U.S. patent application number 15/229276 was filed with the patent office on 2016-12-22 for method for cancer therapy based on co-administration of a parvovirus and a cytokine.
The applicant listed for this patent is Deutsches Krebsforschungszentrum Stiftung Des Oeffentlichen Rechts, Ruprecht-Karls-Universitaet Heidelberg. Invention is credited to Manuel FISCHER, Karsten GELETNEKY, Irina KIPRIJANOVA, Jean ROMMELAERE, Joerg SCHLEHOFER.
Application Number | 20160367609 15/229276 |
Document ID | / |
Family ID | 41212170 |
Filed Date | 2016-12-22 |
United States Patent
Application |
20160367609 |
Kind Code |
A1 |
KIPRIJANOVA; Irina ; et
al. |
December 22, 2016 |
Method for Cancer Therapy Based on Co-Administration of a
Parvovirus and a Cytokine
Abstract
Described is a pharmaceutical composition comprising (a) a
parvovirus and (b) a cytokine, preferably IFN-.gamma., and the use
of said composition for treatment of cancer, e.g., a brain
tumor.
Inventors: |
KIPRIJANOVA; Irina;
(Heidelberg, DE) ; FISCHER; Manuel;
(Neckargemuend, DE) ; ROMMELAERE; Jean;
(Heidelberg, DE) ; SCHLEHOFER; Joerg; (Leimen,
DE) ; GELETNEKY; Karsten; (Heidelberg, DE) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
Deutsches Krebsforschungszentrum Stiftung Des Oeffentlichen
Rechts
Ruprecht-Karls-Universitaet Heidelberg |
Heidelberg
Heidelberg |
|
DE
DE |
|
|
Family ID: |
41212170 |
Appl. No.: |
15/229276 |
Filed: |
August 5, 2016 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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13376150 |
Apr 26, 2012 |
9446099 |
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PCT/EP2010/003070 |
May 19, 2010 |
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15229276 |
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Current U.S.
Class: |
1/1 |
Current CPC
Class: |
A61P 43/00 20180101;
C12N 2750/14333 20130101; C12N 2750/14332 20130101; A61K 35/768
20130101; A61K 38/217 20130101; A61K 38/217 20130101; C12N 7/00
20130101; A61P 35/00 20180101; C12N 2750/14371 20130101; A61K
2300/00 20130101; A61K 35/768 20130101; A61K 2300/00 20130101 |
International
Class: |
A61K 35/768 20060101
A61K035/768; C12N 7/00 20060101 C12N007/00; A61K 38/21 20060101
A61K038/21 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 4, 2009 |
EP |
09007433.7 |
Claims
1. A pharmaceutical composition containing a parvovirus and a
cytokine, wherein said cytokine is IFN-.gamma. and wherein the
parvovirus is H1 (H1-PV) and wherein the cytokine and the
parvovirus are separate entities.
2. The pharmaceutical composition of claim 1, wherein the cytokine
and the parvovirus are contained in separate containers.
3. The pharmaceutical composition of claim 2, wherein the first
container contains about 10.sup.8 to 10.sup.9 pfu H1-PV and the
second container contains about 10.sup.6 to 10.sup.7 U IFN-.gamma..
Description
[0001] This application is a continuation of U.S. application Ser.
No. 13/376,150, filed on Apr. 26, 2012, which is a National Stage
of PCT/EP2010/003070, filed May 19, 2010, which claims priority to
European Application No. 09007433.7 filed Jun. 4, 2009, the
disclosures of which are expressly incorporated by reference
herein.
[0002] The present invention relates to a pharmaceutical
composition comprising (a) a parvovirus and (b) a cytokine,
preferably IFN-.gamma., and the use of said composition for
treatment of cancer, e.g., a brain tumor.
[0003] Malignant human gliomas account for the largest number of
human malignant brain tumors. So far, the treatment of gliomas
includes neurosurgical techniques (resection or stereotactic
procedures), radiation therapy and chemotherapy. However, despite
these therapies gliomas are considered as incurable as they fail to
respond to ionising radiation, chemotherapy and surgical resection.
In other words, with these therapies only a very limited
prolongation of lifespan of patients can be achieved, i.e. despite
these therapies, the average life span after diagnosis is merely 12
to 16 months.
[0004] Cancer therapy using viruses or armed vector derivatives
that specifically kill neoplastically transformed cells (oncolysis)
is a novel approach to the treatment of this lethal disease. Some
autonomous parvoviruses belong to the category of so called
oncolytic viruses. Parvoviruses are small (25-30 nm) non-enveloped
particles containing a 5.1 kb single-stranded DNA genome from which
two nonstructural (NS1, NS2) and two capsid (VP1, VP2) proteins are
expressed. Parvovirus H-1PV has the unique advantage of triggering
a distinct death process, at least in brain and some other tumors,
namely the cytosolic relocation and activation of lysosomal
proteases (cathepsins). Several members of the parvovirus genus
(H-1PV, MVM, LuIII), whose natural hosts are rodents, are presently
under consideration for cancer gene therapy applications due to
their failure to transform host cells, capacity for asymptomatic
infection of humans, and ability to preferentially propagate in
(oncotropism) and kill (oncolysis) neoplastically transformed
cells. MVMp and H-1PV viruses have been shown to exert
oncosuppressive activities in vivo, i.e. they are able to inhibit
the formation of spontaneous, chemically or virally induced tumors
in laboratory animals. Vectors based on a parvoviral expression
cassette retain the oncotropic features of the wild type viruses.
Despite the impressive results achieved, the anticancer efficacy of
the most promising parvovirus candidates for human clinical
applications (including H-1PV) needs to be improved, e.g., as
regards the extension of life span after diagnosis.
[0005] Therefore, it is the object of the present invention to
provide means for an improved parvovirus-based therapy.
[0006] According to the invention this is achieved by the subject
matters defined in the claims. The present invention is based on
the applicant's findings that by the combined treatment with a
parvovirus and a cytokine such as IFN-.gamma. the therapeutic
efficiency can be improved. The observation that the combination of
H-1PV and IFN-.gamma. also shows beneficial effects on
immunodeficient mammals indicates that this effect does not depend
on T cells.
BRIEF DESCRIPTION OF THE DRAWINGS
[0007] FIG. 1: Monitoring of rat tumor (RG2 glioma) growth by MR
imaging
[0008] FIG. 2: Monitoring of human tumor (U87 glioma) growth by MR
imaging
[0009] Thus, the present invention provides a pharmaceutical
composition containing a (a) parvovirus and (b) a cytokine,
preferably (a) a parvovirus and (b) a cytokine as separate
entities, e.g. in separate containers.
[0010] As used herein, the term "cytokine" relates to a category of
signalling molecules that are used extensively in cellular
communication. They comprise proteins, peptides, or glycoproteins.
The term cytokine encompasses a large family of polypeptide
regulators that are produced widely throughout the body by cells of
diverse embryological origin. The action of cytokines may be
autocrine, paracrine, and endocrine. All cytokines are critical to
the development and functioning of both the innate and adaptive
immune response. They are often secreted by immune cells that have
encountered a pathogen, thereby activating and recruiting further
immune cells to increase the system's response to the pathogen.
Relying on the assays shown in Examples 2 and 3 the person skilled
in the art is in a position to select cytokines that show
beneficial effects when administrated according to the present
invention.
[0011] Preferably, the cytokine of the present invention is an
interferon. All interferons (IFNs) are natural cell-signalling
proteins produced by the cells of the immune system of most
vertebrates in response to challenges such as viruses, parasites
and tumor cells. Interferons are produced by a wide variety of
cells in response to the presence of double-stranded RNA, a key
indicator of viral infection. Interferons assist the immune
response by inhibiting viral replication within host cells,
activating natural killer cells and macrophages, increasing antigen
presentation to lymphocytes, and inducing the resistance of host
cells to viral infection. All interferons in general have several
effects in common and, accordingly, the results obtained by use of
IFN-.gamma. in combination with H1-PV might apply to further
interferons. Interferons are antiviral and possess antioncogenic
properties, macrophage and natural killer cell activation, and
enhancement of major histocompatibility complex glycoprotein
classes I and II, and thus presentation of foreign (microbial)
peptides to T cells. The production of interferons is induced in
response to microbes such as viruses and bacteria and their
products (viral glycoproteins, viral RNA, bacterial endotoxin,
bacterial flagella, CpG sites), as well as mitogens and other
cytokines, for example interleukin 1, interleukin 2,
interleukin-12, tumor necrosis factor and colony-stimulating
factor, that are synthesised in the response to the appearance of
various antigens in the body. Their metabolism and excretion take
place mainly in the liver and kidneys. They rarely pass the
placenta but they can cross the blood-brain barrier.
[0012] There are three major classes of interferons that have been
described for humans:
[0013] (a) Interferon type I: The type I interferons present in
humans are IFN-.alpha., IFN-.beta. and IFN-.omega..
[0014] (b) Interferon type II: In humans this is IFN-.gamma..
[0015] (c) Interferon type III: Signal through a receptor complex
consisting of IL10R2 (also called CRF2-4) and IFNLR1 (also called
CRF2-12)
[0016] In a more preferred embodiment of the present invention, the
interferon is interferon-.gamma..
[0017] Preferably, in said pharmaceutical composition the
parvovirus and the cytokine are present in an effective dose and
combined with a pharmaceutically acceptable carrier.
"Pharmaceutically acceptable" is meant to encompass any carrier,
which does not interfere with the effectiveness of the biological
activity of the active ingredients and that is not toxic to the
patient to whom it is administered. Examples of suitable
pharmaceutical carriers are well known in the art and include
phosphate buffered saline solutions, water, emulsions, such as
oil/water emulsions, various types of wetting agents, sterile
solutions etc.. Such carriers can be formulated by conventional
methods and can be administered to the subject at an effective
dose.
[0018] An "effective dose" refers to amounts of the active
ingredients that are sufficient to affect the course and the
severity of the disease, leading to the reduction or remission of
such pathology. An "effective dose" useful for treating and/or
preventing these diseases or disorders may be determined using
methods known to one skilled in the art (see for example, Fingl et
al., The Pharmocological Basis of Therapeutics, Goodman and Gilman,
eds. Macmillan Publishing Co., New York, pp. 1-46 ((1975)).
[0019] Preferred doses for the parvovirus are in the range of about
10.sup.8 to 10.sup.9 pfu (single injection) and for the cytokine,
in particular IFN-.gamma., in the range of about 10.sup.6 to
10.sup.7 U (single injection).
[0020] Additional pharmaceutically compatible carriers can include
gels, bioasorbable matrix materials, implantation elements
containing the therapeutic agent, or any other suitable vehicle,
delivery or dispensing means or material(s).
[0021] Administration of the compounds may be effected by different
ways, e.g. by intravenous, intraperetoneal, subcutaneous,
intramuscular, topical or intradermal administration. The route of
administration, of course, depends on the kind of therapy and the
kind of compounds contained in the pharmaceutical composition. A
preferred route of administration is intravenous administration.
The dosage regimen of the parvovirus and the cytokine is readily
determinable within the skill of the art, by the attending
physician based on patient data, observations and other clinical
factors, including for example the patient's size, body surface
area, age, sex, the particular parvovirus to be administered, the
time and route of administration, the tumor type and
characteristics, general health of the patient, and other drug
therapies to which the patient is being subjected.
[0022] If the parvovirus comprises infectious virus particles with
the ability to penetrate through the blood-brain barrier, treatment
can be performed or at least initiated by intravenous injection of
the parvovirus, e.g., H1 virus. A preferred route of administration
is intratumoral administration.
[0023] Since long-term intravenous treatment is susceptible to
becoming inefficient as a result of the formation of neutralizing
antibodies to the parvovirus, different modes of administration can
be adopted after an initial regimen intravenous viral
administration, or such different administration techniques, e.g.,
intracranial or intratumoral virus administration, can be
alternatively used throughout the entire course of parvoviral
treatment.
[0024] As another specific administration technique, the parvovirus
(virus, vector and/or cell agent) containing composition can be
administered to the patient from a source implanted in the patient.
For example, a catheter, e.g., of silicone or other biocompatible
material, can be connected to a small subcutaneous reservoir
(Rickham reservoir) installed in the patient during tumor removal
or by a separate procedure, to permit the parvovirus containing
composition to be injected locally at various times without further
surgical intervention. The parvovirus or derived vectors containing
composition can also be injected into the tumor by stereotactic
surgical techniques or by neuronavigation targeting techniques.
[0025] Administration of the parvovirus containing compositions can
also be performed by continuous infusion of viral particles or
fluids containing viral particles through implanted catheters at
low flow rates using suitable pump systems, e.g., peristaltic
infusion pumps or convection enhanced delivery (CED) pumps.
[0026] As yet another method of administration of the parvovirus
containing composition is from an implanted article constructed and
arranged to dispense the parvovirus containing composition to the
desired cancer tissue. For example, wafers can be employed that
have been impregnated with the parvovirus containing composition,
e.g., parvovirus H1, wherein the wafer is attached to the edges of
the resection cavity at the conclusion of surgical tumor removal.
Multiple wafers can be employed in such therapeutic intervention.
Cells that actively produce the parvovirus, e.g., parvovirus H1, or
H1 vectors, can be injected into the tumor, or into the tumoral
cavity after tumor removal.
[0027] The combined therapy according to the invention is useful
for the therapeutic treatment of cancer, in particular brain tumors
and can significantly improve the prognosis of said diseases.
Parvovirus H1 infection effects killing of tumor cells but does not
harm normal cells and such infection can, for example, be carried
out by intracerebral use of a suitable parvovirus, e.g., parvovirus
H1 (-1PV), or a related virus or vectors based on such viruses, to
effect tumor-specific therapy without adverse neurological or other
side effects.
[0028] The present invention also relates to the use of (a) a
parvovirus and (b) a cytokine, preferably IFN-.gamma., for the
preparation of a pharmaceutical composition for the treatment of
cancer wherein, preferably, (a) and (b) are sequentially (or
separately) administered.
[0029] In one preferred embodiment of the present invention, the
combination of agents is utilized in the treatment of brain tumors
such as glioma, medulloblastoma and meningioma. Preferred gliomas
are malignant human glioblastomas. However, the therapy according
to the present invention is, in principle, applicable to any tumor
that can be infected with the parvovirus, e.g., parvovirus H1. Such
tumors comprise pancreatic tumors, prostate tumors, lung tumors,
renal tumors, liver tumors, lymphoma, breast cancer and
hepatoma.
[0030] The term "parvovirus" as used herein comprises wild-type or
modified replication-competent derivatives thereof, as well as
related viruses or vectors based on such viruses or derivatives.
Suitable parvoviruses, derivatives, etc. as well as cells which can
be used for actively producing said parvoviruses and which are
useful for therapy, are readily determinable within the skill of
the art based on the disclosure herein, without undue empirical
effort.
[0031] In another preferred embodiment of the present invention,
the parvovirus of the composition includes parvovirus H1 (H1-PV) or
a related parvovirus such as LuIII, Mouse minute virus (MMV), Mouse
parvovirus (MPV), Rat minute virus (RMV), Rat parvovirus (RPV) or
Rat virus (RV).
[0032] Patients treatable by the combination of agents according to
the invention include humans as well as non-human animals. Examples
of the latter include, without limitation, animals such as cows,
sheep, pigs, horses, dogs, and cats.
[0033] Preferably, the parvovirus and the cytokine are administered
as separate compounds. The administration of the cytokine, when
administered separately, can be accomplished in a variety of ways
(see above) including systemically by the parenteral and enteral
routes.
[0034] In a further preferred embodiment, the parvovirus is
administered together with the cytokine.
[0035] The below examples explain the invention in more detail.
EXAMPLE 1
Materials and Methods
(A) Virus Production and Detection
[0036] Wild type H-1 virus was produced by infecting NBK cells,
purified by Iodixanol gradient centrifugation and dialyzed against
Ringer solution. Virus titers were determined as previously
described and expressed as replication center-forming units (cfu).
Briefly, serial dilutions of purified viruses were applied to NBK
cells. At 48 hours post infection, infected cultures were blotted
onto filters and replication centers were detected by
hybridization, using a virus DNA-specific radioactive probe
(Russell et al., J Virol 1992; 66:2821-8).
(B) Animal studies
[0037] (i) Anaesthesia. All surgical and imaging procedures were
performed under gaseous anaesthesia with isoflurane (Aerrane.RTM.,
Baxter, Maurepas, France) in pure oxygen. Isoflurane concentrations
varied between 5% for the initiation of anaesthesia to 2% (+/-0.5%)
during the surgical or imaging procedure.
[0038] (ii) Animals. 6 to 7 weeks old female Wistar rats or
immunodeficient RNU rats (Charles River, Sulzfeld, Germany)
weighing 220-250 g were used for tumor cell implantation. Wistar
rats were implanted with RG-2 glioma cells and RNU-rats were
implanted with human U87 cells. Animals were kept under
conventional conditions (temperature 22.+-.2.degree. C., relative
humidity 55.+-.10%, dark-light rhythm of 12 hr) with unrestricted
access to a balanced pellet diet and water. Animal experiments were
performed according to the French and European Community directives
for animal care (number 86/609/EEC of Nov. 24, 1986).
(C) Magnetic Resonance Imaging
[0039] The animals were examined in a 2.45 Tesla MRI scanner
(Bruker, Ettlingen, Germany) using T1 weighted imaging before and
after injection of 0.4 ml contrast medium (Gadodiamide,
Omniscan.TM., Amersham, Braunschweig, Germany) into the tail vein.
Gadodiamide-enhanced T1 imaging was performed 5 min after
injection. During MR examination, rats were anaesthetized by
Isoflurane insufflations (initial dose 5%, maintenance 2%). Tumor
volumes were determined using MRIcro software.
EXAMPLE 2
Increasing of Treatment Efficiency of Rat Glioma in Immunocompetent
Rats by Combining IFN-.gamma.with Parvovirus H-1 (H-1PV)
[0040] Tumor model: Cells of a rat glioma cell line (RG2 cells)
were implanted (10.sup.4 cells per animal) intracranially in the
right forebrain of Wistar rats. In total, 11 immunocompetent Wistar
rats (6-7 weeks old, 240-250 g, female) were analyzed in the
experiments.
[0041] 3 animals were used as controls, i.e. tumor cells were
implanted but not followed by any treatment, and tumor growth was
monitored by MR imaging. 8 animals were divided in three
groups:
[0042] (a) One group (3 rats, bearing rat glioma tumors) was
treated by i.v. injection of IFN-.gamma. (recombinant rat
IFN-.gamma., 10.sup.3 U per animal), 7 days after tumor cell
implantation.
[0043] (b) The second group (3 rat glioma bearing rats) was treated
by i.v. injection of a combination of H-1PV and rat IFN-.gamma., 7
days after tumor implantation (final dose of H-1PV per animal:
10.sup.8 pfu; final concentration of IFN-.gamma. per animal:
10.sup.3 U).
[0044] (c) The third group (2 rat glioma bearing rats) was treated
by i.v. injection of H-1PV alone (10.sup.8 pfu per animal), 7 days
after tumor implantation.
[0045] Tumor growth was analyzed by MR imaging at intervals of 7
days.
Results
[0046] The results are shown in FIG. 1.
[0047] (a) In all control animals, tumors grew continuously, and
rats were sacrificed after a maximum of 14 days because of signs of
suffering (cachexia, weakening, and difficulty in moving or
eating).
[0048] (b) Complete tumor regression after 7 days post treatment
was observed in 2 rats, in the group treated with the combination
of H-1PV and rat IFN-.gamma.. Tumor growth in one animal ceased
after treatment.
[0049] (c) In the group of animals treated with H-1PV alone, the
tumor volume was reduced 7 days after infection, (not completely
regressed at this time point).
[0050] (d) The tumors in rats, treated only with IFN-.gamma.,
continued to grow after treatment but not as fast as in control
animals. The rats from this group survived to a maximum of 19 days
after tumor cell implantation (i.e. 5 days longer compared to
control animals). Neither tumor regression nor arrest of tumor
growth was observed in this group.
[0051] It can be expected that not only additive but even
synergistic effects can be achieved in vivo by the combined
treatment with H-1PV and a cytokine such as IFN-.gamma..
EXAMPLE 3
Increasing of Treatment Efficiency of Human Glioma Cell
Line-Derived Brain Tumor in Immunodeficient Rats by Combining
IFN-.gamma.with Parvovirus H-1 (H-1PV)
[0052] Tumor model: Cells of a human glioma cell line (U87 cells)
were implanted (10.sup.3 cells per animal) intracranially in the
right forebrain of the rats. In total, 18 immunodeficient RNU rats
(6-7 weeks old, 220-250 g, female) were analyzed in the
experiments.
[0053] Five animals were used as controls, i.e. tumor cells were
implanted but not subject to any treatment, and tumor growth was
monitored by MR imaging. The remaining 13 animals were divided in
three treatment groups:
[0054] (a) One group (5 rats, bearing human glioma cell-derived
tumors) were treated by i.v. injection of IFN-.gamma. (recombinant
human IFN-.gamma., 10.sup.3 U per animal), 7 days after tumor cell
implantation.
[0055] (b) The second group (5 human glioma bearing rats) was
treated by i.v. injection of a combination of H-1PV and human
IFN-.gamma., 7 days after tumor implantation (final dose of H-1PV
per animal: 10.sup.8 pfu; final concentration of IFN-.gamma. per
animal: 10.sup.5 U).
[0056] (c) The third group (3 rats) was treated with i.v. injection
of H-1PV alone (10.sup.8 pfu per animal), 7 days after tumor
implantation.
[0057] Tumour growth was analyzed by MR imaging at intervals of 4
days.
Results
[0058] The results are shown in FIG. 2.
[0059] (a) In all control animals, tumors grew continuously, and
rats were sacrificed after a maximum of 14 days because of signs of
suffering (cachexia, weakening, difficulty in moving or
eating).
[0060] (b) Complete human tumor regression was observed in 3 rats,
in the group treated with the combination of H-1PV and human
IFN-.gamma.. Tumor growth in two animals ceased after treatment,
and tumor volume remained constant until rats were sacrificed for
histological analysis.
[0061] (c) The tumors in rats, treated only with IFN-.gamma.,
continued to grow after treatment but not as fast as in control
animals. The rats from this group survived to a maximum of 3 weeks
post tumor cell implantation (i.e. one week longer compared to
control animals). Neither tumor regression nor arrest of tumor
growth was observed in this group.
[0062] (d) In the group of animals treated with H-1PV alone, the
tumour volume was reduced 7 days after infection (but not
completely regressed at this time point).
* * * * *