U.S. patent application number 15/591922 was filed with the patent office on 2017-08-24 for tumor therapy with an antibody for vascular endothelial growth factor and an antibody for human epithelial growth factor receptor type 2.
This patent application is currently assigned to Hoffmann-La Roche Inc.. The applicant listed for this patent is Hoffmann-La Roche Inc.. Invention is credited to Thomas Friess, Max Hasmann, Werner Scheuer.
Application Number | 20170239353 15/591922 |
Document ID | / |
Family ID | 37998279 |
Filed Date | 2017-08-24 |
United States Patent
Application |
20170239353 |
Kind Code |
A1 |
Friess; Thomas ; et
al. |
August 24, 2017 |
TUMOR THERAPY WITH AN ANTIBODY FOR VASCULAR ENDOTHELIAL GROWTH
FACTOR AND AN ANTIBODY FOR HUMAN EPITHELIAL GROWTH FACTOR RECEPTOR
TYPE 2
Abstract
The present invention provides a method of treating a breast
cancer disease in a patient who has failed prior treatment with an
anti-VEGF antibody, comprising administering to the patient a
therapeutically effective amount of an anti-HER2 antibody while
continuing said anti-VEGF antibody therapy. The invention also
provides corresponding pharmaceutical kits and pharmaceutical
compositions.
Inventors: |
Friess; Thomas;
(Diessen-Dettenhofen, DE) ; Hasmann; Max;
(Muenchen, DE) ; Scheuer; Werner; (Penzberg,
DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Hoffmann-La Roche Inc. |
Little Falls |
NJ |
US |
|
|
Assignee: |
Hoffmann-La Roche Inc.
Little Falls
NJ
|
Family ID: |
37998279 |
Appl. No.: |
15/591922 |
Filed: |
May 10, 2017 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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14745026 |
Jun 19, 2015 |
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15591922 |
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12947264 |
Nov 16, 2010 |
9090700 |
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14745026 |
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11725777 |
Mar 20, 2007 |
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12947264 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 35/04 20180101;
A61K 2039/507 20130101; C07K 16/3015 20130101; C07K 2317/24
20130101; A61P 35/02 20180101; A61K 39/39558 20130101; C07K 16/32
20130101; A61P 35/00 20180101; A61K 39/3955 20130101; C07K 16/22
20130101 |
International
Class: |
A61K 39/395 20060101
A61K039/395; C07K 16/22 20060101 C07K016/22; C07K 16/32 20060101
C07K016/32 |
Foreign Application Data
Date |
Code |
Application Number |
Mar 22, 2006 |
EP |
06111523.4 |
Oct 18, 2006 |
EP |
06021815.3 |
Claims
1. A combined therapy method of treating a breast cancer disease,
comprising administering to the patient a therapeutically effective
amount of an anti-HER2 antibody and an anti-VEGF antibody wherein
the breast cancer disease is characterized by an overexpression of
the HER2 receptor protein.
2. A method of treating a breast cancer disease in a patient who
has failed prior therapy with an anti-VEGF antibody, comprising
administering to the patient a therapeutically effective amount of
an anti-HER2 antibody and an anti-VEGF antibody.
3. The method of claim 2 wherein the anti-VEGF antibody is
bevacizumab.
4. The method of claim 2 wherein the patient is human.
5. The method of claim 2 wherein the anti-HER2 antibody is
trastuzumab.
6. The method of claim 2 wherein the breast cancer disease is
characterized by an overexpression of the HER2 receptor
protein.
7. A pharmaceutical kit comprising a pharmaceutical composition
comprising an anti-VEGF antibody, a pharmaceutical composition
comprising an anti-HE2 antibody, and a package insert instructing
the user of said compositions to administer to a patient having
breast cancer disease, who has failed prior therapy with an
anti-VEGF antibody, said anti-VEGF antibody pharmaceutical
composition and said anti-HER2 antibody pharmaceutical composition,
wherein the anti-VEGF antibody pharmaceutical composition and the
anti-HER2 antibody pharmaceutical composition are packaged either
in a single container or in two separate containers.
8. The pharmaceutical kit of claim 7 wherein the anti-VEGF antibody
is bevacizumab.
9. The pharmaceutical kit of claim 7 wherein the anti-HER2 antibody
is trastuzumab.
10. A pharmaceutical composition comprising an anti-HER2 antibody
and an anti-VEGF antibody useful in the treatment of a breast
cancer disease in a patient which has failed prior therapy with an
anti-VEGF antibody.
11. The pharmaceutical composition of claim 10, wherein the
anti-VEGF antibody is bevacizumab and the anti-HER2 antibody is
trastuzumab.
Description
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a continuation of U.S. patent
application Ser. No. 14/745,026, filed Jun. 19, 2015, which is a
continuation of U.S. patent application Ser. No. 12/947,264, filed
Nov. 16, 2010 (now U.S. Pat. No. 9,090,700), which is a
continuation of U.S. patent application Ser. No. 11/725,777, filed
Mar. 20, 2007 (now abandoned), which claims the benefit of European
Patent Application No. EP 06111523.4, filed Mar. 22, 2006 and
European Patent Application No. EP 06021815.3, filed Oct. 18, 2006,
all of which are hereby incorporated by reference in their
entirety.
FIELD OF THE INVENTION
[0002] The present invention is directed to combined therapy with
anti-HER2 and anti-VEGF antibodies. In particular, the invention
concerns the use of such antibodies to treat breast cancer disease
in a patient who has failed prior breast cancer treatment with an
anti-VEGF antibody.
BACKGROUND OF THE INVENTION
[0003] Angiogenesis is implicated in the pathogenesis of a variety
of disorders which include solid tumors, intraocular neovascular
syndromes such as proliferative retinopathies or age-related
macular degeneration (AMD), rheumatoid arthritis, and psoriasis
(Folkman, J., et al., J. Biol. Chem. 267 (1992) 10931-10934;
Klagsbrun, M., et al., Annu. Rev. Physiol. 53 (1991) 217-239; and
Garner, A, Vascular diseases, In: Pathobiology of ocular disease, A
dynamic approach, (eds.) Garner and A, Klintworth, G K, 2nd Edition
Marcel Dekker, New York, (1994), pp 1625-1710). In the case of
solid tumors, the neovascularization allows the tumor cells to
acquire a growth advantage and proliferative autonomy compared to
the normal cells. Accordingly, a correlation has been observed
between density of microvessels in tumor sections and patient
survival in breast cancer as well as in several other tumors
(Weidner, N., et al., N. Engl. J. Med. 324 (1991) 1-6; Horak, E.
R., et al., Lancet 340 (1992) 1120-1124; and Macchiarini, P., et
al., Lancet 340 (1992) 145-146).
[0004] Vascular endothelial growth factor (VEGF) is involved in the
regulation of normal and abnormal angiogenesis and
neovascularization associated with tumors and intraocular disorders
(Ferrara, N., et al., Endocr. Rev. 18 (1997) 4-25; Berkman, R. A.,
et al., J. Clin. Invest. 91 (1993) 153-159; Brown, L. F., et al.,
Human Pathol. 26 (1995) 86-91; Brown, L. F., et al., Cancer Res. 53
(1993) 4727-4735; Mattern, J., et al., Brit. J. Cancer 73 (1996)
931-934; and Dvorak, H. F., et al., Am. J. Pathol. 146 (1995)
1029-1039). Anti-VEGF neutralizing antibodies suppress the growth
of a variety of human tumor cell lines in mice (Kim, K. J., et al.,
Nature 362 (1993) 841-844; Warren, R. S., et al., J. Clin. Invest.
95 (1995) 1789-1797; Borgstrom, P., et al., Cancer Res. 56 (1996)
4032-4039; and Melnyk, O., et al., Cancer Res. 56 (1996) 921-924).
WO 94/10202, WO 98/45332, WO 2005/00900 and WO00/35956 refer to
antibodies against VEGF. Humanized monoclonal antibody bevacizumab
(sold under the tradename Avastin.RTM.) is an anti-VEGF antibody
used in tumor therapy and is the only anti-angiogenic agent
approved for treatment of cancer (WO 98/45331).
[0005] HER2 is a member of the human epidermal growth factor
receptor family and possesses protein kinase activity in its
cytoplasmic domain. HER2 is over-expressed in tumor cells and is
correlated with poor prognosis and survival. HER2 is therefore a
valuable target of breast cancer therapy. Antibodies against HER2
are known from Takai, N., et al., Cancer 104 (2005) 2701-2708;
Yeon, C. H., et al., Invest. New Drugs 23 (2005) 391-409; Wong, W.
M., et al., Cancer Pract. 7 (1999) 48-50; Albanell, J., et al.,
Drugs Today (Barc). 35 (1999) 931-46.
[0006] Trastuzumab (sold under the tradename Herceptin.RTM.is a
recombinant humanized anti-HER2 monoclonal antibody used for the
treatment of HER2 over-expressed/HER2 gene amplified metastatic
breast cancer. Preclinical studies demonstrated that the antibody
has anti-tumor activity in vivo and in vitro. Moreover, additive or
synergistic enhancement of anti-tumor activity of trastuzumab was
observed in combination with various anti-tumor agents in mouse
models. In clinical studies, extension of survival was observed in
HER2 overexpressing metastatic breast cancer patients.
[0007] According to WO 98/45331, the effectiveness of an anti-VEGF
antibody in preventing or treating disease may be improved by
administering the antibody serially or in combination with another
agent that is effective for those purposes, such as an antibody
capable of binding to HER2 receptor. WO 2005/012531 describes
antibodies that may be combined with an anti-VEGF antibody (e.g.
Avastin.RTM.) and/or anti-ErbB antibodies (e.g. Herceptin.RTM.) in
the treatment of colorectal cancer, metastatic breast cancer and
kidney cancer. According to WO 2005/063816, anti-VEGF antibodies
may be combined with anti-ErbB antibodies in a treatment of
metastatic breast cancer. WO 2005/00090 and WO 2003/077841 also
disclose the combination of anti-VEGF antibodies with anti-ErbB2
antibodies for tumor therapy.
[0008] Clinical oncologists are in agreement that the failure of
cancer treatment is not necessarily caused by the growth of the
primary tumor, which is generally dealt with using surgery, but
rather by the metastatic spread into different organs. The
regression of primary tumors by different cytotoxic drugs is not
always indicative for anti-metastatic activity per se. On the
contrary, enhanced metastasis has been observed in response to
several anti-cancer drugs (Geldof, A. A., et al., Anticancer Res. 8
(1988) 1335-1339; Murphy, S. B., J. Clin. Oncol. 11 (1993) 199-201;
and De Larco, J. E., et al., Cancer Res. 61 (2001) 2857-2861).
Clearly there exists a need to develop treatment therapies that
target not only the primary tumor, but also suppress
metastasis.
SUMMARY OF THE INVENTION
[0009] The invention comprises the use of an anti-HER2 antibody and
an anti-VEGF antibody for the manufacture of a medicament for
treating a breast cancer disease in a patient who has failed prior
cancer therapy with an anti-VEGF antibody, comprising administering
to the patient a therapeutically effective amount of an anti-HER2
antibody and an anti-VEGF antibody.
[0010] In a preferred embodiment, the invention comprises the use
of trastuzumab and bevacizumab for the manufacture of a medicament
for treating a breast cancer disease characterized by an
overexpression of the HER2 receptor protein in a patient who has
failed prior therapy with an anti-VEGF antibody such as
bevacizumab, comprising administering to the patient a
therapeutically effective amount of trastuzumab and
bevacizumab.
[0011] The invention further comprises a method of treating a
breast cancer disease in a patient who has failed prior therapy
with an anti-VEGF antibody, comprising administering to the patient
a therapeutically effective amount of an anti-HER2 antibody while
continuing said anti-VEGF antibody therapy.
[0012] The invention further comprises a method of treating a
breast cancer disease, in a patient who has failed prior therapy
with an anti-VEGF antibody, comprising administering to the patient
a therapeutically effective amount of trastuzumab while continuing
bevacizumab therapy, wherein the breast cancer disease is
characterized by an overexpression of the HER2 receptor
protein.
[0013] The invention further comprises a method for increasing the
duration of survival of a patient having breast cancer disease who
has failed prior therapy with an anti-VEGF antibody, comprising
administering to the patient effective amounts of an anti-VEGF
antibody and an anti-HER2 antibody, whereby the co-administration
of the anti-VEGF antibody and the anti-HER2 antibody effectively
increases the duration of survival.
[0014] The invention further comprises a method for increasing the
progression free survival of a patient having breast cancer disease
who has failed prior therapy with an anti-VEGF antibody, comprising
administering to the patient effective amounts of an anti-VEGF
antibody and an anti-HER2 antibody, whereby the co-administration
of the anti-VEGF antibody and the anti-HER2 antibody effectively
increases the duration of progression free survival.
[0015] The invention further comprises a method for treating a
group of patients, having breast cancer disease and having failed
prior therapy with an anti-VEGF antibody, comprising administering
to the patient effective amounts of an anti-VEGF antibody and an
anti-HER2 antibody, whereby the co-administration of the anti-VEGF
antibody and the anti-HER2 antibody effectively increases the
response rate in the group of patients.
[0016] The invention further comprises a method for increasing the
duration of response of a patient having breast cancer disease who
has failed prior therapy with an anti-VEGF antibody, comprising
administering to the patient effective amounts of an anti-VEGF
antibody and an anti-HER2 antibody, whereby the co-administration
of the anti-VEGF antibody and the anti-HER2 antibody effectively
increases the duration of response.
[0017] The invention further comprises a method of treating a
patient having breast cancer disease who has failed prior therapy
with an anti-VEGF antibody, comprising administering to the patient
effective amounts of an anti-VEGF antibody and an anti-HER2
antibody, whereby the co-administration of the anti-VEGF antibody
and the anti-HER2 antibody results in statistically significant and
clinically meaningful improvement of the treated patient as
measured by the duration of survival, progression free survival,
response rate or duration of response.
[0018] This invention further comprises a method for reducing
metastasis in a patient having breast cancer disease who has failed
prior therapy with an anti-VEGF antibody, comprising administering
to the patient effective amounts of an anti-VEGF antibody and an
anti-HER2 antibody, whereby the co-administration of the anti-VEGF
antibody and the anti-HER2 antibody effectively reduces
metastasis.
[0019] The invention further comprises a method for treating a
group of patients, having breast cancer disease and having failed
prior therapy, with an anti-VEGF antibody, comprising administering
to the patient effective amounts of an anti-VEGF antibody and an
anti-HER2 antibody, whereby the co-administration of the anti-VEGF
antibody and the anti-HER2 antibody effectively reduces metastasis
in the group of patients.
[0020] The invention provides an article of manufacture (e.g.,
pharmaceutical kit) comprising one or more containers, and
preferably at least two containers, a pharmaceutical composition
within a first container comprising an anti-VEGF antibody, a
pharmaceutical composition within a second container comprising an
anti-HER2 antibody and a package insert instructing the user of the
composition to administer to a patient, having breast cancer
disease who has failed prior therapy with an anti-VEGF antibody,
the anti-VEGF antibody within said first container and an anti-HER2
antibody within said second container.
[0021] The invention further provides for a pharmaceutical
composition comprising an anti-HER2 antibody and an anti-VEGF
antibody useful in the treatment of breast cancer disease in a
patient which has failed prior therapy with an anti-VEGF antibody.
Preferably the anti-HER2 antibody is trastuzumab. Also preferably
the anti-VEGF antibody is bevacizumab.
BRIEF DESCRIPTION OF THE DRAWINGS
[0022] FIG. 1 Antitumor activity of combined trastuzumab and
bevacizumab treatment on tumor growth after bevacizumab treatment
failure. Mean values of tumor volume (mm.sup.3) plotted on the
y-axis; number of days after injection of tumor cells plotted on
the x-axis. Vehicle (circles), trastuzumab at loading dose of 30
mg/kg and maintenance dose of 15 mg/kg (squares), bevacizumab at 5
mg/kg until day 55 when treatment also includes trastuzumab at 15
mg/kg (triangles).
[0023] FIG. 2 Effect of combined trastuzumab and bevacizumab
treatment on lung metastasis. Mean value of human Alu DNA sequence
(ng/ml) quantitated from lung tissue using real-time PCR and
plotted on the y-axis.
DETAILED DESCRIPTION OF THE INVENTION
[0024] All references cited herein are hereby incorporated by
reference in their entirety.
Definitions
[0025] The term "VEGF" according to the invention refers to the
vascular endothelial cell growth factor (Swiss-Prot No. P 15692),
alternative splicing forms (see e.g. Leung, D. W., et al., Science,
246 (1989) 1306-1309; and Houck, K. A., et al., Mol. Endocrin. 5
(1991) 1806-1814) and active fragments, preferably N-terminal
fragments thereof.
[0026] The term "anti-VEGF antibody" according to the invention is
an antibody that binds specifically to VEGF. The preferred
humanized anti-VEGF antibody or variant anti-VEGF antibody herein
binds human VEGF with a Kd value of no more than about
1.times.10.sup.8M and preferably no more than about
5.times.10.sup.9M. Preferably the anti-VEGF antibody is a
monoclonal antibody that binds to the same epitope as recombinant
humanized anti-VEGF monoclonal antibody generated according to
Presta, L. G., et al., Cancer Res. 57 (1997) 4593-4599. A preferred
antibody is bevacizumab. Anti-VEGF antibodies and methods for their
manufacture are e.g. described in U.S. Pat. No. 6,054,297, US
2003/0190317, U.S. Pat. No. 6,632,926, U.S. Pat. No. 6,884,879, and
US 2005/0112126.
[0027] Bevacizumab comprises mutated human IgG1 framework regions
and antigen-binding complementarity-determining regions from a
murine anti-hVEGF monoclonal antibody that blocks binding of human
VEGF to its receptors. Approximately 93% of the amino acid sequence
of bevacizumab, including most of the framework regions, is derived
from human IgG1, and about 7% of the sequence is derived from the
murine antibody A4.6.1. Bevacizumab has a molecular mass of about
149,000 Daltons and is glycosylated. Bevacizumab and its method of
preparation are described in EP 1 325 932.
[0028] HER2 is a 185-kDa growth factor receptor also referred to as
neu and c-erbB-2 (Slamon, D. J., et al., Science 235 (1987)
177-182; Swiss-Prot P04626) whose function is related to neoplastic
transformation in human breast cancer cells. Overexpression of this
protein has been identified in 20-30% of breast cancer patients
where it correlates with regionally advanced disease, increased
probability of tumor recurrence, and reduced patient survival. As
many as 30-40% of patients having gastric, endometrial, salivary
gland, non-small cell lung, pancreatic, ovarian, peritoneal,
prostate, or colorectal cancers may also exhibit overexpression of
this protein. Anti-HER2 antibodies and methods for their
manufacture are e.g. described in U.S. Pat. No. 6,054,297, WO
89/06692, U.S. Pat. No. 6,953,842, U.S. Pat. No. 6,949,245, U.S.
Pat. No. 6,399,063, U.S. Pat. No. 6,165,464, U.S. Pat. No.
6,054,297, U.S. Pat. No. 5,772,997, WO 2003/087131, WO 01/00245, WO
01/00238, WO 00/69460, WO 00/52054, WO 99/31140 and WO 98/17797. In
a preferred embodiment of the invention, the anti-HER2 antibody is
trastuzumab. Trastuzumab and its method of preparation are
described in EP 0 590 058.
[0029] The term "overexpression" of the HER2 receptor protein is
intended to indicate an abnormal level of expression of the HER2
receptor protein in a cell from a tumor within a specific tissue or
organ of the patient relative to the level of expression in a
normal cell from that tissue or organ. Patients having a cancer
characterized by overexpression of the HER2 receptor can be
determined by standard assays known in the art. Preferably
overexpression is measured in fixed cells of frozen or
paraffin-embedded tissue sections using immunohistochemical (IHC)
detection. When coupled with histological staining, localization of
the targeted protein can be determined and extent of its expression
within a tumor can be measured both qualitatively and
semi-quantitatively. Such IHC detection assays are known in the art
and include the Clinical Trial Assay (CTA), the commercially
available LabCorp 4D5 test, and the commercially available DAKO
HercepTest.RTM. (DAKO, Carpinteria, Calif.). The latter assay uses
a specific range of 0 to 330 cell staining (0 being normal
expression, 3+ indicating the strongest positive expression) to
identify cancers having overexpression of the HER2 protein (see the
Herceptinitrastuzumab) full prescribing information, September
1998, Genentech Inc., San Francisco, Calif.). Thus, patients having
a cancer characterized by overexpression of the HER2 protein in the
range of 1+, 2+, or 3+, preferably 2+ or 3+, more preferably 3+
would benefit from the methods of therapy of the present
invention.
[0030] The term "breast cancer disease" refers to the uncontrolled
growth of abnormal breast cells. It includes ductal carcinoma in
situ, invasive ductal carcinoma, lobular carcinoma in situ,
invasive lobular carcinoma, medullary carcinoma, Paget's disease of
the nipple and metastatic breast cancer, as well as other cancer
diseases of the breast as known to one of ordinary skill in the
art.
[0031] The term "failed prior therapy with an anti-VEGF antibody"
or "treatment failure" as used herein refers to tumor patients who
failed to respond to previous therapy with an anti-VEGF antibody
("non-responders") or who initially responded to previous therapy,
but in whom the therapeutic response was not maintained (referred
to as "relapsers"). Preferably the term "failed prior therapy with
an anti-VEGF antibody" refers to relapsers. Treatment failure
(respectively Response (RE) and Non-Response (NR)) is established
based on the medical judgment of a practitioner ascertained by the
results from clinical and laboratory data that are generally known
in the art to assess patient treatment. Such data may be obtained,
by way of example, from clinical examination, cytological and
histological techniques, endoscopy and laparoscopy, ultrasound, CT,
PET and MRI scans, chest X-ray and mammography, and measuring the
concentration of tumor markers, such as CEA, Cyfra, CA15-3,
interleukin 8 and soluble HER2. In this context "treatment failure"
is defined as the absence of clinical improvement. Alternatively,
RECIST criteria may be used to determine tumor response (Therasse,
P., et al., J. Nat. Cancer Institute 92 (2000) 205-216) In this
context "treatment failure" is defined as either "incomplete
response/stable disease" or "progressive disease".
[0032] According to these RECIST criteria, tumor response for solid
tumors (Therasse, P., et al., J. Nat. Cancer Institute 92 (2000)
205-216) is categorized in dependency of the volume progression or
regression of the tumors (e.g. measured via CT) into four levels:
complete response (CR) or partial response (PR), stable disease
(SD) and progressive disease (PD) (see Table 1). Furthermore the
European Organization for Research and Treatment of Cancer (EORTC)
proposed a categorization into four levels in dependency of the
metabolism of the tumors measured via
2-[.sup.18F]1-Fluoro-2-deoxyglucose positron emission tomography
(FDG-PET) (Young H., et al., Eur. J. Cancer 35 (1999) 1773-1782 and
Kellof, G. J., et al., Clin. Cancer Res. 11 (2005) 2785-2808):
complete metabolic response (CMR) or partial metabolic response
(PMR), stable metabolic disease (SMD) and progressive metabolic
disease (PMD) (see Table 2).
TABLE-US-00001 TABLE 1 CT-Criteria (acc. to RECIST) CT-measurement:
RECIST Change in sums longest diameters Disappearance; CR conformed
at 4 weeks (after treatment start) 30% decrease; PR confirmed at 4
weeks Neither PR nor PD SD criteria met 20% increase, no CR, PD PR,
SD documented before increased disease
TABLE-US-00002 TABLE 2 Proposed FDG-PET criteria (acc. to EORTC,
see Young H., et al., Eur J Canc 35 (1999) 1773-1782)
PET-measurement Proposed FDG- PET criteria Complete resolution of
2- CMR [.sup.18F]-Fluoro-2-deoxy- glucose (FDG) tumor uptake
Reduction of a minimum PMR of 15-25% of standardized uptake value
(SUV) after one treatment cycle, and of >25% after more than one
treatment cycle Increase of standardized SMD uptake value (SUV)
<25% or decrease of SUV <15% No visible increase the extent
of FDG tumor uptake Increase of SUV >25% PMD Visible increase of
FDG tumor uptake (>20% of longest dimension) Appearance of new
FDG uptake in metastatic lesions
[0033] Thus, preferably, "Response (RE)" and "Non-Response (NR)"
according to this invention are established based on data acquired
by the combination of computer tomography (CT) and
2-.sup.18F1-Fluoro-2-deoxyglucose positron emission tomography
(FDG-PET) (Kellof, G. J., et al., Clin. Cancer Res. 11 (2005)
2785-2808, and Young H., et al., Eur. J. Canc. 35 (1999) 1773-1782)
using both the RECIST and FDG-PET criteria described above.
Accordingly Response (RE) and Non-Response (NR) according to this
invention are determined preferably as follows:
Response (RE): CR or PR is established via CT-RECIST criteria
(Table 1) and at the same time CMR or PMR is established via
FDG-PET (Table 2). Thus Response (RE) means one of the following
four cases for combined CT and PET measurement: CR and CMR, PR and
PMR, CR and PMR, and PR and CMR.
[0034] Non-Response (NR): SD or PD is established via CT-RECIST
criteria (Table 1) and at the same time SMD or PMD is established
via FDG-PET (Table 2). Thus the following four cases for combined
CT and PET measurement signify Non-Response (NR): SD and SMD, SD
and PMD, PD and SMD, and PD and PMD.
[0035] Usually the response is determined at around 3 to 8 weeks,
preferably at around 6 weeks, after treatment start. This response
determination is usually repeated at intervals of 4 to 8 weeks,
preferably of 6 to 8 weeks. When at the first determination a
significant response (RE) was identified, then a relapse (that
means a Non-Response (RE) after the first determination) can be
determined at earliest at the second response determination.
[0036] In this context, the term "patient who has failed prior
therapy with an anti-VEGF antibody" refers to a patient, in whom
either at the first response determination Non-Response (NR) is
established ("Non-Responder") or at the first response
determination Response (RE) is established, and in the second or a
subsequent response determination Non-Response (NR) is established
("Relapser").
[0037] The term "metastasis" according to the invention refers to
the transmission of cancerous cells from the primary tumor to one
or more sites elsewhere in a patient causing secondary tumors. A
tumor formed by cells that have spread is called a "metastatic
tumor" or a "metastasis". The metastatic tumor contains cells that
are like those in the original (primary) tumor. Means to determine
if a cancer has metastasized are known in the art and include tumor
marker tests, bone scan, chest X-ray, computed tomography (CT),
computerized axial tomography (CAT), molecular resonance imaging
(MRI), positron emission tomography (PET), single photon emission
computed tomography (SPECT), fluorescence imaging (FI), and
bioluminescent imaging (BLI) and tumor marker tests (see e.g.
Helms, M. W., et al., Contributions to microbiology 13 (2006)
209-231, and Pantel, K., et al., J. Nat. Cancer Inst. 91 (1999)
1113-1124).
[0038] As used herein, the term "patient" preferably refers to a
human in need of treatment to treat cancer, or a precancerous
condition or lesion. However, the term "patient" can also refer to
non-human animals, preferably mammals such as dogs, cats, horses,
cows, pigs, sheep and non-human primates, among others, that are in
need of treatment.
[0039] The term "group" refers to a group of patients as well as a
sub-group of patients.
[0040] The term "package insert" refers to instructions customarily
included in commercial packages of therapeutic products, which may
include information about the indications, usage, dosage,
administration, contraindications and/or warnings concerning the
use of such therapeutic products.
[0041] The cancer may be, for example, lung cancer, non small cell
lung (NSCL) cancer, bronchioloalviolar cell lung cancer, bone
cancer, pancreatic cancer, skin cancer, cancer of the head or neck,
cutaneous or intraocular melanoma, uterine cancer, ovarian cancer,
rectal cancer, cancer of the anal region, stomach cancer, gastric
cancer, colon cancer, breast cancer, uterine cancer, carcinoma of
the fallopian tubes, carcinoma of the endometrium, carcinoma of the
cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's
Disease, cancer of the esophagus, cancer of the small intestine,
cancer of the endocrine system, cancer of the thyroid gland, cancer
of the parathyroid gland, cancer of the adrenal gland, sarcoma of
soft tissue, cancer of the urethra, cancer of the penis, prostate
cancer, cancer of the bladder, cancer of the kidney or urethra,
renal cell carcinoma, carcinoma of the renal pelvis, mesothelioma,
hepatocellular cancer, biliary cancer, chronic or acute leukemia,
lymphocytic lymphomas, neoplasms of the central nervous system
(CNS), spinal axis tumors, brain stem glioma, glioblastoma
multiforme, astrocytomas, schwannomas, ependymomas,
medulloblastomas, meningiomas, squamous cell carcinomas, pituitary
adenomas, including refractory versions of any of the above
cancers, or a combination of one or more of the above cancers. The
precancerous condition or lesion includes, for example, the group
consisting of oral leukoplakia, actinic keratosis (solar
keratosis), precancerous polyps of the colon or rectum, gastric
epithelial dysplasia, adenomatous dysplasia, hereditary
nonpolyposis colon cancer syndrome (HNPCC), Barrett's esophagus,
bladder dysplasia, and precancerous cervical conditions. In a
preferred embodiment, the cancer to be treated is a breast cancer
disease. Also in a preferred embodiment, the cancer is
characterized by an overexpression of the HER2 receptor
protein.
DETAILED DESCRIPTION
[0042] The invention provides a combined therapy method of treating
a breast cancer disease, comprising administering to the patient a
therapeutically effective amount of an anti-HER2 antibody and an
anti-VEGF antibody wherein the breast cancer disease is
characterized by an overexpression of the HER2 receptor protein.
More specifically, the invention provides a method of treating a
breast cancer disease in a patient who has failed prior therapy
with an anti-VEGF antibody, comprising administering to the patient
a therapeutically effective amount of an anti-HER2 antibody and an
anti-VEGF antibody wherein preferably the anti-VEGF antibody is
bevacizumab, the patient is human; the anti-HER2 antibody is
trastuzumab, and wherein preferably the breast cancer disease is
characterized by an overexpression of the HER2 receptor
protein.
[0043] The invention further comprises a method of treating a
breast cancer disease in a patient who has failed prior therapy
with an anti-VEGF antibody, comprising administering to the patient
a therapeutically effective amount of an anti-HER2 antibody while
continuing said anti-VEGF antibody therapy.
[0044] The term "treating" as used herein, unless otherwise
indicated, means reversing, alleviating, inhibiting the progress
of, or preventing, either partially or completely, the growth of
tumors, tumor metastases, or other cancer-causing or neoplastic
cells in a patient. The term "treatment" as used herein, unless
otherwise indicated, refers to the act of treating.
[0045] The phrase "a method of treating" or its equivalent, when
applied to, for example, cancer refers to a procedure or course of
action that is designed to reduce or eliminate the number of cancer
cells in a patient, or to alleviate the symptoms of a cancer. "A
method of treating" cancer or another proliferative disorder does
not necessarily mean that the cancer cells or other disorder will,
in fact, be eliminated, that the number of cells or disorder will,
in fact, be reduced, or that the symptoms of a cancer or other
disorder will, in fact, be alleviated. Often, a method of treating
cancer will be performed even with a low likelihood of success, but
which, given the medical history and estimated survival expectancy
of a patient, is nevertheless deemed an overall beneficial course
of action.
[0046] The term "patient" as used herein means a mammal, preferably
a human.
[0047] The term "therapeutically effective amount" or "effective
amount" means the amount of the subject compound or combination
that will elicit the biological or medical response of a tissue,
system, animal or human that is being sought by the researcher,
veterinarian, medical doctor or other clinician.
[0048] The invention further comprises the use of an anti-HER2
antibody and an anti-VEGF antibody for the manufacture of a
medicament for treating a breast cancer disease in a patient who
has failed prior therapy with an anti-VEGF antibody, comprising
administering to the patient a therapeutically effective amount of
an anti-HER2 antibody while continuing said anti-VEGF antibody
therapy. The antibodies may be administered separately or
simultaneously.
[0049] The term "method for manufacturing a medicament" relates to
the manufacturing of a medicament for use in the indication as
specified herein and in particular for use in the treatment of
tumors, tumor metastases, or cancer in general. The term relates to
the so-called "Swiss-type" claim format in the indication
specified.
[0050] In the context of this invention, additional other
cytotoxic, chemotherapeutic or anti-cancer agents, or compounds
that enhance the effects of such agents may be used in the
anti-VEGF antibody plus anti-HER2 antibody combination. Such agents
include, for example: alkylating agents or agents with an
alkylating action, such as cyclophosphamide (CTX; e.g.
cytoxan.RTM.), chlorambucil (CHL; e.g. leukeran.RTM.), cisplatin
(CisP; e.g. platino.RTM.) busulfan (e.g. myleran.RTM.), melphalan,
carmustine (BCNU), streptozotocin, triethylenemelamine (TEM),
mitomycin C, and the like; anti-metabolites, such as methotrexate
(MTX), etoposide (VP16; e.g. vepesid.RTM.), 6-mercaptopurine (6MP),
6-thiocguanine (6TG), cytarabine (Ara-C), 5-fluorouracil (5-FU),
capecitabine (e.g. Xeloda.RTM.), dacarbazine (DTIC), and the like;
antibiotics, such as actinomycin D, doxorubicin (DXR; e.g.
adriamycin1, daunorubicin (daunomycin), bleomycin, mithramycin and
the like; alkaloids, such as vinca alkaloids such as vincristine
(VCR), vinblastine, and the like; and other antitumor agents, such
as paclitaxel (e.g. taxal.RTM.) and pactitaxel derivatives, the
cytostatic agents, glucocorticoids such as dexamethasone (DEX; e.g.
decadron.RTM.) and corticosteroids such as prednisone, nucleoside
enzyme inhibitors such as hydroxyurea, amino acid depleting enzymes
such as asparaginase, leucovorin and other folic acid derivatives,
and similar, diverse antitumor agents. The following agents may
also be used as additional agents: arnifostine (e.g. ethyoll,
dactinomycin, mechlorethamine (nitrogen mustard), streptozocin,
cyclophosphamide, lomustine (CCNU), doxorubicin lipo (e.g.
doxil.RTM.), gemcitabine (e.g. gemzar.RTM.), daunorubicin lipo
(e.g. daunoxomel, procarbazine, mitomycin, docetaxel (e.g.
taxotere.RTM.), aldesleukin, carboplatin, oxaliplatin, cladribine,
camptothecin, CPT 11 (irinotecan), 10-hydroxy 7-ethyl-camptothecin
(SN38), floxuridine, fludarabine, ifosfamide, idarubicin, mesna,
interferon beta, interferon alpha, mitoxantrone, topotecan,
leuprolide, megestrol, melphalan, mercaptopurine, plicamycin,
mitotane, pegaspargase, pentostatin, pipobroman, plicamycin,
tamoxifen, teniposide, testolactone, thioguanine, thiotepa, uracil
mustard, vinorelbine, chlorambucil.
[0051] In the context of this invention, an anti-hormonal agent may
be used in the anti-VEGF antibody plus anti-HER2 antibody
combination. As used herein, the term "anti-hormonal agent"
includes natural or synthetic organic or peptidic compounds that
act to regulate or inhibit hormone action on tumors. Antihormonal
agents include, for example: steroid receptor antagonists,
anti-estrogens such as tamoxifen, raloxifene, aromatase inhibiting
4(5)-imidazoles, other aromatase inhibitors, 42-hydroxytamoxifen,
trioxifene, keoxifene, LY 117018, onapristone, and toremifene (e.g.
Fareston.RTM.); anti-androgens such as flutamide, nilutamide,
bicalutamide, leuprolide, and goserelin; and pharmaceutically
acceptable salts, acids or derivatives of any of the above;
agonists and/or antagonists of glycoprotein hormones such as
follicle stimulating hormone (FSH), thyroid stimulating hormone
(TSH), and luteinizing hormone (LH) and LHRH (leuteinizing
hormone-releasing hormone); the LHRH agonist goserelin acetate,
commercially available as Zoladex.RTM. (AstraZeneca); the LHRH
antagonist D-alaninamide
N-acetyl-3-(2-naphthalenyl)-D-alanyl-4-chloro-D-phenylalanyl-3-(3-pyridin-
yl)-D-alanyl-L-seryl-N6-(3-pyridinylcarbonyl)
-L-lysyl-N6-(3-pyridinylcarbonyl)-D-lysyl-L-leucyl-N6-(1-methylethyl)-L-l-
ysyl-L-proline (e.g Antide.RTM., Ares-Serono); the LHRH antagonist
ganirelix acetate; the steroidal anti-androgens cyproterone acetate
(CPA) and megestrol acetate, commercially available as Megace.RTM.
(Bristol-Myers Oncology); the nonsteroidal anti-androgen flutamide
(2-methyl-N-4, 20-nitro-3-(trifluoromethyl) phenylpropanamide),
commercially available as Eulexin.RTM. (Schering Corp.); the
non-steroidal anti-androgen nilutamide,
(5,5-dimethyl-3-4-nitro-3-(trifluoromethyl-4'-nitrophenyl)-4,4-dimethyl-i-
midazolidine-dione); and antagonists for other non-permissive
receptors, such as antagonists for RAR (retinoic acid receptor),
RXR (retinoid X receptor), TR (thyroid receptor), VDR (vitamin-D
receptor), and the like.
[0052] The use of the cytotoxic and other anticancer agents
described above in chemotherapeutic regimens is generally well
characterized in the cancer therapy arts, and their use herein
falls under the same considerations for monitoring tolerance and
effectiveness and for controlling administration routes and
dosages, with some adjustments. For example, the actual dosages of
the cytotoxic agents may vary depending upon the patient's cultured
cell response determined by using histoculture methods. Generally,
the dosage will be reduced compared to the amount used in the
absence of additional other agents.
[0053] Typical dosages of an effective cytotoxic agent can be in
the ranges recommended by the manufacturer, and where indicated by
in vitro responses or responses in animal models, can be reduced by
up to about one order of magnitude concentration or amount. Thus,
the actual dosage will depend upon the judgment of the physician,
the condition of the patient, and the effectiveness of the
therapeutic method based on the in vitro responsiveness of the
primary cultured malignant cells or histocultured tissue sample, or
the responses observed in the appropriate animal models.
[0054] In the context of this invention, additional
antiproliferative agents may be used in the anti-VEGF antibody plus
anti-HER2 antibody combination, induding, for example: Inhibitors
of the enzyme farnesyl protein transferase and inhibitors of the
receptor tyrosine kinase PDGFR, induding the compounds disclosed
and claimed in U.S. Pat. Nos. 6,080,769; 6,194,438; 6,258,824;
6,586,447; 6,071,935; 6,495,564; 6,150,377; 6,596,735 and
6,479,513, and International Publication WO 01/40217.
[0055] In the context of this invention, an effective amount of
ionizing radiation may be carried out and/or a radiopharmaceutical
may be used in addition to the anti-VEGF antibody plus anti-HER2
antibody combination. The source of radiation can be either
external or internal to the patient being treated. When the source
is external to the patient, the therapy is known as external beam
radiation therapy (EBRT). When the source of radiation is internal
to the patient, the treatment is called brachytherapy (BT).
Radioactive atoms for use in the context of this invention can be
selected from the group including, but not limited to, radium,
cesium-137, iridium-192, americium-241, gold-198, cobalt-57,
copper-67, technetium-99, iodine-123, iodine-131, and indium-111.
Where the EGFR kinase inhibitor according to this invention is an
antibody, it is also possible to label the antibody with such
radioactive isotopes.
[0056] Radiation therapy is a standard treatment for controlling
unresectable or inoperable tumors and/or tumor metastases. Improved
results have been seen when radiation therapy has been combined
with chemotherapy. Radiation therapy is based on the principle that
high-dose radiation delivered to a target area will result in the
death of reproductive cells in both tumor and normal tissues. The
radiation dosage regimen is generally defined in terms of radiation
absorbed dose (Gy), time and fractionation, and must be carefully
defined by the oncologist. The amount of radiation a patient
receives will depend on various considerations, but the two most
important are the location of the tumor in relation to other
critical structures or organs of the body, and the extent to which
the tumor has spread. A typical course of treatment for a patient
undergoing radiation therapy will be a treatment schedule over a 1
to 6 week period, with a total dose of between 10 and 80 Gy
administered to the patient in a single daily fraction of about 1.8
to 2.0 Gy, 5 days a week. In a preferred embodiment of this
invention there is synergy when tumors in human patients are
treated with the combination treatment of the invention and
radiation. In other words, the inhibition of tumor growth by means
of the agents comprising the combination of the invention is
enhanced when combined with radiation, optionally with additional
chemotherapeutic or anticancer agents. Parameters of adjuvant
radiation therapies are, for example, contained in International
Publication WO 99/60023.
[0057] The antibodies are administered to a patient according to
known methods, by intravenous administration as a bolus or by
continuous infusion over a period of time, by intramuscular,
intraperitoneal, intracerobrospinal, subcutaneous, intra-articular,
intrasynovial, or intrathecal routes. Intravenous or subcutaneous
administration of the antibodies is preferred.
[0058] The amount of anti-VEGF and anti-HER2 antibody
administration and the timing of administration will depend on the
type (species, gender, age, weight, etc.) and condition of the
patient being treated and the severity of the disease or condition
being treated.
[0059] Dosages for administration of the antibodies according to
the invention are about 1 .mu.g/kg to 50 mg/kg (e.g. 0.1-20 mg/kg)
of antibody by one or more separate administrations, or by
continuous infusion. A typical daily dosage might range from about
1 .mu.g/kg to about 100 mg/kg. In a preferred aspect, the
antibodies are administered every two to three weeks, at a dose
ranged from about 1 mg/kg to about 15 mg/kg. A preferred dose for
bevacizumab is 5 mg/kg once every 14 days as an IV infusion until
disease progression is detected. A preferred dose for trastuzumab
is a loading dose of 4 mg/kg administered over a 90-minute period
and subsequent weekly infusions of 2 mg/kg administered over a
30-minute period.
[0060] The present invention further provides a kit (pharmaceutical
kit) comprising an anti-VEGF antibody (preferably a pharmaceutical
composition thereof), an anti-HE2 antibody (preferably a
pharmaceutical composition thereof) and a package insert
instructing the user of said compositions to administer to a
patient, having breast cancer disease who has failed prior therapy
with an anti-VEGF antibody, the anti-VEGF antibody, preferably
within pharmaceutical composition and the anti-HER2 antibody,
preferably within a pharmaceutical composition. In a preferred
embodiment, the kit containers may further include a
pharmaceutically acceptable carrier. The kit may further include a
sterile diluent, which is preferably stored in a separate
additional container. The kit may further include a package insert
comprising printed instructions directing the use of the combined
treatment as a method for a breast cancer disease. Preferably, the
pharmaceutical kit will include a first container storing a
pharmaceutical composition comprising an anti-VEGF antibody and a
second container storing a pharmaceutical composition comprising an
anti-HER2 antibody.
[0061] Alternatively, the present invention also provides a
pharmaceutical kit comprising a pharmaceutical composition
comprising an anti-VEGF antibody, a pharmaceutical composition
comprising an anti-HE2 antibody, and a package insert instructing
the user of said compositions to administer to a patient having
breast cancer disease, who has failed prior therapy with an
anti-VEGF antibody, said anti-VEGF antibody pharmaceutical
composition and an anti-HER2 antibody pharmaceutical composition,
wherein the anti-VEGF antibody pharmaceutical composition and the
anti-HER2 antibody pharmaceutical composition are packaged either
in a single container or in two separate containers.
[0062] The present invention further provides a pharmaceutical
composition, in particular for use in treating a breast cancer
disease that has failed prior therapy with anti-VEGF antibody,
comprising an anti-HER2 antibody and an anti-VEGF antibody. Such
composition optionally comprises pharmaceutically acceptable
carriers and/or excipients, such as those commonly known to one of
ordinary skill in the art. In a preferred embodiment the anti-VEGF
antibody is bevacizumab and the anti-HER2 antibody is trastuzumab.
The present invention also provides a pharmaceutical kit comprising
said pharmaceutical composition comprising said anti-HER2 antibody
and said anti-VEGF antibody.
[0063] The following Experimental details are provided to aid the
understanding of the present invention, the true scope of which is
set forth in the appended claims. It is understood that the
specific methods and results discussed are merely illustrative of
the invention and are not to be considered in any way limited
thereto.
Introduction
[0064] The current study examined the antitumor activity of the
combination of bevacizumab and trastuzumab after the failure of
bevacizumab treatment alone in human breast xenograft model.
Further aims of the study were to examine the effects of treatment
on metastasis.
Test Agents
[0065] Trastuzumab was provided as a 25 mg/ml stock solution in
Histidine-HCl, alpha-alpha Trehalose (60 mM), 0.01% Polysorb, pH
6.0 (Herceptin.RTM.). Bevacizumab was provided as a 25 mg/ml stock
solution in Na-phosphate, alpha-alpha Trehalose (60 mM), 0.01%
Polysorb, pH 6.0 (Avastin.RTM.). Both solutions were diluted
appropriately in PBS for injections.
Cell Lines and Culture Conditions
[0066] The human breast cancer cell line KPL-4 has been established
from the malignant pleural effusion of a breast cancer patient with
an inflammatory skin metastasis and overexpresses ErbB family
receptors. (Kurebayashi, J., et al., Br. J. Cancer 79 (1999)
707-17) Tumor cells are routinely cultured in DMEM medium (PAA
Laboratories, Austria) supplemented with 10% fetal bovine serum
(PAA) and 2 mM L-glutamine (Gibco) at 37.degree. C. in a
water-saturated atmosphere at 5% CO2. Culture passage is performed
with trypsin/EDTA 1.times. (PAA) splitting twice/week. Cell passage
P6 was used for in vivo study.
Animals
[0067] SCID beige (C.B.-17) mice; age 10-12 weeks; body weight
18-20 g (Charles River, Sulzfeld, Germany) are maintained under
specific-pathogen-free condition with daily cycles of 12 h light 12
h darkness according to international guidelines (GV-Solas; Felasa;
TierschG). After arrival, animals are housed in the quarantine part
of the animal facility for one week to get accustomed to new
environment and for observation. Continuous health monitoring is
carried out on regular basis. Diet food (Alltromin) and water
(acidified pH 2.5-3) are provided ad libitum.
Tumor Growth Inhibition Studies In Vivo
[0068] Tumor cells were harvested (trypsin-EDTA) from culture
flasks (Greiner TriFlask) and transferred into 50 ml culture
medium, washed once and resuspended in PBS. After an additional
washing step with PBS and filtration (cell strainer; Falcon 100
.mu.m) the final cell titer was adjusted to 0.75.times.10.sup.8/ml.
Tumor cell suspension was carefully mixed with transfer pipette to
avoid cell aggregation. Anesthesia was performed using a Stephens
inhalation unit for small animals with preincubation chamber
(plexiglas), individual mouse nose-mask (silicon) and Isoflurane
(Pharmacia-Upjohn, Germany) in a closed circulation system. Two
days before injection the fur of the animals was shaved. For intra
mammary fat pad (i.f.m.p.) injection, cells were injected
orthotopically at a volume of 20 .mu.l into the right penultimate
inguinal mammary fat pad of each anesthetized mouse. For the
orthotopic implantation, the cell suspension was injected through
the skin under the nipple. Tumor cell injection corresponds to day
1 of the experiment.
Monitoring
[0069] Animals were controlled daily for detection of clinical
symptoms of adverse effects. For monitoring throughout the
experiment, the body weight of the animals was documented two times
weekly and the tumor volume was measured by caliper twice weekly.
Primary tumor volume was calculated according to NCI protocol
(TW=1/2ab2, where a and b are long and short diameters of tumor
size in mm, Teicher, B., Anticancer drug development guide, Humana
Press 5 (1997) 92). Calculation values were documented as mean and
standard deviation.
Treatment of Animals
[0070] Tumor-bearing mice were randomized when the tumor volume was
roughly 100 mm.sup.3 (n=10 for each group). Each group was closely
matched before treatment, which began 20 days after tumor cell
injection. Vehicle group (group 1) received 10 ml/kg PBS buffer
intraperitoneally (i.p.) once weekly. Trastuzumab (group 2) was
administered i.p. at a loading dose of 30 mg/kg, followed by once
weekly doses of 15 mg/kg (maintenance dose). The anti-VEGF antibody
bevacizumab was given i.p. at a dosage of 5 mg/kg twice weekly
(group 3). At day 40, treatment for group 3 was switched to a
combination treatment of bevacizumab (5 mg/kg twice weekly i.p.)
with trastuzumab (15 mg/kg once weekly i.p.).
Evaluation of Metastasis
[0071] Spread of tumor cells into the lung was determined in
sacrificed animals. Metastasis was measured according to Schneider,
T., et al., Clin. Exp. Metastasis 19 (2002) 571-582. Briefly, lung
tissue was harvested and human Alu sequences were quantified by
real-time PCR. Higher human DNA levels, quantified by real-time
PCR, correspond to higher levels of metastasis.
Results
[0072] The effect of treatment on primary tumor growth is shown in
FIG. 1 and Table 3. Tumors in the vehicle group (group 1) grew
rapidly and mice were sacrificed 38 days after injection of tumor
cells because of ulceration of tumors and the development of
clinical symptoms. Monotherapy with trastuzumab (group 2) exerted
no significant effect on tumor volume and mice were therefore
sacrificed at day 44. Treatment with bevacizumab suppressed tumor
growth significantly; however, tumors started to regrow around day
44. Combination treatment of bevacizumab and trastuzumab beginning
at day 55 resulted in complete inhibition of tumor growth during
the duration of the experiment (day 99) and treatment was well
tolerated.
TABLE-US-00003 TABLE 3 Antitumor activity of combined trastuzumab
and bevacizumab treatment on tumor growth after bevacizumab
treatment failure (data for FIG. 1). Mean tumor volume in mm.sup.3
is reported and the standard deviation (SD). trastuzumab + Day
Vehicle SD trastuzumab SD bevacizumab SD 20 118 31 120 31 119 35 23
150 30 157 57 126 44 27 209 51 164 77 143 67 30 269 76 169 82 138
65 34 348 114 214 114 167 76 37 431 138 293 162 181 78 42 462 275
172 63 44 547 315 211 65 48 226 68 51 266 78 55 324 103 58 318 100
62 248 81 65 232 75 70 209 69 73 224 56 79 213 68 83 173 57 86 178
80 90 150 73 93 141 74 97 134 67 99 130 76
[0073] The effect of treatment on lung metastasis is shown in FIG.
2 and Table 4. The combination of trastuzumab and bevacizumab after
bevacizumab treatment failure resulted in a sharp decrease in
metastasis. Levels of human Alu sequences (correlating to invasion
of tumor cells into secondary tissue) are significantly lower in
animals treated with combination therapy at 99 days over vehicle
treated animals that were sacrificed at 28 days and over
trastuzumab treated animals sacrificed on day 44. This surprising
effect on metastasis is in contrast with the effect seen with other
cytotoxic drugs (Geldof, A. A., et al., Anticancer Res. 8 (1988)
1335-1339; Murphy, J. Clin. Oncol. 11 (1993) 199-201, and De Larco,
J. E., et al., Cancer Res. 61 (2001) 2857-2861).
TABLE-US-00004 TABLE 4 Effect of treatment on lung metastasis. Alu
DNA was quantified by real-time PCR and is reported for each
animal. trastuzumab + Vehicle trastuzumab bevacizumab human 0.224
1.609 0.010 DNA 0.225 0.084 0.010 [ng/ml] 0.148 0.586 0.014 0.011
0.055 0.009 0.037 2.919 0.012 0.058 0.078 0.010 0.084 2.741 0.041
0.099 0.017 0.010 0.048 0.340 0.016 0.279 0.232 0.027 mean 0.1212*
0.8661** 0.098 median 0.0915 0.2861 0.088 Statistical significance
of combination treatment *p = 0.001 **p = <0.001
* * * * *