WO2006115170A1 - Traitement du cancer de la tete et du cou par le virus hsv1-hf10 - Google Patents

Traitement du cancer de la tete et du cou par le virus hsv1-hf10 Download PDF

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Publication number
WO2006115170A1
WO2006115170A1 PCT/JP2006/308317 JP2006308317W WO2006115170A1 WO 2006115170 A1 WO2006115170 A1 WO 2006115170A1 JP 2006308317 W JP2006308317 W JP 2006308317W WO 2006115170 A1 WO2006115170 A1 WO 2006115170A1
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WIPO (PCT)
Prior art keywords
cancer
neck cancer
mutant
gene
herpes simplex
Prior art date
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PCT/JP2006/308317
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English (en)
Japanese (ja)
Inventor
Yukihiro Nishiyama
Yasushi Fujimoto
Fumi Goshima
Tsutomu Nakashima
Terukazu Mizuno
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National University Corporation Nagoya University
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Priority to JP2007514646A priority Critical patent/JPWO2006115170A1/ja
Publication of WO2006115170A1 publication Critical patent/WO2006115170A1/fr

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    • 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
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • 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
    • 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
    • C12N2710/16661—Methods of inactivation or attenuation
    • C12N2710/16662—Methods of inactivation or attenuation by genetic engineering

Definitions

  • the present invention relates to treatment of human head and neck cancer using attenuated herpesvirus.
  • Head and neck cancer may involve all of the cicada-camilla nerves, easily impair survival functions such as breathing, swallowing, and vocalization, and may involve deformation of the exposed parts of the face and neck There is a feature. Therefore, in the treatment method selection, while organ preservation and function preservation are emphasized, local control is also the biggest issue. Head and neck cancer is difficult to achieve with chemotherapy alone. In addition, dysfunction and deformation have become problems in the treatment of head and neck cancer by surgery, and radiation therapy has problems such as the limitation of irradiation dose and impaired salivation after swallowing, and dysphagia. There are many examples where the effects are limited or the effects are not fully obtained. In addition, clinical trials for the treatment of head and neck cancer using adenovirus mutants have been carried out, but the results are not satisfactory. Therefore, it is desired to develop a new treatment method for treating head and neck cancer.
  • HSV simple herpesvirus
  • the virus has the following useful properties: 1) HSV can infect almost all types of cells, 2) HSV has the ability to infect other viruses, eg, adeno Higher than viruses, adeno-associated virus (AAV), etc. 3) All basic sequences of HSV genome have been elucidated, 4) HSV kills infected cells at low doses 5) Antiviral agents against HSV are available.
  • Patent Document 1 JP 2002-218975
  • Patent Literature 2 Tokuheihei 2004—535798
  • Non-Patent Document 1 J. Surg. Oncol, 85: 42-47, 2004
  • An object of the present invention is to provide a novel therapeutic method and therapeutic agent for head and neck cancer.
  • the present inventors conducted a research clinical trial in which a patient with head and neck cancer was inoculated with an attenuated simple herpesvirus at the subcutaneous tumor site, and the virus had a harmful effect on the patient. It was found that it showed a tumor suppressive effect.
  • the present invention provides a pharmaceutical composition for treating human head and neck cancer, comprising a mutant simple herpesvirus in which the UL56 gene is inactivated.
  • the present invention also provides a pharmaceutical composition for enhancing the effect of radiation therapy for human head and neck cancer, comprising as an active ingredient a mutant herpesvirus in which the UL56 gene is inactivated.
  • the present invention further provides a pharmaceutical composition for enhancing the effect of chemotherapy for human head and neck cancer, comprising as an active ingredient a mutant herpes simplex virus in which the UL56 gene is inactivated. .
  • the present invention provides a method for treating human head and neck cancer by administering a mutant herpes simplex virus in which the UL56 gene is inactivated to a cancer site, Methods for enhancing the effects of radiation therapy for cervical cancer and methods for enhancing the effects of human head and neck cancer chemotherapy are provided.
  • the variant simple herpesvirus is preferably simple herpesvirus type 1 HF10 strain.
  • FIG. 1 shows an illustration of the structure of HSV HF10 used in the present invention.
  • FIG. 2 shows the course of treatment with the pharmaceutical composition of the present invention.
  • FIG. 3 shows the results of HE staining and anti-HSV staining of tissue administered with the pharmaceutical composition of the present invention.
  • FIG. 4 shows the course of treatment with the pharmaceutical composition of the present invention.
  • FIG. 5 shows the results of HE staining of the tissue administered with the pharmaceutical composition of the present invention.
  • FIG. 6 shows the results of anti-HSV staining of tissue administered with the pharmaceutical composition of the present invention.
  • Fig. 7 shows the results of anti-HSV staining of the HSV-uninfected part of the tissue administered with the pharmaceutical composition of the present invention.
  • the herpes simplex virus UL56 protein is a tail anchor type II membrane protein localized in the Golgi apparatus and endonom. The function of UL56 protein is still well elucidated !, but it is thought to be related to neurotoxicity! /
  • Mutant herpesviruses in which the UL56 gene is inactivated are mutated or deleted by a known method, or another gene fragment is inserted into the UL56 gene. It can be produced by insertion.
  • An example of such a method is a homologous gene introduction method in which a gene chain having a target mutation site sandwiched between homologous gene sites is introduced and the gene is introduced. You can also use naturally occurring mutants in which the UL56 gene is inactivated.
  • HF10 strain eg Arch Virol, 148
  • HF10 strain is a clone found as a naturally occurring attenuated mutant virus that has not been genetically modified, and causes extensive cell membrane fusion in infected cells.
  • the genomic structure of HF10 is shown in Figure 1.
  • HF10 is central Although at least four accessory genes, including the gene for neuroinvasiveness (UL56), are deficient and the pathogenicity is markedly reduced, the proliferation ability in tumor cells is extremely high (over 100 times that of the wild type). ).
  • HF10 has been reported to prolong the survival of nude mice inoculated with tumor cells derived from colon cancer, breast cancer, melanoma and sarcoma (J. Surg. Oncol, 85: 42-47, 2004). HF10 was non-toxic when intraperitoneally inoculated into mice.
  • the mutant simple herpesvirus in which the UL56 gene of the present invention is inactivated can be propagated by a known method. For example, it was confirmed that Vero cells derived from African green monkey kidney cells showed a cytopathic effect, collected, and subjected to three consecutive freeze-thaw operations, followed by centrifugation. Precipitate cell components by separation and collect virus-containing supernatant.
  • the grown mutant herpes simplex virus can be formulated as an injection solution by suspending it in sterile water, sterile physiological saline or a suitable buffer. The preparation may be further added with a silkworm, a stabilizer, a preservative and the like.
  • the pharmaceutical composition comprising the mutant herpesvirus of the present invention as an active ingredient is useful as a therapeutic agent for head and neck cancer.
  • the pharmaceutical composition of the present invention can be administered by directly inoculating the tumor site.
  • head and neck cancer that can be treated by the present invention include tongue cancer, oral cavity cancer including oral floor cancer and gingival cancer, lip cancer, nasopharyngeal cancer, oropharyngeal cancer, hypopharyngeal cancer, laryngeal cancer, nasal cavity cancer, thyroid gland.
  • Examples include cancer, parotid gland cancer, submandibular gland cancer and the like.
  • Head and neck cancer is generally characterized by having a lesion near the body surface, or a site that can be easily reached by opening or opening, so that it is easy to administer the virus and the effect of virus therapy is effectively exerted.
  • Endoscopic injection is useful for the nasopharynx, hypopharynx, and larynx that cannot be reached by opening alone. This is a method of injecting a virus under a laryngeal fiber using a local injection needle under clear vision, and it can be administered repeatedly every day because the burden on the patient and the invasiveness are light.
  • the viral therapy of the present invention may be used in combination with radiation therapy or chemotherapy.
  • Synergistic effects can be obtained in combination with virus therapy, and function-preserving therapy can be developed by improving local control rate.
  • combination therapy it is necessary to administer the virus in such a way that the patient is not invaded as much as possible in order to carry out daily radiotherapy without interruption. In that respect, it can be injected only through the opening or under the laryngeal fiber.
  • This treatment method does not require a facility such as an operating room, and is a treatment method that reduces the burden on the patient.
  • HSV for locally advanced recurrent head and neck squamous cell carcinoma As shown in the following examples, HSV for locally advanced recurrent head and neck squamous cell carcinoma
  • the pharmaceutical composition comprising the mutant herpes simplex virus of the present invention as an active ingredient is very attenuated and its neurotoxicity is very low, so there are almost no side effects in patients. It is thought that it will not occur. Even if a few adverse events occur, the attenuated herpesvirus is highly sensitive to anti-herpes drugs such as acyclovir and ganciclovir, It is possible to administer various drugs.
  • SPF chicken fertilized eggs for vaccine production were used to infect the virus using primary embryo cells of chicken embryos as host cells, and cultured in a culture solution supplemented with bovine serum and kanamycin, which have been denied the presence of stray viruses.
  • the virus was infected with the master virus bank with MOIO.Olpfo / cell, cultured for 24 hours under serum-free condition, and the culture supernatant was collected.
  • the cultured virus was subjected to sterility testing by taking over the sterility test method of the Japanese Pharmacopoeia General Test Method and the Mycoplasma Negative Test Method.
  • lxl0 6 pfo / ml virus solution was prepared and stored at -80 ° C. At the time of administration was used by diluting ⁇ -virus solution lxl0 4 pfo / ml.
  • the tumor size increased 45x40x38mm after 7 days, 55x44x34mm after 15 days, and 55x50x33mm after 21 days, and was evaluated as PD (progressive) in the clinical efficacy assessment.
  • Figure 3 shows the pathological examination. 1: HE staining, 2: HSV-1 immunostaining, 3: Negative control. Although the virus antigen has proved powerful, pathologically, the tumor cell necrosis is found in the local injection part, which is more conspicuous than the non-local injection part, which may be effective by HF10. It is considered high.
  • the tumor diameter at the time of excision was 20 X 12 mm.
  • the clinical efficacy assessment was evaluated as NC (unchanged). A large number of necrotic parts were observed in the pathological specimen, where eosin-favored nuclei were obscured or disappeared. In addition, the appearance of multinucleated cells and infiltration of inflammatory cells were observed (Fig. 5). Furthermore, immunostaining with anti-HSV-1 antibody proved the presence of HSV consistent with the necrotic area (Fig. 6). On the other hand, in the uninfected part of HF10, a large number of dividing cells were recognized, and actively proliferating cancer tissues were observed (Fig. 7).
  • HF10 (1 ⁇ 10 5 PFU / 0.5 ml) was injected into the skin metastases once a day for 3 days. There was a mild fever for several days after injection. No other clinical findings changed during the study. The size of the nodule at the injection site was measured and excised 15 days after inoculation.
  • composition comprising the attenuated mutant herpes simplex virus of the present invention as an active ingredient is effective for the treatment of human head and neck cancer.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Virology (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Engineering & Computer Science (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Animal Behavior & Ethology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Genetics & Genomics (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
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  • General Engineering & Computer Science (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé permettant de traiter le cancer humain de la tête et du cou en utilisant un virus mutant de l'herpès simplex présentant le gène UL56 sous forme inactivée. L'invention concerne également une composition pharmaceutique utilisée dans ce procédé. Le virus mutant de l'herpès simplex est de préférence un virus de l'herpès simplex de type 1 et de souche HF10. Un essai clinique est effectué, dans le cadre d'une étude, sur un patient atteint du cancer de la tête et du cou en lui inoculant sur un site tumoral sous-cutané la composition pharmaceutique renfermant une version atténuée du virus de l'herpès simplex. Il est ainsi prouvé que le virus peut produire un effet anti-tumoral sans provoquer d'effets secondaires chez le patient.
PCT/JP2006/308317 2005-04-21 2006-04-20 Traitement du cancer de la tete et du cou par le virus hsv1-hf10 WO2006115170A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2007514646A JPWO2006115170A1 (ja) 2005-04-21 2006-04-20 頭頸部癌に対するｈｓｖ１−ｈｆ１０ウイルス療法

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JP2005-123776 2005-04-21
JP2005123776 2005-04-21

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002218975A (ja) * 2001-01-26 2002-08-06 Yukihiro Nishiyama 弱毒化単純ヘルペスウイルス
JP2004535798A (ja) * 2001-05-09 2004-12-02 株式会社エムズサイエンス ヘルペスウイルスを用いた癌処置のための組成物および方法

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002218975A (ja) * 2001-01-26 2002-08-06 Yukihiro Nishiyama 弱毒化単純ヘルペスウイルス
JP2004535798A (ja) * 2001-05-09 2004-12-02 株式会社エムズサイエンス ヘルペスウイルスを用いた癌処置のための組成物および方法

Non-Patent Citations (10)

* Cited by examiner, † Cited by third party
Title
DATABASE CAPLUS [online] KIM S.H. ET AL.: "COMBINATION OF MUTATED HERPES SIMPLEX VIRUS TYPE 1 (G207 VIRUS) WITH RADIATION FOR THE TREATMENT OF SQUAMOUS CELL CARCINOMA OF THE HEAD AND NECK", XP003006869, accession no. STN Database accession no. (2005:69114) *
DATABASE MEDLINE [online] WONG R.J. ET AL.: "EFFECTIVE TREATMENT OF HEAD AND NECK SQUAMOUS CELL CARCINOMA BY AN ONCOLYTIC HERPES SIMPLEX VIRUS", XP003006870, accession no. STN Database accession no. (2001388262) *
EUR. J. CANCE, vol. 41, no. 2, January 1995 (1995-01-01), pages 313 - 322 *
J. AM. COLL. SURG., vol. 193, no. 1, 2001, pages 12 - 21 *
KIMATA H. ET AL.: "Hen'i Tanjun Herpes virus "HF10" o Mochiita Gan Chiryo", JOURNAL OF JAPAN SURGICAL SOCIETY, vol. 105, no. SPECIAL EXTRA ISSUE, 15 March 2004 (2004-03-15), pages 305, SF-098-1, XP003006867 *
KIMATA H. ET AL.: "Hen'i Tanjun Herpes virus HF10 o Mochiita Daichogan Fukumaku Hashu ni Taisuru Chiryo", JOURNAL OF JAPANA SURGICAL SOCIETY, vol. 104, no. SPECIAL EXTRA TISSUE, 30 April 2003 (2003-04-30), pages 343, PS1014-3, XP003006868 *
KIMATA H. ET AL.: "Herpesvirus HF10 ni yoru Nyugan Chiryo o Saishin Kenkyu", JAPANESE JOURNAL OF CLINICAL MEDICINE, vol. 62, no. 5, 2004, pages 987 - 993, XP003006866 *
KIMATA H. ET AL: "Tokushu Virus Vector: Saishin no Wadai 2. Herpesvirus no Igakuteki Riyo - Idenshi Chiryo to Gan Chiryo eno Oyo -", VIRUS, vol. 53, no. 2, 2003, pages 155 - 162, XP003006865 *
SUGIURA A. ET AL.: "Tanjun Herpesvirus Hen'i Kabu o Mochiita Kokei Shuyo ni Taisuru oncolytic virus therapy", THE OTO-RHINO-LARYNGOLOGICAL SOCIETY OF JAPAN, INC. KAIHO, vol. 107, no. 9 SPECIAL EXTRA ISSUE, 2004, pages 909 - 910, XP003006864 *
WONG R.J. ET AL.: "CYTOKINE GENE TRANSFER ENHANCES HERPES ONCOLYTIC THERAPY IN MURINE SQUAMOUS CARCINOMA", HUMAN GENE THERAPY, vol. 12, no. 3, 2001, pages 253 - 265, XP002971611 *

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