxol.
46. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein peripheral blood monocytes and/or tumor
associated macrophages are stimulated to express cytocidal activity
in a manner that is insensitive to the inhibitory effects of
prostaglandins.
47. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein suitable modulation of the immune system is
elicited in a patient in need of such modulation by activating
macrophages and/or monocytes to produce and/or release cytokines or
promote activity to seek and remove or destroy cancerous cells.
48. The method of any one of claims 29, 30, 31, 32, 33, 34, 35, 36,
37 or 38, wherein the release of TNF, Il-1.beta. and GM-CSF is
stimulated.
Description
FIELD OF THE INVENTION
[0001] The present invention relates to anticancer biological
response modifier combinations, pharmaceutical compositions
comprising the same, and the use thereof in the treatment of
cancer.
BACKGROUND OF THE INVENTION
[0002] There are a number of therapies directed towards the
treatment of cancer, including chemotherapeutic drugs, radiation,
gene therapy and antisense oligonucleotides. One drawback to
current therapies is the toxicity associated with most treatments.
Moreover, oftentimes large dosages must be administered over an
extended period of time in order to attain therapeutic benefit.
Thus, a need remains for more effective treatments.
[0003] A bile extract has been prepared that is known to be able to
modify the biological response of cells of the immune system. The
production and characterization of this bile-derived Biological
Response Modifier (BD-BRM) has been described in International
Patent Application Serial No. PCT/CA94/00494, published Feb. 16,
1995 as WO 95/07089, International Patent Application Serial No.
PCT/CA96/00152, published Sep. 19, 1996 as WO 96/28175 and U.S.
Pat. No. 6,280,774. The use of this immunomodulatory composition as
an anti-viral has been described in International Patent
Application Serial No. PCT/CA98/00494, published Nov. 26, 1998 as
WO 98/52585. These applications are herein incorporated by
reference in their entirety.
[0004] The BD-BRM composition is composed of small molecular weight
components of less than 3000 daltons, and has one or more of the
following properties:
[0005] a) is extracted from bile of animals;
[0006] b) is capable of stimulating monocytes and/or macrophages in
vitro and/or in vivo:
[0007] c) is capable of modulating tumor necrosis factor production
and/or release;
[0008] d) contains no measurable level of IL-1.alpha., IL-1.beta.,
TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-.gamma.;
[0009] e) shows no cytotoxicity to human peripheral blood
mononuclear cells or lymphocytes; and
[0010] f) is not an endotoxin.
[0011] The bile-derived biologic response modifier (BD-BRM) is a
composition that has been hypothesized to exert anti-tumour
activity via the activation of macrophages, with subsequent
enhancement of cell-mediated immune response to tumours. Its
precise mechanism of action remains unknown.
[0012] The cumulative results of studies with BD-BRM revealed
following:
[0013] (1) BD-BRM does not directly stimulate lymphocytes to
synthesize DNA or undergo blastogenesis and cell division. BD-BRM
does not directly stimulate the development of lymphocyte-mediated
cytotoxicity.
[0014] (2) BD-BRM can stimulate normal peripheral blood monocytes
to express cytocidal activity in a dose-dependent manner. The
activity elicited by BD-BRM is equal to or greater than the
activity produced in response to more conventional macrophage
activators that are currently under investigation in cancer
patients including: Gamma Interferon; Granulocyte-Monocyte Colony
Stimulating Factor; Monocyte Colony Stimulating Factor; and
Interleukin-12.
[0015] (3) BD-BRM can stimulate both the peripheral blood monocytes
and regional, tumour-associated macrophages from cancer patients to
express significant cytocidal activity. This included peritoneal
macrophages from women with gynaecological malignancies and
alveolar macrophages from patients with lung cancer. BD-BRM has
been found to stimulate macrophages from cancer patients to kill
autologous and heterologous tumour cells obtained from surgical
specimens of patients. Of potentially greater importance is the
finding that BD-BRM can often stimulate cancer patient macrophages
that are unresponsive to stimulation with conventional activators
such as gamma interferon+endotoxin.
[0016] (4) The hypersecretion of prostaglandins, both by
macrophages and by tumor cells from cancer patients has been shown
to be a principal cause of the immunosuppression seen in patients
with advanced malignant disease. One determinant of the biological
activity of different macrophage activators in cancer patients
PBMs, therefore, is the sensitivity of the activator to arachidonic
acid metabolism and the secretion by the cell of prostaglandins.
The development of macrophage cytocidal function in response to
BD-BRM was found to be insensitive to the inhibitory effects of
prostaglandins. This is considered important therapeutically
because the effectiveness of many other biological activators is
limited by prostaglandins.
[0017] (5) BD-BRM can stimulate cytocidal function in macrophages
obtained from cancer patients (including pancreatic cancer) who are
undergoing cytotoxic therapy. Of note is the fact that BD-BRM was
more effective in stimulating tumouricidal function than
conventional activators such as gamma interferon plus
endotoxin.
[0018] (6) BD-BRM can also stimulate cytocidal function in
macrophages obtained from patients with Kaposi's sarcoma even at
very late stages of the disease. Thus, the action of BD-BRM appears
to be independent of the need for collaboration with other immune
cell types including helper T-lymphocytes.
[0019] (7) The macrophage cytocidal function that develops in
response to BD-BRM may be associated with the expression of
TNF.alpha. by the macrophages. However, other mechanisms for
cytotoxicity may also be involved. The BD-BRM composition from
bovine sources promotes the release of TNF from human peripheral
blood mononuclear cells and from the pre-monocyte cell line U-937
in what appears to be physiological quantities. Because TNF is
known to initiate a cascade of inflammatory and antitumor cytokine
effects, the composition could exert its antineoplastic effect by
stimulating human leukocytes to release TNF (and possibly other
cytokines).
[0020] (8) Demonstrates anti-tumour activity in a mouse tumour
(plasmacytoma) model.
[0021] (9) Exhibits no toxicity in animals at doses up to 125 X the
doses used in human toxicity studies with no LD.sup.50 yet reached
in toxicity studies.
[0022] (10) Induces the phenomenon of apoptosis in some continuous
cell lines.
[0023] (k) Is non-cytotoxic to human PBMNs and lymphocytes. The
survival of human peripheral blood mononuclear cells (PBMNs) and
lymphocytes is not affected by BD-BRM.
[0024] The central hypothesis guiding investigations of the BD-BRM
composition is that the therapeutic efficacy of a powerful
biological stimulator can depend on its ability to elicit suitable
modulation of the immune system, such as by activating macrophages
and/or monocytes to produce certain cytokines or promote activity
to seek and remove or destroy disease-causing viruses or cells
negatively affected by such viral infections. Such function could
be generated by direct stimulation of resident immune cells in
tumour microenvironments. Alternatively, this function could be
generated by stimulation of circulating immune cells if those cells
were then able to home in on tumour sites and to function in that
environment.
[0025] This background information is provided for the purpose of
making known information believed by the applicant to be of
possible relevance to the present invention. No admission is
necessarily intended, nor should be construed, that any of the
preceding information constitutes prior art against the present
invention. Publications referred to throughout the specification
are hereby incorporated by reference in their entireties in this
application.
SUMMARY OF THE INVENTION
[0026] An object of the present invention is to provide anticancer
biological response modifier combinations. In accordance with an
aspect of the present invention, there is provided a combination
comprising: (i) a composition comprising small molecular weight
components of less than 3000 daltons, and having the following
properties: is extracted from bile of animals; is capable of
stimulating monocytes and/or macrophages in vitro and/or in vivo;
is capable of modulating tumor necrosis factor production and/or
release; contains no measurable level of IL-1.alpha., IL-1.beta.,
TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma; is not cytotoxic to
human peripheral blood mononuclear cells; is not an endotoxin; and
(ii) one or more anticancer agent(s), wherein said combination has
therapeutic synergy or improves the therapeutic index in the
treatment of cancer over the composition or the anticancer agent(s)
alone. Another aspect of the present invention provides the use of
this combination in the manufacture of a medicament or a
pharmaceutical kit.
[0027] In accordance with another aspect of the invention, there is
provided a pharmaceutical kit comprising: (i) a dosage unit of a
composition and a pharmaceutically acceptable carrier wherein the
composition comprises small molecular weight components of less
than 3000 daltons, and has the following properties: is extracted
from bile of animals; is capable of stimulating monocytes and/or
macrophages in vitro and/or in vivo; is capable of modulating tumor
necrosis factor production and/or release; contains no measurable
level of IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or
IFN-gamma; is not cytotoxic to human peripheral blood mononuclear
cells; is not an endotoxin; and (ii) a dosage unit of one or more
chemotherapeutic drug(s) and a pharmaceutically acceptable carrier,
(i) and (ii) being provided in amounts that are effective, in
combination, for killing tumour or metastatic cells.
[0028] In accordance with another aspect of the invention, there is
provided a pharmaceutical composition comprising: (i) a composition
comprising small molecular weight components of less than 3000
daltons, and having the following properties: is extracted from
bile of animals; is capable of stimulating monocytes and/or
macrophages in vitro and/or in vivo; is capable of modulating tumor
necrosis factor production and/or release; contains no measurable
level of IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or
IFN-gamma; is not cytotoxic to human peripheral blood mononuclear
cells; is not an endotoxin; (ii) one or more chemotherapeutic
drug(s); and (iii) a pharmaceutically acceptable carrier; wherein
said pharmaceutical composition has therapeutic synergy or improves
the therapeutic index in the treatment of cancer over the
composition or the chemotherapeutic drug(s) alone.
[0029] In accordance with another aspect of the invention, there is
provided a combination for use in the treatment of cancer,
comprising: (i) a composition comprising small molecular weight
components of less than 3000 daltons, and having the following
properties: is extracted from bile of animals; is capable of
stimulating monocytes and/or macrophages in vitro and/or in vivo;
is capable of modulating tumor necrosis factor production and/or
release; contains no measurable level of IL-1.alpha., IL-1.beta.,
TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma; is not cytotoxic to
human peripheral blood mononuclear cells; is not an endotoxin; and
(ii) one or more anticancer agent(s), wherein said combination has
therapeutic synergy or improves the therapeutic index in the
treatment of cancer over the composition or the anticancer agent(s)
alone.
[0030] In accordance with another aspect of the invention, there is
provided a method for treating cancer, comprising the step of
administering a therapeutically effective amount of a combination
comprising: (i) a composition comprising small molecular weight
components of less than 3000 daltons, and having the following
properties: is extracted from bile of animals; is capable of
stimulating monocytes and/or macrophages in vitro and/or in vivo;
is capable of modulating tumor necrosis factor production and/or
release; contains no measurable level of IL-1.alpha., IL-1.beta.,
TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma; is not cytotoxic to
human peripheral blood mononuclear cells; is not an endotoxin; and
(ii) one or more anticancer agent(s), wherein said combination has
therapeutic synergy or improves the therapeutic index in the
treatment of cancer over the composition or the anticancer agent(s)
alone.
BRIEF DESCRIPTION OF THE DRAWINGS
[0031] Further details of the invention are described below with
the help of the examples illustrated in the accompanying drawings
in which:
[0032] FIG. 1 is a graph showing dose response of the composition
of the invention in stimulating peripheral blood monocyte
function.
[0033] FIG. 2 illustrates the growth of Human Pancreatic
Adenocarcinoma (BxPC-3) in CD-1 Nude Mice.
[0034] FIG. 3 illustrates the weight of Human Pancreatic
Adenocarcinoma (BxPC-3) in CD-1 Nude Mice.
[0035] FIG. 4 illustrates the growth of Human Pancreatic Carcinoma
(SU.86.86.) in CD-1 Nude Mice.
[0036] FIG. 5 illustrates the weight of Human Pancreatic Carcinoma
(SU.86.86.) in CD-1 Nude Mice.
[0037] FIG. 6 illustrates the growth of Human Melanoma(A2058) in
CD-1 Nude Mice.
[0038] FIG. 7 illustrates the weight of Human Melanoma(A2058) in
CD-1 Nude Mice.
[0039] FIG. 8 illustrates the growth of Human Melanoma(C8161) in
CD-1 Nude Mice.
[0040] FIG. 9 illustrates the weight of Human Melanoma(C8161) in
CD-1 Nude Mice.
[0041] FIG. 10 illustrates the growth of Human Breast
Adenocarcinoma(MDA-MB-231) in CD-1 Nude Mice.
[0042] FIG. 11 illustrates the weight of Human Breast
Adenocarcinoma (MDA-MB-231) in CD-1 Nude Mice.
[0043] FIG. 12 illustrates the growth of Human Breast
Adenocarcinoma (MDA-MB-231) in CD-1 Nude Mice.
[0044] FIG. 13 illustrates the weight of Human Breast
Adenocarcinoma (MDA-MB-231) in CD-1 Nude Mice.
[0045] FIG. 14 illustrates the growth of Human Prostate Carcinoma
(PC-3) in SCID Mice.
[0046] FIG. 15 illustrates the weight of Human Prostate Carcinoma
(PC-3) in SCID Mice.
[0047] FIG. 16 illustrates the growth of Human Pancreatic Carcinoma
(BxPC-3) in CD-1 Nude Mice.
[0048] FIG. 17 illustrates the weight of Human Pancreatic Carcinoma
(SU.86.86) in CD-1 Nude Mice.
[0049] FIG. 18 illustrates the growth of Human Prostate Carcinoma
(DU145) in SCID Mice.
[0050] FIG. 19 illustrates the weight of Human Prostate Carcinoma
(DU145) in SCID Mice.
[0051] FIG. 20 illustrates the growth of Human Ovary Adenocarcinoma
(SK-OV-3) in CD-1 Nude Mice.
[0052] FIG. 21 illustrates the growth of Human Ovary Adenocarcinoma
(SK-OV-3) in CD-1 Nude Mice.
[0053] FIG. 22 illustrates the growth of Human Lung Adenocarcinoma
(H460) in CD-1 Nude Mice.
[0054] FIG. 23 illustrates the weight of Human Lung Adenocarcinoma
(H460) in CD-1 Nude Mice.
[0055] FIG. 24 illustrates the growth of Human Small Cell Lung
Carcinoma (H209) in SCID Mice.
[0056] FIG. 25 illustrates the weight of Human Small Cell Lung
Carcinoma (H209) in SCID Mice.
DETAILED DESCRIPTION OF THE INVENTION
[0057] The present invention provides anticancer biological
response modifier (BD-BRM) combinations. The combination comprises
(i) a composition comprising small molecular weight components of
less than 3000 daltons, and having the following properties: is
extracted from bile of animals; is capable of stimulating monocytes
and/or macrophages in vitro and/or in vivo; is capable of
modulating tumor necrosis factor production and/or release;
contains no measurable level of IL-1.alpha., IL-1.beta., TNF, IL-6,
IL-8, IL-4, GM-CSF or IFN-gamma; is not cytotoxic to human
peripheral blood mononuclear cells; is not an endotoxin; and (ii)
one or more anticancer agent(s), wherein the BD-BRM combination has
therapeutic synergy or improves the therapeutic index in the
treatment of cancer over the composition or the anticancer agent(s)
alone. The present invention further provides the use of the
combination in the manufacture of a medicament or a pharmaceutical
kit and in the treatment of cancer.
[0058] Components of the Combination
[0059] BD-BRM Composition
[0060] Experimental evidence to date indicates that the unique
immunomodulatory properties of BD-BRM activity are associated with
low molecular weight material derived from bile. The BD-BRM
composition of the present invention comprises small molecular
weight components of less than 3000 daltons, and having at least
one of the following properties:
[0061] a) is extracted from the bile of animals;
[0062] b) is capable of stimulating or activating monocytes and/or
macrophages in vitro and/or in vivo;
[0063] c) is capable of modulating tumor necrosis factor production
and/or release;
[0064] d) contains no measurable level of IL-1.alpha., IL-1.beta.,
TNF, IL-6, IL-8, IL-4, GN-CSF or IFN-.gamma.;
[0065] e) shows no cytotoxicity to human peripheral blood
mononuclear cells or lymphocytes; and
[0066] f) is not an endotoxin.
[0067] As mentioned above, the production and characterization of
the BD-BRM composition has been described in preceding patent
applications, and is also summarized in Example 1. The composition
can be produced in a consistently reproducible form using the
method as generally described above with demonstrated identity,
potency and purity from batch to batch.. Identity and purity are
determined using reverse-phase high pressure liquid chromatography.
(See Example 1). The compositions have a consistently reproducible
pattern on reverse-phase HPLC. The composition may be used in a
concentrated form. The composition may also be lyophilized. The
composition may be used without further modification by simply
packaging it in vials and sterilizing
[0068] The compositions are also characterized by the properties
hereinbefore mentioned, for example their ability to stimulate
monocytes and macrophages in vitro and in vivo, etc. The
compositions activate PBMNs to release TNF in vitro as measured by
the Monocyte/Macrophage Activation Assay (TNF-Release) as described
in Example 2.
[0069] Anticancer Agents
[0070] This invention provides for a BD-BRM composition in
combination with one or more other anticancer agents. An
"anticancer agent" is any compound, composition or treatment that
prevents or delays the growth and/or metastasis of cancer cells.
Such anticancer agents include but are not limited to
chemotherapeutic drug treatment, radiation, gene therapy, hormonal
manipulation, immunotherapy and antisense oligonucleotide therapy.
It is to be understood that anticancer agents for use in the this
invention also include novel compounds or treatments developed in
the future that can be used to generate therapeutic combinations as
described herein.
[0071] Examples of candidate anti-cancer compounds that may be
useful in the combinations of this invention are: antisense
sequences, Drugs for Promyelocytic Leukemia: Tretinoin
(Vesanoid.RTM.); Drugs for Chronic Myeloid Leukemia: Low-dose
Interferon (IFN)-alpha; Drugs Used in Gastric Cancer: Antibiotics,
Antineoplastics; Acute Lymphoblastic Leukemia: Pegaspargase
(Oncaspar.RTM.), Rhone-poulenc Rorer, L-asparaginase, Il-2; Drugs
for Colon Cancer: Edatrexate or 10-ethyl-10deaza-aminopterin or
10-edam, 5-fluorouracil (5-FU) and Levamisole, Methyl-ccnu
(Methyl-chloroethyl-cyclohexyl-nitrosourea), Fluorodeoxyuridine
(Fudr), Vincristine; Drugs for Esophageal Cancer: Porfimer Sodium
(Photofrin.RTM.), Quadra Logic Technologies, or Treatment with a
Neodymium:yag (Nd:yag.RTM.) Laser; Drugs Used in Colorectal Cancer:
Irinotecan (Camptosar.RTM.), Pharmacia & Upjohn, Topotecan
(Hycamtin.RTM.), Loperamide (Imodium.RTM.), 5-fluorouracil (5-FU);
Drugs For Advanced Head and Neck Cancers: Docetaxel
(Taxotere.RTM.); Drugs for Non-hodgkin's Lymphoma: Rituximab,
Etoposide; Drugs for Non-small-cell Lung Cancer: A Vinca Alkaloid,
Vinorelbine Tartrate (Navelbine.RTM.), Wellcome, Paitaxel,
(Taxol.RTM.), Docetaxel (Taxotere.RTM.), Topotecan, Irinotecan,
Gemcitabine; Drugs for Ovarian Cancer: Docetaxel (Taxotere.RTM.),
Gemcitabine, (Gemzar.RTM.), Irinotecan (Camptosar.RTM.), Paclitaxel
(Taxol.RTM.), Topotecan (Hycamtin.RTM.), Amifostine (Ethyol.RTM.),
Us Bioscience (For Reducing the Cumulative Renal Toxicity
Associated with Repeated Cisplatin Therapy in Patients with
Advanced Ovarian.degree. Cancer); Drugs to Prevent Melanoma (Sun
Screens): 2-ethylhexyl-p-methoxy-cinnamate (2-ehmc),
Octyl-N-dimethyl-paminobenzoat- e (O-paba), Benzophenone-3 (Bp-3);
Drugs for Prostate Cancer: Flutamide (Eulexin.RTM.), Finasteride
(Proscar.RTM.), Terazosin (Hytrin.RTM.), Doxazosin (Cardura.RTM.),
Goserelin Acetate (Zoladex.RTM.), Liarozole, Nilutamide
(Nilandron.RTM.), Mitoxantrone (Novantrone.RTM.), Prednisone
(Deltasone.RTM.); Drugs for Pancreatic Cancer:
[0072] Gemcitabine (Gemzar.RTM.), 5-fluorouracil; Drugs for
Advanced Renal Cancer: Interleukin-2 (Proleukin.RTM.), Chiron
Corp.; Additional Anti-neoplastic Drugs: Porfimer Sodium, Axcan,
Dacarbazine, Faulding, Etoposide, Faulding, Procarbazine HCl,
Sigma-tau, Rituximab, Roche, Paclitaxel (Taxol.RTM.), Bristol-myers
Squibb, Trastuzumab (Herceptin.RTM.), Roche, Temozolomide
(Temodal.RTM.), Schering; Alkylating Agents Used in Combination
Therapy for Different Cancers: Cyclophosphamide, Cisplatin,
Melphalan.
[0073] Antisense Compounds
[0074] The specificity and sensitivity of antisense compounds makes
them useful in diagnostics, therapeutics, prophylaxis, as research
reagents and in kits. In the context of the present invention, the
terms "antisense compound" and "antisense oligonucleotide" each
refer to an oligomer or polymer of ribonucleic acid (RNA), or
deoxyribonucleic acid (DNA), or mimetics thereof. These terms also
include chimeric antisense compounds, which are antisense compounds
that contain two or more chemically distinct regions, each made up
of at least one monomer unit. In accordance with the present
invention, the terms "antisense compound" and "antisense
oligonucleotide" further include oligonucleotides composed of
naturally occurring nucleobases, sugars and covalent
internucleoside (backbone) linkages, as well as oligonucleotides
comprising non-naturally-occurring moieties that function
similarly. Such modified or substituted oligonucleotides are well
known to workers skilled in the art and often preferred over native
forms because of desirable properties such as, for example,
enhanced cellular uptake, enhanced affinity for nucleic acid target
and increased stability in the presence of nucleases. The antisense
compounds in accordance with the present invention comprise from
about 7 to about 50 nucleobases, or from about 7 to about 30.
Alternatively, the antisense compounds comprise a mixture of short
oligomers which will bind to the target nucleic acid in tandem
(i.e. they are complementary to sequences that are adjacent to one
another in the target nucleic acid).
[0075] Examples of antisense compounds useful in the present
invention include oligonucleotides containing modified backbones or
non-natural internucleoside linkages. In accordance with the
present invention, oligonucleotides having modified backbones
include those that retain a phosphorus atom in the backbone and
those that do not have a phosphorus atom in the backbone.
[0076] For the purposes of the present invention, and as sometimes
referenced in the art, modified oligonucleotides that do not have a
phosphorus atom in their intemucleoside backbone can also be
considered to be oligonucleosides.
[0077] The antisense compounds used in accordance with this
invention may be conveniently and routinely made through the
well-known technique of solid phase synthesis. Equipment for such
synthesis is sold by several vendors including, for example,
Applied Biosystems (Foster City, Calif.). Any other means for such
synthesis known in the art may be additionally or alternatively
employed. Similar techniques using phosphorothioates and alkylated
derivatives have been employed to produce oligonucleotides.
[0078] Antisense oligonucleotides have been successfully employed
as therapeutic moieties in the treatment of disease states such as
cancer. Antisense compounds exert their effects by specifically
modulating expression of a gene implicated in a specific disease
state. Thus, the present invention contemplates the therapeutic
administration of an effective amount of a combination of the
BD-BRM composition of the present invention and an appropriate
antisense compound to a mammal suspected of having a disease or
disorder which can be treated by specifically modulating gene
expression. The present invention further contemplates the
prophylactic use of a combination of the BD-BRM composition and an
antisense compound in the prevention of a cancer which is related
to over- or under-expression of a specific gene.
[0079] Pharmaceutical Compositions
[0080] The combinations of the present invention may be converted
using customary methods into pharmaceutical compositions. The
pharmaceutical composition contain the combination of the invention
either alone or together with other active or inactive substances.
Such pharmaceutical compositions can be for oral, topical, rectal,
parenteral, local, inhalant, or intracerebral use. They are
therefore in solid or semisolid form, for example pills, tablets,
creams, gelatin capsules, capsules, suppositories, soft gelatin
capsules, gels, membranes, and tubelets. For parenteral and
intracerebral uses, those forms for intramuscular or subcutaneous
administration can be used, or forms for infusion or intravenous or
intracerebral injection can be used, and can therefore be prepared
as solutions of the combinations or as powders of the combinations
to be mixed with one or more pharmaceutically acceptable excipients
or diluents, suitable for the aforesaid uses and with an osmolarity
that is compatible with the physiological fluids. For local use,
those preparations in the form of creams or ointments for topical
use or in the form of sprays may be considered; for inhalant uses,
preparations in the form of sprays, for example nose sprays, may be
considered. Preferably, the BD-BRM composition of the combination
is administered intramuscularly.
[0081] The pharmaceutical compositions can be prepared by per se
known methods for the preparation of pharmaceutically acceptable
compositions which can be administered to patients, and such that
an effective quantity of the active substance is combined in a
mixture with a pharmaceutically acceptable vehicle. Suitable
vehicles are described, for example, in Remington's Pharmaceutical
Sciences (Nack Publishing Company, Easton, Pa., USA 1985).
[0082] On this basis, the pharmaceutical compositions include,
albeit not exclusively, the combination of the invention in
association with one or more pharmaceutically acceptable vehicles
or diluents, and are contained in buffered solutions with a
suitable pH and iso-osmotic with the physiological fluids.
[0083] The compositions and agents of the invention are intended
for administration to humans or animals.
[0084] The dosage requirements of the pharmaceutical compositions
according to the present invention will vary with the particular
combinations employed, the route of administration and the
particular cancer and cancer patient being treated. Treatment will
generally be initiated with small dosages less than the optimum
dose of the compound. Thereafter the dosage is increased until the
optimum effect under the circumstances is reached. In general, the
pharmaceutical compositions according to the present invention are
most administered at a concentration that will generally afford
effective results without causing any harmful or deleterious side
effects. The compounds can be administered either as a single unit
dose, or if desired, the dosage can be divided into convenient
subunits administered at suitable times throughout the day. The
amount of the pharmaceutical composition that will be effective in
treatment can be determined by standard clinical techniques, known
to a worker skilled in the art [for example, see Remington's
Pharmaceutical Sciences, 18.sup.th Edition, Mack Publishing Co.,
Easton, Pa. (1990)].
[0085] Therapeutic Activity of the Combination
[0086] The combination of the present invention has a net
anticancer effect that is greater than the anticancer effect of the
individual components of the combination when administered alone.
The anticancer effect is increased without a concomitant increased
toxic effect. Without being limited by mechanism, by combining one
or more anticancer agents with a BD-BRM composition it is possible
to:
[0087] (i) increase the therapeutic effect of the anticancer
agent(s);
[0088] (ii) increase the therapeutic effect of the BD-BRM
composition;
[0089] (iii) decrease or delay the toxicity phenomena associated
with the anticancer agent(s); and/or
[0090] (iv) decrease or delay the toxicity phenomena associated
with the BD-BRM composition,
[0091] in comparison to treatment with the individual components of
the combination.
[0092] In one embodiment the combination of the present invention
provides an improved efficacy, over treatment using the components
of the combination alone, that may be demonstrated by determination
of the therapeutic synergy.
[0093] A combination manifests therapeutic synergy if it is
therapeutically superior to one or other of the constituents used
at its optimum dose [T. H. Corbett et al., (1982) Cancer Treatment
Reports, 66, 1187]. To demonstrate the efficacy of a combination,
it may be necessary to compare the maximum tolerated dose of the
combination with the maximum tolerated dose of each of the separate
constituents in the study in question. This efficacy may be
quantified using technique and equations commonly known to workers
skilled in the art. [T. H. Corbett et al., (1977) Cancer, 40,
2660.2680; F. M. Schabel et al., (1979) Cancer Drug Development,
Part B, Methods in Cancer Research, 17, 3-51, New York, Academic
Press Inc.].
[0094] The combination, used at its own maximum tolerated dose, in
which each of the constituents will be present at a dose generally
not exceeding its maximum tolerated dose, will manifest therapeutic
synergy when the efficacy of the combination is greater than the
efficacy of the best constituent when it is administered alone.
[0095] In another embodiment the combination of the present
invention improves the therapeutic index in the treatment of cancer
over that of the BD-BRM composition or the anticancer agent(s) when
administered to a patient alone.
[0096] A median effective dose (ED.sub.50) of a drug is the dose
required to produce a specified effect in 50% of the population.
Similarly, the median lethal dose (LD.sub.50) of a drug, as
determined in preclinical studies, is the dose that has a lethal
effect on 50% of experimental animals. The ratio of the LD.sub.50
to the ED.sub.50 can be used as an indication of the therapeutic
index. Alternatively the therapeutic index can be determined based
on doses that produce a therapeutic effect and doses that produce a
toxic effect (e.g. ED.sub.90 and LD.sub.10, respectively). During
clinical studies, the dose, or the concentration (e.g. solution,
blood, serum, plasma), of a drug required to produce toxic effects
can be compared to the concentration required for the therapeutic
effects in the population to evaluate the clinical therapeutic
index. Methods of clinical studies to evaluate the clinical
therapeutic index are well known to workers skilled in the art.
[0097] In one embodiment the combination of the present invention
provides an improved therapeutic index, in comparison to that of
the individual components of the combination when administered
alone, by decreasing the observed LD.sub.50 of at least one of the
one or more anticancer agents in the combination.
[0098] In a related embodiment the combination of the present
invention provides an improved therapeutic index, in comparison to
that of the individual components of the combination when
administered alone, by increasing the observed ED.sub.50 of at
least one of the one or more anticancer agents in the combination.
In a further embodiment the combination of the present invention
provides an improved therapeutic index, in comparison to that of
the individual components of the combination when administered
alone, by increasing the observed ED.sub.50 of the bile-derived
biological response modifier.
[0099] In another embodiment the efficacy of a combination
according to the present invention may also be characterized by
adding the actions of each constituent.
[0100] In order to prepare a combination according to the present
invention one first selects one or more candidate anticancer
agent(s) and measure its efficacy in a model of a cancer of
interest, as would be well understood by one skilled in the art.
The next step may be to perform a routine analysis to compare the
efficacy of the one or more anticancer agent(s) alone to the
efficacy of the one or more anticancer agent(s) in combination with
varying amounts of the BD-BRM composition. Successful candidates
for use in the combinations of the present invention will be those
that demonstrate a therapeutic synergy with the BD-BRM or that
improve the therapeutic index in comparison to the therapeutic
index of the candidate agent(s).
[0101] The efficacy of the combinations of the present invention
may be determined experimentally using standard techniques using
cancer models well known to workers skilled in the art. Such cancer
models allow the activity of combinations to be tested in vitro and
in vivo in relation to the cancer of interest. Exemplary methods of
testing activity are described in the Examples provided herein,
although, it should be understood that these methods are not
intended to limit the present invention.
[0102] One example of a method for studying the efficacy of the
combinations on solid tumors in vivo involves the use of subject
animals, generally mice, that are subcutaneously grafted
bilaterally with 30 to 60 mg of a tumor fragment on day 0. The
animals bearing tumors are mixed before being subjected to the
various treatments and controls. In the case of treatment of
advanced tumors, tumors are allowed to develop to the desired size,
animals having insufficiently developed tumors being eliminated.
The selected animals are distributed at random to undergo the
treatments and controls. Animals not bearing tumors may also be
subjected to the same treatments as the tumor-bearing animals in
order to be able to dissociate the toxic effect from the specific
effect on the tumor. Chemotherapy generally begins from 3 to 22
days after grafting, depending on the type of tumor, and the
animals are observed every day. The different animal groups are
weighed 3 or 4 times a week until the maximum weight loss is
attained, and the groups are then weighed at least once a week
until the end of the trial.
[0103] The tumors are measured 2 or 3 times a week until the tumor
reaches approximately 2 g, or until the animal dies if this occurs
before the tumor reaches 2 g. The animals are autopsied when
sacrificed. The antitumour activity is determined in accordance
with various recorded parameters.
[0104] For a study of the combinations on leukaemias, the animals
are grafted with a particular number of cells, and the antitumour
activity is determined by the increase in the survival time of the
treated mice relative to the controls.
[0105] Administration of the Combination
[0106] The uses and methods of the present invention comprise
administering to a subject in need thereof an effective amount of a
BD-BRM composition in combination with one or more anticancer
agents to a subject. As used herein, combination components are
said to be administered in combination when the two or more
components are administered simultaneously or are administered
independently in a fashion such that the components will act at the
same time.
[0107] Components administered independently can, for example, be
administered separately (in time) or concurrently. Separately in
time means at least minutes apart, and potentially hours, days or
weeks apart. The period of time elapsing between the administration
of the components of the combination of the invention can be
determined by a worker of skill in the art, and will be dependent
upon, for example, the age, health, and weight of the recipient,
nature of the combination treatment, side effects associated with
the administration of other component(s) of the combination,
frequency of administration(s), and the nature of the effect
desired. Components of the combinations of the invention may also
be administered independently with respect to location and, where
applicable, route of administration.
[0108] In another embodiment, an effective amount of a therapeutic
composition comprising a BD-BRM composition and one or more
anticancer agents, and a pharmaceutically acceptable carrier is
administered to a subject. The combination or the pharmaceutical
composition of the invention can be administered before during or
after other anticancer treatment(s), or treatments for other
diseases or conditions. For example a drug to treat adverse side
effects of the anticancer treatment(s) can be administered
concurrently with a combination of the invention or a
pharmaceutical composition of the invention.
[0109] As indicated above the components of the combination of the
present invention may be administered separately, concurrently, or
simultaneously. In the case of separate administration the BD-BRM
composition may be administered before, during or after
administration of the anticancer agent(s). Furthermore, it would be
readily apparent to a worker skilled in the art that the route of
administration of each component of the combination is selected in
order to maximize the therapeutic benefit of the component and it
is not necessary that each component be delivered via the same
route. The BD-BRM composition and/or the anticancer agent(s) of the
combination may be administered via a single dose or via continuous
perfusion.
[0110] The agents, compounds and compositions of this invention can
be utilised in vivo, ordinarily in mammals, such as humans, sheep,
horses, cattle, pigs, dogs, cats, rats and mice, or in vitro to
treat cancer or cancer cells.
[0111] Cancers
[0112] As used herein, "cancer" refers to all types of cancer or
neoplasm or malignant tumors found in mammals, including carcinomas
and sarcomas. Examples of cancers are cancer of the brain, breast,
cervix, colon, head and neck, kidney, lung, non-small cell lung,
melanoma, mesothelioma, ovary, sarcoma, stomach, uterus and
Medulloblastoma.
[0113] The term "leukemia" refers broadly to progressive, malignant
diseases of the blood-forming organs and is generally characterized
by a distorted proliferation and development of leukocytes and
their precursors in the blood and bone marrow. Leukemia is
generally clinically classified on the basis of (1) the duration
and character of the disease--acute or chronic; (2) the type of
cell involved; myeloid (myelogenous), lymphoid (lymphogenous), or
monocytic; and (3) the increase or non-increase in the number of
abnormal cells in the blood--leukemic or aleukemic (subleukemic).
Leukemia includes, for example, acute nonlymphocytic leukemia,
chronic lymphocytic leukemia, acute granulocytic leukemia, chronic
granulocytic leukemia, acute promyelocytic leukemia, adult T-cell
leukemia, aleukemic leukemia, a leukocythemic leukemia, basophylic
leukemia, blast cell leukemia, bovine leukemia, chronic myelocytic
leukemia, leukemia cutis, embryonal leukemia, eosinophilic
leukemia, Gross' leukemia, hairy-cell leukemia, hemoblastic
leukemia, hemocytoblastic leukemia, histiocytic leukemia, stem cell
leukemia, acute monocytic leukemia, leukopenic leukemia, lymphatic
leukemia, lymphoblastic leukemia, lymphocytic leukemia,
lymphogenous leukemia, lymphoid leukemia, lymphosarcoma cell
leukemia, mast cell leukemia, megakaryocytic leukemia,
micromyeloblastic leukemia, monocytic leukemia, myeloblastic
leukemia, myelocytic leukemia, myeloid granulocytic leukemia,
myelomonocytic leukemia, Naegeli leukemia, plasma cell leukemia,
plasmacytic leukemia, promyelocytic leukemia, Rieder cell leukemia,
Schilling's leukemia, stem cell leukemia, subleukemic leukemia, and
undifferentiated cell leukemia.
[0114] The term "sarcoma" generally refers to a tumor which is made
up of a substance like the embryonic connective tissue and is
generally composed of closely packed cells embedded in a fibrillar
or homogeneous substance. Sarcomas include chondrosarcoma,
fibrosarcoma, lymphosarcoma, melanosarcoma, myxosarcoma,
osteosarcoma, Abemethy's sarcoma, adipose sarcoma, liposarcoma,
alveolar soft part sarcoma, ameloblastic sarcoma, botryoid sarcoma,
chloroma sarcoma, chorio carcinoma, embryonal sarcoma, Wilms' tumor
sarcoma, endometrial sarcoma, stromal sarcoma, Ewing's sarcoma,
fascial sarcoma, fibroblastic sarcoma, giant cell sarcoma,
granulocytic sarcoma, Hodgkin's sarcoma, idiopathic multiple
pigmented hemorrhagic sarcoma, immunoblastic sarcoma of B cells,
lymphoma, immunoblastic sarcoma of T-cells, Jensen's sarcoma,
Kaposi's sarcoma, Kupffer cell sarcoma, angiosarcoma, leukosarcoma,
malignant mesenchymoma sarcoma, parosteal sarcoma, reticulocytic
sarcoma, Rous sarcoma, serocystic sarcoma, synovial sarcoma, and
telangiectaltic sarcoma.
[0115] The term "melanoma" is taken to mean a tumor arising from
the melanocytic system of the skin and other organs. Melanomas
include, for example, acral-lentiginous melanoma, amelanotic
melanoma, benign juvenile melanoma, Cloudman's melanoma, S91
melanoma, Harding-Passey melanoma, juvenile melanoma, lentigo
maligna melanoma, malignant melanoma, nodular melanoma subungal
melanoma, and superficial spreading melanoma.
[0116] The term "carcinoma" refers to a malignant new growth made
up of epithelial cells tending to infiltrate the surrounding
tissues and give rise to metastases. Exemplary carcinomas include,
for example, acinar carcinoma, acinous carcinoma, adenocystic
carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum,
carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell
carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid
carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma,
bronchiolar carcinoma, bronchogenic carcinoma, cerebriform
carcinoma, cholangiocellular carcinoma, chorionic carcinoma,
colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform
carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical
carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma
durum, embryonal carcinoma, encephaloid carcinoma, epiermoid
carcinoma, carcinoma epitheliale adenoides, exophytic carcinoma,
carcinoma ex ulcere, carcinoma fibrosum, gelatiniform carcinoma,
gelatinous carcinoma, giant cell carcinoma, carcinoma
gigantocellulare, glandular carcinoma, granulosa cell carcinoma,
hair-matrix carcinoma, hematoid carcinoma, hepatocellular
carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypemephroid
carcinoma, infantile embryonal carcinoma, carcinoma in situ,
intraepidermal carcinoma, intraepithelial carcinoma, Krompecher's
carcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma,
lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma,
lymphoepithelial carcinoma, carcinoma medullare, medullary
carcinoma, melanotic carcinoma, carcinoma molle, mucinous
carcinoma, carcinoma muciparum, carcinoma mucocellulare,
mucoepidermoid carcinoma, carcinoma mucosum, mucous carcinoma,
carcinoma myxomatodes, naspharyngeal carcinoma, oat cell carcinoma,
carcinoma ossificans, osteoid carcinoma, papillary carcinoma,
periportal carcinoma, preinvasive carcinoma, prickle cell
carcinoma, pultaceous carcinoma, renal cell carcinoma of kidney,
reserve cell carcinoma, carcinoma sarcomatodes, schneiderian
carcinoma, scirrhous carcinoma, carcinoma scroti, signet-ring cell
carcinoma, carcinoma simplex, small-cell carcinoma, solanoid
carcinoma, spheroidal cell carcinoma, spindle cell carcinoma,
carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma,
string carcinoma, carcinoma telangiectaticum, carcinoma
telangiectodes, transitional cell carcinoma, carcinoma tuberosum,
tuberous carcinoma, verrucous carcinoma, and carcinoma
villosum.
[0117] Additional cancers include, for example, Hodgkin's Disease,
Non-Hodgkin's Lymphoma, multiple myeloma, neuroblastoma, breast
cancer, ovarian cancer, lung cancer, rhabdomyosarcoma, primary
thrombocytosis, primary macroglobulinemia, small-cell lung tumors,
primary brain tumors, stomach cancer, colon cancer, malignant
pancreatic insulanoma, malignant carcinoid, urinary bladder cancer,
premalignant skin lesions, testicular cancer, lymphomas, thyroid
cancer, neuroblastoma, esophageal cancer, genitourinary tract
cancer, malignant hypercalcemia, cervical cancer, endometrial
cancer, adrenal cortical cancer, and prostate cancer.
[0118] Pharmaceufical Kits
[0119] The present invention additionally provides for therapeutic
kits containing (i) a dosage unit of a composition and a
pharmaceutically acceptable carrier wherein the composition
comprises small molecular weight components of less than 3000
daltons, and has the following properties: is extracted from bile
of animals; is capable of stimulating monocytes and/or macrophages
in vitro and/or in vivo; is capable of modulating tumor necrosis
factor production and/or release; contains no measurable level of
IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or
IFN-gamma; is not cytotoxic to human peripheral blood mononuclear
cells; is not an endotoxin; and (ii) dosage unit of one or more
chemotherapeutic drug(s) and a pharmaceutically acceptable carrier,
said (i) and (ii) being provided in amounts that are effective, in
combination, for selectively killing tumor or metastatic cells.
[0120] As used herein, a "dosage unit" is a pharmaceutical
composition or formulation comprising at least one active
ingredient and optionally one or more inactive ingredient(s). The
dosage unit can be unitary, such as a single pill or liquid,
containing all of the desired active ingredients and the inactive
ingredients necessary and desired for making a dosage suitable for
administration (e.g., tabletting compounds such as binders,
fillers, and the like); the dosage unit can consist of a number of
different dosage forms (e.g., pill(s) and/or liquid(s)) designed to
be taken simultaneously as a dosage unit.
[0121] The contents of the kit can be lyophilized and the kit can
additionally contain a suitable solvent for reconstitution of the
lyophilized components. Individual components of the kit would be
packaged in separate containers and, associated with such
containers, can be a notice in the form prescribed by a
governmental agency regulating the manufacture, use or sale of
pharmaceuticals or biological products, which notice reflects
approval by the agency of manufacture, use or sale for human
administration.
EXAMPLES
[0122] A worker skilled in the art can produce BD-BRM compositions,
and assay BD-BRM compositions for activities such as in vitro
and/or in vivo monocyte and/or macrophage stimulation, modulation
of tumor necrosis factor production and/or release, content of
IL-1.alpha., IL-1.beta., TNF, IL-6, IL-8, IL-4, GM-CSF or IFN-gamma
and endotoxin and cytotoxicity to human peripheral blood
mononuclear cells, using the methods described in International
Patent Application Serial No. PCT/CA94/00494, published Feb. 16,
1995 as WO 95/07089.
Example 1
In Vivo Evaluation Of Efficacy Of BD-BRM In The Treatment Of Human
Pancreatic Adenocarcinoma In Cd-1 Nude Mice
[0123] The mouse xenograft model of neoplasia was used in these
studies to demonstrate the effect of treatment with a BD-BRM
composition on tumor growth in mice. For comparison, separate
groups of mice were treated with saline (control), a conventional
chemotherapeutic drug or concurrently with a combination of a
BD-BRM composition and a chemotherapeutic drug.
[0124] A human carcinoma cell line was grown as monolayer culture
in Minimum essential medium (.alpha.-MEM) supplemented with 10%
fetal bovine serum (FBS), 0.1 mM non-essential amino acid, 1.0 mM
sodium pyruvate, 100 U/ml penicillin, 100 .mu.g/ml streptomycin and
0.25 .mu.g/ml amphotericin B and 2 mM L-alanyl-1-glutamine at
37.degree. C. in an atmosphere of 5% CO.sub.2 in air. The tumor
cells were routinely subcultured twice weekly by trypsin-EDTA
treatment. The cells were harvested from subconfluent
logarithmically growing culture by treatment with trypsin-EDTA and
counted for tumor inoculation. The cell lines used in the
experiments herein are listed hereafter, though any carcinoma cell
line capable of tumor formation upon inoculation could be used:
[0125] pancreatic adenocarcinoma (BxPC-3) (a gemcitabine-resistant
cell line)
[0126] melanoma (A2058)
[0127] melanoma(C8161)
[0128] breast adenocarcinoma(MDA-MB-231)
[0129] prostate carcinoma (PC-3)
[0130] ovary adenocarcinoma (SK-OV-3)
[0131] large cell lung adenocarcinoma (H460)
[0132] small cell lung carcinoma (H209).
[0133] Tumor Inoculation: An acclimation period of at least 7 days
was allowed between receipt of the immunocompromised animal and its
inoculation. Typically CD-1 or SCID mice were used. When the female
mice were 6-9 (most typically 6-7) weeks of age, each mouse was
subcutaneously injected in the right flank with 3-10 million human
carcinoma cells in 0.1 ml of PBS. Inoculated animals were divided
into equal sized treatment groups of 9-20 (typically about 10) mice
each and treated daily with saline (0.2 ml/mouse/day, i.p.), BD-BRM
(0.2 ml/mouse/day, i.p.), a chemotherapeutic drug, or concurrently
with BD-BRM (0.2 ml/mouse/day, i.p.) and a chemotherapeutic drug.
The drug doses used in the experiments herein are listed hereafter,
though any chemotherapeutic drug(s) or other anticancer agent(s)
could be used:
[0134] gemcitabine (100 mg/kg in 0.1 ml saline/mouse/3 day,
i.v.)
[0135] dacarbazine (DTIC) (80 mg/kg in 0.1 ml saline/mouse/day,
i.p.)
[0136] taxol (10 mg/kg/week, i.v.)
[0137] 5-fluorouracil
[0138] taxotere
[0139] cisplatin
[0140] mitoxanthrone (i.v.)
[0141] Tumour sizes were measured every other day in two dimensions
using a caliper, and the volume was expressed in mm.sup.3 using the
formula: V=0.5 a.times.b.sup.2, where a and b are the long and
short diameters of the tumor, respectively. Mean tumor volumes
calculated from each measurement were then plotted in a standard
graph to compare the anti-tumor efficacy of drug treatments to that
of control. A day after the last treatment, tumors were excised
from the animals and their weights were measured. The data are
displayed as a tumour growth curve, and a bar graph showing mean
tumor weights.
1 Mouse xenograft experiments with BD-BRM compositions and BD-BRM
combinations Figure Human Mouse combination # mice with total #
carcinoma cell line strain drug expt tumor regression 2, 3
pancreatic CD-1 gemcitabine -- BRM: 4 (of 9) 4, 5 pancreatic
SU.86.86 CD-1 gemcitabine gemcitabine 6, 7 melanoma A2058 CD-1
dacarbazine dacarbazine 8, 9 melanoma C8161 CD-1 -- dacarbazine
comb: 5 (of 10) 10, 11 breast MDA-MB-231 CD-1 Taxol Taxol 12, 13
breast MDA-MB-231 CD-1 Taxol Taxol BRM: 2; comb: 5 (of 10) 14, 15
prostate PC-3 SCID mitoxantrone -- 16 pancreatic BxPC-3 CD-1
5-fluorouracil 5-fluorouracil comb: (5 of 10) 17 pancreatic
SU.86.86 CD-1 5-fluorouracil 5-fluorouracil 18, 19 prostate DU145
SCID mitoxantrone -- 20 ovarian SK-OV-3 CD-1 cisplatin cisplatin 21
ovarian SK-OV-3 CD-1 taxol taxol lung, large 22, 23 cell H460 CD-1
taxotere taxotere 24, 25 lung, small H209 SCID -- --
[0142] The results of the mouse xenograft experiments outlined in
the table above are shown in FIGS. 2-25. BD-BRM treatments always
resulted in significant delay of tumor growth compared to saline
control. Where a chemotherapeutic drug treatment group was
included, the delay in tumor growth achieved with BD-BRM was
typically superior to the inhibitory effects observed with the
chemotherapeutic drug. As indicated in the above table, total
regression of the tumor was also observed in some of the animals,
when the animals were treated with a BRM composition alone or with
a combination of the BD-BRM composition and a chemotherapeutic drug
was used. In the remaining animals treated with a combination,
significantly enhanced antitumor effects were observed.
[0143] The efficacy of the combinations of the invention can also
be determined experimentally using other protocols to study animal
models grafted with cancerous cells. The animals subjected to the
experiment, can be grafted with a tumor fragment, and the graft may
be placed subcutaneously. In the case of the treatment of advanced
tumors, tumors are allowed to develop to the desired size, animals
having insufficiently developed tumors being eliminated. Animals
not bearing tumors may also be subjected to the same treatments as
the tumor-bearing animals in order to b able to dissociate the
toxic effect from the specific effect on the tumor. Treatment
generally begins 3 days to 4 weeks after grafting, depending on the
type of tumor, and the animal are observed and animal weight change
recorded, and the tumors measured regularly, for example daily, or
2 or 3 times per week until the tumor reaches a defined size (e.g.
2 g in a mouse), or until the animal dies if this occurs before hte
tumor reaches 2 g. The animals are autopsied when sacrificed. To
study leukemia, cancerous cells can be injected intravenously.
Antitumor activity is determined by the increase in the survival
time of the treated animals relative to the controls. The efficacy
of the treatment with the combination of the invention is assessed
in terms of changes in the mean survival time of the animal.
Alternative methods of assessing efficacy, and therapeutic synergy,
can also be used.
[0144] These animal models are recognized in the art to be
predictive tests for anticancer effects in humans.
[0145] From the foregoing, it will be appreciated that, although
specific embodiments of the invention have been described herein
for purposes of illustration, various modifications may be made
without deviating from the spirit and scope of the invention.
Accordingly, the invention is not limited except as by the appended
claims.
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