[go: up one dir, main page]

WO1997005898A1 - Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer - Google Patents

Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer Download PDF

Info

Publication number
WO1997005898A1
WO1997005898A1 PCT/US1996/012837 US9612837W WO9705898A1 WO 1997005898 A1 WO1997005898 A1 WO 1997005898A1 US 9612837 W US9612837 W US 9612837W WO 9705898 A1 WO9705898 A1 WO 9705898A1
Authority
WO
WIPO (PCT)
Prior art keywords
epithelium
ductal
ductal epithelium
csf
vector
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/US1996/012837
Other languages
English (en)
Inventor
Saraswati Vaidyanathan Sukumar
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Johns Hopkins University
Original Assignee
Johns Hopkins University
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 Johns Hopkins University filed Critical Johns Hopkins University
Priority to AU68959/96A priority Critical patent/AU6895996A/en
Publication of WO1997005898A1 publication Critical patent/WO1997005898A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/045—Hydroxy compounds, e.g. alcohols; Salts thereof, e.g. alcoholates
    • 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
    • A61K35/768—Oncolytic viruses not provided for in groups A61K35/761 - A61K35/766
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19—Cytokines; Lymphokines; Interferons
    • A61K38/193—Colony stimulating factors [CSF]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/45—Transferases (2)
    • 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
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24132—Use of virus as therapeutic agent, other than vaccine, e.g. as cytolytic agent

Definitions

  • the present invention relates to the use of an epithelium-destroying agent to destroy the epithelium of an exocrine gland, particularly the mammary gland, in the prophylactic and therapeutic treatment of disease, in particular cancer.
  • Exocrine glands are glands that release a secretion external to or at the surface of an organ by means of a canal or duct.
  • exocrine glands include, among others, the mammary glands, prostate, liver, gall bladder, pancreas, kidneys, sweat glands, and salivary glands.
  • Cancers of exocrine glands pose a major health problem, frequently resulting in death.
  • cancers of the breast and prostate are among the leading causes of death among women and men, respectively.
  • the mature human breast comprises from six to nine major ducts, which emanate from the nipple, serially branch into ducts and terminate in lobuloalveolar structures (Russo et al., Lab. Invest. 62(3) : 244-278 (1990) ) .
  • This branching network of ducts is composed of epithelial cells in a supporting matrix of connective tissue and endothelial cells.
  • Tissues removed from the human female breast during surgery and autopsy have been examined in numerous studies directed to the nature and site of origin of neoplastic growth. Subgross sampling and histological confirmation have enabled pathological characterization of entire breasts, leading to the postulation of the existence of four major possible sites of origin of mammary carcinomas, namely ducts, terminal ducts, ductules, and acini (Russo et al. , supra ) . Ductal origin is supported by the presence of more extensive epithelial proliferations, which are presumed to be preneoplastic, in surgically removed cancerous breasts as compared to nonmalignant breasts removed during autopsies (Russo et al. , supra) .
  • an object of the present invention to provide a method of locally treating an exocrine gland prophylactically for disease. It is another object of the present invention to provide a method of locally treating an exocrine gland, in particular the mammary gland, prophylactically for cancer. Another object of the present invention is to provide a method of locally treating an exocrine gland therapeutically for disease. Yet another object of the present invention is to provide a method of locally treating a mammary gland therapeutically for cancer. Still yet another object of the present invention is to provide a method of locally treating a mammary gland both therapeutically and prophylactically for cancer.
  • the present invention provides prophylactic and therapeutic methods of treating the ductal epithelium of an exocrine gland, in particular the mammary gland, for disease, in particular cancer.
  • the method comprises contacting the ductal epithelium of the exocrine gland, in particular the mammary gland, with an epithelial- destroying agent.
  • the ductal epithelium is preferably contacted with the agent by ductal cannulation.
  • the epithelium-destroying agent is preferably a vector comprising a thymidine kinase gene, which is used in combination with ganciclovir (GCV) , which can be systemically administered.
  • GCV ganciclovir
  • Another preferred epithelium- destroying agent is a vector comprising a hypoxanthine phosphoribosyl transferase (HPRT) gene, which is used in combination with hypoxanthine aminopterin thymidine (HAT) nucleotide, which can be systemically administered.
  • HPRT hypoxanthine phosphoribosyl transferase
  • HAT hypoxanthine aminopterin thymidine
  • a vector comprising a gene, which, upon transformation of a cell of the ductal epithelium and expression therein, induces apoptosis or death of the transformed cell.
  • a preferred apoptosis-inducing gene is bclxs .
  • Other preferred epithelial-destroying agents include a cytolytic virus, such as Vaccinia virus, and ethanol.
  • the preceding methods can additionally comprise contacting the ductal epithelium with a cytokine or hematopoietic growth factor, such as GM-CSF.
  • a method of treating the ductal epithelium of a mammary gland both therapeutically and prophylactically for cancer comprises treating the mammary gland therapeutically by surgery, radiation and/or chemotherapy and contacting the ductal epithelium of the mammary gland, either concomitantly or subsequently, with an epithelium-destroying agent, which does not specifically target cancerous cells.
  • the combined therapeutic/prophylactic method can additionally comprise contacting the ductal epithelium with a cytokine or hematopoietic growth factor, such as GM-CSF.
  • Figure 1 is a graph of the ratio of cell no. in the presence of GCV over cell no. in the absence of GCV (Cell No. + GCV/Cell No. - GCV) versus viral multiplicity of infection (viral MOI) , which shows the effect of the addition of 10 ⁇ g/ml GCV on NMU68 and RBA rat tumor cell lines 6 hrs after transduction with adenoviral-J ⁇ erpes simplex thymidine kinase (AdHS-tk) at titers of 0, 100, 500 and 1,000 moi.
  • AdHS-tk simplex thymidine kinase
  • the present invention is based on the observation that the overwhelming majority of breast cancers arise from epithelial cells, particularly those epithelial cells which line the ducts of the mammary gland and are collectively referred to as the ductal epithelium. The present invention is also based on the exocrine nature of the mammary gland.
  • the central canal or duct could provide a means of directly accessing the ductal epithelium for localized prophylactic and therapeutic treatment of cancer. Based on these observations, the prophylactic and therapeutic methods of the present invention were developed.
  • the prophylactic method of the present invention is a method of treating the ductal epithelium of an exocrine gland prophylactically for a disease that affects the ductal epithelium of the exocrine gland.
  • the method comprises contacting, preferably by ductal cannulation, the ductal epithelium of the exocrine gland with an epithelium-destroying agent so as to destroy cells of the ductal epithelium affected by the disease.
  • the ductal epithelium of a mammary gland is treated prophylactically for cancer so as to inhibit the formation of cancer of ductal epithelial origin.
  • the method comprises contacting, preferably by ductal cannulation, the ductal epithelium of the mammary gland with an epithelium-destroying agent.
  • the agent preferably is a vector comprising a thymidine kinase gene, such as a Herpes simplex thymidine kinase gene, and ganciclovir, a vector comprising a HPRT gene and HAT nucleotide, a cytolytic virus, such as a Vaccinia virus, or ethanol.
  • the method can additionally comprise contacting the ductal epithelium with a cytokine or hematopoietic growth factor, such as GM-CSF.
  • the ductal epithelium of an exocrine gland such as a mammary gland
  • the method comprises contacting, preferably by ductal cannulation, the ductal epithelium of the exocrine gland with an epithelium-destroying agent to destroy less than all of the ductal epithelium so as to inhibit the formation of cancer of ductal epithelial origin.
  • an epithelium-destroying agent to destroy less than all of the ductal epithelium so as to inhibit the formation of cancer of ductal epithelial origin.
  • up to about 70%, 80%, 85%, 90% or 95% of the ductal epithelium is destroyed.
  • the epithelium-destroying agent is preferably a vector comprising a thymidine kinase gene, such as that from Herpes simplex, and ganciclovir, which can be brought into contact with the ductal epithelium by any suitable means, preferably by ductal cannulation or by systemic administration, a vector comprising a HPRT gene and HAT nucleotide, which can be brought into contact with the ductal epithelium by any suitable means, preferably by ductal cannulation or by systemic administration, a vector comprising a gene, which upon transformation of a cell of the ductal epithelium and expression therein, induces apoptosis or death of the transformed cell, such as bclxs, ethanol, or a cytolytic virus, such as Vaccinia virus.
  • a thymidine kinase gene such as that from Herpes simplex, and ganciclovir
  • the above method can additionally comprise the administration of a cytokine or hematopoietic growth factor, such as GM-CSF.
  • GM-CSF can be brought into contact with the ductal epithelium of the mammary gland by any suitable means, such as by ductal cannulation of GM-CSF or a vector comprising a gene encoding GM-CSF, in which case the vector can be the same vector as the one encoding the thymidine kinase, HPRT or apopotosis- inducing gene, or it can be systemically administered.
  • This embodiment of the prophylactic method can be used to treat any exocrine gland. However, it is particularly useful in the treatment of the mammary gland.
  • the above-described prophylactic method of treating a mammary gland is particularly useful in treating a mammary gland in a mammal at risk for developing breast cancer.
  • the mammary gland can be characterized as one that has never had a tumor, one that had a tumor previously but the tumor is no longer detectable due to other prior therapeutic treatment, or one that has an incipient or occult tumor, preneoplasia or ductal hyperplasia. Normally, hyperplasias and incipient and occult tumors are not detectable by means of physical examination or radiography.
  • the prophylactic method will find use in cases where there is reason to take some prophylactic measures, such as when there are known inherited factors predisposing to cancers, where there are suspicious lesions present in a breast with the potential for developing into a malignancy, where there has been exposure to carcinogenic agents in the environment, where age predisposes to a cancer, where cancer of another gland, e.g., the mammary gland of the contralateral breast, suggests a propensity for developing cancer, or where there is a fear or suspicion of metastasis.
  • some prophylactic measures such as when there are known inherited factors predisposing to cancers, where there are suspicious lesions present in a breast with the potential for developing into a malignancy, where there has been exposure to carcinogenic agents in the environment, where age predisposes to a cancer, where cancer of another gland, e.g., the mammary gland of the contralateral breast, suggests a propensity for developing cancer, or where there is a fear or suspicion of metasta
  • the therapeutic method of the present invention is a method of treating the ductal epithelium of an exocrine gland therapeutically for a disease that affects the ductal epithelium of the exocrine gland.
  • the method comprises contacting, preferably by ductal cannulation, the ductal epithelium of the exocrine gland with an epithelium-destroying agent so as to destroy cells of the ductal epithelium affected by the disease.
  • the ductal epithelium of a mammary gland is locally treated therapeutically for cancer so as to destroy cancerous and noncancerous cells of the ductal epithelium and inhibit the spread of cancer.
  • the method comprises contacting, preferably by ductal cannulation, the ductal epithelium of the mammary gland with an epithelium-destroying agent, which need not, and preferably does not, specifically target cancerous cells.
  • the agent preferably is a vector comprising a thymidine kinase gene, such as a Herpes simplex thymidine kinase gene, and ganciclovir, a vector comprising a HPRT gene and HAT nucleotide, a cytolytic virus, such as a Vaccinia virus, or ethanol.
  • the method can additionally comprise contacting the ductal epithelium with a cytokine or hematopoietic growth factor, such as GM-CSF.
  • the epithelial-destroying agent should destroy all of the diseased or malignant epithelium.
  • the ductal epithelium immediately surrounding the diseased/malignant epithelium also preferably should be destroyed.
  • the present invention also provides a method of treating the ductal epithelium of a mammary gland both therapeutically and prophylactically for cancer.
  • the method comprises treating the mammary gland therapeutically with any given therapeutic method, such as those currently known and used in the art. Examples of such methods include surgical removal of the cancerous tissue, radiation therapy and chemotherapy.
  • the method further comprises contacting, either concomitantly with or subsequently to the therapeutic treatment, the ductal epithelium of the mammary gland, e.g., by ductal cannulation, with an epithelium-destroying agent, which preferably does not specifically target cancerous cells, so as to destroy any remaining cancerous cells and noncancerous cells and inhibit the spread of cancer.
  • the epithelium-destroying agent is preferably a vector comprising a thymidine kinase gene, such as a Herpes simplex thymidine kinase gene, combined with ganciclovir, a vector comprising a HPRT gene combined with HAT nucleotide, a cytolytic virus, such as a Vaccinia virus, or ethanol.
  • the method can additionally comprise contacting the ductal epithelium with a cytokine or hematopoietic growth factor, such as GM-CSF.
  • the methods of the present invention can be used to treat the ductal epithelium of any exocrine gland.
  • exocrine glands other than the mammary gland, which can be treated with the present inventive methods include, among others, the prostate, liver, gall bladder, pancreas, kidneys, sweat glands, and salivary glands. The methods are especially useful in the prophylactic and therapeutic treatment of the ductal epithelium of mammary glands.
  • the methods can be used to treat an exocrine gland for any disease that affects the exocrine ductal epithelium.
  • the methods are particularly useful in the treatment of cancer, including the stages of hyperplasia, adenoma, carcinoma in situ, and carcinoma, of ductal epithelial origin.
  • Any method can be used to destroy the ductal epithelium. It is preferred, however, that the destruction is limited to the ductal epithelium or a part thereof.
  • ductal cannulation is any method of contacting the ductal epithelium.
  • ductal cannulation is used.
  • any duct or lobule can be cannulated, it is preferred that the central canal or duct be cannulated.
  • Ductal cannulation also enables direct injection of a tumor mass, if desired.
  • Any epithelium-destroying agent can be used to destroy the ductal epithelium of an exocrine gland.
  • the agent preferably should not destroy cells other than cells of the ductal epithelium and preferably should not result in side effects, the adversity of which outweigh the benefits of destruction of the ductal epithelium.
  • the methods be used to destroy completely the ductal epithelium of an exocrine gland in the prophylactic/therapeutic treatment of a given disease, wherein the complete destruction of the ductal epithelium, in and of itself, would result in death of the mammal so treated.
  • agents that can be used in the context of the prophylactic and therapeutic methods of the present invention include cytotoxic agents .
  • cytotoxic agent known in the art and suitable for contacting the ductal epithelium of an exocrine gland of a mammal can be used.
  • cyotoxic agents and their prodrugs include genistein, okadaic acid, 1- ⁇ -D- arabinofuranosyl-cytosine, arabinofuranosyl-5-aza- cytosine, cisplatin, carboplatin, actinomycin D, asparaginase, bis-chloro-ethyl-nitroso-urea, bleomycin, chlorambucil, cyclohexyl-chloro-ethyl-nitroso-urea, cytosine arabinoside, daunomycin, etoposide, hydroxyurea, melphalan, mercaptopurine, mitomycin C, nitrogen mustard, procarbazine, teniposide, thioguanine, thiot
  • the agent is locally administered, especially if administration of the agent is accompanied by toxic side effects. Otherwise, the agent can be administered by any suitable route, such as systemic administration.
  • a cytolytic virus also can be used as an agent. Any cytolytic virus can be used as long as the organism mounts a rapid immunological response to it such that the virus cannot cause disease if it escapes the ductal epithelium. Examples of cytolytic viruses include Vaccinia viruses and Sindbis viruses, which can also be used as vectors. Preferably, a Vaccinia virus is used.
  • Vaccinia infectious particles enter and lyse the breast epithelial cells, yet stromal immunity destroys the particles as soon as they leave the ductile tree of the exocrine gland, thereby preventing cytolysis beyond the ductal epithelium.
  • the advantages of Vaccinia administration are that it eliminates the need for high titer virus, the need to induce cell division in the breast, and the need to administer a drug to effect cell death.
  • a vector comprising a suicide gene also can be used as an agent, in conjunction with an agent that destroys the ductal epithelium of an exocrine gland.
  • the vector comprising a suicide gene upon transformation of a cell of the ductal epithelium and expression therein, renders the transformed cell sensitive to the epithelium- destroying agent, increases the sensitivity of the transformed cell to the agent, converts the agent from a prodrug to an active drug, activates the conversion of the agent from a prodrug to an active drug, enhances the effect of the agent or, itself, produces a protein that is cytotoxic.
  • a preferred suicide gene for use in the present inventive methods is the one described above, i.e., a thymidine kinase, such as the one from Herpes simplex, which phosphorylates GCV, which, in turn, inhibits DNA replication.
  • a thymidine kinase such as the one from Herpes simplex
  • Another example of a suicide gene is cytosine deaminase, which is used in conjunction with 5-fluorocytosine . If the vector comprising the suicide gene is administered locally to the ducts, the cytotoxic agent or precursor can be administered systemically, since only transfected cells will be affected.
  • the bystander effect i.e., the death of neighboring uninfected cells, presumably due to transfer of toxic byproducts through gap junctions between cells in the same compartment, obviates the need for every cell in the ductal epithelium, which is to be destroyed, to be infected.
  • sufficient time must be allowed between contacting the ductal epithelium with the suicide gene and the prodrug, for example, to achieve efficient killing of the breast epithelial cells.
  • a vector comprising an apoptosis-inducing gene also can be used as an agent that destroys the ductal epithelium of an exocrine gland (Vaux, Cell 76: 777-779 (1994)) .
  • apoptosis-inducing genes include ced genes, myc genes (overexpressed) , the bclxs gene, the bax gene, and the bak gene.
  • the apoptosis-inducing gene causes death of transfected cells, i.e., by inducing programmed cell death.
  • the bclxs gene, bax gene, or bak gene can be used to inhibit bcl -2 or bcl -x , leading to apoptosis.
  • a vector comprising an apoptosis-inducing gene can be used in combination with an agent that inactivates apoptosis inhibitors such as bcl -z, p35, IAP, NAIP, DAD1 and A20 proteins .
  • Suicide and apopotosis genes can be administered by way of a viral vector, such as an adenoviral or retroviral vector.
  • a viral vector such as an adenoviral or retroviral vector.
  • Adenoviral vectors enable the generation of high titer recombinant viruses (10 11 /ml) and the efficient transduction of postmitotic cells because adenoviral DNA exists as an episome in the nucleus (Verma, Molecular Medicine 1: 2-3 (1994) ) .
  • the gene can be under the transcriptional regulation of a Rous sarcoma viral promoter. Alternatively, it can be under the control of an epithelial tissue-specific or cell-specific promoter.
  • Uptake of recombinant virus can be facilitated by pretreatment or simultaneous treatment with polybrene or, for example, in the case of a retrovirus, attachment of the functional fragment of an antibody to the viral particle.
  • the apoptosis gene or suicide gene can be present in a recombinant microorganism, which will express the gene.
  • a particularly preferred microorganism for this purpose is the bacterium Listeria monocytogenes .
  • liposomes complexes between polypeptide ligands for receptors on mammary ductal epithelial cells, including complexes of antibodies and functional fragments thereof, and plasmids can be used (Mulligan, Science 260: 926-931 (1993)) .
  • Epithelial cell-specific promoters such as whey acidic protein ( wap)
  • wap whey acidic protein
  • Use can also be made of wild-type tumor suppressor genes, such as p53 or Mcs-1 (rat) , homeobox genes expressed in normal cells but not in cancerous cells, and the maspin gene.
  • the ductal epithelium can be contacted with an agent to effect the scavenging of epithelial cells destroyed in accordance with the present invention, e.g., a cytokine/growth factor.
  • a cytokine/growth factor include GM-CSF, G-CSF, IL-2, IL- 4, IL-6, IL-7, hCG, TNF- ⁇ , INF- ⁇ and INF- ⁇ .
  • Such factors can be contacted with the ductal epithelium directly or by expression of a vector comprising a gene encoding the factor, in which case the vector can be the same one that comprises a suicide gene, for example.
  • the factors stimulate a potent, long-lasting, and specific cell immunity, requiring both CD4 and CD8 cells.
  • the immune response is designed to scavenge destroyed ductal epithelial cells by generating autoimmunity towards epithelial cell antigens.
  • the ductal epithelium is preferably contacted with the agent by introduction of the agent through the central canal or duct of the exocrine ductal epithelium, such as by ductal cannulation.
  • ductal cannulation enables intratumoral injection.
  • the methods of the present invention can be combined with other methods of prophylactic and therapeutic treatment in addition to those cited above, such as methods that target destruction of cancer cells, e.g., by targeting of cell-surface markers, receptor ligands, e.g., ligands to gastrin-releasing peptide-like receptors, tumor-associated antigens, e.g., the 57 kD cytokeratin or the antigen recognized by the monoclonal antibody GB24, the extracellular matrix glycoprotein tamascin, antisense oncogenes such as c-fos, homeobox genes that are expressed in cancer cells but not normal cells, tumor-infiltrating lymphocytes that express cytokines, RGD-containing peptides and proteins, which are administered following surgery, lipophilic drug- containing liposomes to which are covalently conjugated monoclonal antibodies for targeting to cancer cells, low fat diet, moderate physical exercise and hormonal modulation.
  • methods that target destruction of cancer cells e.g.
  • anti-testosterone agents can be used as well as an inhibitor of cell proliferation produced by prostatic stromal cells and C- CAM, an epithelial cell adhesion molecule.
  • the following examples are presented to illustrate the present invention, not to limit its scope.
  • the examples make use of the rat mammary tumor model, which has been deemed an appropriate experimental model for understanding breast cancer in humans (Sukumar et al. , Mutation Res. 333 (1-2) : 37-44 (1995); Russo et al. , supra) .
  • 90-100% of female rats develop mammary tumors in this model when they are administered the carcinogen NMU at 55 days of age (Sukumar, Cancer Cells 4: 199-204 (1990) ) .
  • This example demonstrates the successful delivery of virus and other agents into the mammary ductile tree by a single injection through the teat.
  • ADV/CMV- ⁇ -gal (from Dr. William Burns, Johns Hopkins University) is an adenoviral 5 vector constructed with a ⁇ -galactosidase gene controlled by a cytomegaloviral promoter. It was delivered into the mammary gland by injection of a viral suspension in 20 ⁇ l of 0.2% trypan blue in Tris buffer through the teat of a rat. The nipple was extruded, and the sphincter removed by excising the nipple. In the rat, the muscle prevents fluid from regurgitating into the breast and had to be excised in order to visualize the ductal opening and administer the agent. Trypan blue was used as a tracking dye to ensure correct delivery to the ductile tree. Injection about 30 days postpartum resulted in the mammary epithelial tree being clearly visible. Ethyl alcohol (70%) was also successfully delivered by a single injection through the teat. Exampl e 2
  • adenovirus can efficiently transduce human mammary epithelial cells in vitro .
  • ADV/CMV- ⁇ -gal was used to transduce HBL100 mammary epithelial cells in vi tro .
  • the ⁇ -gal enzyme in this construct contains a nuclear localization signal and results in dense nuclear staining.
  • HBL100 cells IO 2
  • IO 2 HBL100 cells
  • This example demonstrates that infection with an AdHS-tk construct followed by GCV treatment effectively kills mammary tumor cells in vitro .
  • the mammary glands (6 on each side) of six virgin 50 day old Sprague Dawley rats were injected with AdHS-tk on the left side and trypan blue on the right side or left untreated.
  • One rat remained completely untreated with the virus or GCV and served as a positive control for NMU-induced tumorigenesis.
  • rats were given an intramuscular injection of 5 ⁇ g estradiol valerate and were anesthetized with an isofluorane/0 2 mixture.
  • the nipples were cannulated with a 33 gauge needle.
  • AdHS-tk diluted in trypan blue carrier (1 mM MgCl 2 , 20 ⁇ g/ml polybrene in 10% glycerol, 0.4% trypan blue in saline) at a concentration of 5 x IO 7 particles/ ⁇ l were injected into the duct.
  • Carrier control mammary glands received 20 ⁇ l of trypan blue carrier alone.
  • An animal was considered treated when at least three glands were successfully injected with trypan blue and another three glands were successfully injected with AdHS-tk. The remaining glands were left untreated. Twelve hrs later, the rats were injected with 125 mg/kg body weight GCV twice daily for three days.
  • the rats were then given a second intramuscular injection of 5 ⁇ g estradiol valerate and intraperitoneal injections of 100 ⁇ g/kg body weight GCV once daily for three days.
  • the rats were given an intravenous injection of NMU dissolved in 0.05% acetic acid (Ash Stevens, CO; 50 mg NMU/kg body weight) and were subsequently monitored for general health and the appearance of tumors at weekly intervals for 8 months.
  • NMU dissolved in 0.05% acetic acid
  • This example demonstrates the efficient transfection of mammary epithelial cells in vivo.
  • Lytic Vaccinia virus (10 Vaccinia-HA, which also carries lacZ, in 20 ⁇ l 0.2% trypan blue) was injected into the mammary glands through the teat of 45 day old virgin rats. Contralateral control glands were injected with 0.2% trypan blue. The glands were excised after 3 days. Frozen mammary gland sections were stained with X- gal and counterstained with eosin. When Vaccinia-HA was injected via the rat teat, it was able to infect the epithelial cells. At 3 days post-infection, the X-gal staining was confined primarily to the epithelial cells.
  • This example demonstrates the cytotoxicity of Vaccinia/HA on HBL100 cells in vitro .
  • HBL100 cells (from ATCC, Rockville, MD) were plated in DME:F12 medium (50% Dulbecco's Modified Eagles Medium: 50%Ham's F12 Supplement) containing 10% fetal bovine serum and 10 ⁇ g/ml insulin at a density of 5 x 10 cells/well and incubated at 37°C overnight.
  • laccinia/HA at concentrations of 0 moi, 0.1 moi, or 1.0 moi, was added to the culture medium, and the cells were incubated at 37°C for at least 3 days. More than 90% of the cells were dead within 72 hrs of infection at 0.1 moi.
  • This example shows the death of rat mammary tumor cells in culture by infection with Vaccinia/HA .
  • Vaccinia virus engineered to express ⁇ -galactosidase and hemagglutinin genes was added to the culture medium at concentrations of 0 moi, 0.1 moi, or 1.0 moi and incubated at 37°C for at least 3 days. Up to 90% of the cells were lysed within 72 hours of injection at 1.0 moi.
  • This example demonstrates the destruction of mammary epithelium by transfection with Vaccinia/HA in vivo .
  • Vaccinia/HA in 20 ⁇ l 0.2% trypan blue (tracking dye) .
  • Contralateral control glands were injected with 0.2% trypan blue. The glands were excised after 3 days, fixed in chloroform:methanol:acetic acid and stained in iron- hematoxylin. Branching structures of a whole-mounted mammary gland injected with tracking dye alone were visible up to the end buds and alveoli. Also visible as brown bodies were the mammary lymph nodes. Examination of a whole-mounted mammary gland of the same rat receiving Vaccinia/HA in trypan blue on the contralateral side revealed that only about 30% of the ducts remained. In addition, the lymph nodes were considerably enlarged, denoting the mounting of an immune response to clear the Vaccinia from the vicinity.

Landscapes

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

Abstract

Méthodes de traitement prophilactique et thérapeutique de l'épithélium ductal d'une glande exocrine, en particulier d'une glande mammaire, contre une maladie, en particulier le cancer. Ces méthodes consistent à soumettre l'épithélium ductal de la glande exocrine au contact d'un agent destructeur de l'épithélium, de préférence par cannulation ductale, pour obtenir un effet prophilactique ou thérapeutique.
PCT/US1996/012837 1995-08-03 1996-08-02 Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer Ceased WO1997005898A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU68959/96A AU6895996A (en) 1995-08-03 1996-08-02 Delivery of an agent to the ductal epithelium in the prophylactic and therapeutic treatment of cancer

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US51062395A 1995-08-03 1995-08-03
US2892995P 1995-08-03 1995-08-03
US08/510,623 1995-08-03

Publications (1)

Publication Number Publication Date
WO1997005898A1 true WO1997005898A1 (fr) 1997-02-20

Family

ID=26704264

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1996/012837 Ceased WO1997005898A1 (fr) 1995-08-03 1996-08-02 Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer

Country Status (1)

Country Link
WO (1) WO1997005898A1 (fr)

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6168779B1 (en) 1997-09-16 2001-01-02 The Regents Of The University Of California Methods and kits for identifying ductal orifices
US6221622B1 (en) 1998-04-28 2001-04-24 The Regents Of The University Of California Method and kit for obtaining fluids and cellular material from breast ducts
US6314315B1 (en) 1999-01-13 2001-11-06 Pro Duct Health, Inc. Ductal orifice identification by characteristic electrical signal
US6391026B1 (en) 1998-09-18 2002-05-21 Pro Duct Health, Inc. Methods and systems for treating breast tissue
US6413228B1 (en) 1998-12-28 2002-07-02 Pro Duct Health, Inc. Devices, methods and systems for collecting material from a breast duct
US6610484B1 (en) 1999-01-26 2003-08-26 Cytyc Health Corporation Identifying material from a breast duct
US6638727B1 (en) 1999-01-26 2003-10-28 Cytyc Health Corporation Methods for identifying treating or monitoring asymptomatic patients for risk reduction or therapeutic treatment of breast cancer
AU774637B2 (en) * 1998-10-02 2004-07-01 Atossa Genetics, Inc. Methods for identification, diagnosis, and treatment of breast cancer
US6890311B2 (en) 1997-09-16 2005-05-10 The Regents Of The University Of California Methods for performing medical procedures within a breast duct
WO2009129352A3 (fr) * 2008-04-15 2009-12-10 Windy Hill Medical Dispositif et procédé pour accéder aux canaux des glandes mammaires et les traiter

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0118365A2 (fr) * 1983-03-04 1984-09-12 Health Research, Inc. Anticorps monoclonaux contre les cellules carcinomateuses mammaires humaines et leur application en diagnostic et en thérapie
EP0352722A2 (fr) * 1988-07-25 1990-01-31 Lloyd H. Smith Procédés de liaison d'antigène d'adènocarcinome et réactifs
WO1993002556A1 (fr) * 1991-07-26 1993-02-18 University Of Rochester Therapie du cancer faisant appel a des cellules malignes
WO1994009160A1 (fr) * 1992-10-13 1994-04-28 La Jolla Cancer Research Foundation Procede permettant d´inhiber la replication de cellules hyperproliferatives
WO1994026915A1 (fr) * 1993-05-10 1994-11-24 The Regents Of The University Of Michigan Transfert de genes dans les cellules epitheliales pancreatiques et biliaires
EP0657174A1 (fr) * 1993-12-06 1995-06-14 Mayo Foundation For Medical Education And Research Utilisation de la somatostatine

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0118365A2 (fr) * 1983-03-04 1984-09-12 Health Research, Inc. Anticorps monoclonaux contre les cellules carcinomateuses mammaires humaines et leur application en diagnostic et en thérapie
EP0352722A2 (fr) * 1988-07-25 1990-01-31 Lloyd H. Smith Procédés de liaison d'antigène d'adènocarcinome et réactifs
WO1993002556A1 (fr) * 1991-07-26 1993-02-18 University Of Rochester Therapie du cancer faisant appel a des cellules malignes
WO1994009160A1 (fr) * 1992-10-13 1994-04-28 La Jolla Cancer Research Foundation Procede permettant d´inhiber la replication de cellules hyperproliferatives
WO1994026915A1 (fr) * 1993-05-10 1994-11-24 The Regents Of The University Of Michigan Transfert de genes dans les cellules epitheliales pancreatiques et biliaires
EP0657174A1 (fr) * 1993-12-06 1995-06-14 Mayo Foundation For Medical Education And Research Utilisation de la somatostatine

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
DATABASE MEDLINE FILE SERVER STN KARLSRUHE; FRYKBERG ET AL: "OVERVIEW OF THE BIOLOGY AND MANAGEMENT OF DUCTAL CARCINOMA IN SITU OF THE BREAST", XP002020885 *
RUSSO ET AL: "BIOLOGY OF DISEASE. COMPARATIVE STUDY OF HUMAN AND RAT MAMMARY TUMORIGENESIS", LABORATORY INVESTIGATION, vol. 62, 1990, pages 244 - 278, XP000612052 *
TAMAKI ET AL: "A HUMAN MONOCLONAL ANTIBODY DERIVED FROM AXILLARY LYMPH NODES OF A BREAST CANCER PATIENT", HYBRIDOMA, vol. 8, 1989, pages 293 - 302, XP000612021 *
TAMM ET AL: "INTERLEUKIN 6 DECREASES CELL-CELL ASSOCIATION AND INCREASES MOTILITY OF DUCTAL BREAST CARCINOMA CELLS", THE JOURNAL OF EXPERIMENTAL MEDICINE, vol. 170, 1989, pages 1649 - 1669, XP000612051 *
ZENKLUSEN ET AL: "THE IMMUNOHISTOCHEMICAL REACTIVITY OF A NEW ANTI-EPITHELIAL MONOCLONAL ANTIBODY (MAB B-12) AGAINST BREAST CARCINOMA AND OTHER NORMAL AND NEOPLASTIC HUMAN TISSUES", VIRCHOWS ARCHIV A PATHOLOGICAL ANATOMY AND HISTOPATHOLOGY, vol. 413, 1988, pages 3 - 10, XP000612025 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6455027B1 (en) 1997-09-16 2002-09-24 The Regents Of The University Of California Methods and kits for identifying ductal orifices in a nipple
US6890311B2 (en) 1997-09-16 2005-05-10 The Regents Of The University Of California Methods for performing medical procedures within a breast duct
US6168779B1 (en) 1997-09-16 2001-01-02 The Regents Of The University Of California Methods and kits for identifying ductal orifices
US6221622B1 (en) 1998-04-28 2001-04-24 The Regents Of The University Of California Method and kit for obtaining fluids and cellular material from breast ducts
US6494859B2 (en) 1998-04-28 2002-12-17 The Regents Of The University Of California Methods using pressure to obtain fluids and cellular material from breast ducts
US6391026B1 (en) 1998-09-18 2002-05-21 Pro Duct Health, Inc. Methods and systems for treating breast tissue
US6712816B2 (en) 1998-09-18 2004-03-30 Cytyc Health Corporation Methods and systems for treating breast tissue
AU774637B2 (en) * 1998-10-02 2004-07-01 Atossa Genetics, Inc. Methods for identification, diagnosis, and treatment of breast cancer
US6413228B1 (en) 1998-12-28 2002-07-02 Pro Duct Health, Inc. Devices, methods and systems for collecting material from a breast duct
US6314315B1 (en) 1999-01-13 2001-11-06 Pro Duct Health, Inc. Ductal orifice identification by characteristic electrical signal
US6610484B1 (en) 1999-01-26 2003-08-26 Cytyc Health Corporation Identifying material from a breast duct
US6638727B1 (en) 1999-01-26 2003-10-28 Cytyc Health Corporation Methods for identifying treating or monitoring asymptomatic patients for risk reduction or therapeutic treatment of breast cancer
US6642010B2 (en) 1999-01-26 2003-11-04 Cytyc Health Corporation Identifying, monitoring, and treating women for breast precancer or cancer
WO2009129352A3 (fr) * 2008-04-15 2009-12-10 Windy Hill Medical Dispositif et procédé pour accéder aux canaux des glandes mammaires et les traiter
US20130046171A1 (en) * 2008-04-15 2013-02-21 Jerald A. Johansen Device and method for accessing and treating ducts of mammary glands

Similar Documents

Publication Publication Date Title
US6153184A (en) Destruction of the epithelium of an exocrine gland in the prophylactic and therapeutic treatment of cancer
Wakimoto et al. Human glioblastoma–derived cancer stem cells: Establishment of invasive glioma models and treatment with oncolytic herpes simplex virus vectors
Bouvet et al. Suppression of the immune response to an adenovirus vector and enhancement of intratumoral transgene expression by low-dose etoposide
O’Leary et al. Novel oncolytic chimeric orthopoxvirus causes regression of pancreatic cancer xenografts and exhibits abscopal effect at a single low dose
US6326356B1 (en) Suppression of neu overexpression using a mini-E1A gene
JP2003277272A (ja) ＤＮＡ損傷剤およびр５３を含有する組成物
JPH08504095A (ja) 腫瘍抑制遺伝子を有するレトロウイルスベクターによる遺伝子療法
AU3001799A (en) Producer cells for replication selective viruses in the treatment of malignancy
US20200289591A1 (en) Isolated Recombinant Oncolytic Poxvirus, Pharmaceutical Composition, and Use Thereof in Treatment of Tumors and/or Cancer
WO1997005898A1 (fr) Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer
WO2005009287A2 (fr) Administration de medicament dans l'oreille interne et methodes d'utilisation correspondantes
Rosenfeld et al. Pancreatic carcinoma cell killing via adenoviral mediated delivery of the herpes simplex virus thymidine kinase gene
CN101918035A (zh) 在对抗癌剂有抗性的癌中具有抗癌剂增强活性的癌细胞死亡诱导剂
Li et al. Efficacy of multiple administrations of a recombinant adenovirus expressing wild-type p53 in an immune-competent mouse tumor model
US20040265277A1 (en) Agents for treating malignant diseases using the protein YB-1
Zhuang et al. Allelotype analysis of 2′, 3′-dideoxycytidine-and 1, 3-butadiene-induced lymphomas in B6C3F1 mice
Shibata et al. Massive apoptotic cell death in chemically induced rat urinary bladder carcinomas following in situ HSVtk electrogene transfer
Behbakht et al. Adenovirus-mediated gene therapy of ovarian cancer in a mouse model
US11951182B2 (en) Combination therapy for treatment of thoracic cancer using Ad-REIC/Dkk-3 and a checkpoint inhibitor
Kolodkin-Gal et al. Herpes simplex virus delivery to orthotopic rectal carcinoma results in an efficient and selective antitumor effect
CA2239616A1 (fr) Destruction de l'epithelium d'une glande exocrine dans le traitement prophilactique et therapeutique du cancer
Lim et al. Adenovirus-mediated p53 treatment enhances photodynamic antitumor response
Todryk et al. Cell death associated with genetic prodrug activation therapy of colorectal cancer
CN111500632B (zh) 表达st13和trail的溶瘤腺病毒构建及其应用
Huang et al. Biodistribution and kinetics of the novel selective oncolytic adenovirus M1 after systemic administration

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH 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
ENP Entry into the national phase

Ref document number: 2239616

Country of ref document: CA

Ref country code: CA

Ref document number: 2239616

Kind code of ref document: A

Format of ref document f/p: F

122 Ep: pct application non-entry in european phase