+

WO1999045783A1 - Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite - Google Patents

Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite Download PDF

Info

Publication number
WO1999045783A1
WO1999045783A1 PCT/US1999/005466 US9905466W WO9945783A1 WO 1999045783 A1 WO1999045783 A1 WO 1999045783A1 US 9905466 W US9905466 W US 9905466W WO 9945783 A1 WO9945783 A1 WO 9945783A1
Authority
WO
WIPO (PCT)
Prior art keywords
cell
tumor
cells
producer
hsv
Prior art date
Application number
PCT/US1999/005466
Other languages
English (en)
Inventor
Katherine L. Molnar-Kimber
David J. Caparrelli
Larry R. Kaiser
Steven M. Albelda
George Coukos
Original Assignee
The Trustees Of The University Of Pennsylvania
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by The Trustees Of The University Of Pennsylvania filed Critical The Trustees Of The University Of Pennsylvania
Priority to AU30017/99A priority Critical patent/AU3001799A/en
Priority to CA002323067A priority patent/CA2323067A1/fr
Priority to EP99911363A priority patent/EP1061806A4/fr
Publication of WO1999045783A1 publication Critical patent/WO1999045783A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/76—Viruses; Subviral particles; Bacteriophages
    • A61K35/763—Herpes virus
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/16011—Herpesviridae
    • C12N2710/16611—Simplexvirus, e.g. human herpesvirus 1, 2
    • C12N2710/16632—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Definitions

  • the field of the invention is treatment of malignancy using oncolytic virus agents.
  • BACKGROUND OF THE INVENTION Cancer remains one of the leading causes of morbidity and mortality of humans worldwide. Although certain tumors remain localized at discrete locations within the body, at least during certain stages of their growth, other tumors are dispersed from their earliest stages or arise in tissues which line body cavities or organs. For example, epithelial cancers arise in tissues which line the lungs, the ovaries, the exterior of the body, and various body cavities. Epithelial ovarian cancer (EOC) is one such epithelial cancer.
  • EOC epithelial ovarian cancer
  • EOC Despite the aggressive surgical approaches and combination chemotherapeutic regimens investigated over the past two decades, EOC remains a disease with a grim prognosis. For example, recent statistics indicate that 25,000 new patients afflicted with EOC are diagnosed yearly in the U.S.; 15,000 deaths occur there from this disease yearly. Unfortunately, due to the lack of symptoms, the majority of patients afflicted with EOC are diagnosed at a late stage. In addition, although 70% of EOC patients initially respond to cisplatin-based chemotherapy, the majority of these patients relapse and develop chemotherapy-resistant disease. As a result, the overall five-year survival rate is approximately 20% for advanced-stage EOC.
  • anti-cancer gene therapy must involve use of replication defective viruses to administer the desired transgene in order to prevent systemic spread of virus and its associated complications, toxicity, or both. Therefore, until recently much of gene therapy for malignant disease has centered on the delivery
  • HIV-1716 is a replication-competent herpes simplex virus type 1 which has a 759-bp deletion in both copies of the RL1 gene which encodes for the protein ICP34.5, a major determinant of herpes pathogenicity. Viruses with this mutation exhibit drastically reduced neurovirulence. These viruses do not cause encephalitis when inoculated either intracerebrally or peripherally into a host. Moreover, these mutants replicate as well as their wild-type parental strain (e.g. 17+) in a variety of dividing cells lines, but replicate poorly in cells not undergoing mitosis. These characteristics make HSV-1716 and other RL1 mutants attractive as vectors for cancer gene therapy.
  • wild-type parental strain e.g. 17+
  • Replication selective adenovirus strains have also been studied. Several groups have shown efficacy in both immunocompromised and immunocompetent mouse models of intracranial malignancies. Further, it has been shown that HSV antigen staining is restricted to the tumor mass with no spread to adjacent normal tissue.
  • malignant mesothelioma lends itself to study because of its location in the lining of the pleural cavity, there is interest in other, more prevalent, thoracic malignancies which have poor prognoses unless identified early.
  • Other malignancies in which morbidity is associated with localized disease include, for example, bronchoalveolar cell, bladder, endometrial, cervical, and ovarian cancers.
  • Endothelial ovarian cancer for example, remains localized within the peritoneal cavity in a large proportion of patients, ultimately causing local morbidity and lethal complications. Because of its localized nature, EOC lends itself to intraperitoneal approaches of therapy.
  • One such approach is gene therapy.
  • Gene therapy comprising either delivering the herpes simplex virus- 1 thymidine kinase (HSVtk) suicide gene to diseased cells followed by administration of ganciclovir to the patient or delivering tumor suppressor genes and/or oncogenes to cells has been tested in experimental ovarian cancer models in vitro and in vivo (Tong et al., 1996, Gynecol. Oncol.
  • Replication-competent and replication-restricted viral agents provide a feasible alternative for cancer therapy.
  • Replication-restricted recombinant attenuated forms of herpes simplex virus- 1 (HSV-1) represent one family of such agents (Chambers et al., 1995, Proc. Natl. Acad. Sci. USA 92:1411-1415; Jia et al., 1994, J. Natl. Cancer Inst. 86: 1209-1215; Glorioso et al., 1995, Annu. Rev. Microbiol.
  • HSV-1 mutants have been generated that harbor alterations in genes such as thymidine kinase (tk) or ribonucleotide reductase (RR) and exhibit decreased viral replication in non-dividing neuronal cells.
  • tk thymidine kinase
  • RR ribonucleotide reductase
  • HSV- 1 mutants Another series of HSV- 1 mutants has been produced by making alterations in both copies of the RL1 gene, a diploid fragment of the HSV-1 genome (Chambers et al., 1995, Proc. Natl. Acad. Sci. USA 92:1411-1415; Kramm et al., 1997,
  • ICP34.5-null HSV-1 mutants have been shown to replicate preferentially in tumor cells, causing a direct oncolytic effect, but appear to spare normal differentiated tissues (Randazzo et al., 1996, Virology 223:392-395; Brown et al., 1994, J. Gen. Virol. 75:3767-3686). These strains have been successfully used to reduce or cure tumors of the central nervous system (CNS) in experimental models (Chambers et al., 1995, Proc. Natl. Acad. Sci. USA ; Jia et al., 1994, J. Natl.
  • CNS central nervous system
  • HSV- 1716 an ICP34.5 null mutant of HSV- 1
  • SCID severe combined immunodeficient mice
  • extra-CNS administration of replication-restricted HSV-1 is safe.
  • HSV-1716 administered intraperitoneally to SCID mice appeared to be completely avirulent. In fact, there was no viral spread outside the tumors, as was documented in Kucharczuk et al. (1997, Cancer Res.
  • a ribonuclease reductase-deleted mutant was used in an experimental animal model of metastatic colorectal carcinoma of the liver (Carroll et al., 1996, Ann. Surg. 224:323-329).
  • a replication-restricted ICP34.5 mutant was used to treat experimental metastatic and subcutaneous melanoma (Randazzo et al., 1995, Virology 211:94-101; Randazzo et al., 1997, J. Invest. Dermatol. 108:933-937).
  • HSV-G207 a multi-attenuated mutant, HSV-G207, was efficacious for treatment of breast cancer (Toda et al., 1998, Human Gene Ther. 9:2173-2185).
  • these studies demonstrate that use of various oncolytic viruses to kill tumor cells is well accepted, even if prior art uses of such oncolytic vectors have been plagued with shortcomings such as low efficacy, low tissue specificity, rapid clearing of oncolytic viruses, and inability to deliver a sufficiently high or prolonged doses of virus to the desired tumor tissue.
  • the present invention includes producer cells and methods of using them that overcome the shortcomings of the prior art, thereby permitting efficacious delivery of oncolytic viruses to tumor tissue and effective treatment of cancers such as EOC.
  • the invention relates to a producer cell for administration to a subject having tumor cells.
  • the producer cell comprises an oncolytic virus which is capable of replicating in the producer cell.
  • the producer cell is incapable of sustained survival in the body of the subject.
  • the oncolytic virus is cytotoxic with respect to the producer cell in the body of the subject.
  • the producer cell is rendered incapable of sustained survival in the body of the subject by exposing the producer cell to a lethal dose of radiation.
  • the lethal dose of radiation may be a dose which enhances the burst size of the producer cell (e.g. about 3 Gray).
  • the producer cell is rendered incapable of sustained survival in the body of the subject by incorporating a suicide gene (e.g. thymidine kinase or cytosine deaminase) into the producer cell.
  • a suicide gene e.g. thymidine kinase or cytosine deaminase
  • the producer cell exhibits binding affinity for a tumor cell in the subject, such as an epithelial tumor cell (e.g. an epithelial ovarian cancer cell).
  • a tumor cell in the subject such as an epithelial tumor cell (e.g. an epithelial ovarian cancer cell).
  • the oncolytic virus is capable of replicating in a tumor cell of the subject.
  • the oncolytic virus may be less capable of replicating in a non-tumor cell of the subject than in the tumor cell.
  • the oncolytic virus may incapable of replicating in a non-tumor cell of the subject, or it may be incapable of replicating in any cell of the subject.
  • Replication of the oncolytic virus may, for example, be under the control of a
  • tumor-associated transcriptional promoter such as the prostate specific antigen promoter or the tumor growth factor- ⁇ promoter.
  • the producer cell may, for example, be selected from the group consisting of a PA- 1 cell, an REN cell, a PER.C6 cell a 293 cell, a melanoma cell, a glioma cell, and a teratocarcinoma cell.
  • the producer cell is a PA-1 cell.
  • the oncolytic virus may, for example, be selected from the group consisting of a herpes simplex virus- 1, a herpes simplex virus-2, an adenovirus, a vesicular stomatitis virus, a Newcastle disease virus, and a vaccinia virus.
  • a herpes simplex virus- 1 When the oncolytic virus is a herpes simplex virus- 1, it preferably does not express functional ICP34.5.
  • Suitable herpes simplex virus-1 include, but are not limited to, HSV-1716,
  • Suitable herpes simplex virus- 2 include, but are not limited to, strain 2701, strain 2616, and strain 2604.
  • Suitable adenoviruses include, but are not limited to, ONYX-15, Ad5dl520, Ad5dl312, CN706, Addll 10, Addll 11, Addll 18, and Addl004.
  • the producer cell further comprises a composition selected from the group consisting of an immunomodulatory molecule, a cytokine, a targeting molecule, a cell growth receptor, an immunoglobulin which is specific for the tumor, a nucleic acid encoding an immunomodulatory molecule, a nucleic acid encoding a cytokine, a nucleic acid encoding a targeting molecule, a nucleic acid encoding a cell growth receptor, and a nucleic acid encoding an immunoglobulin which is specific for the tumor.
  • the invention also relates to an anti-tumor agent comprising a mammalian cell which comprises thymidine kinase.
  • the mammalian cell exhibits binding affinity for a tumor cell in a human patient and is incapable of sustained survival in the body of the patient.
  • the mammalian cell binds with a tumor cell in the patient.
  • gancyclovir is thereafter administered to the patient, the mammalian cell metabolizes gancyclovir to generate a cytotoxic metabolite which is provided to the tumor cell with which the mammalian cell has bound.
  • the invention further relates to a method of killing tumor cells in a mammal. This method comprises administering to the mammal a producer cell.
  • the producer cell comprising a oncolytic virus which is capable of replicating in the producer cell.
  • the producer cell is incapable of sustained survival in the body of the mammal.
  • the mammal may, for example, be a human afflicted with an epithelial cancer or a human afflicted with an tumor.
  • the invention still further relates to use of a producer cell for manufacture of a medicament for administration to a patient having tumor cells.
  • the producer cell comprises a oncolytic virus which is capable of replicating in the producer cell.
  • the producer cell is incapable of sustained survival in the body of the patient.
  • the invention relates to a producer cell for administration to a subject (e.g. a human patient) having tumor cells.
  • the producer cell comprises any of a wide variety of oncolytic viruses (e.g. the herpes simplex virus-1 mutant designated HSV-
  • the oncolytic virus is capable of replicating in the producer cell, and may also be capable of replicating in tumor cells in the subject.
  • the producer cell is not capable of sustained survival in the body of the subject. Because the producer cell supports replication of the oncolytic virus, it may contain many (e.g. tens, hundreds, or thousands) of copies of the virus.
  • the copies of the virus escape from the cell and are delivered (e.g. by fluid-mediated dispersion or by producer cell-to-tumor cell contact) to tumor cells in the subject. Once delivered to tumor cells in the subject, the oncolytic viruses kill the tumor cells.
  • oncolytic viruses using producer cells has advantages over prior art direct injection methods of delivering such viruses to tumor cells.
  • the virus is, in some embodiments, capable of replicating in the producer cell of the invention, the amount of virus which can be administered in a given volume of fluid can be greatly increased, since cells in which a virus has replicated may contain tens, hundreds, or even thousands of copies of the virus.
  • delivery of a virus within a producer cell may enable the virus to elude the subject's immune system, increasing the likelihood that the virus will reach and kill a tumor cell.
  • the producer cell used to deliver the oncolytic virus exhibits binding affinity for tumor cells in a subject
  • delivery of the virus using the producer cell will increase localization of the virus to the tumor cells in the patient. This may be particularly important for treatment of tumors that are not discretely localized.
  • an element means one element or more than one element.
  • a “subject” is an animal, preferably a mammal such as a human.
  • a subject “has tumor cells” if the subject comprises or is suspected to comprise tumor cells in any form (i.e. in the form of a solid tumor, a dispersed tumor, a metastatic tumor cell, or the like).
  • a tumor cell is "killed” if it is induced to lyse, if it is induced to undergo apoptosis, or if it is rendered incapable of growing or dividing.
  • an "oncolytic virus” is any virus which is able to kill a tumor cell by infecting the tumor cell.
  • a virus is "cytotoxic with respect to” a cell if the virus is able to kill the cell after infecting the cell.
  • An “anti-tumor agent” is a composition of matter which, when applied to a tumor cell, kills the tumor cell.
  • a “transcriptional promoter” is a nucleic acid which, when operably linked with a second nucleic acid encoding a gene product such as an RNA or a protein, enables the gene product to be expressed in a cell by virtue of permitting an RNA polymerase enzyme to transcribe the second nucleic acid.
  • two polynucleotides as "operably linked" as used herein is meant that a single-stranded or double-stranded nucleic acid moiety comprises each of the two polynucleotides and that the two polynucleotides are arranged within the nucleic acid moiety in such a manner that at least one of the two nucleic acid sequences is able to exert a physiological effect by which it is characterized upon the other.
  • a "suicide gene” is a gene which, when expressed in a cell, induces lysis or apoptosis of the cell or renders the cell incapable of growth or division.
  • Replication of a virus is "under the control of a promoter" if at least one gene product required for replication for replication of the virus is operably linked with the promoter.
  • a cell "exhibits binding affinity" for a tumor cell if the cell binds to the tumor cell with greater affinity than the affinity with which it binds to a non-tumor cell.
  • a "functional" biological molecule is a biological molecule in a form in which it exhibits a property by which it is characterized.
  • a functional enzyme for example, is one which exhibits the characteristic catalytic activity by which the enzyme is characterized.
  • An oncolytic virus is "replication-selective" if it is more capable of replicating in an tumor cell of a subject than in a non-tumor cell of the subject.
  • a goal of viral gene therapy for treatment of malignant disease has been delivery of a therapeutic/suicide gene (e.g. thymidine kinase, cytosine deaminase, p53, etc.) to tumor cells in a subject.
  • a therapeutic/suicide gene e.g. thymidine kinase, cytosine deaminase, p53, etc.
  • a replication defective virus such as an adenovirus.
  • Use of replication competent or replication selective viruses for the treatment of neoplastic disease provides certain advantages. Such viruses deliver the desired gene to a larger percentage of tumor cells. Furthermore, replication of these viruses in vivo may well be oncolytic in their own right.
  • HSV-1 replication competent herpes simplex type 1 viruses having deletions in the RL1 gene, which encodes the protein designated ICP34.5. This protein is a major determinant of
  • Viruses having mutations in this gene exhibit reduced neurovirulence 105 . For example, they do not cause encephalitis when inoculated either intracerebrally or peripherally. Moreover, these mutants replicate as well as their wild-type parental strain (e.g. 17+) in a variety of dividing cells lines, but replicate poorly in cells not undergoing mitosis. These characteristics make these HSV-1 mutant viruses attractive for use as oncolytic viruses.
  • the present inventors have devised a virus delivery method whereby cells that are infected with the mutant herpesvirus serve as "producer cells" for an oncolytic virus.
  • Use of producer cells to deliver oncolytic viruses in vivo provides several advantages. For example, rapidly dividing cell lines have the ability to replicate viruses very efficiently, producing as many as about 6,000 copies of a virus per infected cell.
  • the producer cells are, in effect, viral factories which can increase the effective dose of virus administered to the patient.
  • Administering producer cells may protect, at least initially, the virus from neutralizing host immunity.
  • Producer cell lines may be engineered to enhance tumor killing, for example, by selecting or designing producer cells which produce cytokines which enhance the oncolytic effects of the virus with which they are infected.
  • Oncolytic gene therapy has, until now, focused on localized malignancies where tumors can be directly injected with vector (e.g. glioma) or where vector can be instilled into a discrete body cavity (e.g. the pleural cavity for malignant mesothelioma).
  • the producer cell of the invention may be used to treat both localized and diffuse or disseminated malignancies.
  • the presence of producer cells may have a positive effect on tumor killing by inducing an immune response against the tumor cells.
  • the invention includes a producer cell for administration to a subject having tumor cells.
  • the producer cell comprises a suicide gene (e.g. thymidine kinase) and exhibits binding affinity for a tumor cell.
  • a suicide gene e.g. thymidine kinase
  • the producer cell binds with a tumor cell in the subject, if one is present.
  • Expression of the suicide gene optionally coupled with administration to the subject of a substrate of an enzyme encoded by the suicide gene (e.g. ganciclovir when the suicide
  • the producer cell comprises an oncolytic virus.
  • the oncolytic virus is capable of replicating in the producer cell, but the producer cell is incapable of sustained survival in the body of the patient.
  • the producer cell makes many copies of the oncolytic virus, and releases them into the subject's body upon the death of the producer cell.
  • the oncolytic virus is not capable of replicating in the producer cell, but is nonetheless carried into and released within the body of the subject by the producer cell.
  • the producer cell of the invention is preferably one of many types of cells which are known to exhibit binding affinity for tumor cells, but it is not necessary that the producer cell exhibit such affinity.
  • the producer cell may be a PA-1 cell, an REN cell, a PER C6 cell, a 293 cell, a melanoma cell, a glioma cell, or a teratocarcinoma cell.
  • the producer cell is obtained from an animal of the same species (and strain, if applicable) as the subject or from a cell line derived from such an animal.
  • the producer cell is well tolerated by the subject (i.e. is not rejected by the immune system of the subject) when the producer cell is injected into the patient.
  • acceptable producer cells for use in human patients include any human cell line which is derived from a human source and which does not induce hyperacute rejection when injected into the patient.
  • acceptable producer cells for use in humans include any cell line which has been engineered to not induce hyperacute rejection.
  • the producer cell is not capable of sustained survival in the subject's body, by which is meant that the producer cell does not endure more than several months, several weeks, or several days in the body of the subject following administration of the producer cell to the subject.
  • the producer cell is not capable of replicating in the subject's body. Numerous methods are known in the art
  • the oncolytic virus of the invention may be selected such that it is cytotoxic with respect to the producer cell in the body of the subject.
  • the virus kills not only tumor cells in the patient, but also the producer cells which are used to deliver the virus.
  • the producer cell may also be rendered incapable of sustained survival in the body of the patient by exposing the producer cell to a lethal dose of radiation prior to providing the producer cell to the subject.
  • a radiation dose of 20 Gray will kill nearly all known cells which might be used as producer cells.
  • certain lethal doses of radiation enhance the burst size of the oncolytic virus in the producer cell.
  • the producer cell may be rendered incapable of sustained survival in the body of the patient by incorporating a suicide gene (e.g. thymidine kinase or cytosine deaminase) into the producer cell.
  • a suicide gene e.g. thymidine kinase or cytosine deaminase
  • the suicide gene is derived from the body of the patient by incorporating a suicide gene (e.g. thymidine kinase or cytosine deaminase) into the producer cell.
  • a suicide gene e.g. thymidine kinase or cytosine deaminase
  • the producer cell is killed. Potential undesirable immune reactions may be minimized or avoided by using producer cells which are incapable of sustained survival in the subject.
  • the producer cell of the invention preferably exhibits binding affinity for a tumor cell in the patient.
  • Use of such producer cells has a number of benefits.
  • binding of a producer cell to a tumor cell necessarily brings the oncolytic virus(es) in or on the producer cell into close association with the tumor cell, increasing the likelihood that the virus will infect and kill the tumor cell.
  • at least certain viruses may be transmitted by cell-to-cell contact.
  • binding of a producer cell and a tumor cell may enhance targeting of virus to the tumor cell in this manner as well.
  • formation of a producer cell-tumor cell complex may generate or expose antigenic regions which can be recognized by the subject's immune system, leading to generation of an immune response against the tumor cells. If the
  • the producer cells are engineered to contain, display, or express various immune modulators, growth factors, or suicide genes, binding of the producer cell to a tumor cell in the subject localized the biological activity of these molecules to the tumor site.
  • the producer cell may comprising an immunomodulatory molecule, a cytokine, a targeting molecule, a cell growth receptor, an immunoglobulin which is specific for the tumor, or a nucleic acid encoding one of these.
  • the cell may either naturally comprise one of these molecules or be engineered to comprise the molecule.
  • the producer cell exhibits binding affinity for an epithelial tumor cell, such as an epithelial ovarian cancer cell.
  • epithelial tumors like dispersed tumors such as various leukemias
  • prior art gene therapy methods have experienced difficulty delivering the gene vector to all tumor sites. If the producer cell binds with one of these dispersed or widely spread tumor types, then the oncolytic virus of the invention will be delivered to the tumor cells wherever they are dispersed or spread.
  • the oncolytic virus of the invention may be substantially any virus which is known to exhibit oncolytic activity and which is capable of replicating in, or at least being carried by, a producer cell of the invention without ablating the oncolytic activity of the virus.
  • the oncolytic virus of the invention is also able to replicate in a tumor cell of the patient, and is preferably less capable of replicating in a non-tumor cell of the patient than in a tumor cell of the patient.
  • the oncolytic virus may incapable of replicating in a non-tumor cell of the patient.
  • the virus may be incapable of replicating in any cell of the subject.
  • An oncolytic virus may be made replication-selective if replication of the virus is placed under the control of a regulator of gene expression such as, for example, a minimal enhancer/promoter region derived from the 5'-flank of the human PSA gene (e.g. see Rodriguez et al., 1997, Cancer Res. 57:2559-2563).
  • a regulator of gene expression such as, for example, a minimal enhancer/promoter region derived from the 5'-flank of the human PSA gene (e.g. see Rodriguez et al., 1997, Cancer Res. 57:2559-2563).
  • the main transcriptional unit of an oncolytic virus may be placed under transcriptional control of the tumor growth factor- ⁇ (TGF- ⁇ ) promoter by operably linking virus genes to the TGF- ⁇ promoter.
  • TGF- ⁇ tumor growth factor- ⁇
  • an oncolytic virus wherein replication is subject to transcriptional control of the TGF- ⁇ promoter is replication-selective, in that it is more capable of replicating in the certain tumor cells than in non-tumor cells of the same type.
  • Similar replication-selective oncolytic viruses may be made using any regulator of gene expression which is known to selectively cause overexpression in an affected cell.
  • the replication-selective oncolytic virus may, for example, be an HSV strain in which a gene encoding ICP34.5 is mutated.
  • the oncolytic virus of the invention may also be one which exhibits binding affinity for a tumor cell of the subject.
  • the oncolytic virus of the invention may, for example, be a herpes simplex virus- 1, a herpes simplex virus-2, an adenovirus, a vesicular stomatitis virus, a Newcastle disease virus, or a vaccinia virus.
  • oncolytic viruses are described, for example, in Kirn (1999, In: Gene Therapy of Cancer. Academic Press, San Diego, CA, pp. 235-248).
  • the oncolytic virus of the invention is a herpes simplex virus- 1 , it is preferably one which does not express functional ICP34.5 protein (e.g. HSV-1716) or one of the HSV-1 viruses described in Coukos et al., (1998, Gene Ther. Mol. Biol.
  • HSV-1 viruses include HSV-1716, HSV-3410, HSV-3616, and HSV-4009.
  • Other replication selective HSV-1 virus strains which may be used as the oncolytic virus of the invention include, by way of example and not limitation, HSV-
  • R3616 in which the gene encoding ICP34.5 is deleted
  • HSV-R47 in which genes encoding proteins R3616 and ICP47 are deleted
  • HSV-G207 in which genes encoding ICP34.5 and ribonucleotide reductase are deleted
  • HSV-7020, HSV-NVR10 in which genes encoding 7020 and ICP47 are deleted
  • HSV-3616-UB in which genes encoding ICP34.5 and uracil DNA glycosylase are deleted
  • HSV-G92A in which the albumin promoter is a transcriptional regulated promoter
  • HSV-3616-IL-4 in which the gene encoding ribonucleotide reductase is deleted
  • HSV strains which do not express functional ICP34.5 and which express a cytokine such as interleukin-2, interleukin-4, or GM-CSF.
  • virus when the virus is a herpes simplex virus-2, it may, for example, be selected from the group consisting of strain 2701, strain 2616, and strain 2604.
  • the virus when it is an adenovirus, it may, for example, be selected from the group consisting of ONYX-15, Ad5dl520, Ad5dl312, CN706, Addll 10, Addll 11, Addll 18, and Addl004.
  • the producer cells of the invention may be used in a method of killing tumor cells in a mammal.
  • the method comprising administering to the mammal a producer cell comprising a oncolytic virus which is capable of replicating in the producer cell.
  • the mammal may be substantially any mammal having tumor cells, and is preferably a human patient afflicted with a localized cancer such as malignant mesothelioma, EOC, bladder cancer, or the like.
  • the producer cells of the invention may be selectively engineered to enhance their organ- or cell-specificity as well as their ability to induce tumor killing.
  • Known gene products may be used enhance the oncolytic effect of viruses delivered to tumor cells by the producer cells of the invention. For example, increased levels of certain cytokines (e.g. interleukins 2, 4, and 6, ⁇ -interferon, and tumor necrosis factor ⁇ ) inhibit tumor growth and metastasis.
  • cytokines e.g. interleukins 2, 4, and 6, ⁇ -interferon, and tumor necrosis factor ⁇
  • Producer cells which secrete such a cytokine in conjunction with virus production will have a synergistic effect on tumor killing.
  • Producer cell lines may similarly be modified to enhance immune recruitment to the area of administration or decrease the effect of pre-existing immunity to the vector.
  • the invention also includes use as producer cells of genetically- modified cells which express a suicide gene such as HSV-7jfv and exhibit binding
  • compositions are sensitive to ganciclovir because they convert ganciclovir into various toxic metabolites. These cells exhibit therapeutic effect on tumor cells, based on the "bystander effect," which is death of cells adjacent to the genetically-modified cells. The bystander effect is thought to be caused by transport of one or more toxic metabolites of ganciclovir from cell to cell through gap junctions as well as by induction of an immune response.
  • the invention encompasses the preparation and use of medicaments and pharmaceutical compositions comprising the producer cell of the invention as an active ingredient.
  • a pharmaceutical composition may consist of the active ingredient alone, in a form suitable for administration to a subject, or the pharmaceutical composition may comprise the active ingredient and one or more pharmaceutically acceptable carriers, one or more additional ingredients, or some combination of these.
  • Administration of one of these pharmaceutical compositions to a subject is useful for killing tumor cells or arresting tumor growth or spread in the subject, as described elsewhere in the present disclosure.
  • the term "pharmaceutically acceptable carrier” means a chemical composition with which the active ingredient may be combined and which, following the combination, can be used to administer the active ingredient to a subject.
  • the formulations of the pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit.
  • compositions are principally directed to pharmaceutical compositions which are suitable for ethical administration to humans, it will be understood by the skilled artisan that such compositions are generally suitable for administration to animals of all sorts.
  • compositions suitable for administration to humans in order to render the compositions suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation.
  • Subjects to which administration of the pharmaceutical compositions of the invention is contemplated include, but are not limited to, humans, primates, and other mammals.
  • compositions that are useful in the methods of the invention may be prepared, packaged, or sold in formulations suitable for intraperitoneal, oral, rectal, vaginal, parenteral, topical, pulmonary, intranasal, buccal, ophthalmic, or another route of administration.
  • Other contemplated formulations include projected nanoparticles, liposomal preparations, and immunologically-based formulations.
  • a pharmaceutical composition of the invention may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses.
  • a "unit dose" is discrete amount of the pharmaceutical composition comprising a predetermined amount of the active ingredient.
  • the amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.
  • Controlled- or sustained-release formulations of a pharmaceutical composition of the invention may be made using conventional technology.
  • Liquid suspensions may be prepared using conventional methods to achieve suspension of the active ingredient in an aqueous or oily vehicle.
  • Aqueous vehicles include, for example, water and isotonic saline.
  • Oily vehicles include, for example, almond oil, oily esters, ethyl alcohol, vegetable oils such as arachis, olive, sesame, or coconut oil, fractionated vegetable oils, and mineral oils such as liquid paraffin.
  • Liquid suspensions may further comprise one or more additional ingredients including, but not limited to, suspending agents, dispersing or wetting agents, emulsifying agents, demulcents, preservatives, buffers, salts, flavorings,
  • Oily suspensions may further comprise a thickening agent.
  • suspending agents include, but are not limited to, sorbitol syrup, hydrogenated edible fats, sodium alginate, polyvinylpyrrolidone, gum tragacanth, gum acacia, and cellulose derivatives such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose.
  • Known dispersing or wetting agents include, but are not limited to, naturally-occurring phosphatides such as lecithin, condensation products of an alkylene oxide with a fatty acid, with a long chain aliphatic alcohol, with a partial ester derived from a fatty acid and a hexitol, or with a partial ester derived from a fatty acid and a hexitol anhydride (e.g. polyoxyethylene stearate, heptadecaethyleneoxycetanol, polyoxyethylene sorbitol monooleate, and polyoxyethylene sorbitan monooleate, respectively).
  • Known emulsifying agents include, but are not limited to, lecithin and acacia.
  • a pharmaceutical composition of the invention may also be prepared, packaged, or sold in the form of oil-in-water emulsion or a water-in-oil emulsion.
  • the oily phase may be a vegetable oil such as olive or arachis oil, a mineral oil such as liquid paraffin, or a combination of these.
  • compositions may further comprise one or more emulsifying agents such as naturally occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soybean or lecithin phosphatide, esters or partial esters derived from combinations of fatty acids and hexitol anhydrides such as sorbitan monooleate, and condensation products of such partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate.
  • emulsions may also contain additional ingredients including, for example, sweetening or flavoring agents.
  • a pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for rectal administration.
  • a composition may be in the form of, for example, a suppository, a retention enema preparation, and a solution for rectal or colonic irrigation.
  • Suppository formulations may be made by combining the active ingredient with a non-irritating pharmaceutically acceptable excipient which is solid at
  • Suitable pharmaceutically acceptable excipients include, but are not limited to, cocoa butter, polyethylene glycols, and various glycerides. Suppository formulations may further comprise various additional ingredients including, but not limited to, antioxidants and preservatives.
  • Retention enema preparations or solutions for rectal or colonic irrigation may be made by combining the active ingredient with a pharmaceutically acceptable liquid carrier.
  • enema preparations may be administered using, and may be packaged within, a delivery device adapted to the rectal anatomy of the subject.
  • Enema preparations may further comprise various additional ingredients including, but not limited to, antioxidants and preservatives.
  • a pharmaceutical composition of the invention may be prepared, packaged, or sold in a formulation suitable for vaginal administration.
  • a composition may be in the form of, for example, a suppository, an impregnated or coated vaginally-insertable material such as a tampon, a douche preparation, or a solution for vaginal irrigation.
  • Methods for impregnating or coating a material with a chemical composition include, but are not limited to methods of depositing or binding a chemical composition onto a surface, methods of incorporating a chemical composition into the structure of a material during the synthesis of the material (i.e. such as with a physiologically degradable material), and methods of absorbing an aqueous or oily solution or suspension into an absorbent material, with or without subsequent drying.
  • Douche preparations or solutions for vaginal irrigation may be made by combining the active ingredient with a pharmaceutically acceptable liquid carrier.
  • douche preparations may be administered using, and may be packaged within, a delivery device adapted to the vaginal anatomy of the subject.
  • Douche preparations may further comprise various additional ingredients including, but not limited to, antioxidants, antibiotics, antifungal agents, and preservatives.
  • parenteral administration of a pharmaceutical composition includes any route of administration characterized by physical breaching of a tissue of a subj ect and administration of the pharmaceutical composition through the breach in the tissue.
  • Parenteral administration thus includes, but is not limited to, administration of a pharmaceutical composition by injection of the composition, by application of the composition through a surgical incision, by application of the composition through a tissue-penetrating non-surgical wound, and the like.
  • parenteral administration is contemplated to include, but is not limited to, subcutaneous, intraperitoneal, intravenous, intraarterial, intramuscular, or intrasternal injection and intravenous, intraarterial, or kidney dialytic infusion techniques.
  • Formulations of a pharmaceutical composition suitable for parenteral administration comprise the active ingredient combined with a pharmaceutically acceptable carrier, such as sterile water or sterile isotonic saline. Such formulations may be prepared, packaged, or sold in a form suitable for bolus administration or for continuous administration. Injectable formulations may be prepared, packaged, or sold in unit dosage form, such as in ampules, in multi-dose containers containing a preservative, or in single-use devices for auto-injection or injection by a medical practitioner. Formulations for parenteral administration include, but are not limited to, suspensions, solutions, emulsions in oily or aqueous vehicles, pastes, and implantable sustained-release or biodegradable formulations.
  • Such formulations may further comprise one or more additional ingredients including, but not limited to, suspending, stabilizing, or dispersing agents.
  • the active ingredient is provided in dry (i.e. powder or granular) form for reconstitution with a suitable vehicle (e.g. sterile pyrogen-free water) prior to parenteral administration of the reconstituted composition.
  • compositions may be prepared, packaged, or sold in the form of an injectable aqueous or oily suspension. This suspension or solution may
  • injectable formulations may be prepared using a non-toxic parenterally-acceptable diluent or solvent, such as water or 1,3 -butane diol, for example.
  • a non-toxic parenterally-acceptable diluent or solvent such as water or 1,3 -butane diol, for example.
  • Other acceptable diluents and solvents include, but are not limited to, Ringer's solution, isotonic sodium chloride solution, and fixed oils such as synthetic mono- or di-glycerides.
  • Other parentally-administrable formulations which are useful include those which comprise the active ingredient in microcrystalline form, in a liposomal preparation, or as a component of a biodegradable polymer systems.
  • Compositions for sustained release or implantation may comprise pharmaceutically acceptable polymeric or hydrophobic materials such as an emulsion, an ion exchange resin, a sparingly soluble polymer, or a sparingly soluble salt.
  • additional ingredients include, but are not limited to, one or more of the following: excipients; surface active agents; dispersing agents; inert diluents; granulating and disintegrating agents; binding agents; lubricating agents; sweetening agents; flavoring agents; coloring agents; preservatives; physiologically degradable compositions such as gelatin; aqueous vehicles and solvents; oily vehicles and solvents; suspending agents; dispersing or wetting agents; emulsifying agents, demulcents; buffers; salts; thickening agents; fillers; emulsifying agents; antioxidants; antibiotics; antifungal agents; stabilizing agents; and pharmaceutically acceptable polymeric or hydrophobic materials.
  • kits comprising a pharmaceutical composition of the invention and an instructional material.
  • an "instructional material” includes a publication, a recording, a diagram, or any other medium of expression which is used to communicate the usefulness of the pharmaceutical composition of the invention for killing tumor cells in a subject, for
  • the instructional material of the kit of the invention may, for example, be affixed to a container which contains a pharmaceutical composition of the invention or be shipped together with a container which contains the pharmaceutical composition. Alternatively, the instructional material may be shipped separately from the container with the intention that the instructional material and the pharmaceutical composition be used cooperatively by the recipient.
  • the invention also includes a kit comprising a pharmaceutical composition of the invention and a delivery device for delivering the composition to a subject.
  • the delivery device may be a squeezable spray bottle, a metered-dose spray bottle, an aerosol spray device, an atomizer, a dry powder delivery device, a self-propelling solvent/powder-dispensing device, a syringe, a needle, a tampon, or a dosage measuring container.
  • the kit may further comprise an instructional material as described herein.
  • Example 1 In order to develop an effective method of vector delivery, EJ-6-2-Bam-6a cells, an NIH 3T3 fibroblast cell line, were used as producer cells for HSV-1716 in a immunocompetent murine model of lung cancer. It was demonstrated that Lewis Lung Carcinoma (LLC), a spontaneous non-immunogenic murine lung cancer, is sensitive to treatment using HSV-1716 in vitro as well as in vivo.
  • LLC Lewis Lung Carcinoma
  • LLC supported replication of and was efficiently lysed by HSV-1716 at a multiplicity of infection (MOI) of 1.0 ( ⁇ 20% of cells were viable four days following infection). HSV-1716 replicated in LLC cells with a burst size of 20.
  • MOI multiplicity of infection
  • HSV-1716 is useful as a new gene therapy vector for, at least, lung cancer and that the use of allogeneic producer cells such as EJ-6-2 can enhance the oncolytic efficacy of virus both by increasing the effective MOI of the virus and by inducing an inflammatory response within the tumor milieu.
  • the rsHSV-1 used in the experiments in this Example, HSV-1716 is a replication-competent attenuated strain of HSV- 1 which does not express ICP34.5.
  • a single intraperitoneal administration of 5 x 10" plaque-forming units (pfu) of HSV- 1716 resulted in significant reduction of tumor volume and tumor spread and increase in survival in a mouse xenograft model of EOC.
  • PA-1 cells support HSV-1716 replication in vitro and bind preferentially to human ovarian carcinoma surfaces, relative to mesothelial surfaces, both in vitro and in vivo.
  • intraperitoneal administration of irradiated PA-1 cells infected with HSV-1716 induced significantly greater tumor reduction in the two xenograft models tested.
  • Histologic evaluation indicated the presence of extensive necrosis at tumor sites infected with HSV- 1716.
  • Immunohistochemistry to detect HSV- 1716 indicated that areas of viral infection were present within tumor nodules, and that such infection persisted for several weeks following treatment.
  • Administration of HSV-1716-infected producer cells induced more widespread infection of tumor tissue by the virus.
  • HSV-1716 Isolation of HSV-1716 has been described previously (MacLean et al., 1991, J. Gen. Virol. 72:631-639).
  • the genome of this virus contains a 759-base-pair deletion located within each copy of the BamHl fragment of the long repeat region of the genome. These deletions encompass most of the gene encoding ICP34.5. The mutant therefore does not express this protein. Passage of the virus has also been described (MacLean et al., 1991, J. Gen. Virol. 72:631-639; Kucharczuk et al., 1997, Cancer Res. 57:466-471).
  • ovarian cancer cell lines SKOV3, NIH:OVCAR3, CaOV3, and human ovarian teratocarcinoma line PA-1 have been previously described, and were obtained from the American Tissue Culture Collection (Plainview, MD; Tainsky et al., 1988, Anticancer Res. 8:899-913).
  • the A2780 EOC cell line was obtained from the Fox Chase Cancer Center (Philadelphia, PA).
  • Primary ovarian cultures were obtained from patients afflicted with advanced EOC, at stages III or IV, according to the criteria set by the International Federation of Gynecologists and Obstetricians (DiSaia et al., 1993, In: Clinical Gynecologic Oncology. Mosby-Year Book, Inc., St. Louis, MO). Malignant effusions, obtained at the time of exploratory laparotomy or diagnostic/therapeutic paracentesis,
  • Normal peritoneal mesothelial cells were obtained from intraoperative pelvic peritoneal lavages carried out using normal saline in patients undergoing laparotomy for benign pelvic pathology (e.g. pelvic relaxation, uterine myomata). Lavage fluids were centrifuged at 300 g for 10 minutes at room temperature and the cell pellets were collected and seeded in standard media (see below). These cells assumed an epithelial-like phenotype and grew in a cobblestone pattern. Their doubling time was longer than that of primary EOC cultures (average 36 hours), and their non-malignant nature was confirmed by the fact that they propagated for only a few (4-5) passages even in the presence of growth factors.
  • benign pelvic pathology e.g. pelvic relaxation, uterine myomata
  • culture media were aspirated 30 min following plating, and suspended cells were re-seeded in new culture flasks. All cell lines and EOC primary isolates were cultured under standard conditions (37 °C in a 5% CO 2 atmosphere) in RPMI media comprising 10% (v/v) heat-inactivated fetal calf serum (FCS) and antibiotics.
  • FCS heat-inactivated fetal calf serum
  • media were supplemented with a mixture of growth factors (SerXtendTM, Irvine
  • ⁇ t Cells were incubated in 96-well plates at a density of 3 10 3 cells per well. The cells were incubated in the presence of HSV-1716 at multiplicities of infection (MOI) of 0.1 and 1 in serum-free media for one hour. Serum-enriched media was subsequently added and cultures were observed for four days. Cell proliferation assays were performed using a chromogenic kit (CellTiter AQueous96TM, Promega,
  • CPE Cytopathic effects
  • PA-1 cells were cultured in T25 flasks until they reached 60% confluence. The cells were then infected with 0.5-2.5 MOI of HSV-1716, as described above. Cells were harvested at 16 hours using a 0.05% trypsin solution, washed once with phosphate buffered saline (PBS) and fixed and permeabilized using 70% methanol at -20 °C for 20 minutes.
  • PBS phosphate buffered saline
  • Cells were labeled using a polyclonal antibody (obtained from American Qualex, La Mirada, CA) which specifically binds with HSV-1 proteins, which was used at a dilution of 1 :250, and a secondary anti-rabbit fluorescein isothiocyanate (F ⁇ TC)-conjugated antibody (obtained from Jackson Immunoresearch Laboratories Inc., West Groove, PA), which was used at a dilution of 1 :250.
  • F ⁇ TC secondary anti-rabbit fluorescein isothiocyanate
  • Flow cytometric analysis was performed using an EPICSTM XL flow cytometer (Coulter Corporation, Hialeah, FL).
  • PA-1 teratocarcinoma cells were incubated overnight in 6-well plates at a density of 4 10- cells per well under standard culture conditions and were then infected with HSV-1716 at 0.3 MOI.
  • PA-1 cells were subjected to a single 20 Gray dose of ionizing radiation one hour prior to infection with HSV-1716.
  • Cells were harvested in the accompanying medium by mechanical scraping one hour after infection with the virus (designated '0 hours') as well as 6, 20, 24, and 48 hours later and stored at -80°C.
  • One-step growth curves were generated as described (Kucharczuk et al., 1997, Cancer Res. 57:466-471).
  • PA-1 teratocarcinoma cells were labeled using a rhodamine fluorescent dye (PKH26 Red Fluorescent Cell Linker Kit, Sigma Chemical Co., St. Louis, MO) as recommended by the manufacturer. Briefly, cells were harvested using a 0.05% (w/v) trypsin solution, re-suspended in PBS, and incubated with a 1:250 dilution of the dye at for 8 minutes. The labeling reaction was termination by addition of 100% FCS.
  • PA-1 cells were then seeded at 5x10 ⁇ per well on cell monolayers and allowed to interact with adherent mesothelial or ovarian cancer cells for 30 minutes.
  • PA-1 teratocarcinoma cells were radio-labeled using JJ S-methionine, as described above, and subjected to ionizing radiation (20
  • A2780 and SKOV3 cells were produced using standard culture conditions until they reached 70% confluence. The cells were harvested using 0.05% (w/v) trypsin solution, washed with serum-enriched media, and centrifuged at 300xg for 5 minutes at 4°C. About 5xl0 6 SKOV3 cells or 1 x10" A2780 cells per animal were injected intraperitoneally in 0.5 milliliter of RPMI medium containing 10% FCS and 1% SerXtendTM (Irvine Scientific). Five mice were sacrificed at selected times to confirm the presence of intraperitoneal tumors prior to administering treatment in each experiment. At the end of the experiments, animals were sacrificed and intraperitoneal tumors were assessed for spread and volume. A semi-quantitative scoring system was devised to assess tumor spread.
  • Each site was assigned a score of 0-3 as following: "0" if no microscopic tumor was seen; “1” if one or more microscopic tumors were visible with the aid of a dissecting microscope at 2.5 x magnification; “2” if tumor nodules less than 5 millimeters in diameter were visible; and “3” if tumor nodules equal to or more than 5 millimeters in diameter were present.
  • the presence of ascites resulted in addition of one point in the scoring system. The maximum score accumulated in this scale was therefore 16. All visible tumor nodules were dissected from surrounding normal peritoneum and viscera, and the total weight of intra-abdominal tumor was determined for each animal.
  • PA-1 human teratocarcinoma cells were exposed to ionizing radiation at a single dose of 20 Gray and were then infected with HSV-1716 at an MOI of 2. Two hours after infection, cells were washed with PBS and harvested using a 0.05% (w/v) trypsin solution. Cells were washed twice in media comprising 10%) heat-inactivated
  • Tumors obtained from treated and control animals were immediately fixed in formalin (3.7% (v/v) formaldehyde in PBS) and embedded in paraffin.
  • Six- micrometer-thick sections were de-paraffinized and stained using hematoxylin-eosine (H&E).
  • H&E hematoxylin-eosine
  • the slides were subjected to antigen retrieval at 105°C for 10 minutes in 0.1 normal citric acid and incubated with a monoclonal antibody which binds specifically with HSV-1 (DAKO, Carpenteria, California) at a dilution of 1 :4 for 30 minutes at 47 °C.
  • a horseradish peroxidase-conjugated anti-mouse antibody (Vectastain, Vector Laboratories, Burlingame, CA) was used at a dilution of 1 :400.
  • HSV-1716 Exerts an Oncolytic Effect on Epithelial Ovarian Cancer in vitro
  • HSV-1716 In order to assess the oncolytic effect of HSV-1716 on EOC cells in vitro, primary EOC cultures and established EOC cell lines were exposed to HSV-1716 at 0.1 and 1 MOI. Cell survival was assessed by proliferation colorimetric assays and evaluation of cytopathic effect (CPE), as assessed by phase-contrast microscopy.
  • CPE cytopathic effect
  • HSV-1716 exerted a direct dose-dependent cytolytic effect on all EOC cell lines and on human teratocarcinoma PA-1 cell line.
  • EOC cell line A2780 was the most sensitive of the cell lines tested, and SKOV3 was the least sensitive of those tested.
  • Teratocarcinoma PA-1 cells were highly sensitive to HSV-1716-induced cytolysis. The sensitivity of PA-1 cells was comparable to that of primary EOC cultures.
  • HSV-1716 Exerts an Oncolytic Effect on Epithelial Ovarian Cancer in vivo
  • a murine xenograft model of EOC was used together with two well characterized EOC cell lines, namely cell lines SKOV3 and A2780. These two cell lines were selected because, in addition to being well characterized, these two cell lines exhibited the least and greatest, respectively, sensitivity to cytolysis by HSV-1716
  • HSV-1716 was evaluated by examining xenografted mice having different tumor burdens, by varying the duration of treatment, and by using the two aforementioned cell lines to generate the tumors, thereby yielding intraperitoneal tumors having different volumes.
  • the virus was administered one week following the injection of SKOV3 ovarian cancer cells into mice. Intraperitoneal injection of 5x10" SKOV3 cells resulted in formation of microscopic tumor(s) at the diaphragm and occasionally at the omentum, mesentery, or lesser omentum within one week.
  • the tumor score (as described above) for these mice was 2/16.
  • the A2780 cell line was injected into mice.
  • Administration of a single dose of HSV-1716 three weeks following cell injection resulted in arrest of tumor growth at 6 weeks (three weeks later), relative to the pre-treatment counterpart mice.
  • a difficulty of comparing the effect of virus administered alone and virus administered by way of producer cells relates to controlling the number of particles inoculated.
  • the lowest MOI at which 100% of PA-1 cells were infected with HSV-1716 was determined.
  • the viral burst size (i.e. the number of infectious virus particles released from a PA-1 cell upon cytolysis following infection of the cell with the virus and incubation) was determined.
  • the burst size of HSV-1716 in PA-1 teratocarcinoma cells was 200 in the absence of ionizing radiation. Because administration of HSV- 1 infected producer cells into humans will most likely be performed after eliminate the risk of administering potentially uninfected producer cells (i.e. by irradiating the cells), PA-1 cells were subjected to a single (lethal) dose of 20 Gray one hour prior to infection, and viral replication in the irradiated cells was assessed.
  • PA-1 Producer Cells Preferentially Bind to Epithelial Ovarian Cancer Surface Compared to Normal Peritoneum in vivo
  • PA-1 Producer Cells Preferentially Bind to Epithelial Ovarian Cancer Surface Compared to Normal Peritoneum in vitro
  • PA-1 cell binding to tumor and non- tumor cells in vivo could have been related to molecular factors controlling cell-cell interactions between PA-1 cells and murine mesothelial cells.
  • these differences have been related to physical forces governing peritoneal fluid circulation, which might direct both SKOV3 tumor cells and PA-1 cells towards the same peritoneal sites.
  • these differences have been caused by decreased affinity
  • PA-1 teratocarcinoma cells displayed significantly higher binding to ovarian cancer cells than to normal human mesothelium (pO.OOl for each mesothelial culture versus each ovarian cancer primary culture or cell line except A2780).
  • adhesion of PA-1 teratocarcinoma cells to different ovarian cancer cell lines following radiation was compared with adhesion following infection with
  • HSV-1716 and adhesion after both radiation treatment and infection with virus did not significantly affect PA-1 adhesion to cancer cells or normal mesothelial cells.
  • PA-1 Producer Cells Effectively Deliver HSV-1716 in vivo In order to assess the suitability of producer cells for delivering
  • HSV-1716 particles to tumor cells in vivo direct administration of HSV-1716 (i.e. via injection of a suspension of virus particles) was compared with administration of PA-1 cells infected with HSV-1716.
  • SCID mice received a single intraperitoneal administration of either virus alone or HSV-infected PA-1 cells. Control animals received medium or non-infected, irradiated PA-1 cells, respectively.
  • mice Similar results were obtained in experiments performed using SKOV3 cells injected intraperitoneally (5 10 ⁇ cells per animal) into SCID mice, as indicated in Table 3.
  • Control animals from group 2 exhibited a 6.5-fold increase in tumor weight (pO.OOl versus pre-treatment) and extensive tumor spread (tumor score 15.9/16) at 4 weeks.
  • Treatment Group Number 4 Weeks Post-Treatment 7 Weeks Post-Treatment
  • Animals receiving HSV-infected producer cells (group 4) also had significantly smaller tumors (pO.OOl) and less tumor spread (score 2/16) at 4 and 7 weeks than their corresponding controls (group 2).
  • mice of group 4 exhibited a significantly smaller tumor burden at 7 weeks than HSV-1716-treated animals (group 3, p ⁇ .05).
  • PA-1 cells infected with HSV-1716 at 2 MOI were observed for 16 weeks and then sacrificed in order to evaluate the presence of intraperitoneal tumors. No microscopic or gross tumor could be detected.
  • HSV- 1716 survival experiments performed using animals bearing SKOV3 tumors confirmed the oncolytic activity of HSV- 1716 against EOC cells.
  • a single dose of HSV- 1716 was administered four weeks following the inj ection of tumor cells, a time at which there was a sizable intraperitoneal tumor burden.
  • HSV-1716 Penetrates Deeply in Tumor Nodules and Causes Extensive Necrosis in a Dose-Dependent Manner
  • HSV-treated animals i.e. those treated with virus alone and those treated with virus-infected producer cells
  • the virus was present deep within solid tumor nodules. In distinct areas of the tumor nodules, viral spread was detectable several weeks following a single intraperitoneal injection.
  • Immunohistochemical evaluation of tumors obtained from animals treated with HSV-infected PA-1 cells indicated that even larger areas of the tumor nodules were subject to viral infection, relative to animals treated with HSV-1716 alone. Areas of viral infection were primarily found in the interface between uninfected tumor and areas of necrosis, suggesting that, by replicating, the virus advanced into the tumor leaving behind an area of necrosis.
  • the virus could not be detected in normal murine tissues such as liver, pancreas, kidney, adrenal, spleen, small bowel myenteric plexus, or brain.
  • producer cells may have promoted direct cell-to-cell viral infection of tumor cells by binding to EOC cells.
  • PA-1 cells appeared to adhere predominantly to areas of peritoneum that were covered by established tumor and not to close-by areas of normal peritoneum. There was a close correlation between the presence of tumor and PA-1 cells, as calculated by odds ratios. It is possible that the physical forces governing fluid migration within the peritoneal cavity were partially responsible for this effect or that species-related differences in the affinity of interaction might have influenced these findings.
  • the in vitro studies performed using human normal mesothelial cells demonstrated that PA-1 cells bound predominantly to EOC surfaces compared to normal human mesothelium. This suggests that cell-cell interactions promote adhesion of PA-1 producer cells to tumor cells, and that such adhesion promotes transfer of virus from producer cells to tumor cells.
  • HSV- 1716 a replicating herpes simplex type 1 virus with a mutation in the gamma 34.5 gene, is non-neurovirulent and has shown efficacy as an oncolytic treatment of mesothelioma in immunodeficient mice.
  • an immunocompetent animal model was investigated. EJ-62, a r ⁇ s-mutated murine fibroblast, grows well in the peritoneum of immunocompetent BALB/c mice and, unlike most murine cells, is sensitive to herpesvirus infection.
  • HSV-1716 in this syngeneic intraperitoneal model, which resembles mesothelioma and ovarian cancer, were studied, and the efficacy of both single and multiple virus injections of virus were evaluated. Use of producer cells in this model was also investigated.
  • Producer cells were infected ex vivo with the virus, irradiated, and then injected intraperitoneally. Cell viability studies were performed on EJ-62 at varying MOI values. At an MOI of 1 or greater, greater than 90% cell death, relative to control cells, was observed. MOI values as low as 0.01 resulted in approximately 75% cell death, relative to control cells.
  • Established intraperitoneal tumors were treated with virus injections in a single (4 10 particles) dose, multiple doses (three doses given every fourth day), or a single comparable dose of producer cells. Prolonged survival was observed in all treated groups, relative to control cells. The median survival of control cells was about 30 days. The median survival of cells treated with a single viral injection was about 62 days. Cells treated with multiple virus injections exhibited >60% survival at 70 days. Cells treated with a single dose of producer cells exhibited >65% survival at 70 days.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Virology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention se rapporte à des cellules productrices qu'il est possible d'administrer à un patient porteur de cellules cancéreuses dans le but de détruire lesdites cellules cancéreuses ou d'éviter la croissance ou la propagation de la tumeur. Une telle cellule productrice comporte un virus oncolytique qui est apte à la réplication dans ladite cellule productrice. La cellule productrice n'est pas capable de survivre de manière prolongée dans le corps du patient. L'invention se rapporte également à des méthodes d'utilisation de telles cellules productrices pour traiter un patient porteur de cellules cancéreuses et à des méthodes de fabrication d'un médicament pouvant être utilisé dans ce type de traitement.
PCT/US1999/005466 1998-03-12 1999-03-12 Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite WO1999045783A1 (fr)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU30017/99A AU3001799A (en) 1998-03-12 1999-03-12 Producer cells for replication selective viruses in the treatment of malignancy
CA002323067A CA2323067A1 (fr) 1998-03-12 1999-03-12 Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite
EP99911363A EP1061806A4 (fr) 1998-03-12 1999-03-12 Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US7768198P 1998-03-12 1998-03-12
US60/077,681 1998-03-12

Publications (1)

Publication Number Publication Date
WO1999045783A1 true WO1999045783A1 (fr) 1999-09-16

Family

ID=22139477

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/005466 WO1999045783A1 (fr) 1998-03-12 1999-03-12 Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite

Country Status (4)

Country Link
EP (1) EP1061806A4 (fr)
AU (1) AU3001799A (fr)
CA (1) CA2323067A1 (fr)
WO (1) WO1999045783A1 (fr)

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001035970A1 (fr) * 1999-11-12 2001-05-25 Oncolytics Biotech Inc. Virus pour le traitement des troubles de la proliferation cellulaire
WO2001019380A3 (fr) * 1999-09-17 2001-11-22 Pro Virus Inc Virus oncolytique
WO2002043647A3 (fr) * 2000-12-01 2003-01-03 Univ Ottawa Virus oncolytique
EP1385466A2 (fr) * 2001-05-11 2004-02-04 Wellstat Biologics Corporation Traitement par virus oncolytique
WO2003090676A3 (fr) * 2002-04-26 2004-07-08 Wellstat Biologics Corp Traitement a base d'agents immunogenes mettant en oeuvre la plasmapherese ou l'exsanguinotransfusion
WO2005037321A1 (fr) * 2003-10-15 2005-04-28 The New Industry Research Organization Medicament de traitement anticancereux
WO2008009115A1 (fr) * 2006-07-18 2008-01-24 Ottawa Health Research Institute Cellules porteuses suicidaires disparates pour le ciblage tumoral de virus oncolytiques ubiquistes
US7361354B1 (en) 1999-11-25 2008-04-22 Viralytics Limited Methods for treating malignancies expressing ICAM-1 using coxsackie a group viruses
US7431932B2 (en) 2000-05-03 2008-10-07 Oncolytics Biotech Inc. Reovirus clearance of ras-mediated neoplastic cells from mixed cellular compositions
US7470426B1 (en) 1997-10-09 2008-12-30 Wellstat Biologics Corporation Treatment of neoplasms with viruses
US7485292B2 (en) 2002-12-18 2009-02-03 Viralytics Limited Method of treating a malignancy in a subject via direct picornaviral-mediated oncolysis
US7780962B2 (en) 1997-10-09 2010-08-24 Wellstat Biologics Corporation Treatment of neoplasms with RNA viruses
US8043612B2 (en) 1997-10-09 2011-10-25 Wellstat Biologics Corporation Infection and treatment of neoplasms with vesicular stomatitis virus
US8105578B2 (en) 1997-10-09 2012-01-31 Wellstat Biologics Corporation Treatment of neoplasms with viruses
US8147822B1 (en) 1999-09-17 2012-04-03 Wellstat Biologics Corporation Oncolytic virus
US8222036B2 (en) 2000-05-03 2012-07-17 Oncoltyics Biotech Inc. Oncolytic viruses as phenotyping agents for neoplasms
US8491884B2 (en) 2000-05-03 2013-07-23 Oncolytics Biotech Inc. Virus clearance of neoplastic cells from mixed cellular compositions
US20140154216A1 (en) * 2000-01-21 2014-06-05 Amgen Inc. Virus strains
US9101658B2 (en) 2005-09-30 2015-08-11 The Board Of Trustees Of The Leland Stanford Junior University Immune effector cells pre-infected with oncolytic virus
CN104958324A (zh) * 2015-05-29 2015-10-07 黄波 一种溶瘤病毒制剂及其制备方法
JP2015195741A (ja) * 2014-03-31 2015-11-09 国立大学法人愛媛大学 新規細胞、それを用いた抗腫瘍効果の誘導剤、癌の遺伝子治療用医薬および抗腫瘍効果の誘導方法
US10078725B2 (en) 2011-11-17 2018-09-18 Bayer Healthcare Llc Methods and techniques for collecting, reporting and managing ionizing radiation dose
JP2021507718A (ja) * 2017-12-29 2021-02-25 ジーンメディスン・カンパニー・リミテッド 遺伝子伝達用細胞シート
US11430558B2 (en) 2017-07-07 2022-08-30 Bayer Healthcare Llc System, method, and computer program product for peer exchange of data between injection systems

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5585096A (en) * 1994-06-23 1996-12-17 Georgetown University Replication-competent herpes simplex virus mediates destruction of neoplastic cells
US5601818A (en) * 1991-07-26 1997-02-11 University Of Rochester Cancer therapy utilizing malignant cells expressing HSV-TK

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0723460A4 (fr) * 1993-10-01 1998-09-30 Us Health Therapie genique concernant le systeme nerveux
EP1314430A1 (fr) * 1996-01-25 2003-05-28 The University Court Of The University Of Glasgow Traitement de cancer non neuronal à l'aide d'un mutant de hsv

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5601818A (en) * 1991-07-26 1997-02-11 University Of Rochester Cancer therapy utilizing malignant cells expressing HSV-TK
US5585096A (en) * 1994-06-23 1996-12-17 Georgetown University Replication-competent herpes simplex virus mediates destruction of neoplastic cells

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of EP1061806A4 *

Cited By (58)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7780962B2 (en) 1997-10-09 2010-08-24 Wellstat Biologics Corporation Treatment of neoplasms with RNA viruses
US8043612B2 (en) 1997-10-09 2011-10-25 Wellstat Biologics Corporation Infection and treatment of neoplasms with vesicular stomatitis virus
US7470426B1 (en) 1997-10-09 2008-12-30 Wellstat Biologics Corporation Treatment of neoplasms with viruses
US8105578B2 (en) 1997-10-09 2012-01-31 Wellstat Biologics Corporation Treatment of neoplasms with viruses
AU782402B2 (en) * 1999-09-17 2005-07-28 Wellstat Biologics Corporation Oncolytic virus
WO2001019380A3 (fr) * 1999-09-17 2001-11-22 Pro Virus Inc Virus oncolytique
US8147822B1 (en) 1999-09-17 2012-04-03 Wellstat Biologics Corporation Oncolytic virus
JP2004525855A (ja) * 1999-09-17 2004-08-26 ウェルスタット バイオロジクス コーポレイション 腫瘍崩壊性ウイルス
US7252817B2 (en) 1999-11-12 2007-08-07 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7799329B2 (en) 1999-11-12 2010-09-21 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
AU782020B2 (en) * 1999-11-12 2005-06-30 Oncolytics Biotech, Inc. Viruses for the treatment of cellular proliferative disorders
US8491886B2 (en) 1999-11-12 2013-07-23 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7582289B2 (en) 1999-11-12 2009-09-01 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7731951B2 (en) 1999-11-12 2010-06-08 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US8080241B2 (en) 1999-11-12 2011-12-20 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US6649157B2 (en) 1999-11-12 2003-11-18 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US6596268B1 (en) 1999-11-12 2003-07-22 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
US7964186B2 (en) 1999-11-12 2011-06-21 Oncolytics Biotech Inc. Viruses for the treatment of cellular proliferative disorders
WO2001035970A1 (fr) * 1999-11-12 2001-05-25 Oncolytics Biotech Inc. Virus pour le traitement des troubles de la proliferation cellulaire
US7361354B1 (en) 1999-11-25 2008-04-22 Viralytics Limited Methods for treating malignancies expressing ICAM-1 using coxsackie a group viruses
US8722036B2 (en) 1999-11-25 2014-05-13 Viralytics Limited Methods for treating malignancies using coxsackieviruses
US10301600B2 (en) 2000-01-21 2019-05-28 Biovex Limited Virus strains
US20140154216A1 (en) * 2000-01-21 2014-06-05 Amgen Inc. Virus strains
US8491884B2 (en) 2000-05-03 2013-07-23 Oncolytics Biotech Inc. Virus clearance of neoplastic cells from mixed cellular compositions
US8222036B2 (en) 2000-05-03 2012-07-17 Oncoltyics Biotech Inc. Oncolytic viruses as phenotyping agents for neoplasms
US9365823B2 (en) 2000-05-03 2016-06-14 Oncolytics Biotech Inc. Virus clearance of neoplastic cells from mixed cellular compositions
US8147845B2 (en) 2000-05-03 2012-04-03 Oncolytics Biotech Inc. Reovirus clearance of ras-mediated neoplastic cells from mixed cellular compositions
US7431932B2 (en) 2000-05-03 2008-10-07 Oncolytics Biotech Inc. Reovirus clearance of ras-mediated neoplastic cells from mixed cellular compositions
US8512712B2 (en) 2000-05-03 2013-08-20 Oncolytics Biotech Inc. Reovirus clearance of ras-mediated neoplastic cells from mixed cellular compositions
US7727534B2 (en) 2000-05-03 2010-06-01 Oncolytics Biotech Inc. Reovirus clearance of ras-mediated neoplastic cells from mixed cellular compositions
WO2002043647A3 (fr) * 2000-12-01 2003-01-03 Univ Ottawa Virus oncolytique
EP1385466A2 (fr) * 2001-05-11 2004-02-04 Wellstat Biologics Corporation Traitement par virus oncolytique
US7595042B2 (en) 2001-05-11 2009-09-29 Wellstat Biologics Corporation Oncolytic virus therapy
US8137663B2 (en) 2001-05-11 2012-03-20 Wellstat Biologics Corporation Oncolytic virus therapy
EP1385466A4 (fr) * 2001-05-11 2005-10-12 Wellstat Biologics Corp Traitement par virus oncolytique
WO2003090676A3 (fr) * 2002-04-26 2004-07-08 Wellstat Biologics Corp Traitement a base d'agents immunogenes mettant en oeuvre la plasmapherese ou l'exsanguinotransfusion
US7485292B2 (en) 2002-12-18 2009-02-03 Viralytics Limited Method of treating a malignancy in a subject via direct picornaviral-mediated oncolysis
EP1676589A1 (fr) * 2003-10-15 2006-07-05 The New Industry Research Organization Medicament de traitement anticancereux
EP1676590A1 (fr) * 2003-10-15 2006-07-05 The New Industry Research Organization Médicament de traitement anticancereux
EP1676590A4 (fr) * 2003-10-15 2007-08-15 New Ind Res Organization Médicament de traitement anticancereux
EP1676589A4 (fr) * 2003-10-15 2007-08-15 New Ind Res Organization Medicament de traitement anticancereux
WO2005037322A1 (fr) * 2003-10-15 2005-04-28 The New Industry Research Organization Médicament de traitement anticancéreux
WO2005037321A1 (fr) * 2003-10-15 2005-04-28 The New Industry Research Organization Medicament de traitement anticancereux
CN100438920C (zh) * 2003-10-15 2008-12-03 财团法人新产业创造研究机构 癌基因治疗药物
KR100798996B1 (ko) * 2003-10-15 2008-01-28 자이단호우진 신산교소우조우 겐큐키코 암 유전자 치료약
KR100842658B1 (ko) * 2003-10-15 2008-06-30 자이단호우진 신산교소우조우 겐큐키코 암 유전자 치료약
CN100488567C (zh) * 2003-10-15 2009-05-20 财团法人新产业创造研究机构 癌基因治疗药物
US9101658B2 (en) 2005-09-30 2015-08-11 The Board Of Trustees Of The Leland Stanford Junior University Immune effector cells pre-infected with oncolytic virus
US10064893B2 (en) 2005-09-30 2018-09-04 The Board Of Trustees Of The Leland Stanford Junior University Immune effector cells pre-infected with oncolytic virus
WO2008009115A1 (fr) * 2006-07-18 2008-01-24 Ottawa Health Research Institute Cellules porteuses suicidaires disparates pour le ciblage tumoral de virus oncolytiques ubiquistes
US10078725B2 (en) 2011-11-17 2018-09-18 Bayer Healthcare Llc Methods and techniques for collecting, reporting and managing ionizing radiation dose
JP2015195741A (ja) * 2014-03-31 2015-11-09 国立大学法人愛媛大学 新規細胞、それを用いた抗腫瘍効果の誘導剤、癌の遺伝子治療用医薬および抗腫瘍効果の誘導方法
CN104958324A (zh) * 2015-05-29 2015-10-07 黄波 一种溶瘤病毒制剂及其制备方法
US10548853B2 (en) 2015-05-29 2020-02-04 Hubei Soundny Biotechnology Co., Ltd. Oncolytic virus formulation and preparation method thereof
US11430558B2 (en) 2017-07-07 2022-08-30 Bayer Healthcare Llc System, method, and computer program product for peer exchange of data between injection systems
US11984213B2 (en) 2017-07-07 2024-05-14 Bayer Healthcare Llc System, method, and computer program product for peer exchange of data between injection systems
JP2021507718A (ja) * 2017-12-29 2021-02-25 ジーンメディスン・カンパニー・リミテッド 遺伝子伝達用細胞シート
JP2022095954A (ja) * 2017-12-29 2022-06-28 ジーンメディスン・カンパニー・リミテッド 遺伝子伝達用細胞シート

Also Published As

Publication number Publication date
EP1061806A4 (fr) 2001-09-12
CA2323067A1 (fr) 1999-09-16
EP1061806A1 (fr) 2000-12-27
AU3001799A (en) 1999-09-27

Similar Documents

Publication Publication Date Title
WO1999045783A1 (fr) Cellules productrices pour virus aptes a la replication utilisees dans le traitement de la malignite
Coukos et al. Use of carrier cells to deliver a replication-selective herpes simplex virus-1 mutant for the intraperitoneal therapy of epithelial ovarian cancer
Fountzilas et al. Oncolytic virotherapy, updates and future directions
Watanabe et al. Oncolytic virotherapy by HSV
Wakimoto et al. Human glioblastoma–derived cancer stem cells: Establishment of invasive glioma models and treatment with oncolytic herpes simplex virus vectors
Blank et al. Replication-selective herpes simplex virus type 1 mutant therapy of cervical cancer is enhanced by low-dose radiation
US6428968B1 (en) Combined therapy with a chemotherapeutic agent and an oncolytic virus for killing tumor cells in a subject
Ning et al. Oncolytic herpes simplex virus-based strategies: toward a breakthrough in glioblastoma therapy
Campadelli‐Fiume et al. Rethinking herpes simplex virus: the way to oncolytic agents
Lun et al. Myxoma virus virotherapy for glioma in immunocompetent animal models: optimizing administration routes and synergy with rapamycin
Cinatl Jr et al. Potent oncolytic activity of multimutated herpes simplex virus G207 in combination with vincristine against human rhabdomyosarcoma
Adusumilli et al. Imaging and therapy of malignant pleural mesothelioma using replication‐competent herpes simplex viruses
Coukos et al. Multi-attenuated herpes simplex virus-1 mutant G207 exerts cytotoxicity against epithelial ovarian cancer but not normal mesothelium and is suitable for intraperitoneal oncolytic therapy
Delwar et al. Oncolytic virotherapy blockade by microglia and macrophages requires STAT1/3
Jefferson et al. The mechanisms of genetically modified vaccinia viruses for the treatment of cancer
Malhotra et al. Use of an oncolytic virus secreting GM-CSF as combined oncolytic and immunotherapy for treatment of colorectal and hepatic adenocarcinomas
Wong et al. Effective intravenous therapy of murine pulmonary metastases with an oncolytic herpes virus expressing interleukin 12
WO2000054795A1 (fr) Therapie combinee consistant a administrer un agent chimiotherapeutique et un virus oncolytique pour tuer des cellules cancereuses chez un sujet
Xu et al. Oncolytic vaccinia virus and cancer immunotherapy
Todo et al. Corticosteroid administration does not affect viral oncolytic activity, but inhibits antitumor immunity in replicationcompetent herpes simplex virus tumor therapy
EP1173598A1 (fr) Facteur derive de l'herpes simplex virus de type 1(hsv-1) utile pour inhiber de maniere selective des cellules malignes et procedes d'utilisation de ce dernier pour traiter des cancers et pour exprimer des caracteristiques desirees dans des cellules malignes et non malignes de mammiferes
Hu et al. Advances in oncolytic herpes simplex virus and adenovirus therapy for recurrent glioma
US20010006633A1 (en) Adenovirus-chemotherapeutic combination for treating cancer
EP0862445B1 (fr) Combinaison de virus herpes simplex avec la chimithérapie pour le traitement du cancer
Jiang et al. Temozolomide resistant human brain tumor stem cells are susceptible to recombinant vesicular stomatitis virus and double-deleted Vaccinia virus in vitro

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
WWE Wipo information: entry into national phase

Ref document number: 09649372

Country of ref document: US

ENP Entry into the national phase

Ref document number: 2323067

Country of ref document: CA

Ref country code: CA

Ref document number: 2323067

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 30017/99

Country of ref document: AU

WWE Wipo information: entry into national phase

Ref document number: 1999911363

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1999911363

Country of ref document: EP

WWW Wipo information: withdrawn in national office

Ref document number: 1999911363

Country of ref document: EP

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载