EP0695312A1 - Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation - Google Patents

Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation

Info

Publication number
EP0695312A1
EP0695312A1 EP94914433A EP94914433A EP0695312A1 EP 0695312 A1 EP0695312 A1 EP 0695312A1 EP 94914433 A EP94914433 A EP 94914433A EP 94914433 A EP94914433 A EP 94914433A EP 0695312 A1 EP0695312 A1 EP 0695312A1
Authority
EP
European Patent Office
Prior art keywords
immunonanoparticles
anticoφs
nanoparticles
monoclonal
antibodies
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
EP94914433A
Other languages
German (de)
English (en)
French (fr)
Inventor
Nicole Bru-Magnierz
Jean-Claude 8 route de la Treille CHERMANN
François LESCURE
Jean-Marie Teulon
Pascal Breton
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.)
Institut National de la Sante et de la Recherche Medicale INSERM
UPSA SAS
Original Assignee
Institut National de la Sante et de la Recherche Medicale INSERM
UPSA SAS
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
Priority claimed from FR9304750A external-priority patent/FR2704227B1/fr
Application filed by Institut National de la Sante et de la Recherche Medicale INSERM, UPSA SAS filed Critical Institut National de la Sante et de la Recherche Medicale INSERM
Publication of EP0695312A1 publication Critical patent/EP0695312A1/fr
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/44—Antibodies bound to carriers
    • 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
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2812—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K17/00—Carrier-bound or immobilised peptides; Preparation thereof
    • C07K17/02—Peptides being immobilised on, or in, an organic carrier
    • C07K17/08—Peptides being immobilised on, or in, an organic carrier the carrier being a synthetic polymer
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • 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

Definitions

  • the invention relates to immunoparticles consisting of polymerized methylidene malonate nanoparticles coated with anti-CD4 antibodies and their use for the prophylaxis and / or treatment of pathologies due to infection by the HIV virus, or as biological reagents.
  • HIV virus human immunodeficiency virus
  • anti-CD4 monoclonal antibodies to inhibit the formation of syncytia (giant cells formed by the joining of healthy T lymphocytes and infected T lymphocytes) has been reported in F. Rey et al, Virology, 1991, 181. 165 -171.
  • Nanoparticle-monoclonal antibody conjugates in which the nanoparticle consists of a hexylcyano-acrylate polymer and the monoclonal antibody is an anti-alpha fetoprotein antibody (for application to the determination of alpha fetoprotein) of on the one hand, or an anti-sarcoma antibody (for anti-tumor application) have been described respectively in C. Kubiak et al, Int. J. Pharmaceutics, 1988, 41, 181-187 and L. Illum et al, J. Pharmacol. Exp. Therapeutics, 1984, 230. 733-736. However, these conjugates based on cyano-acrylic polymer exhibit toxicity with respect to T4 lymphocytes.
  • Liposomes coated with anti CD4 LEU3-A antibodies have been described in N. Phillips et al, Cancer Detect Prev., 1990, .14, 383-390. However, these liposomes have had the disadvantage of causing the appearance of anti-liposome antibodies in mice (N. Philipps, Second European Symposium on Controlled Drug Delivery, 1992, Noorduijk aan Zee, Netherlands, Abstract book, p. 172- 175).
  • immunonanoparticles carrying anti-CD4 monoclonal antibodies in the prophylaxis and / or treatment of pathologies due to infection with the HIV virus, in particular AIDS.
  • These immunonanoparticles have the advantages of giving said antibodies better stability, greater activity and better immunoreactivity. They are also completely biodegradable and have very low toxicity, their degradation products being themselves non-toxic. In addition, they can advantageously be sterilized and lyophilized.
  • the invention therefore relates to immunonanoparticles consisting of nanoparticles of polymer of a methylidene malonate compound of formula (I):
  • R ⁇ and R 2 identical or different, represent a linear or branched C -C5 alkyl group and n is an integer from 1 to 5, said nanoparticles being coated with anti-CD4 antibodies.
  • the compounds of formula (I) polymerize in the form of a polymer network to form the nanoparticles having a diameter between 100 and 400 nm, preferably of the order of 300 nm.
  • the average molecular weight (Mp) of the methylidene malonate polymers of formula (I) is of the order of 600 expressed in polystyrene equivalents.
  • the immunonanoparticles according to the invention can advantageously contain within their polymer network a biologically active substance, preferably an antiviral agent.
  • an antiviral agent preferably an antiviral agent.
  • antiviral agents non-limiting mention may be made of didanosine, ribavirin, zidovudine, aciclovir, vidarabine and zalcitabine.
  • the anti-CD4 monoclonal antibody with which the nanoparticle is coated to constitute the immunonanoparticle is chosen from the antibodies OKT4, OKT4-A, and the antibody IOT4a derived from hybridoma 13 B8-2, the antibody IOT4a being preferred.
  • monoclonal antibodies will be used in the form of a solution containing bovine serum albumin as supplied commercially.
  • anti-CD4 monoclonal antico ⁇ s marked with a fluorescent substance such as fluorescein isothiocyanate, or, preferably, unlabeled anti-CD4 antico ⁇ s.
  • the anti-CD4 monoclonal antico ⁇ s are fixed on the nanoparticles to constitute the immunonanoparticles by techniques known per se, preferably by passive adsorption, in particular in a phosphate buffer.
  • Said antico ⁇ s can also be attached to the nanoparticles by specific adsorption in particular via protein A of Staphylococcus aureus or a biotin-avidin bond.
  • the invention relates to the use of the immunonanoparticles according to the invention for the prophylaxis and / or the treatment of pathologies due to infection with the HIV virus and in particular for the manufacture of a medicament for the prophylaxis and / or the treatment of pathologies due to infection with the HIV virus.
  • the immunonanoparticles described above have properties of inhibiting the formation of syncytia in the experimental model of F. Rey et al, Virology, 1991, 181, 165-171.
  • the invention also relates to a method for detecting the presence of CD4 antigens in a sample of biological fluid originating from a mammal, human or animal, consisting in bringing a sample of this biological fluid into contact with immunonanoparticles described above, consisting of nanoparticles of polymer of a methylidene malonate compound of formula (I):
  • H 2 C C (I) X COO- (CH 2 ) n -COOR 2 in which Ri and R 2 , identical or different, represent a linear or branched C1-C5 alkyl group and n is an integer from 1 to 5 , said nanoparticles being coated with anti-CD4 antico ⁇ s, in sufficient quantity to form an antigen-antico ⁇ s complex capable of being detected.
  • the invention also relates to the use of the immunanoparticles coated with anti-CD4 antibodies described above as biological reagents, in particular as controls for the evaluation of the activity of compounds in a model inhibition of syncytia formation, or in immunoseparation, immunoprecipitation, immunostaining or immunopurification techniques.
  • EXAMPLE 1 Preparation of 1-ethoxycarbonyl-1-ethoxycarbonylmethyleneoxycarbonylethene immunonanoparticles coated with anti-CD4 IOT4a antibodies. 1) Preparation of nanoparticles:
  • the nanoparticles are prepared under sterile conditions at 25 ° C by emulsion-polymerization of monomer as described in F. Lescure et al,
  • nanoparticles are recovered by filtration on 8 ⁇ m filter paper (Whatman, Great Britain), then divided into aliquots, lyophilized and stored at room temperature.
  • the nanoparticles are resuspended in water for injection.
  • the average size of the nanoparticles is measured before and after lyophilization using a device for analyzing submicrometric particles (Coulter
  • the average particle diameter is around 320 nm.
  • the molecular masses of the constituent polymers of nanoparticles are determined by steric exclusion chromatography (SEC) (Polymer laboratories, Great Britain). For the analysis, the nanoparticulate suspension is ultracentrifuged at 200,000 g for 5 minutes, then the pellet taken up in THF to which 0.5% toluene is added, is injected. Detection is done by refractometry. 2) Attachment of the antico ⁇ s to the nanoparticles:
  • the nanoparticles are diluted in a phosphate buffer pH 7.4 to 2.7 mg / ml.
  • 100 ⁇ ⁇ of monoclonal antico4s IOT4a labeled or not with fluorescein isothiocyanate (clone 13B8-2, solution at 100 ⁇ g / ml containing 0.2% bovine serum albumin, IMMUNOTECH, France) is added to a suspension of 450 ⁇ l of nanoparticles.
  • the mixture is incubated at 20 ° C for 2 hours with shaking.
  • the immunonanoparticles are added at a polymer concentration of 50 ⁇ g ml to an RPMI 1640 medium free of phenol red (GIBCO, USA) containing 1% glutamine, 1% penicillin, 1% streptomycin, 1% neomycin, and 10% fetal calf serum.
  • the nanoparticles coated with antico ⁇ s are separated from the free antico ⁇ s by ultracentrifugation at 40,000 g for 5 min, immediately and 2 h after the start of incubation at 37 C.
  • nanoparticles of isohexylcyano acrylate were also prepared as described in the patents.
  • the nanoparticles coated with antico ⁇ s are separated from the free antico ⁇ s by centrifugation at 80,000 g for 5 min.
  • the concentration of free antico ⁇ s is calculated by reference to a calibration curve, and the percentage of fixed antico ⁇ s is deducted by difference.
  • Table 1 reports the results obtained with the immunonanoparticles of Example 1;
  • Table 2 reports, by way of comparative example, the results obtained with PIHCA nanoparticles coated with the same antico ⁇ s as the immunonanoparticles of Example 1.
  • the nanoparticles prepared according to step 1) of Example 1 are diluted in a phosphate buffer pH 7.4 at 2.7 mg / ml.
  • 100 ⁇ l of OKT4-A monoclonal antico ⁇ s 50 ⁇ g / ml solution containing 0.1% bovine serum albumin, ORTHO DIAGNOSTIC SYSTEMS, USA
  • fluorescein isothiocyanate labeled with fluorescein isothiocyanate are added to a suspension of 450 ⁇ l of nanoparticles.
  • the mixture is incubated at 20 ° C for 2 hours with shaking.
  • the stability of these immunonanoparticles was tested under the experimental conditions of Example 2.
  • EXAMPLE 4 Preparation of 1-ethoxycarbonyl methylenoxycarbonylethene immunonanoparticles coated with OKT4 anti-CD4 antico ⁇ s and study of their stability.
  • the immunonanoparticles are prepared according to the protocol described in Example 3 using 100 ⁇ l of monoclonal antico ⁇ s OKT4 (ORTHO DIAGNOSTIC SYSTEMS, USA) labeled with fluorescein isothiocyanate.
  • the CEM and MT4 cell lines are used.
  • Infection with HIV-1 BRU virus (F. Barre-Sinoussi et al., Science, 1983, 220, 868-871) is performed using a cell suspension at 2.10 cells / ml incubated at 37 ° C for lh using an equal volume of virus stock dilution. After infection, the cells are washed by centriguation 4 times with PBS buffer to remove the unbound viral particles.
  • the cells are then cultured at a concentration of 5 ⁇ 10 ⁇ cells / ml in RPMI 1640 medium (GIBCO, USA) containing phenol red supplemented with 10% fetal calf serum, 1% antibiotics and 1% glutamine. Every 3 or 4 days, the cells are diluted 3 times (MT4 cells) or 4 times (CEM cells) and returned to culture in a new culture medium. Lymphocyte infection is demonstrated using the reverse transcriptase test.
  • the syncytia are generally observed at 1 or 2 days of incubation after mixing a volume of chronically infected CEM cells with 2 volumes of MT4 cells at 2.10 5 cells / ml.
  • the ability of anti-CD4 monoclonal antibodies and immunonanoparticles carrying anti-CD4 antibodies to inhibit the formation of syncytia is evaluated by incubating the MT4 cells with the various preparations for 2 h and then adding the infected CEM cells.
  • Table 5 relates to the results obtained with the immunonanoparticles according to the invention carrying antico ⁇ s IOT4a;
  • Table 6 reports, by way of comparative example, the results obtained with PIHCA nanoparticles coated with IOT4a antico ⁇ s;
  • Table 7 reports, by way of comparative example, the results obtained using the antico fixés IOT4a which is not fixed.
  • Example 1 coated with anti-CD4 IOT4a monoclonal antibodies were tested according to the experimental protocol of Example 5, using instead of the HIV-1 BRU virus, the viral strain PAS 246 (F. Rey et al., Virology, 1991, 181, 165-171).

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • Communicable Diseases (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Oncology (AREA)
  • Virology (AREA)
  • Engineering & Computer Science (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)
  • Medicinal Preparation (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
EP94914433A 1993-04-22 1994-04-22 Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation Ceased EP0695312A1 (fr)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
FR9304750 1993-04-22
FR9304750A FR2704227B1 (fr) 1993-04-22 1993-04-22 Immunonanoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation pour la prophylaxie et/ou le traitemeent de pathologies dues a une infection par le virus hiv.
US10170093A 1993-08-04 1993-08-04
US101700 1993-08-04
PCT/FR1994/000459 WO1994024168A1 (fr) 1993-04-22 1994-04-22 Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation

Publications (1)

Publication Number Publication Date
EP0695312A1 true EP0695312A1 (fr) 1996-02-07

Family

ID=26230266

Family Applications (1)

Application Number Title Priority Date Filing Date
EP94914433A Ceased EP0695312A1 (fr) 1993-04-22 1994-04-22 Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation

Country Status (10)

Country Link
EP (1) EP0695312A1 (ja)
JP (1) JPH08512287A (ja)
CN (1) CN1121725A (ja)
AU (1) AU6681494A (ja)
CA (1) CA2160983A1 (ja)
CZ (1) CZ276295A3 (ja)
FI (1) FI954827A0 (ja)
HU (1) HUT73392A (ja)
SK (1) SK130395A3 (ja)
WO (1) WO1994024168A1 (ja)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FR2722411B1 (fr) * 1994-07-18 1996-10-04 Union Pharma Scient Appl Immunonanoparticules revetues d'anticorps monoclonaux anti-beta2 microglobuline et leur utilisation pour la prophylaxie et/ou le traitement de pathologies dues a une infection par le virus hiv
JP4611561B2 (ja) * 2000-04-12 2011-01-12 Ｄｉｃ株式会社 新規なビニル系樹脂およびその製造方法
GB2381792B (en) * 2000-04-12 2004-08-04 Dainippon Ink & Chemicals Antifouling coating composituion
JP3961312B2 (ja) 2002-02-26 2007-08-22 株式会社デンソー 内燃機関の制御装置
WO2012113348A1 (zh) * 2011-02-25 2012-08-30 厦门大学 抗cd4蛋白的单克隆抗体及其活性片段及用途
CN110903394B (zh) * 2019-12-27 2021-09-07 源道隆（苏州）医学科技有限公司 可结合cd4的多肽及其应用

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0329184A3 (en) * 1988-02-19 1990-05-23 Neorx Corporation Antimers and antimeric conjugation

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9424168A1 *

Also Published As

Publication number Publication date
FI954827A (fi) 1995-10-11
WO1994024168A1 (fr) 1994-10-27
CN1121725A (zh) 1996-05-01
CZ276295A3 (en) 1996-06-12
CA2160983A1 (en) 1994-10-27
JPH08512287A (ja) 1996-12-24
HU9503028D0 (en) 1995-12-28
FI954827A0 (fi) 1995-10-11
SK130395A3 (en) 1996-06-05
HUT73392A (en) 1996-07-29
AU6681494A (en) 1994-11-08

Similar Documents

Publication Publication Date Title
Knight et al. Non-adherent, low-density cells from human peripheral blood contain dendritic cells and monocytes, both with veiled morphology.
CA2144425C (fr) Microparticules portant des antigenes et leur utilisation pour l'introduction de reponses humorales ou cellulaires
FR2615394A1 (fr) Utilisation de certains 17-cetosteroides, eventuellement avec un immunomodulateur et/ou un agent antiviral, pour fabriquer un medicament pour le traitement et la prevention des infections retrovirales
JPH10513053A (ja) 未分化細胞の製造方法
JP4216076B2 (ja) 性的に伝達される病気の予防および治療のための薬剤−ｉ
WO1994024168A1 (fr) Immunoparticules porteuses d'anticorps monoclonaux anti-cd4 et leur utilisation
EP1851321B1 (fr) Epitopes de vih et composition pharmaceutique les contenant
AU783250B2 (en) Autologous adoptive immunotherapy with primed antigen-specific T cells or B cells
EP0542630A2 (fr) Utilisation de dérivés d'amphotéricine B, comme inhibiteurs de protéases
Esparza et al. Inhibition of macrophage tumoricidal activity by immune complexes and altered erythrocytes.
FR2704227A1 (fr) Immunonanoparticules porteuses d'anticorps monoclonaux anti-CD4 et leur utilisation pour la prophylaxie et/ou le traitemeent de pathologies dues à une infection par le virus HIV.
WO1996002278A1 (fr) Immunonanoparticules revetues d'anticorps monoclonaux anti-beta-2 microglobuline
Dore-Duffy Differential effect of prostaglandins and other products of arachidonic acid metabolism on measles virus replication in Vero cells
FR2626472A1 (fr) Utilisation des proteines du lait pour la fabrication d'un medicament antiviral
EP1663146B1 (fr) Procede de preparation de microparticules bioresorbables, microparticules obtenues et utilisation
JP2022537967A (ja) 同種異系ｃａｒ－ｔ細胞療法
JPH08501803A (ja) アミノ糖および糖蛋白、その製法、それらアミノ糖または糖蛋白を含む医薬製剤およびそれらの製剤の用途
JP2007525473A (ja) 単球由来の自己寛容誘導性自家細胞および薬学的組成物におけるその使用
US5669396A (en) Methods of detecting and treating vaso-occlusive crisis in sickle cell disease
EP2537880A2 (en) Carbosilane dendrimers and the use thereof as antiviral agents
WO2002079298A1 (en) Agents for the prevention and treatment of sexually transmitted diseases -ii
EP0804408B1 (fr) Nouveaux composes 2,6-di-tert-butylphenols substitues en position 4, les compositions pharmaceutiques les contenant et leur procede de preparation
EP0149581A2 (fr) Nouvelles compositions contenant des oligo-muramylpeptides et leur application, notamment pour l'activation des macrophages
Breton et al. Biocompatible poly (methylidene malonate)-made materials for pharmaceutical and biomedical applications
CA2215414A1 (en) Compositions and methods for reducing adhesiveness of defective red blood cells

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 19951017

AK Designated contracting states

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FR GB GR IE IT LI LU MC NL PT SE

AX Request for extension of the european patent

Free format text: SI PAYMENT 951017

RAX Requested extension states of the european patent have changed

Free format text: SI PAYMENT 951017

RIN1 Information on inventor provided before grant (corrected)

Inventor name: BRETON, PASCAL

Inventor name: TEULON, JEAN-MARIE

Inventor name: LESCURE, FRANCOIS

Inventor name: CHERMANN, JEAN-CLAUDE, 8,ROUTE DE LA TREILLE

Inventor name: BRU-MAGNIERZ, NICOLE

GRAG Despatch of communication of intention to grant

Free format text: ORIGINAL CODE: EPIDOS AGRA

17Q First examination report despatched

Effective date: 19960415

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION HAS BEEN REFUSED

18R Application refused

Effective date: 19961204