+

WO1992004903A1 - Procede d'inhibition de la production virale - Google Patents

Procede d'inhibition de la production virale Download PDF

Info

Publication number
WO1992004903A1
WO1992004903A1 PCT/US1991/006734 US9106734W WO9204903A1 WO 1992004903 A1 WO1992004903 A1 WO 1992004903A1 US 9106734 W US9106734 W US 9106734W WO 9204903 A1 WO9204903 A1 WO 9204903A1
Authority
WO
WIPO (PCT)
Prior art keywords
oligonucleotide
antigen
viral
ebv
expression
Prior art date
Application number
PCT/US1991/006734
Other languages
English (en)
Inventor
Ian Trevor Magrath
Kishor Gopaldas Bhatia
Walter Leon Goldschmidts
Original Assignee
The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce
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 United States Of America, As Represented By The Secretary, U.S. Department Of Commerce filed Critical The United States Of America, As Represented By The Secretary, U.S. Department Of Commerce
Publication of WO1992004903A1 publication Critical patent/WO1992004903A1/fr

Links

Classifications

    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof

Definitions

  • the present invention relates, in general, to a method of treating viral infection, and, in particular, to a method of treating Epstein-Barr virus (EBV) associated diseases.
  • the invention further relates to oligomers (oligonucleotides) suitable for use in such a method, and to pharmaceutical compositions comprising same.
  • Latent antigens are viral antigens not directly associated with the replication cycle of the virus, but in the case of EBV, are required for the maintenance of the viral genome within the infected cell.
  • Lytic antigens are antigens directly associated with virus replication, and can be further subdivided into early versus late. Early antigens differ from late in that they are expressed early in the viral replication cycle, and require no new viral DNA synthesis for their expression. These early antigens (EA) most likely function in the early stages of viral DNA replication, although the specific function of each component of this group remains unknown.
  • EA-R The major restricted early antigen of EBV (EA-R) has been found to be a viral specific ribonucleotide reductase (Goldschmidts et al. Virology 157:220-226
  • This enzyme is responsible for mediating the first unique step of DNA synthesis by reducing all four ribonucleotides to their corresponding deoxyribonucleotides.
  • the present invention relates to an antisense oligonucleotide that selectively inhibits expression of a viral antigen.
  • the present invention relates to a method of inhibiting expression of a viral antigen.
  • the method comprises contacting the virus with the above-described oligonucleotide under conditions such that inhibition of expression is effected.
  • the present invention relates to a method of treating viral infection.
  • the method comprises contacting a virally infected cell with the above-described oligonucleotide under conditions such that the treatment is effected.
  • the present invention relates to a pharmaceutical composition, in dosage unit form.
  • the composition comprises the above-described oligonucleotide in an amount sufficient to effect the selective inhibition, together with a pharmaceutically acceptable carrier.
  • Figure 1 BamHl map of B95-8 strain of EBV.
  • Figure 2 Prevention of progression of EBV into the lytic cycle by treatment of EBV infected cells with anti-EA-R oligonucleotides.
  • Figure 3 Sequence specific inhibition of EA-R and EA-D antigens of EBV within dosage and time window. DETAILED DESCRIPTION OF THE INVENTION
  • the present invention relates to a method of treating a viral infection utilizing antisense oligonucleotides that specifically inhibit expression of genes involved in virus replication.
  • the invention also relates to antisense oligomers suitable for use in such a method and to pharmaceutical compositions comprising same.
  • the antisense oligonucleotides of the invention are designed so as to be sufficiently complementary to a specific viral messenger RNA to form a stable hybrid therewith (see in this regard Heikkila et al. Nature 7:445 (1987); Holt et al, Mol. Cell. Biol. 8:963 (1988); see also McManaway et al. Lancet 335:808-811 (1990)).
  • the oligonucleotides target lytic antigens, preferably, early lytic antigens.
  • the oligonucleotides of the invention are specific for the EA-R antigen of EBV.
  • One example of such an oligonucleotide has the sequence 3 « TGG-TGT-GTC-GTA-CCG-TTG-CTG 5' (see Figure 1).
  • Oligonucleotides suitable for use in the present invention can be selected by choosing accessible regions in the messenger RNA, specifically regions flanking the start codon. Other regions including the CAP site may also be useful.
  • the oligomers of the invention can be synthesized using methods known in the art (Schott, M.E. Am. Biotech. Lab. 3:20-23 (1985)).
  • the oligonucleotides of the invention can be formulated together with an appropriate carrier, diluent or excipient, into a pharmaceutical composition. Such compositions can be administered intraperitoneally, intravenously, intrathecally or by injection at the site of the lesion.
  • the concentration of oligonucleotide present in the composition is, for example, about 100 ⁇ M to about 1 mM.
  • concentration level is achieved at the lesion site.
  • Actual concentrations can be adjusted so as to be applicable to the particular disease state.
  • Optimum doses can be determined using methods known in the art.
  • the pharmaceutical composition of the invention is formulated so as to be suitable for topical application.
  • Topical application of oligomers obviates oligonucleotide degradation which can be a problem with parenterally administered oligonucleotides. Further, topical application ensures adequate concentrations of oligomer at the target site.
  • the oligonucleotides of the invention can be used in methods of treating viral infection, including EBV infection and in treating viral associated tumors. Application of the oligomers to either treatment methodology can have a variety of effects, including obliteration of virus replication within infected cells.
  • the application of the antisense oligonucleotides of the invention is expected to prevent the spread of the virus to other cells and thereby control the disease.
  • This treatment can be achieved by local or topical application of the oligonucleotide in a form suitable for such application.
  • EBV "latent" genes such as EBNA-1 and LMP may be linked to the maintenance of the neoplastic state.
  • the viral genes can be expected to be inhibitable with greatity, since they are foreign to the cell and, therefore, not necessary for any normal cellular function.
  • the method of the present invention should not result in impairment of immunity against the viral infection, unless it results in elimination of all EBV containing cells from the body (a possibility if the gene which is inhibited by antisense is responsible for maintenance of the latent infection, i.e. EBNA-1). However, where this is not the case, cells are likely to continue to express other viral genes responsible for immunity. If elimination of infection were to occur, however, the present method would be of great value in patients being prepared for bone marrow transplantation, since EBV lymphoproliferative states may occur in such individuals.
  • Antisense oligomers can also be applied to other conditions shown to be due, at least in part, to expression of viral genes.
  • P3HR-1 an EBV genome positive Burkitt's lymphoma cell line
  • TPA tumor-promoting agent
  • oligonucleotides were added to individual wells at a final concentration of 200 ⁇ M, and incubated for 30 hours (see Figure 1 for oligomer sequences) .
  • EA-R The decreased expression of EA-R observed in the cells was accompanied by a parallel decrease in the production of a 50-52 kDa diffuse early antigen (EA-D) , suggesting the expression of the two early antigens may be coupled. Inhibition of these early antigens dramatically inhibited the progression of the induced cells to virus production, as monitored by the expression of viral capsid proteins (antigens) (VCA) .
  • VCA viral capsid proteins
  • Example 2 In Inhibition of EA-R with Dosage and Time Individual wells of induced P3HR-1 cells were expose at various time points to a final concentration of 300 ⁇ M antisense or nonsense oligonucleotides (see Figure 1) , and harvested 30 hours after induction of the viral lytic cycle. After determining cell viability, equal numbers of cells were harvested, extracted, and exposed to SDS-PAGE. The gels were subsequently transferred to nitrocellulose paper, and then incubated with a mixture of monoclonal antibodies directed toward the EA-R and EA-D antigenes of EBV. The results are shown in Figure 3:
  • Lane 2 Induced cells which received nonsense oligonucleotides one hour prior to and two hours post induction.
  • Lane 3 Induced cells which received antisense oligonucleotides one hour prior to induction.
  • Lane 4 Induced cells which received nonsense oligonucleotides one hour prior to induction.
  • Lane 7 Induced cells which received antisense oligonucleotides twelve hours post induction.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Procédé de traitement d'une infection virale utilisant des oligonucléotides non codants inhibant spécifiquement l'expression de gènes impliqués dans la réplication de virus. L'invention concerne également des oligomères non codants s'utilisant emploi dans ledit procédé ainsi que des compositions pharmaceutiques les contenant.
PCT/US1991/006734 1990-09-21 1991-09-20 Procede d'inhibition de la production virale WO1992004903A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US58608790A 1990-09-21 1990-09-21
US586,087 1990-09-21

Publications (1)

Publication Number Publication Date
WO1992004903A1 true WO1992004903A1 (fr) 1992-04-02

Family

ID=24344258

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1991/006734 WO1992004903A1 (fr) 1990-09-21 1991-09-20 Procede d'inhibition de la production virale

Country Status (2)

Country Link
AU (1) AU8872191A (fr)
WO (1) WO1992004903A1 (fr)

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993011267A1 (fr) * 1991-12-05 1993-06-10 The Institute Of Cancer Research Agent antiviral comprenant des oligonucleotides non codants complementaires au gene bhrf1 du virus epstein-barr
US5550047A (en) * 1994-02-18 1996-08-27 University Of Massachusetts Oligonucleotides with anti-Epstein-Barr virus activity
US5837854A (en) * 1996-04-05 1998-11-17 University Of Massachusetts Oligonucleotides with anti-Epstein-Barr virus activity

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4689320A (en) * 1983-10-17 1987-08-25 Akira Kaji Method for inhibiting propagation of virus and anti-viral agent
US5023243A (en) * 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5023243A (en) * 1981-10-23 1991-06-11 Molecular Biosystems, Inc. Oligonucleotide therapeutic agent and method of making same
US4689320A (en) * 1983-10-17 1987-08-25 Akira Kaji Method for inhibiting propagation of virus and anti-viral agent

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
PROC. NATL. ACAD. SCI., Vol. 82, issued December 1985, SPECK et al., "Analysis of the transcript encoding the latent Epstein-Barr virus nuclear antigen I: A potentially polycistronic message generated by long-range splicing several exons", pages 8305-8309. *
VIROLOGY, Vol. 157, issued 1987, GOLDSCHMIDTS et al., "A Restricted Component of the Epstein-Barr Virus Early Antigen Complex is Structurally Related to Ribonucleotide Reductase", pages 220-226. *

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993011267A1 (fr) * 1991-12-05 1993-06-10 The Institute Of Cancer Research Agent antiviral comprenant des oligonucleotides non codants complementaires au gene bhrf1 du virus epstein-barr
US5550047A (en) * 1994-02-18 1996-08-27 University Of Massachusetts Oligonucleotides with anti-Epstein-Barr virus activity
US5837854A (en) * 1996-04-05 1998-11-17 University Of Massachusetts Oligonucleotides with anti-Epstein-Barr virus activity

Also Published As

Publication number Publication date
AU8872191A (en) 1992-04-15

Similar Documents

Publication Publication Date Title
US11834649B2 (en) CRISPR/CAS-related methods and compositions for treating herpes simplex virus
Huang et al. Ribonucleic acid synthesis of vesicular stomatitis virus IV. Transcription by standard virus in the presence of defective interfering particles
US4689320A (en) Method for inhibiting propagation of virus and anti-viral agent
Rose et al. Identification of two homologs of the Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) in retroperitoneal fibromatosis of different macaque species
Yaron et al. Stimulation of prostaglandin E production in cultured human fibroblasts by poly (I)· poly (C) and human interferon
US5594122A (en) Antisense oligonucleotides targeted against human immunodeficiency virus
JP2708960B2 (ja) サイトメガウイルス感染の作用を変調するオリゴヌクレオチド
EP0625192B1 (fr) Nucleoenzymes
Williams et al. Activation of a nuclease by pppA2′ p5′ A2′ p5′ A in intact cells
US6310044B1 (en) Oligonucleotide therapies for modulating the effects of herpesviruses
Baron et al. Role of interferon during viremia: II. Protective action of circulating interferon
Kleinschmidt et al. Interferon stimulation by a double stranded RNA of a mycophage in statolon preparations
JP2020531465A (ja) 単純ヘルペスウイルス糖タンパク質をコードする修飾ｍＲＮＡワクチン及びその使用
JP2001500471A (ja) プロテアーゼインヒビターの生物学的及び抗ウイルス活性を改善する方法
BR9712138A (pt) Vìrus de rna isolado, vacina, processo para imunizr um indivìduo para induzir proteção contra um vìrus de rna não segmentado, sentido negativo, de filamento único, da ordem mononegavirales e para produzir vìrus de rna, molécula de ácido nucleico isolada e composição.
JPS6181792A (ja) 水痘−帯状疱疹ウイルスに対するワクチン
Zaia et al. Cytomegalovirus infection in the bone marrow transplant recipient
Kaplan et al. Isolation of simian virus 40-transformed inbred hamster cell lines heterogeneous for virus induction by chemicals or radiation
Karpov et al. A plasmid-expressed CRISPR/Cas9 system suppresses replication of HSV type I in a vero cell culture
WO1992004903A1 (fr) Procede d'inhibition de la production virale
RADO et al. Herpes zoster house epidemic in steroid-treated patients: A clinical and viral study
Klein et al. Effect of acyclovir on latent herpes simplex virus infections in trigeminal ganglia of mice
BR9707400A (pt) Vacinas contra produto do gene 63 de vìrus varicella zoster.
US20190330635A1 (en) Antiviral agent and method for treating viral infection
Bausek et al. Two mechanisms of cell resistance to repeated stimulation of interferon

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 FR GB GR IT LU NL SE

122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

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