CN1339318A - Method for treating AIDS and its preparing method - Google Patents

Method for treating AIDS and its preparing method Download PDF

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Publication number
CN1339318A
CN1339318A CN00123483A CN00123483A CN1339318A CN 1339318 A CN1339318 A CN 1339318A CN 00123483 A CN00123483 A CN 00123483A CN 00123483 A CN00123483 A CN 00123483A CN 1339318 A CN1339318 A CN 1339318A
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Prior art keywords
epitope
medicine
aids
eldkwa
immune deficiency
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CN00123483A
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CN1172717C (en
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陈应华
肖翌
田海军
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Tsinghua University
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Tsinghua University
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Priority to CNB001234838A priority Critical patent/CN1172717C/en
Priority to PCT/CN2001/001192 priority patent/WO2002032452A1/en
Priority to AU2002214920A priority patent/AU2002214920A1/en
Publication of CN1339318A publication Critical patent/CN1339318A/en
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Publication of CN1172717C publication Critical patent/CN1172717C/en
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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12Antivirals
    • A61P31/14Antivirals for RNA viruses
    • A61P31/18Antivirals for RNA viruses for HIV
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K16/00Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/08Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
    • C07K16/10Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
    • C07K16/1036Retroviridae, e.g. leukemia viruses
    • C07K16/1045Lentiviridae, e.g. HIV, FIV, SIV
    • C07K16/1063Lentiviridae, e.g. HIV, FIV, SIV env, e.g. gp41, gp110/120, gp160, V3, PND, CD4 binding site
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00Medicinal preparations containing antigens or antibodies
    • A61K2039/505Medicinal preparations containing antigens or antibodies comprising antibodies
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2740/00Reverse transcribing RNA viruses
    • C12N2740/00011Details
    • C12N2740/10011Retroviridae
    • C12N2740/16011Human Immunodeficiency Virus, HIV
    • C12N2740/16111Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Genetics & Genomics (AREA)
  • Oncology (AREA)
  • Biophysics (AREA)
  • AIDS & HIV (AREA)
  • General Chemical & Material Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Communicable Diseases (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Hematology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The AIDS treating medicine of the present invention includes at least a kind of monoclonal antibody, the antigen of which is ELDKWA neutralizing epitope and its variation epitopes ELEKWA, ELNKWA and ELDEWAF of human immune deficiency virus transmembrane protein gp41. It is prepared through the steps of: artificial synthesis of at least one polypeptide containing ELDKWA neutralizing epitope and/or its variation epitopes of human immune deficiency virus transmembrane protein gp41; coupling the said polypeptide to carrier protein or carrier polypeptide; immunizing animal with the said conjugate; preparing hybrid tumor; and compounding the obtained antibody with medicinal adjuvant to obtain the medicine.

Description

A kind of medicine for the treatment of acquired immune deficiency syndrome (AIDS) and preparation method thereof
The present invention relates to medicine with the treatment acquired immune deficiency syndrome (AIDS) of biotechnology preparation and preparation method thereof, particularly relate to medicine with the treatment acquired immune deficiency syndrome (AIDS) of the Monoclonal Antibody of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) memebrane protein and preparation method thereof.
Acquired immune deficiency syndrome (AIDS) claims acquired immune deficiency syndrome (AIDS) again, is a kind of immune disease that is caused by HIV (human immunodeficiency virus) (HIV).Because it is propagated rapidly, case fatality rate is high, and does not still have the specific treatment method at present, does not more have vaccine and can prevent, thereby the title of " 20 century plague " is arranged.
Existingly be used for only deferrable period of disease and prolong patient's life of clinical anti-AIDS drug, and cost an arm and a leg, toxicity is big.Recently external anti-AIDS drug clinical research result proves, the mucosa that can suppress HIV-1 virus at neutralizing antibody (comprising the monoclonal antibody 2F5 of ELDKWA-epitope specificity) being used in combination in passive immunotherapy of the several specific neutralizing epitopes on the HIV-1 memebrane protein gp160 (precursor protein of memebrane protein gp120 and transmembrane protein gp41) infects and mother-to-baby transmission, and can remove HIV-1 virus (Nature Medicine 1999,5:204 in the blood; Nature Medicine 2000,6:200; Nature Medicine1999,5:211); The epitope specificity monoclonal antibody of main neutralizing epitope ELDKWA on human immunodeficiency virus (HIV-1) the transmembrane protein gp41 can the multiple HIV-1 Strain of vitro inhibition target cell infection (J.Virology 1993,67:6642; AIDS Res.Human Retroviruses 1994,10:1651; AIDS1996,10:587).
The medicine that the purpose of this invention is to provide a kind of effective treatment acquired immune deficiency syndrome (AIDS), this medicine can tackle the variation of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus).
Another object of the present invention provides a kind of method for preparing the medicine of above-mentioned treatment acquired immune deficiency syndrome (AIDS).
For achieving the above object, the present invention takes following design: a kind of medicine for the treatment of acquired immune deficiency syndrome (AIDS), and it consists essentially of at least a monoclonal antibody, and the antigen of described antibody is ELDKWA epi-position and the variant epitope thereof on human immunodeficiency virus's transmembrane protein gp41.
Described variant epitope is ELEKWA, ELNKWA or ELDEWA.
It is preferably composed of the following components that the present invention treats the medicine of acquired immune deficiency syndrome (AIDS):
The monoclonal antibody of ELDKWA-epitope specificity
The monoclonal antibody of ELEKWA-epitope specificity
The monoclonal antibody of ELNKWA-epitope specificity
The monoclonal antibody of ELDEWA-epitope specificity
A kind of method for preparing the medicine of above-mentioned treatment acquired immune deficiency syndrome (AIDS) consists essentially of following steps:
(1), synthetic at least one contains the epitope polypeptide of the neutralizing epitope on human immunodeficiency virus's transmembrane protein gp41 respectively, described neutralizing epitope be selected from the ELDKWA epi-position or/and its variant epitope or at least once multiple ELDKWA epi-position or/and its variant epitope;
(2), above-mentioned epitope polypeptide is coupled to respectively on carrier protein or the carrier polypeptide forms coupling matter;
(3), be equipped with acceptable adjuvant immunity animal respectively with above-mentioned coupling matter;
(4), adopt conventional cell-fusion techniques to prepare hybridoma respectively;
(5), the antibody purification that will obtain from above-mentioned different hybridoma cell lines, be mixed and made into the medicine of treatment acquired immune deficiency syndrome (AIDS).
Wherein, at least one polypeptide of described synthetic is preferably: contain at least once multiple neutralizing epitope ELDKWA or its variant epitope on human immunodeficiency virus's transmembrane protein gp41 respectively.
The variant epitope of described ELDKWA epi-position is ELEKWA, ELNKWA or ELDEWA.
Studies show that, in treating AIDS, can reduce the carrying capacity of virus, delay the carrying out of disease at the antibody of neutralizing epitope on the HIV-1 memebrane protein.The medicine that the present invention treats acquired immune deficiency syndrome (AIDS) is the medicine that contains the multiple antibody of main neutralizing epitope ELDKWA epi-position on the anti human immune deficiency virus transmembrane protein gp41 and variant epitope (as ELEKWA, ELNKWA, ELDEWA) thereof, even produce in HIV (human immunodeficiency virus) (Human Immunodeficiency Virus) under the situation of variation, injected in the human body of this medicine and also had the virus that corresponding antibody is killed variation.
Medicine of the present invention is avirulence not only, and in the immunization therapy effect that improves acquired immune deficiency syndrome (AIDS), also can reduce the treating AIDS cost.
The present invention adopts the method for synthetic epitope polypeptide to induce, prepare the monoclonal antibody of predetermined epitope specificity, overcome and need utilize native protein or recombiant protein immunity, a large amount of then screenings and evaluation just can obtain many complex work of the monoclonal antibody of predetermined epitope specificity.
According to the present invention, can produce the medicine of its respective type very soon according to the variation situation of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), need not test for a long time, reduce production costs.This technology will produce significant impact to world's preventive medicine research, and will bring huge economic benefit and social benefit.
The invention will be further described below in conjunction with non-limiting specific embodiment.
Embodiment one: by the variant epitope ELNKWA-epitope specificity antibody of the neutralizing epitope ELDKWA on the anti-HIV-1 gp41 is the medicine of the treatment acquired immune deficiency syndrome (AIDS) made of main active, by following steps production:
(1), one of synthetic contains the epitope polypeptide of 4 multiple ELNKWA neutralizing epitopes on human immunodeficiency virus's transmembrane protein gp41
CELNKWAGELNKWAGELNKWAGELNKWA;
(2), utilize MBS (m-maleimidobenzoyl-N-hydoxy succinimide ester) with above-mentioned epitope polypeptide and carrier protein BSA coupling connection;
(3), above-mentioned coupling matter is mixed with freund adjuvant that (volume ratio of two kinds of materials is coupling matter: immune Balb/c mice freund adjuvant=1: 1).Use for the first time complete freund adjuvant, later per two all immunity once, full freund adjuvant toos many or too much for use.The antigen dose of each immunity is: contain the coupling matter of 10 microgram epitope polypeptides/time/only, immunity is 3 times altogether;
(4), adopt conventional cell-fusion techniques to merge the mouse boosting cell after the immunity and mouse myeloma cell line, and the preparation hybridoma;
(5), from above-mentioned hybridoma cell line, filter out the clone that can produce the monoclonal antibody of predefined epitope specificity, the antibody of purifying is made the medicine of treatment acquired immune deficiency syndrome (AIDS).
This medicine using dosage is in actual applications determined according to patient's the state of an illness.
Embodiment two: by the antibody of the neutralizing epitope ELDKWA on the anti-HIV-1 gp41 and variant epitope ELNKWA, ELEKWA and ELDEWA is the medicine of the treatment acquired immune deficiency syndrome (AIDS) made of main active, by following steps production:
(1), 4 of synthetic contain neutralizing epitope ELDKWA on human immunodeficiency virus's transmembrane protein gp41 and the polypeptide of variant epitope ELNKWA, ELEKWA or ELDEWA thereof respectively:
C?ELDKWA?G?ELDKWA?G?ELDKWA?G?ELDKWA
C?ELNKWA?G?ELNKWA?G?ELNKWA?G?ELNKWA
C?ELEKWA?G?ELEKWA?G?ELEKWA?G?ELEKWA
C?ELDEWA?G?ELDEWA?G?ELDEWA?G?ELDEWA
(2), utilize glutaraldehyde or MBS that aforementioned polypeptides is coupled to respectively on the carrier protein bovine serum albumin and form coupling matter;
(3), above-mentioned coupling matter is mixed with freund adjuvant respectively (volume ratio of two kinds of materials is coupling matter: freund adjuvant=1: 1), immune Balb/c mice respectively.Use for the first time complete freund adjuvant, later per two all immunity once, full freund adjuvant toos many or too much for use.The antigen dose of each immunity is: contain the coupling matter of 10 microgram epitope polypeptides/time/only, immunity is 3 times altogether;
(4), adopt conventional cell-fusion techniques to merge the mouse boosting cell after the immunity and mouse myeloma cell line, and the preparation hybridoma;
(5), filter out the clone that can produce the monoclonal antibody of predefined 4 kinds of epitope specificities from above-mentioned hybridoma cell line.
(6), obtain 4 kinds of monoclonal antibodies of neutralizing epitope ELDKWA on the anti-HIV-1 gp41 and variant epitope ELNKWA, ELEKWA, ELDEWA respectively from above-mentioned hybridoma cell line, above-mentioned antibody is mixed, make the medicine of treatment acquired immune deficiency syndrome (AIDS), wherein, the probability of occurrence of the various epi-positions that drawn by the statistical procedures that is suitable for the crowd of the addition of various antibody is adjusted.
Use medicine of the present invention, can obviously resist the variation of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), reduce the carrying capacity of HIV (human immunodeficiency virus) (Human Immunodeficiency Virus), delay the carrying out of disease, reach the purpose of healing at last.

Claims (6)

1, a kind of medicine for the treatment of acquired immune deficiency syndrome (AIDS), it consists essentially of at least a monoclonal antibody, and the antigen of described antibody is ELDKWA epi-position and the variant epitope thereof on human immunodeficiency virus's transmembrane protein gp41.
2, a kind of medicine for the treatment of acquired immune deficiency syndrome (AIDS) according to claim 1, it is characterized in that: described variant epitope is ELEKWA, ELNKWA or ELDEWA.
3, a kind of medicine for the treatment of acquired immune deficiency syndrome (AIDS) according to claim 1 and 2, it is characterized in that: it is composed of the following components: the monoclonal antibody of ELDKWA-epitope specificity
The monoclonal antibody of ELEKWA-epitope specificity
The monoclonal antibody of ELNKWA-epitope specificity
The monoclonal antibody of ELDEWA-epitope specificity
4, a kind of method for preparing the medicine of the described treatment acquired immune deficiency syndrome (AIDS) of claim 1-3 consists essentially of following steps:
(1), synthetic at least one contains the epitope polypeptide of the neutralizing epitope on human immunodeficiency virus's transmembrane protein gp41 respectively, described neutralizing epitope be selected from the ELDKWA epi-position or/and its variant epitope or at least once multiple ELDKWA epi-position or/and its variant epitope;
(2), above-mentioned epitope polypeptide is coupled to respectively on carrier protein or the carrier polypeptide forms coupling matter;
(3), be equipped with acceptable adjuvant immunity animal respectively with above-mentioned coupling matter;
(4), adopt conventional cell-fusion techniques to prepare hybridoma respectively;
(5), the antibody purification that will obtain from above-mentioned different hybridoma cell lines, be mixed and made into the medicine of treatment acquired immune deficiency syndrome (AIDS).
5, the method for the medicine of preparation treatment acquired immune deficiency syndrome (AIDS) according to claim 4, it is characterized in that: at least one polypeptide of described synthetic contains at least once multiple neutralizing epitope ELDKWA or its variant epitope on human immunodeficiency virus's transmembrane protein gp41 respectively.
6, according to the method for the medicine of claim 4 or 5 described preparations treatment acquired immune deficiency syndrome (AIDS), it is characterized in that: the variant epitope of described ELDKWA epi-position is ELEKWA, ELNKWA or ELDEWA.
CNB001234838A 2000-08-18 2000-08-18 Method for treating AIDS and its preparing method Expired - Fee Related CN1172717C (en)

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Application Number Priority Date Filing Date Title
CNB001234838A CN1172717C (en) 2000-08-18 2000-08-18 Method for treating AIDS and its preparing method
PCT/CN2001/001192 WO2002032452A1 (en) 2000-08-18 2001-07-20 Composition for aids and method producing it
AU2002214920A AU2002214920A1 (en) 2000-08-18 2001-07-20 Composition for aids and method producing it

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Application Number Priority Date Filing Date Title
CNB001234838A CN1172717C (en) 2000-08-18 2000-08-18 Method for treating AIDS and its preparing method

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CN1172717C CN1172717C (en) 2004-10-27

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AU (1) AU2002214920A1 (en)
WO (1) WO2002032452A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1327897C (en) * 2002-09-24 2007-07-25 重庆前沿生物技术有限公司 Peptide derivative fusion inhibitors of HIV infection

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108779168A (en) 2015-12-05 2018-11-09 沃迪奥斯大学医院中心 HIV bonding agents
WO2020012435A1 (en) 2018-07-13 2020-01-16 Lausanne University Hospital Hiv binding agents
US20220267416A1 (en) 2019-07-15 2022-08-25 Lausanne University Hospital Hiv binding agents

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5459060A (en) * 1989-08-24 1995-10-17 Bioclonetics Incorporated Human monoclonal antibodies directed against the transmembrane glycoprotein (gp41) of human immunodeficiency virus-1 (HIV-1)
WO1994012533A1 (en) * 1992-11-23 1994-06-09 President And Fellows Of Harvard College Gp41 mutants and their use as hiv therapeutics
CN1111540C (en) * 1993-06-09 2003-06-18 康诺特实验室有限公司 Placed in-line synthetic HIV-1 peptide class
US5380668A (en) * 1993-07-06 1995-01-10 University Of Utah Research Foundation Compounds having the antigenicity of hCG
JPH09502348A (en) * 1993-09-11 1997-03-11 ポリムン・シエンティフィーク・イムノビオロギッシェ・フォルシュング・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング Peptides for inducing neutralizing antibodies against genetically branched HIV-1 strains

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1327897C (en) * 2002-09-24 2007-07-25 重庆前沿生物技术有限公司 Peptide derivative fusion inhibitors of HIV infection

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WO2002032452A1 (en) 2002-04-25
CN1172717C (en) 2004-10-27

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