WO2011055398A1 - A process for the production of immunoglobulines extracted from human plasma for therapeutic use, for the neutralization of the epstein-barr virus, and the medicine containing said immunoglobulins" - Google Patents

A process for the production of immunoglobulines extracted from human plasma for therapeutic use, for the neutralization of the epstein-barr virus, and the medicine containing said immunoglobulins" Download PDF

Info

Publication number
WO2011055398A1
WO2011055398A1 PCT/IT2010/000434 IT2010000434W WO2011055398A1 WO 2011055398 A1 WO2011055398 A1 WO 2011055398A1 IT 2010000434 W IT2010000434 W IT 2010000434W WO 2011055398 A1 WO2011055398 A1 WO 2011055398A1
Authority
WO
WIPO (PCT)
Prior art keywords
immunoglobulins
epstein
barr virus
antibodies
virus
Prior art date
Application number
PCT/IT2010/000434
Other languages
French (fr)
Inventor
Michele Pitaro
Marco Salvetti
Original Assignee
Michele Pitaro
Marco Salvetti
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 Michele Pitaro, Marco Salvetti filed Critical Michele Pitaro
Publication of WO2011055398A1 publication Critical patent/WO2011055398A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/06—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies from serum
    • C07K16/065—Purification, fragmentation
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
    • C07K16/085—Herpetoviridae, e.g. pseudorabies virus, Epstein-Barr virus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/21—Immunoglobulins specific features characterized by taxonomic origin from primates, e.g. man
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Definitions

  • the present invention concerns the field of immunoglobulins and of drugs deriving therefrom.
  • the present invention concerns a process for the production of immunoglobulins extracted from human plasma, for the neutralization of the Epstein-Barr virus.
  • Epstein-Barr virus has been considered the pathogen associated with many and different pathologies, from the infective mononucleosis to severe neoplastic pathologies like the Burkitt lymphoma, the Hodgkin lymphoma, other rare blood diseases like the H-linked lymphoproliferative disorder, carcinomas like the nasopharyngeal and the breast carcinoma, hepatic carcinoma, mesenchymal cancer and other.
  • Epstein-Barr virus is associated with the beginning of neoplastic and autoimmune pathologies that occur when a genetically predisposed person develops the infection, adequate antiviral medicines for fighting its presence in the human body might allow the treatment of the pathologies connected to said virus.
  • pathogen viruses there are no specific antivirals for preventing the spread of the Epstein-Barr virus or for successfully counter the syntomatology once the infection has occurred; furthermore, among all the herpesviruses, the Epstein-Barr is the most resistant to therapy with guanosine-like compounds.
  • the aim of the present invention is reached by means of a process for the production of immunoglobulins extracted from human plasma, for therepeutic use, for the neutralization of the Epstein-Barr virus, comprising the following steps:
  • the process for the production of immunoglobulins according to the present invention comprises the further step of:
  • B-cell lymphomas associated to Aids posttransplantation lymphoproliferative disease, lymphomatoid granulomatosis, B-cell lymphoma associated with methotrexate therapy, B-cell lymphomas associated with severe combined immunodeficiency, with Wiskott Aldrich syndrome, with X-linked lymphoproliferative disorder, leiomyosarcoma).
  • the production method object of the present invention comprises performing diagnostic exams arranged for the unfailing identification of blood units positive to the Epstein-Barr virus infection, by finding in the same antibodies specific to said virus. Successively, by means of exams based on the Rocchi-Hewetson quantitative micro-test, on the Elisa technique or on the flow cytometry quick tests, performed on the blood units positive to the virus, those blood units provided with the greater capacity of neutralizing said virus are identified.
  • Said neutralizing capacity is due to the presence, in the blood units, of antibodies specific to the antigens of the virus, localized onto the viral envelope and on the surface of the B-lymphocytes that have been infected during the lytic phase of the same, i.e. glycoproteins classified , according to their molecular weight, as gp350, gp340, gp270, gp85, gp42, the antigen BMRF-2 and other antigens important for their capacity of neutralizing their relative immunoglobulins.
  • those blood units characterized in the presence of high neutralizing antibodies values, are selected and hyper-immune immunoglobulins with high concentration of antibodies neutralizing the Epstein-Barr virus are extracted from said blood units, through cold ethanol fractionation as provided by the Cohn-Oncley method.
  • fractionation methods like cryoprecipitation, or chromatographic techniques like gel precipitation, ion exchange chromatography and affinity chromatography can be used.

Abstract

The present invention concerns a process for the production of immunoglobulins extracted from human plasma, for therapeutic use, for the neutralization of the Epstein-Barr virus. From said process it is possible to obtain hyper-immune immunoglobulins containing high titles of antibodies neutralizing the Epstein-Barr virus, and specific immunoglobulins, containing almost solely antibodies neutralizing said virus. Such drugs may be given to patients, as usual, by intravenous, intramuscular or hypodermic injection or by local treatment of the infections due to the Epstein-Barr virus and of the pathologies associated thereto.

Description

A PROCESS FOR THE PRODUCTION OF IMMUNOGLOBULINES EXTRACTED FROM HUMAN PLASMA FOR THERAPEUTIC USE, FOR THE NEUTRALIZATION OF THE EPSTEIN-BARR VIRUS, AND THE MEDICINE CONTAINING SAID IMMUNOGLOBULINES
The present invention concerns the field of immunoglobulins and of drugs deriving therefrom.
More in detail, the present invention concerns a process for the production of immunoglobulins extracted from human plasma, for the neutralization of the Epstein-Barr virus.
It is well known that since the '60 the Epstein-Barr virus has been considered the pathogen associated with many and different pathologies, from the infective mononucleosis to severe neoplastic pathologies like the Burkitt lymphoma, the Hodgkin lymphoma, other rare blood diseases like the H-linked lymphoproliferative disorder, carcinomas like the nasopharyngeal and the breast carcinoma, hepatic carcinoma, mesenchymal cancer and other.
It is further known that, in recent times, the presence of the Epstein-Barr virus in the human body has been put into relation with the beginning of autoimmune diseases like rheumatoid arthritis, lupus erythematous, the Sjogren's syndrome, myasthenia gravis, multiple sclerosis and other.
Therefore, it would appear logic to suppose that, as the Epstein-Barr virus is associated with the beginning of neoplastic and autoimmune pathologies that occur when a genetically predisposed person develops the infection, adequate antiviral medicines for fighting its presence in the human body might allow the treatment of the pathologies connected to said virus. Unlike other pathogen viruses, there are no specific antivirals for preventing the spread of the Epstein-Barr virus or for successfully counter the syntomatology once the infection has occurred; furthermore, among all the herpesviruses, the Epstein-Barr is the most resistant to therapy with guanosine-like compounds.
It is the aim of the present invention to overcome said inconveniences. The aim set forth is reached by means of a process for the production of immunoglobulins extracted from human plasma, for therepeutic use, for the neutralization of the Epstein-Barr virus, comprising the following steps:
- performing early diagnostic exams for the identification of blood units positive to the Epstein-Barr virus infection;
performing further diagnostic exams on the blood units positive to the virus, for identifying the antibodies that neutralize it; performing a selective process of the blood units containing high titles of said neutralizing antibodies;
realizing a process for the extraction of hyper-immune immunoglobulins, from said blood units, for the neutralization of the Epstein-Barr virus.
According to a further aspect of the present invention, the process for the production of immunoglobulins according to the present invention comprises the further step of:
realizing a process for the extraction of specific immunoglobulins from said hyper-immune immunoglobulins, containing almost solely antibodies neutralizing the virus.
The present invention has the advantages listed hereinbelow:
it allows to make available immunoglobulins specific for the treatment of the pathologies connected to the Epstein-Barr virus; it allows the realization of drugs for the prevention of the Epstein-Barr virus infection in those people who are at risk of infection;
it allows to produce drugs for the treatment of infective mononucleosis and of its complications, like Epstein-Barr virus hepatitis;
- it allows the production of drugs for the treatments of pathologies associated to the Epstein-Barr virus (rheumatoid arthritis, systemic lupus erythematous, the Sjogren's syndrome, myasthenia gravis, multiple sclerosis);
it allows the realization of drugs for the treatment of neoplasias associated with the Epstein-Barr virus, both in immunocompetent persons (Burkitt lymphoma, Hodgkin lymphoma, nasopharyngeal carcinoma, lymphoepithelial carcinomas of the salivary glands, of the thymus gland, of the lungs and of the stomach, breast carcinoma, hepatocellular carcinoma, follicular dendritic cell sarcoma), and in patients with congenital and acquired immunodeficiencies (B-cell lymphomas associated to Aids, posttransplantation lymphoproliferative disease, lymphomatoid granulomatosis, B-cell lymphoma associated with methotrexate therapy, B-cell lymphomas associated with severe combined immunodeficiency, with Wiskott Aldrich syndrome, with X-linked lymphoproliferative disorder, leiomyosarcoma).
According to the present invention, the production method object of the present invention comprises performing diagnostic exams arranged for the unfailing identification of blood units positive to the Epstein-Barr virus infection, by finding in the same antibodies specific to said virus. Successively, by means of exams based on the Rocchi-Hewetson quantitative micro-test, on the Elisa technique or on the flow cytometry quick tests, performed on the blood units positive to the virus, those blood units provided with the greater capacity of neutralizing said virus are identified.
Said neutralizing capacity is due to the presence, in the blood units, of antibodies specific to the antigens of the virus, localized onto the viral envelope and on the surface of the B-lymphocytes that have been infected during the lytic phase of the same, i.e. glycoproteins classified , according to their molecular weight, as gp350, gp340, gp270, gp85, gp42, the antigen BMRF-2 and other antigens important for their capacity of neutralizing their relative immunoglobulins.
Now those blood units, characterized in the presence of high neutralizing antibodies values, are selected and hyper-immune immunoglobulins with high concentration of antibodies neutralizing the Epstein-Barr virus are extracted from said blood units, through cold ethanol fractionation as provided by the Cohn-Oncley method.
As an alternative, fractionation methods like cryoprecipitation, or chromatographic techniques like gel precipitation, ion exchange chromatography and affinity chromatography can be used.
As far as safety is concerned, the recommendations of the World Health Organization concerning human plasma fractionation should be observed.
If necessary, specific immunoglobulins, containing nearly solely antibodies neutralizing said virus, may be obtained, exploiting the viral proteins gp350, gp340, gp270, gp85, gp42, BMRF-2 (or other antigens important for the capacity of neutralizing the immunoglobulins), as binders of the same in chromatographic columns. Extractive hyper-immune immunoglobulins, and specific extractive immunoglobulins, produced by means of above mentioned process, and provided with efficiency levels proportional to the amount of neutralizing antibodies contained therein, may be given to patients as usual by intravenous, intramuscular or hypodermic injection or by local treatment of the infections due to the Epstein-Barr virus and of the pathologies associated thereto.
Naturally, it being understood that the embodiments of the invention can widely vary with respect to the description, without departing from the scope of the present invention.

Claims

1. A process for the production of immunoglobulins extracted from human plasma, for therapeutic use, for the neutralization of the Epstein-Barr virus, comprising the following steps:
performing early diagnostic exams for the identification of blood units positive to the Epstein-Barr virus infection;
performing further diagnostic exams, on the blood units positive to the virus, for identifying the antibodies that neutralize it; performing a selective process of the blood units containing high titles of said neutralizing antibodies;
realizing a process for the extraction of hyper-immune immunoglobulins, from said blood units, for the neutralization of the Epstein-Barr virus.
2. A process according to claim 1 , characterized in that the early diagnostic exams for the identification of blood units positive to the Epstein-Barr virus infection are based on the detection of the specific antibodies of the lag phase and of the lithic phase of said virus.
3. A process according to claim 1 , characterized in that the successive diagnostic exams for the detection of the antibodies neutralizing the Epstein-Barr virus comprise a Rocchi-Hewetson quantitative micro-test, the Elisa technique and the flow cytometry quick tests.
4. A process according to claim 1 , characterized in that the process used for extracting hyper-immune immunoglobulins from the blood units is the cold ethanol fractionation as provided by the Cohn-Oncley method.
5. A process according to claim 4, characterized in that the process for the extraction of hyper-immune immunoglobulins from the blood units may be alternatively performed by cryo-precipitation, or by chromatographic techniques as gel-precipitation, ionic-exchange chromatography and affinity chromatography.
6. A process according to claims 4 or 5, characterized in that the process for extracting hyper-immune immunoglobulins is performed only on blood units with a high title of antibodies neutralizing the Epstein-Barr virus.
7. A process according to claim 1 , characterized in that it comprises a further extraction phase - from said hyper-immune immunoglobulins - of specific immunoglobulins, containing almost solely antibodies neutralizing said virus.
8. A process according to claim 7, characterized in that the process for the extraction of the specific immunoglobulins from said hyperimmune immunoglobulins, practically containing solely antibodies neutralizing the virus, comprises methods able to exploit glycoproteins gp350, gp340, gp270, gp85, gp42 and BMRF-2 (or other antigens important for their capacity of neutralizing immunoglobulins), as binders of the same in chromatographic columns.
9. A drug containing antibodies neutralizing the Epstein-Barr virus, characterized in that said antibodies are obtained by means of a process according to at least one of the preceding claims.
PCT/IT2010/000434 2009-11-03 2010-10-27 A process for the production of immunoglobulines extracted from human plasma for therapeutic use, for the neutralization of the epstein-barr virus, and the medicine containing said immunoglobulins" WO2011055398A1 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ITRM2009A000558 2009-11-03
IT000558A ITRM20090558A1 (en) 2009-11-03 2009-11-03 PROCEDURE FOR THE PRODUCTION OF IMMUNOGLOBULINES EXTRACTED FROM HUMAN PLASMA FOR THERAPEUTIC USE NEUTRALIZING THE VIRUS OF EPSTEIN BARR AND DRUG CONTAINING DETTE IMMUNOGLOBULINE

Publications (1)

Publication Number Publication Date
WO2011055398A1 true WO2011055398A1 (en) 2011-05-12

Family

ID=42238826

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IT2010/000434 WO2011055398A1 (en) 2009-11-03 2010-10-27 A process for the production of immunoglobulines extracted from human plasma for therapeutic use, for the neutralization of the epstein-barr virus, and the medicine containing said immunoglobulins"

Country Status (2)

Country Link
IT (1) ITRM20090558A1 (en)
WO (1) WO2011055398A1 (en)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0764658A1 (en) * 1995-09-22 1997-03-26 Rotkreuzstiftung Zentrallaboratorium Blutspendedienst Srk Process for obtaining immunoglobulins from fractions originating from the fractionation of human blood plasma
WO2007017859A1 (en) * 2005-08-11 2007-02-15 Omrix Biopharmaceuticals Ltd. Intravenous immunoglobulin composition
WO2007078202A1 (en) * 2006-01-03 2007-07-12 Tto Nord As Preparation useful for, and method for treatment of neonatal alloimmune thrombocytopenia (nait)

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0764658A1 (en) * 1995-09-22 1997-03-26 Rotkreuzstiftung Zentrallaboratorium Blutspendedienst Srk Process for obtaining immunoglobulins from fractions originating from the fractionation of human blood plasma
WO2007017859A1 (en) * 2005-08-11 2007-02-15 Omrix Biopharmaceuticals Ltd. Intravenous immunoglobulin composition
WO2007078202A1 (en) * 2006-01-03 2007-07-12 Tto Nord As Preparation useful for, and method for treatment of neonatal alloimmune thrombocytopenia (nait)

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
ABEDI M R ET AL: "Preventive effect of IgG from EBV-seropositive donors on the development of human lympho-proliferative disease in SCID mice", INTERNATIONAL JOURNAL OF CANCER 1997 US LNKD- DOI:10.1002/(SICI)1097-0215(19970516)71:4<624::AID-IJC19>3.0.CO;2-B, vol. 71, no. 4, 1997, pages 624 - 629, XP002588394, ISSN: 0020-7136 *
ALLEN U ET AL: "Epstein-Barr virus and posttransplant lymphoproliferative disorder in solid organ transplant recipients", AMERICAN JOURNAL OF TRANSPLANTATION 2009 BLACKWELL PUBLISHING LTD GBR LNKD- DOI:10.1111/J.1600-6143.2009.02898.X, vol. 9, no. SUPPL. 4, December 2009 (2009-12-01), pages S87 - S96, XP002588398 *
ANONYMUS: "Immunoglobulin preparations - updated July 2009", July 2009 (2009-07-01), XP002588400, Retrieved from the Internet <URL:http://www.health.gov.au/internet/immunise/publishing.nsf/Content/Handbook-immunoglobulin> [retrieved on 20100621] *
COHEN J I: "Optimal treatment for chronic active Epstein-Barr virus disease: Editorial", PEDIATRIC TRANSPLANTATION - BLACKWELL PUBLISHING LTD GBR LNKD-, vol. 13, no. 4, June 2009 (2009-06-01), pages 393 - 396, XP002588397, DOI: 10.1111/J.1399-3046.2008.01095.X *
CUMMINS L M ET AL: "Preparation and characterization of an intravenous solution of IgG from human immunodeficiency virus-seropositive donors", BLOOD, AMERICAN SOCIETY OF HEMATOLOGY, US, vol. 77, no. 5, 1 March 1991 (1991-03-01), pages 1111 - 1117, XP002511596, ISSN: 0006-4971 *
GREEN M ET AL: "The role of antiviral and immunoglobulin therapy in the prevention of Epstein-Barr virus infection and post-transplant lymphoproliferative disease following solid organ transplantation.", TRANSPLANT INFECTIOUS DISEASE : AN OFFICIAL JOURNAL OF THE TRANSPLANTATION SOCIETY JUN 2001 LNKD- PUBMED:11395975, vol. 3, no. 2, June 2001 (2001-06-01), pages 97 - 103, XP002588395, ISSN: 1398-2273 *
KRAUSE I ET AL: "In vitro antiviral and antibacterial activity of commercial intravenous immunoglobulin preparations - A potential role for adjuvant intravenous immunoglobulin therapy in infectious diseases", TRANSFUSION MEDICINE 2002 GB LNKD- DOI:10.1046/J.1365-3148.2002.00360.X, vol. 12, no. 2, 2002, pages 133 - 139, XP002588396, ISSN: 0958-7578 *
NADAL DAVID ET AL: "Human immunoglobulin preparations suppress the occurrence of Epstein-Barr virus-associated lymphoproliferation", EXPERIMENTAL HEMATOLOGY (CHARLOTTESVILLE), vol. 25, no. 3, 1997, pages 223 - 231, XP008123558, ISSN: 0301-472X *

Also Published As

Publication number Publication date
ITRM20090558A1 (en) 2011-05-04

Similar Documents

Publication Publication Date Title
Das et al. Hepatitis B vaccine and immunoglobulin: key concepts
Hou et al. Epidemiology and prevention of hepatitis B virus infection
Krishnan et al. Developments in vaccination for herpes simplex virus
Friedman et al. Binding of complement component C3b to glycoprotein gC of herpes simplex virus type 1: mapping of gC-binding sites and demonstration of conserved C3b binding in low-passage clinical isolates
Malhi et al. Immunization with a self-assembling nanoparticle vaccine displaying EBV gH/gL protects humanized mice against lethal viral challenge
Jacobs Neutralizing antibodies mediate virus-immune pathology of COVID-19
CN113876938A (en) Construction and application of fusion protein vaccine platform
Jung et al. Immune control of oncogenic γ-herpesviruses
Pol Immunotherapy of chronic hepatitis B by anti HBV vaccine
Louthrenoo Treatment considerations in patients with concomitant viral infection and autoimmune rheumatic diseases
Niu et al. Synergistic and additive effects of cimetidine and levamisole on cellular immune responses to hepatitis B virus DNA vaccine in mice
WO2011055398A1 (en) A process for the production of immunoglobulines extracted from human plasma for therapeutic use, for the neutralization of the epstein-barr virus, and the medicine containing said immunoglobulins&#34;
CN102199217B (en) Hepatitis B virus multi-epitope fusion protein and preparation method and application thereof
Levine et al. Epstein-Barr virus and human disease
Deng et al. Recombinant vaccinia vector-based vaccine (Tiantan) boosting a novel HBV subunit vaccine induced more robust and lasting immunity in rhesus macaques
CN105061591A (en) Anti-hepatitis B virus surface antigen completely humanized human antibody and use thereof
Huang et al. New advances in hepatitis B vaccination for adults
EP0896545B1 (en) Method for producing drug containing hcv hyperimmune globulins
Sogut et al. Monoclonal antibodies specific for hepatitis B e antigen and hepatitis B core antigen
Gomes Post COVID vaccination reactive arthritis: a contemporary assailant
Grigorova Overview of the neutralizing antibody and memory B cell response kinetics in SARS-CoV-2 convalescent and/or mRNA vaccinated individuals
CN103627714A (en) Novel synthesized molecules of EBV (Epstein-Barr Virus) consensus DNA (Deoxyribonucleic Acid) sequence and vaccine formed thereby
Tamura et al. Genotypic analysis of Epstein‐Barr virus associated with nasopharyngeal carcinoma of Japanese patients
Cohen The Efficacy of a Peptide-Nucleic Acid Solution (RETICULOSE™*) for the Treatment of Hepatitis A and Hepatitis B-A Preliminary Controlled Human Clinical Trial
Foung et al. Uses of human monoclonal antibodies to human cytomegalovirus and varicella-zoster virus

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 10790681

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 10790681

Country of ref document: EP

Kind code of ref document: A1