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EP0688432A1 - Kit for diagnosing an hcg- or hcg fragment-secreting cancer and immunotherapeutical means therefor - Google Patents

Kit for diagnosing an hcg- or hcg fragment-secreting cancer and immunotherapeutical means therefor

Info

Publication number
EP0688432A1
EP0688432A1 EP94909172A EP94909172A EP0688432A1 EP 0688432 A1 EP0688432 A1 EP 0688432A1 EP 94909172 A EP94909172 A EP 94909172A EP 94909172 A EP94909172 A EP 94909172A EP 0688432 A1 EP0688432 A1 EP 0688432A1
Authority
EP
European Patent Office
Prior art keywords
hcg
polypeptides
polypeptide
primary sequence
fragments
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.)
Withdrawn
Application number
EP94909172A
Other languages
German (de)
French (fr)
Inventor
Dominique Bellet
Frédéric TROALEN
Nathalie Rouas-Freiss
Jean-Michel Bidart
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.)
Cephalon France SAS
Original Assignee
Laboratoire L Lafon SA
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 FR9302820A external-priority patent/FR2702494B1/en
Priority claimed from FR9312039A external-priority patent/FR2710845B1/en
Application filed by Laboratoire L Lafon SA filed Critical Laboratoire L Lafon SA
Publication of EP0688432A1 publication Critical patent/EP0688432A1/en
Withdrawn legal-status Critical Current

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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
    • A61K39/0005—Vertebrate antigens
    • A61K39/0006—Contraceptive vaccins; Vaccines against sex hormones
    • 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
    • A61K39/0005—Vertebrate antigens
    • A61K39/0011—Cancer antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/575—Hormones
    • C07K14/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
    • G01N33/76—Human chorionic gonadotropin including luteinising hormone, follicle stimulating hormone, thyroid stimulating hormone or their receptors

Definitions

  • Diagnostic kit for a cancer secreting hCG or hCG fragments and means intended for the therapy of such cancer.
  • the present invention relates to a method for the treatment of cancer secreting hCG or fragments of hCG and means for implementing this method.
  • Human gonadotropic chorionic hormone Human gonadotropic chorionic hormone
  • hCG is an oligomeric glycoprotein consisting of the non-covalent association of two subunits ⁇ (hCG- ⁇ ) and ⁇ (hCG- ⁇ ). It belongs to the family of gonadothyreotropic hormones also comprising folliculotropic hormone (FSH), luteotropic hormone (LH) and thyreotropic hormone (TSH). HCG is secreted physiologically by the trophoblast from the start of pregnancy and stimulates the corpus luteum to maintain the synthesis of steroid hormones essential for implantation of the embryo.
  • FSH folliculotropic hormone
  • LH luteotropic hormone
  • TSH thyreotropic hormone
  • hCG and / or its subunits are also secreted by a majority of trophoblastic tumors and certain non-trophoblastic tumors, thus constituting biological markers which can help in the diagnosis or prognosis and in the monitoring of these tumors (Bellet and al. Rev. Prat. 40: 1677-1990).
  • hCG- ⁇ is detectable in healthy men ( ⁇ 1 ng / ml) and women over 45 years of age without menopause ( ⁇ 3 ng / ml). In contrast, hCG- ⁇ is not detectable in the normal population.
  • trophoblastic tumors In the case of trophoblastic tumors, it has been shown that benign (hydatidiform moles) and malignant (choriocarcinomas) placental tumors are characterized by an important serum hCG / hCG- ⁇ ratio: the percentage of free hCG which is 0.05 to 1% during normal pregnancies amounts to 1-5% in the case of moles and exceeds 5% in choriocarcinomas.
  • This serum hCG / hCG- ⁇ ratio is used as a control for the development of testicular tumors: a percentage of hCG- ⁇ becoming less than 5% is often characteristic of a favorable development of the tumor during treatment. This ratio will rise quickly in the event of an unfavorable development or a relapse.
  • hCG- ⁇ not detectable in healthy individuals, is found in many patients with certain non-trophoblastic tumors (tumors of the bladder, pancreas, cervix, tumors without primitive found, endometrial and lung tumors).
  • the subject of the present invention is a method of treating cancer secreting hCG or fragments of hCG which consists in
  • the polypeptides are formed by a chain of 9 to 40 amino acids corresponding to a part of the primary sequence of hCG. They may in particular be polypeptides comprising from 15 to 40 amino acids or, in the case of polypeptides capable of being recognized by cytotoxic T lymphocytes, shorter polypeptides having in particular from 9 to 11 amino acids.
  • set of polypeptides covering at least part of the primary sequence of hCG denotes a set of polypeptides as defined above which correspond to successive sequences of hCG or of its fragments or better which correspond to sequences overlapping.
  • At least one set of polypeptides will be used which covers the primary sequence of the ⁇ subunit of hCG or at least part of this sequence.
  • the synthesis of the ⁇ subunit is dependent on an ultigenic group comprising 7 genes (Talmadge et al., Nucl. Acids Res. 12, 8415, 1984). Analysis of the nucleotic sequences shows that three of these genes (1, 3, 5) have an identical structure, four others of these genes (2, 6, 7 and 8) encode a variant at codon 117 with substitution. alanine instead of an aspartic acid.
  • a study of gene expression shows that genes 1, 3 and 5 are expressed by the normal placenta.
  • tumourogenesis processes involving non-trophoblastic cells the expression of several genes leads to both the secretion of the normal protein and of a protein mutated to 117.
  • ⁇ subunit denotes both the normal protein and the protein mutated at 117.
  • polypeptide corresponding to the antigen responsible for the state of immunization means a polypeptide which has the same sequence as one of the polypep ⁇ tides responsible for the state of immunization or a sequence resulting from the combination of sequences at least partial of the polypeptides responsible for the state of immunization.
  • the state of immunization with respect to one of the polypeptides can be identified by a test for stimulating the proliferation of T lymphocytes by the polypeptides, or also by a test for cytotoxicity of T lymphocytes against vis the polypeptides.
  • the recognized polypeptides are formed by a sequence of at least 9 amino acids corresponding to a part of the primary sequence of 1'hCG.
  • the polypeptides interact with the class I molecules of the major histocompatibility complex of the patient. Each class I molecule interacts with peptides with specific motifs.
  • the present invention also relates to a diagnostic kit for a cancer secreting hCG or hCG fragments comprising a set of polypeptides covering at least part of the primary sequence of 1'hCG.
  • the subject of the present invention is also a composition intended for the immunotherapy of cancers secreting hCG or hCG fragments comprising an effective dose of a polypeptide corresponding to at least part of the primary sequence of hCG and in particular a polypeptide corresponding to at least part of the primary sequence of the hCG ⁇ subunit and a pharmaceutically acceptable vehicle.
  • the present invention also relates to the use of a polypeptide corresponding to at least a part of the primary sequence of 1'hCG and in particular of a polypeptide corresponding to at least a part of the primary sequence of the ⁇ subunit hCG for the manufacture of a composition for the immunotherapy of cancers secreting hCG or fragments of hCG.
  • poly ⁇ peptides can be used in free form or in a more immunogenic form, in particular a form in which the peptide is coupled to a polylysine matrix according to the MAP technique (Posnett et al. J. Biol. Chem. 263, 17, 1988).
  • the polypeptide is advantageously administered with an adjuvanting immunity.
  • adjuvants of immunity we may cite muramyldipeptide and SAF-1 (Allison et al., J. Immunol. Methods 45, 157, 1986).
  • polypeptide can be protected at its NH 2 and COOH terminal ends in order to reduce the risks of degradation.
  • the BCG vaccine (Bacillus Calmette Guérin) constituted by living attenuated strains of tuberculous bacillus (see in particular D. Lamm et al, New England Journal of Medicine 325, 17, 1205, 1991).
  • the present invention therefore further relates to a composition intended for the immunotherapy of cancers secreting hCG or hCG fragments comprising an effective dose of a polypeptide corresponding to at least part of the primary sequence of hCG and an effective dose of BCG in a pharmaceutically acceptable vehicle.
  • immunotherapy can be carried out with doses of polypeptides from 1 to 100 mg administered in particular at the level of the tumor.
  • the polypeptides can be administered in the case of bladder cancer, as is already practiced in the case of immunotherapy with BCG, that is to say by intravesical instillation one week after resection. tumor, then this administration is repeated every week for 6 weeks and additional intravesical administration is performed 3, 6, 12, 18 and 24 months later.
  • Percutaneous or general administrations are also possible. Description of the figures.
  • Fig. 1 shows the proliferation of lymphocytes from a patient with bladder cancer in the presence of hCG, free ⁇ and ⁇ subunits and various polypeptides.
  • Fig. 2 shows the proliferation of lymphocytes from another patient with bladder cancer in the presence of free ⁇ and ⁇ subunits and different polypeptides.
  • Fig. 3A shows the proliferation of lymphocytes from a patient with bladder cancer in the presence of different polypeptides.
  • Fig. 3B shows the proliferation of lymphocytes from a patient with testicular cancer in the presence of different polypeptides.
  • PBMC mononuclear cells
  • the PBMCs are cultured (2 ⁇ 10 5 cells per well) in RPMI 1640 supplemented with 10% human AB serum, 2 mM L-glutamine, 100 IU / ml of penicillin and 100 ⁇ g / ml of streptomycin, in the presence of polypeptide at different concentrations.
  • the cells are then incubated at 37 ° C with 5% C0 2 for 5 days.
  • a ⁇ Ci of tritiated thymidine is then added to each well.
  • the cells are collected 18 hours later and The incorporation of the radioactivity is analyzed by spectrometry in a scintillation liquid using a ⁇ counter.
  • FIG. 1 We have given in FIG. 1 the results of proliferation of lymphocytes from a patient with bladder cancer in the presence
  • results demonstrate a significant proliferation with the polypeptide corresponding to the sequence 95-115 of the ⁇ subunit of hCG as well as with the polypeptide corresponding to the sequence 25-59 of the ⁇ subunit of the hCG.
  • the 95-115 hCG- ⁇ and 25-59 hCG- ⁇ polypeptides can be used for immunotherapy in the patient tested.
  • FIG. 2 results of the same type on lymphocytes from another patient with bladder cancer.
  • the results demonstrate a significant proliferation with the polypeptide corresponding to the sequence 31 - 50 of the ⁇ subunit of hCG as well as with the polypeptide corresponding to the sequence 41 - 61 of the ⁇ subunit of the hCG.
  • hCG- ⁇ and 41-61 hCG- ⁇ polypeptides 31 - 50 can be used for immunotherapy.
  • FIG. 3A We have given in FIG. 3A the results of PBMC proliferation tests of another patient with bladder cancer secreting the ⁇ subunit of hCG in the presence of different polypeptides covering the entire ⁇ subunit of hCG (50 ⁇ m).
  • T cells from a patient with testicular cancer secreting hCG- ⁇ , proliferated in the presence of the peptide hCG- ⁇ (20-47) (Fig. 3B).
  • PBMCs isolated by Ficoll gradient are tested for their cytotoxic action against autologous targets (patient B lymphocytes immortalized by Epstein-Barr virus) labeled with chromium 51 and serving as cells presenting the hCG polypeptides. . Cytotoxicity is evaluated by counting the radioactivity due to the release of 51 Cr from the lysed target.

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Abstract

A kit for diagnosing an hCG- or hCG fragment-secreting cancer including an assembly of polypeptides covering at least a part of the primary sequence of hCG. The use of a polypeptide corresponding to at least one portion of the primary sequence of hCG for producing a composition useful in hCG- or hCG fragment-secreting cancer immunotherapy is also disclosed.

Description

Trousse de diagnostic d'un cancer sécrétant l'hCG ou des fragments d'hCG et moyens destinés à l'iππiunothérapie d'un tel cancer. Diagnostic kit for a cancer secreting hCG or hCG fragments and means intended for the therapy of such cancer.
La présente invention concerne une méthode de traitement d'un cancer sécrétant l'hCG ou des fragments d'hCG et des moyens pour la mise en oeuvre de cette méthode. L'hormone chorionique gonadotrope humaineThe present invention relates to a method for the treatment of cancer secreting hCG or fragments of hCG and means for implementing this method. Human gonadotropic chorionic hormone
(hCG) est une glycoprotéine oligomérique constituée de l'association non covalente de deux sous-unités α(hCG-α) et β(hCG-β). Elle appartient à la famille des hormones gonadothyréotropes comprenant également l'hormone folliculotrope (FSH), l'hormone lutéotrope (LH) et 1'hormone thyréotrope (TSH) . L'hCG est sécrétée physiolo- giquement par le trophoblaste dès le début de la gros¬ sesse et stimule le corps jaune pour maintenir la synthèse d'hormones stéroïdiennes essentielle à l'implan- tation de l'embryon. Par ailleurs, l'hCG et/ou ses sous- unités sont également sécrétées par une majorité de tumeurs trophoblastiques et certaines tumeurs non trophoblastiques, constituant ainsi des marqueurs biologiques pouvant aider au diagnostic ou au pronostic et à la surveillance de ces tumeurs (Bellet et al. Rev. Prat. 40:1677-1990).(hCG) is an oligomeric glycoprotein consisting of the non-covalent association of two subunits α (hCG-α) and β (hCG-β). It belongs to the family of gonadothyreotropic hormones also comprising folliculotropic hormone (FSH), luteotropic hormone (LH) and thyreotropic hormone (TSH). HCG is secreted physiologically by the trophoblast from the start of pregnancy and stimulates the corpus luteum to maintain the synthesis of steroid hormones essential for implantation of the embryo. Furthermore, hCG and / or its subunits are also secreted by a majority of trophoblastic tumors and certain non-trophoblastic tumors, thus constituting biological markers which can help in the diagnosis or prognosis and in the monitoring of these tumors (Bellet and al. Rev. Prat. 40: 1677-1990).
La séquence des acides aminés de 1'hCG a été décrite par Morgan F. J. et al. The amino acid séquence of Human chorionic gonadotropin, J. Biol. Chem. 250, 5247, 1975.The amino acid sequence of h'hCG has been described by Morgan F. J. et al. The amino acid sequence of Human chorionic gonadotropin, J. Biol. Chem. 250, 5247, 1975.
Au sein de la population normale, des taux sériques très faibles d'hCG dimérique sont retrouvés chez l'homme (<0,05 ng/ml correspondant à la limite de détection de la méthode utilisée) et chez la femme non ménopausée (<0,1 ng/ml). L'hCG est principalement détectable chez 30% des femmes de plus de 45 ans (de 0,1 à 1 ng/ml) . Plusieurs études suggèrent que 1'hCG présente dans le sérum des sujets sains serait produite par des cellules hypophysaires. 'hCG-α est détectable chez l'homme sain (<1 ng/ml) et la femme de plus de 45 ans non ménopausée (<3 ng/ml). En revanche, l'hCG-β n'est pas détectable dans la population normale. Dans le cas de tumeurs trophoblastiques on a montré que les tumeurs placentaires bénignes (môles hydatiformes) et malignes (choriocarcinomes) sont caractérisées par un rapport hCG/hCG-β sérique important: le pourcentage d'hCG-libre qui est de 0,05 à 1% lors de grossesses normales s'élève à 1-5% en cas de môle et dépasse 5% dans les choriocarcinomes. Ce rapport hCG/hCG- β sérique est utilisé comme un témoin d'évolution des tumeurs testiculaires : un pourcentage d'hCG-β devenant inférieur à 5% est souvent caractéristique d'une évolu- tion favorable de la tumeur au cours du traitement. Ce rapport s'élèvera rapidement en cas d'évolution défavora¬ ble ou de rechute.In the normal population, very low serum levels of dimeric hCG are found in men (<0.05 ng / ml corresponding to the detection limit of the method used) and in premenopausal women (<0 , 1 ng / ml). HCG is mainly detectable in 30% of women over 45 (0.1 to 1 ng / ml). Several studies suggest that the hCG present in the serum of healthy subjects would be produced by pituitary cells. hCG-α is detectable in healthy men (<1 ng / ml) and women over 45 years of age without menopause (<3 ng / ml). In contrast, hCG-β is not detectable in the normal population. In the case of trophoblastic tumors, it has been shown that benign (hydatidiform moles) and malignant (choriocarcinomas) placental tumors are characterized by an important serum hCG / hCG-β ratio: the percentage of free hCG which is 0.05 to 1% during normal pregnancies amounts to 1-5% in the case of moles and exceeds 5% in choriocarcinomas. This serum hCG / hCG- β ratio is used as a control for the development of testicular tumors: a percentage of hCG-β becoming less than 5% is often characteristic of a favorable development of the tumor during treatment. This ratio will rise quickly in the event of an unfavorable development or a relapse.
Dans le cas de tumeurs non trophoblastiques, Marcillac et al. Cancer Res. 52:3901, 1992 ont montré que l'hCG-β, non détectable chez les individus sains, est retrouvée chez de nombreux patients atteints de certaines tumeurs non trophoblastiques (tumeurs de la vessie, du pancréas, du col de l'utérus, les tumeurs sans primitif retrouvé, les tumeurs de l'endomètre et du poumon). La présente invention a pour objet une méthode de traitement d'un cancer sécrétant 1'hCG ou des fragments d'hCG qui consiste àIn the case of non-trophoblastic tumors, Marcillac et al. Cancer Res. 52: 3901, 1992 have shown that hCG-β, not detectable in healthy individuals, is found in many patients with certain non-trophoblastic tumors (tumors of the bladder, pancreas, cervix, tumors without primitive found, endometrial and lung tumors). The subject of the present invention is a method of treating cancer secreting hCG or fragments of hCG which consists in
- identifier dans le sérum d'un patient à 1'aide d'un ensemble de polypeptides recouvrant au moins une partie de la séquence primaire de l'hCG, un état d'immunisation vis-à-vis d'un de ces polypeptides.- Identify in the serum of a patient using a set of polypeptides covering at least a part of the primary sequence of hCG, a state of immunization with respect to one of these polypeptides.
- administrer au patient au moins un polypep¬ tide correspondant à 1'antigène responsable de 1'état d'immunisation. Dans le cadre de la présente invention les polypeptides sont formés par un enchaînement de 9 à 40 aminoacides correspondant à une partie de la séquence primaire de l'hCG. Il peut s'agir en particulier de polypeptides comportant de 15 à 40 aminoacides ou, dans le cas de polypeptides susceptibles d'être reconnus par des lymphocytes T cytotoxiques, de polypeptides plus courts ayant notamment de 9 à 11 aminoacides.- Administer to the patient at least one polypep¬ tide corresponding to the antigen responsible for the immunization state. In the context of the present invention, the polypeptides are formed by a chain of 9 to 40 amino acids corresponding to a part of the primary sequence of hCG. They may in particular be polypeptides comprising from 15 to 40 amino acids or, in the case of polypeptides capable of being recognized by cytotoxic T lymphocytes, shorter polypeptides having in particular from 9 to 11 amino acids.
On désigne par ensemble de polypeptides recouvrant au moins une partie de la séquence primaire de l'hCG un ensemble de polypeptides tels que définis ci- dessus qui correspondent à des séquences successives de 1'hCG ou de ses fragments ou mieux qui correspondent à des séquences qui se chevauchent. On peut par exemple utiliser un ensemble de polypeptides correspondant aux séquences suivantes : hCG-α 1-20The term “set of polypeptides covering at least part of the primary sequence of hCG” denotes a set of polypeptides as defined above which correspond to successive sequences of hCG or of its fragments or better which correspond to sequences overlapping. One can for example use a set of polypeptides corresponding to the following sequences: hCG-α 1-20
15-3515-35
30-50 45-6530-50 45-65
60-8060-80
75-92 hCG-β 1-2075-92 hCG-β 1-20
15-35 30-5015-35 30-50
45-65 60-80 75-95 90-110 105-12545-65 60-80 75-95 90-110 105-125
120-145 De préférence on utilisera au moins un ensemble de polypeptides qui recouvre la séquence primaire de la sous-unité β de l'hCG ou au moins une partie de cette séquence. Il est à noter que la synthèse de la sous- unité β est sous la dépendance d'un groupe ultigénique comportant 7 gènes (Talmadge et al., Nucl. Acids Res. 12, 8415, 1984). L'analyse des séquences nucléotiques montre que trois de ces gènes (1, 3, 5) ont une structure identique, quatre autres de ces gènes (2, 6, 7 et 8) codent un variant au niveau du codon 117 avec substitu¬ tion de l'alanine à la place d'un acide aspartique. Une étude de l'expression des gènes (Bo et al., J. Biol. Chem., 267, 3179, 1992) montre que les gènes 1, 3 et 5 sont exprimés par le placenta normal.120-145 Preferably, at least one set of polypeptides will be used which covers the primary sequence of the β subunit of hCG or at least part of this sequence. It should be noted that the synthesis of the β subunit is dependent on an ultigenic group comprising 7 genes (Talmadge et al., Nucl. Acids Res. 12, 8415, 1984). Analysis of the nucleotic sequences shows that three of these genes (1, 3, 5) have an identical structure, four others of these genes (2, 6, 7 and 8) encode a variant at codon 117 with substitution. alanine instead of an aspartic acid. A study of gene expression (Bo et al., J. Biol. Chem., 267, 3179, 1992) shows that genes 1, 3 and 5 are expressed by the normal placenta.
On pense que dans les processus de tumorogé- nèse intéressant les cellules non trophoblastiques l'expression de plusieurs gènes conduit à la fois à la sécrétion de la protéine normale et d'une protéine mutée en 117.It is believed that in the tumourogenesis processes involving non-trophoblastic cells, the expression of several genes leads to both the secretion of the normal protein and of a protein mutated to 117.
Dans la présente invention on désigne donc par sous-unité β à la fois la protéine normale et la protéine mutée en 117.In the present invention, therefore, the term β subunit denotes both the normal protein and the protein mutated at 117.
Par "polypeptide correspondant à l'antigène responsable de 1'état d'immunisation on désigne un polypeptide qui a la même séquence que 1'un des polypep¬ tides responsable de l'état d'immunisation ou une séquence résultant de la combinaison de séquences au moins partielles des polypeptides responsables de l'état d'immunisation.By "polypeptide corresponding to the antigen responsible for the state of immunization means a polypeptide which has the same sequence as one of the polypep¬ tides responsible for the state of immunization or a sequence resulting from the combination of sequences at least partial of the polypeptides responsible for the state of immunization.
L'état d'immunisation vis-à-vis d'un des polypeptides peut être identifié par un test de stimula- tion de la prolifération des lymphocytes T par les polypeptides, ou encore par un test de cytotoxicité des lymphocytes T vis-à-vis des polypeptides.The state of immunization with respect to one of the polypeptides can be identified by a test for stimulating the proliferation of T lymphocytes by the polypeptides, or also by a test for cytotoxicity of T lymphocytes against vis the polypeptides.
Dans le cas de lymphocytes T cytotoxiques, les polypeptides reconnus sont formés par un enchaînement d'au moins 9 aminoacides correspondant à une partie de la séquence primaire de 1'hCG. Pour être reconnus par les lymphocytes T cytotoxiques, les polypeptides interagis¬ sent avec les molécules de classe I du complexe majeur d'histocompatibilité du patient. Chaque molécule de classe I interagit avec des peptides comportant des motifs précis. On peut par exemple utiliser les polypeptides correspondant aux séquences suivantes de 1'hCG-β :In the case of cytotoxic T lymphocytes, the recognized polypeptides are formed by a sequence of at least 9 amino acids corresponding to a part of the primary sequence of 1'hCG. To be recognized by cytotoxic T lymphocytes, the polypeptides interact with the class I molecules of the major histocompatibility complex of the patient. Each class I molecule interacts with peptides with specific motifs. One can for example use the polypeptides corresponding to the following sequences of 1'hCG-β:
3 - 12 4 - 123 - 12 4 - 12
8 - 168 - 16
10 - 1810 - 18
11 - 2011 - 20
17 - 2517 - 25
23 - 3323 - 33
24 - 3324 - 33
28 - 3728 - 37
32 - 4132 - 41
40 - 4840 - 48
40 - 4940 - 49
44 - 5244 - 52
47 - 5547 - 55
48 - 5648 - 56
51 - 5951 - 59
55 - 6255 - 62
61 - 6961 - 69
68 - 7668 - 76
71 - 8071 - 80
72 - 8072 - 80
75 - 8475 - 84
77 - 8677 - 86
78 - 8678 - 86
83 - 9283 - 92
100 - - 108100 - - 108
119 - - 128 125 - 134 134 - 143.119 - - 128 125 - 134 134 - 143.
On peut également mettre en évidence la présence éventuelle d'anticorps sériques spécifiques de 1'hCG ou de ses fragments. Ces anticorps peuvent être mis en évidence notamment par une méthode immuno-enzymatique telle qu'une méthode ELISA.It is also possible to demonstrate the possible presence of serum antibodies specific for the hCG or its fragments. These antibodies can be demonstrated in particular by an immunoenzymatic method such as an ELISA method.
La présente invention a également pour objet une trousse de diagnostic d'un cancer sécrétant l'hCG ou des fragments d'hCG comprenant un ensemble de polypepti¬ des recouvrant au moins une partie de la séquence primaire de 1'hCG.The present invention also relates to a diagnostic kit for a cancer secreting hCG or hCG fragments comprising a set of polypeptides covering at least part of the primary sequence of 1'hCG.
La présente invention a également pour objet une composition destinée à 1'immunothérapie de cancers sécrétant l'hCG ou des fragments d'hCG comprenant une dose efficace d'un polypeptide correspondant à au moins une partie de la séquence primaire de l'hCG et notamment un polypeptide correspondant à au moins une partie de la séquence primaire de la sous-unité β de l'hCG et un véhicule pharmaceutiquement acceptable.The subject of the present invention is also a composition intended for the immunotherapy of cancers secreting hCG or hCG fragments comprising an effective dose of a polypeptide corresponding to at least part of the primary sequence of hCG and in particular a polypeptide corresponding to at least part of the primary sequence of the hCG β subunit and a pharmaceutically acceptable vehicle.
La présente invention a également pour objet l'utilisation d'un polypeptide correspondant à au moins une partie de la séquence primaire de 1'hCG et notamment d'un polypeptide correspondant à au moins une partie de la séquence primaire de la sous-unité β de l'hCG pour la fabrication d'une composition destinée à 1'immunothérapie de cancers sécrétant l'hCG ou des fragments d'hCG.The present invention also relates to the use of a polypeptide corresponding to at least a part of the primary sequence of 1'hCG and in particular of a polypeptide corresponding to at least a part of the primary sequence of the β subunit hCG for the manufacture of a composition for the immunotherapy of cancers secreting hCG or fragments of hCG.
Pour la thérapie on peut utiliser les poly¬ peptides sous forme libre ou sous une forme plus immuno- gène, notamment une forme dans laquelle le peptide est couplé à une matrice de polylysine selon la technique du MAP (Posnett et al. J. Biol. Chem. 263, 17, 1988).For therapy, the poly¬ peptides can be used in free form or in a more immunogenic form, in particular a form in which the peptide is coupled to a polylysine matrix according to the MAP technique (Posnett et al. J. Biol. Chem. 263, 17, 1988).
En outre, pour la thérapie on administre le polypeptide avantageusement avec un agent adjuvant de l'immunité. Comme exemples d'adjuvants de l'immunité on peut citer le muramyldipeptide et le SAF-1 (Allison et al., J. Immunol. Methods 45, 157, 1986).In addition, for therapy, the polypeptide is advantageously administered with an adjuvanting immunity. As examples of adjuvants of immunity we may cite muramyldipeptide and SAF-1 (Allison et al., J. Immunol. Methods 45, 157, 1986).
On peut également administrer le polypeptide couplé à des nanosphères ou à des nanoparticules telles que celles décrites dans EP 02 40 424.It is also possible to administer the polypeptide coupled to nanospheres or to nanoparticles such as those described in EP 02 40 424.
Enfin, le polypeptide peut être protégé à ses extrémités NH2 et COOH terminales afin de réduire les risques de dégradation.Finally, the polypeptide can be protected at its NH 2 and COOH terminal ends in order to reduce the risks of degradation.
Dans le cas du traitement de certains cancers et notamment dans le cas du cancer de la vessie, on peut également administrer en plus de façon concomitante ou en mélange du vaccin BCG (Bacille de Calmette Guérin) constitué par des souches vivantes atténuées de bacille tuberculeux (voir notamment D. Lamm et al, New England Journal of Medicine 325, 17, 1205, 1991).In the case of the treatment of certain cancers and in particular in the case of bladder cancer, it is also possible to administer, concomitantly or as a mixture, the BCG vaccine (Bacillus Calmette Guérin) constituted by living attenuated strains of tuberculous bacillus ( see in particular D. Lamm et al, New England Journal of Medicine 325, 17, 1205, 1991).
La présente invention a donc en outre pour objet une composition destinée à l'immunothérapie de cancers sécrétant 1'hCG ou des fragments d'hCG comprenant une dose efficace d'un polypeptide correspondant à au moins une partie de la séquence primaire de 1'hCG et une dose efficace de BCG dans un véhicule pharmaceutiquement acceptable.The present invention therefore further relates to a composition intended for the immunotherapy of cancers secreting hCG or hCG fragments comprising an effective dose of a polypeptide corresponding to at least part of the primary sequence of hCG and an effective dose of BCG in a pharmaceutically acceptable vehicle.
En pratique 1'immunothérapie peut être réalisée avec des doses de polypeptides de 1 à 100 mg administrés notamment au niveau de la tumeur. Par exemple, les polypeptides peuvent être administrés dans le cas d'un cancer de la vessie, comme cela est pratiqué déjà dans le cas d'une immunothérapie par le BCG, c'est- à-dire par instillation intravésicale une semaine après la résection de la tumeur, puis cette administration est répétée chaque semaine pendant 6 semaines et des admi¬ nistrations intravésicales supplémentaires sont effec¬ tuées 3, 6, 12, 18 et 24 mois après. Des administrations percutanées ou par voie générale sont également possi- blés. Description des figures.In practice, immunotherapy can be carried out with doses of polypeptides from 1 to 100 mg administered in particular at the level of the tumor. For example, the polypeptides can be administered in the case of bladder cancer, as is already practiced in the case of immunotherapy with BCG, that is to say by intravesical instillation one week after resection. tumor, then this administration is repeated every week for 6 weeks and additional intravesical administration is performed 3, 6, 12, 18 and 24 months later. Percutaneous or general administrations are also possible. Description of the figures.
La Fig. 1 représente la prolifération des lymphocytes provenant d'un patient ayant un cancer de la vessie en présence d'hCG, des sous-unités α et β libres et de différents polypeptides.Fig. 1 shows the proliferation of lymphocytes from a patient with bladder cancer in the presence of hCG, free α and β subunits and various polypeptides.
La Fig. 2 représente la prolifération des lymphocytes provenant d'un autre patient ayant un cancer de la vessie en présence des sous-unités α et β libres et de différents polypeptides. La Fig. 3A représente la prolifération des lymphocytes provenant d'un patient ayant un cancer de la vessie en présence de différents polypeptides.Fig. 2 shows the proliferation of lymphocytes from another patient with bladder cancer in the presence of free α and β subunits and different polypeptides. Fig. 3A shows the proliferation of lymphocytes from a patient with bladder cancer in the presence of different polypeptides.
La Fig. 3B représente la prolifération des lymphocytes provenant d'un patient ayant un cancer du testicule en présence de différents polypeptides.Fig. 3B shows the proliferation of lymphocytes from a patient with testicular cancer in the presence of different polypeptides.
On donnera ci-après le compte-rendu d'essais de diagnostic.The diagnostic test reports are given below.
1) Isolement des cellules mononuclées (PBMC) par gradient de ficoll : Le sang total héparine prélevé sur des patients présentant un cancer est déposé sur une solution de ficoll-hypaque (1,077 g/1) dans un tube conique. Le tube est centrifugé 30 mn à 900 g, 18-20PC. La couche correspondant aux cellules mononuclées est récupérée à l'aide d'une pipette. Les cellules sont lavées dans du tampon de HANKs.1) Isolation of mononuclear cells (PBMC) by ficoll gradient: Whole heparin blood drawn from patients with cancer is deposited on a ficoll-hypaque solution (1.077 g / 1) in a conical tube. The tube is centrifuged for 30 min at 900 g, 18-20 P C. The layer corresponding to the mononuclear cells is recovered using a pipette. The cells are washed in HANKs buffer.
2) Test de prolifération lymphocvtaire2) Lymphocyte proliferation test
Les PBMC sont cultivés (2xl05 cellules par puits) dans du RPMI 1640 additionné de 10% de sérum humain AB, 2mM de L-glutamine, 100 Ul/ml de pénicilline et 100 μg/ml de streptomycine, en présence de polypeptide à différentes concentrations. Les cellules sont alors mises à incuber à 37°C avec 5% de C02 pendant 5 jours. Un μCi de thymidine tritiée est ensuite ajouté dans chaque puits. Les cellules sont collectées 18 heures après et 1'incorporation de la radioactivité est analysée par spectrométrie dans un liquide de scintillation utilisant un compteur β.The PBMCs are cultured (2 × 10 5 cells per well) in RPMI 1640 supplemented with 10% human AB serum, 2 mM L-glutamine, 100 IU / ml of penicillin and 100 μg / ml of streptomycin, in the presence of polypeptide at different concentrations. The cells are then incubated at 37 ° C with 5% C0 2 for 5 days. A μCi of tritiated thymidine is then added to each well. The cells are collected 18 hours later and The incorporation of the radioactivity is analyzed by spectrometry in a scintillation liquid using a β counter.
On a donné sur la Fig. 1 les résultats de prolifération des lymphocytes provenant d'un patient ayant un cancer de la vessie en présenceWe have given in FIG. 1 the results of proliferation of lymphocytes from a patient with bladder cancer in the presence
- du milieu de culture (M)- culture medium (M)
- de l'hCG et de ses sous-unités α et β- hCG and its α and β subunits
- de différents polypeptides appartenant aux sous-unités α et β.- different polypeptides belonging to the α and β subunits.
Les résultats mettent en évidence une prolifération significative avec le polypeptide corres¬ pondant à la séquence 95-115 de la sous unité β de l'hCG ainsi qu'avec le polypeptide correspondant à la séquence 25-59 de la sous unité α de l'hCG.The results demonstrate a significant proliferation with the polypeptide corresponding to the sequence 95-115 of the β subunit of hCG as well as with the polypeptide corresponding to the sequence 25-59 of the α subunit of the hCG.
Les polypeptides 95-115 hCG-β et 25-59 hCG-α peuvent être utilisés pour une immunothérapie chez le patient testé.The 95-115 hCG-β and 25-59 hCG-α polypeptides can be used for immunotherapy in the patient tested.
On a donné sur la Fig. 2 des résultats du même type sur des lymphocytes d'un autre patient ayant un cancer de la vessie. Les résultats mettent en évidence une prolifération significative avec le polypeptide correspondant à la séquence 31 - 50 de la sous-unité β de l'hCG ainsi qu'avec le polypeptide correspondant à la séquence 41 - 61 de la sous-unité α de l'hCG. Chez ce patient les polypeptides 31 - 50 de l'hCG-β et 41-61 de l'hCG-α peuvent être utilisés pour une immunothérapie.We have given in FIG. 2 results of the same type on lymphocytes from another patient with bladder cancer. The results demonstrate a significant proliferation with the polypeptide corresponding to the sequence 31 - 50 of the β subunit of hCG as well as with the polypeptide corresponding to the sequence 41 - 61 of the α subunit of the hCG. In this patient, hCG-β and 41-61 hCG-α polypeptides 31 - 50 can be used for immunotherapy.
On a donné sur la Fig. 3A les résultats de tests de prolifération de PBMC d'un autre patient ayant un cancer de la vessie sécrétant la sous-unité β de l'hCG en présence de différents polypeptides recouvrant la totalité de la sous-unité β de l'hCG (50 μm).We have given in FIG. 3A the results of PBMC proliferation tests of another patient with bladder cancer secreting the β subunit of hCG in the presence of different polypeptides covering the entire β subunit of hCG (50 μm).
Les résultats montrent que pour ce patient le polypeptide correspondant à la région β (20-47) de 1'hCG est le plus efficace pour induire une prolifération des cellules T.The results show that for this patient the polypeptide corresponding to the β (20-47) region of 1'hCG is the most effective in inducing proliferation of T cells
Par ailleurs, d'autres tumeurs (pancréatiques et testiculaires) ont été étudiées. Les cellules T d'un patient, présentant un cancer du testicule, sécrétant de l'hCG-β, ont proliféré en présence du peptide hCG-β (20- 47) (Fig. 3B).In addition, other tumors (pancreatic and testicular) have been studied. T cells from a patient with testicular cancer, secreting hCG-β, proliferated in the presence of the peptide hCG-β (20-47) (Fig. 3B).
Ces résultats montrent que des peptides correspondant à une partie de la séquence de l'hCG-β sont immunogéniques chez des malades ayant une tumeur produc¬ trice d'hCG-β et peuvent ainsi être utilisés pour développer une immunothérapie antitumorale.These results show that peptides corresponding to part of the hCG-β sequence are immunogenic in patients with a hCG-β producing tumor and can thus be used to develop anti-tumor immunotherapy.
A l'inverse, aucune réponse contre l'hCG, ses sous-unités ou des peptides de structure analogue n'a été observée chez 21 femmes enceintes.Conversely, no response against hCG, its subunits or peptides of similar structure was observed in 21 pregnant women.
On donnera ci-après les résultats ainsi obtenus.The results thus obtained will be given below.
Réponse immune cellulaire contre l'hCG ou ses sous-unités.Cellular immune response against hCG or its subunits.
Populations testées Total RéponsesPopulations tested Total Responses
Tumeurs vésicales 21 5 Tumeurs hCGβ+ 14 5 Tumeurs hCGβ- 7 0Bladder tumors 21 5 hCGβ + tumors 14 5 hCGβ- 7 tumors 0
Femmes enceintes 21 0Pregnant women 21 0
Sujets sains 13 0Healthy subjects 13 0
3) Test de cvtotoxicité3) Test for toxicity
Les PBMC isolés par gradient de Ficoll sont testés pour leur action cytotoxique vis-à-vis de cibles autologues (lymphocytes B du patient immortalisés par 1'Epstein-Barr virus) marquées du chrome 51 et servant de cellules présentatrices des polypeptides de l'hCG. La cytotoxicité est évaluée par le comptage de la radioacti¬ vité due au relargage du 51Cr de la cible lysée. PBMCs isolated by Ficoll gradient are tested for their cytotoxic action against autologous targets (patient B lymphocytes immortalized by Epstein-Barr virus) labeled with chromium 51 and serving as cells presenting the hCG polypeptides. . Cytotoxicity is evaluated by counting the radioactivity due to the release of 51 Cr from the lysed target.

Claims

REVENDICATIONS
1. Trousse de diagnostic d'un cancer sécré¬ tant l'hCG ou des fragments d'hCG comprenant un ensemble de polypeptides recouvrant au moins une partie de la séquence primaire de 1'hCG.1. A diagnostic kit for cancer secreted both by hCG or by hCG fragments comprising a set of polypeptides covering at least part of the primary sequence of 1'hCG.
2. Trousse de diagnostic selon la revendica¬ tion 1, comprenant un ensemble de polypeptides qui recouvre la séquence primaire de la sous-unité β de l'hCG. 2. Diagnostic kit according to claim 1, comprising a set of polypeptides which covers the primary sequence of the β subunit of hCG.
3. Trousse de diagnostic selon la revendica¬ tion 1, comprenant un ensemble de polypeptides qui recouvre au moins une partie de la séquence primaire de la sous-unité β de l'hCG.3. A diagnostic kit according to claim 1, comprising a set of polypeptides which covers at least part of the primary sequence of the hCG β subunit.
4. Trousse de diagnostic selon 1'une quelcon- que des revendications 1 à 3, comprenant des polypeptides ayant de 9 à 40 aminoacides.4. A diagnostic kit according to any one of claims 1 to 3, comprising polypeptides having 9 to 40 amino acids.
5. Trousse de diagnostic selon 1'une quelcon¬ que des revendications 1 à 4, comprenant des polypeptides dont les séquences se chevauchent. 5. A diagnostic kit according to any one of claims 1 to 4, comprising polypeptides whose sequences overlap.
6. Utilisation d'un polypeptide correspondant à au moins une partie de la séquence primaire de 1'hCG pour la fabrication d'une composition destinée à 1'immu¬ nothérapie de cancers sécrétant 1'hCG ou des fragments d'hCG. 6. Use of a polypeptide corresponding to at least part of the primary sequence of 1'hCG for the manufacture of a composition intended for the immu¬ notherapy of cancers secreting 1'hCG or fragments of hCG.
7. Utilisation selon la revendication 6, dans laquelle le polypeptide correspond à au moins une partie de la séquence primaire de la sous unité β de l'hCG.7. Use according to claim 6, in which the polypeptide corresponds to at least part of the primary sequence of the β subunit of hCG.
8. Utilisation selon la revendication 6 ou la revendication 7, dans laquelle le ou les polypeptides ont de 9 à 40 aminoacides.8. Use according to claim 6 or claim 7, wherein the polypeptide (s) have from 9 to 40 amino acids.
9. Utilisation selon l'une quelconque des revendications 6 à 8, dans laquelle le polypeptide est couplé à une matrice de polylysine.9. Use according to any one of claims 6 to 8, wherein the polypeptide is coupled to a polylysine matrix.
10. Utilisation selon 1'une quelconque des revendications 6 à 9, comprenant en outre un agent adjuvant de l'immunité.10. Use according to any of claims 6 to 9, further comprising an agent adjuvanted immunity.
11. Utilisation selon l'une quelconque des revendications 6 à 10, dans laquelle le polypeptide est protégé aux extrémités NH2 et COOH terminales. 11. Use according to any one of claims 6 to 10, in which the polypeptide is protected at the NH 2 and COOH terminal ends.
12. Composition destinée à l'immunothérapie de cancers sécrétant 1'hCG ou des fragments d'hCG comprenant une dose efficace d'un polypeptide correspon¬ dant à au moins une partie de la séquence primaire de 1'hCG et une dose efficace de BCG dans un véhicule pharmaceutiquement acceptable.12. Composition intended for the immunotherapy of cancers secreting 1'hCG or hCG fragments comprising an effective dose of a polypeptide corresponding to at least part of the primary sequence of 1'hCG and an effective dose of BCG in a pharmaceutically acceptable vehicle.
13. Méthode de diagnostic in vitro en vue du traitement d'un cancer sécrétant l'hCG ou des fragments d'hCG qui consiste à identifier dans le sang circulant d'un patient à 1'aide d'un ensemble de polypeptides recouvrant au moins une partie de la séquence primaire de l'hCG, un état d'immunisation vis-à-vis d'un de ces polypeptides.13. Method of in vitro diagnosis for the treatment of a cancer secreting hCG or fragments of hCG which consists in identifying in the circulating blood of a patient using a set of polypeptides covering at least part of the primary sequence of hCG, a state of immunization with respect to one of these polypeptides.
14. Méthode selon la revendication 13, dans laquelle l'état d'immunisation vis-à-vis d'un des polypeptides est identifié par un test de stimulation des lymphocytes T par les polypeptides.14. The method of claim 13, wherein the state of immunization vis-à-vis one of the polypeptides is identified by a test for stimulation of T lymphocytes by the polypeptides.
15. Méthode selon la revendication 13, dans laquelle on met en évidence la présence éventuelle d'anticorps sériques spécifiques de l'hCG ou de ses fragments.15. Method according to claim 13, in which the possible presence of serum antibodies specific for hCG or of its fragments is demonstrated.
16. Méthode selon la revendication 13, dans laquelle 1'état d'immunisation vis-à-vis d'un des polypeptides est identifié par un test de cytotoxicité des lymphocytes T vis-à-vis des polypeptides. 16. The method of claim 13, wherein the state of immunization with respect to one of the polypeptides is identified by a test of cytotoxicity of the T lymphocytes with respect to the polypeptides.
17. Méthode de traitement d'un cancer sécré¬ tant l'hCG ou des fragments d'hCG qui consiste à17. Method of treatment of a cancer secreting both hCG or fragments of hCG which consists in
- identifier dans le sérum d'un patient à 1'aide d'un ensemble de polypeptides recouvrant au moins une partie de la séquence primaire de l'hCG, un état d'immunisation vis-à-vis d'un de ces polypeptides. - administrer au patient au moins un polypep¬ tide correspondant à 1'antigène responsable de 1'état d'immunisation.- Identify in the serum of a patient using a set of polypeptides covering at least a part of the primary sequence of hCG, a state of immunization with respect to one of these polypeptides. - Administer to the patient at least one polypep¬ tide corresponding to the antigen responsible for the immunization state.
18. Méthode selon la revendication 17, dans laquelle l'ensemble des polypeptides recouvre au moins une partie de la séquence primaire de la sous unité β de l'hCG.18. The method of claim 17, wherein the set of polypeptides covers at least part of the primary sequence of the β subunit of hCG.
19. Méthode selon la revendication 17 ou la revendication 18, dans laquelle les polypeptides ont de 9 à 40 aminoacides.19. The method of claim 17 or claim 18, wherein the polypeptides have from 9 to 40 amino acids.
20. Méthode selon la revendication 19, dans laquelle les séquences des différents polypeptides se chevauchent.20. The method of claim 19, wherein the sequences of the different polypeptides overlap.
21. Méthode selon l'une quelconque des revendications 17 à 20, dans laquelle l'état d'immunisa¬ tion vis-à-vis d'un des polypeptides est identifié par un test de stimulation des lymphocytes T par les polypep¬ tides.21. Method according to any one of claims 17 to 20, in which the state of immunization with respect to one of the polypeptides is identified by a test for stimulation of T lymphocytes by the polypepides.
22. Méthode selon l'une quelconque des revendications 1 à 20, dans laquelle on met en évidence la présence éventuelle d'anticorps sériques spécifiques de l'hCG ou de ses fragments.22. Method according to any one of claims 1 to 20, in which the possible presence of serum antibodies specific for hCG or of its fragments is demonstrated.
23. Méthode selon la revendication 13, dans laquelle l'état d'immunisation vis-à-vis d'un des polypeptides est identifié par un test de cytotoxicité des lymphocytes T vis-à-vis des polypeptides.23. The method of claim 13, wherein the state of immunization vis-à-vis one of the polypeptides is identified by a test of cytotoxicity of T lymphocytes vis-à-vis the polypeptides.
24. Méthode selon l'une quelconque des revendications 17 à 23, dans laquelle on administre avec le polypeptide un agent adjuvant de l'immunité. 24. Method according to any one of claims 17 to 23, in which an adjuvanting agent of immunity is administered with the polypeptide.
25. Méthode selon l'une quelconque des revendications 17 à 24, dans laquelle on administre de façon concomitante à l'administration du polypeptide ou en mélange avec le polypeptide, du BCG. 25. Method according to any one of claims 17 to 24, in which BCG is administered concomitantly with the administration of the polypeptide or as a mixture with the polypeptide.
EP94909172A 1993-03-11 1994-03-10 Kit for diagnosing an hcg- or hcg fragment-secreting cancer and immunotherapeutical means therefor Withdrawn EP0688432A1 (en)

Applications Claiming Priority (5)

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FR9302820A FR2702494B1 (en) 1993-03-11 1993-03-11 Diagnostic kit for cancer secreting hCG or fragments of hCG and vaccine intended for immunotherapy of such cancer.
FR9302820 1993-03-11
FR9312039 1993-10-08
FR9312039A FR2710845B1 (en) 1993-10-08 1993-10-08 Composition intended for the immunotherapy of a cancer secreting hCG or fragments of hCG.
PCT/FR1994/000265 WO1994020859A1 (en) 1993-03-11 1994-03-10 Kit for diagnosing an hcg- or hcg fragment-secreting cancer and immunotherapeutical means therefor

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US5677275A (en) * 1994-08-05 1997-10-14 The United States Of America As Represented By The Department Of Health And Human Services Treatment of cancer with human chorionic gonadotropin
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US6319504B1 (en) * 1996-06-24 2001-11-20 University Of Maryland Biotechnology Institute Treatment and prevention of HIV infection by administration of derivatives of human chorionic gonadotropin
US5997871A (en) * 1996-06-24 1999-12-07 University Of Maryland Biotechnology Insitute Treatment and prevention of cancer by administration of derivatives of human chorionic gonadotropin
US5968513A (en) * 1996-06-24 1999-10-19 University Of Maryland Biotechnology Institute Method of promoting hematopoiesis using derivatives of human chorionic gonadotropin
US6583109B1 (en) 1997-06-24 2003-06-24 Robert C. Gallo Therapeutic polypeptides from β-hCG and derivatives
US7994278B1 (en) 1999-08-06 2011-08-09 Nobel Biosciences Llc Biologically active polypeptides derived from a novel early stage pregnancy factor designated maternin (MA)

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