EP2148694A2 - Médicament utilisé pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques et pour une transplantation, appliqué à la médecine vétérinaire, notamment chez les chevaux, et procédé de fabrication et de contrôle thé - Google Patents

Médicament utilisé pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques et pour une transplantation, appliqué à la médecine vétérinaire, notamment chez les chevaux, et procédé de fabrication et de contrôle thé

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Publication number
EP2148694A2
EP2148694A2 EP08759974A EP08759974A EP2148694A2 EP 2148694 A2 EP2148694 A2 EP 2148694A2 EP 08759974 A EP08759974 A EP 08759974A EP 08759974 A EP08759974 A EP 08759974A EP 2148694 A2 EP2148694 A2 EP 2148694A2
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EP
European Patent Office
Prior art keywords
ecg
seq
treatment
disorders
pro
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EP08759974A
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German (de)
English (en)
Inventor
Henry Alexander
Gerolf Zimmermann
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Universitaet Leipzig
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Universitaet Leipzig
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/24—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g. HCG; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • 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
    • C07K16/26—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against hormones ; against hormone releasing or inhibiting factors

Definitions

  • the invention relates to agents for the treatment of fertility and pregnancy disorders (pregnancy disorders) and immunologically-related diseases (autoimmune diseases) and transplants, and to processes for their preparation and therapy control for use in veterinary medicine, in particular in horses.
  • the object of the invention is therefore to provide an agent for the treatment of pregnancy disorders (pregnancy disorders), in particular for the treatment of fertility disorders, implantation disorders, early pregnancy losses, imminent pregnancy losses and habitual abortions as well as premature birth and growth retardation, for use in veterinary medicine, in particular in horses.
  • pregnancy disorders pregnancy disorders
  • fertility disorders in particular for the treatment of fertility disorders, implantation disorders, early pregnancy losses, imminent pregnancy losses and habitual abortions as well as premature birth and growth retardation
  • the object of the invention is further to provide agents for the treatment of infectological and immunological diseases of the intestine and the lungs and the joints or for transplantation (cornea, skin) for use in veterinary medicine, in particular in horses.
  • this object is achieved by a medicament containing at least one protein component of equine chorionic gonadotropin (eCG) or a nucleic acid sequence (eCG) coding for a protein component of equine chorionic gonadotropin.
  • eCG equine chorionic gonadotropin
  • eCG nucleic acid sequence
  • the protein component of equine chorionic gonadotropin is a prehormonal subunit (pre- ⁇ -eCG or pre- ⁇ -eCG), an alpha or beta subunit of equine chorionic gonadotropin ( ⁇ -eCG or ⁇ -eCG) or a Fragment of the alpha or beta subunit.
  • the fragments of the alpha unit preferably have a length of 10 to 95 amino acids.
  • Fragments of the beta unit preferably have a length of 20 to 148 Amino acids. Particular preference is given in each case to fragments having a length of from 25 to 50 amino acids.
  • the medicament is advantageously suitable for the treatment of pregnancy disorders, in particular for the treatment of fertility disorders, implantation disorders, early pregnancy loss, imminent and habitual abortion and premature birth, and growth retardation in veterinary medicine, especially in Perissodactyla and Equinae, such as horses.
  • the equine chorionic gonadotropin preferably consists of a non-covalently linked heterodimer of an ⁇ -subunit ( ⁇ -eCG) and a ⁇ -subunit ( ⁇ -eCG).
  • ⁇ -eCG ⁇ -subunit
  • ⁇ -eCG ⁇ -subunit
  • ⁇ -eCG ⁇ -subunit
  • the ⁇ -subunits including the CTP sections ("C-terminal repeats") of the human ( ⁇ -hCG amino acids 121-145) and the equine ⁇ subunit ( ⁇ -eCG amino acids 121-149) differ in about 50% of the amino acids and in the number of N- and O-glycosidic side chains (1 N- and 12-O-glycosidic side chains in ⁇ -eCG and 2 N and 4 O-glycosidic side chains in ⁇ -hCG) .beta.-eCG In contrast to ⁇ -hCG, only one gene is read in.
  • eCG expressed in the placenta (ie in the Girdle cells of the chorio-villous cup structure of the conceptus (embryo)) and the hypophysis-expressed el_H (luteinizing hormone
  • the el_H and the eCG differ in the glycan side chain and thus by their biological activity at the receptor and in the half-life.
  • the equine chorionic gonadotropin (eCG) is formed by a further glycosylation of equine LH with a carbohydrate moiety of From 40% to 45%, eCG is the most abundant glycosylated mammalian hormone in the placenta.
  • the placental eCG is more glycosylated than eLH.
  • the eCG also has a lower sulphin group content and a higher sialic acid content than the eLH.
  • eCG is not only expressed in embryonic (or fetal) tissue, but also in the endometrium and the decidua of the non-pregnant or pregnant mare.
  • the endometrial (maternal) eCG of the non-pregnant horse in the normal luteal phase as detected in the invention differs from the pituitary el_H, fetal (placental, chorio-villous) eCG and deciduous eCG of the pregnant horse in terms of the structure of the glycan side chains and may hCG and chorio-villous eCG can be used as drugs.
  • the equine chorionic gonadotropin has a very extensive and diverse spectrum of activity.
  • eCG supports the corpus luteum function to generate progesterone, which in turn stimulates eCG secretion in the endometrium.
  • eCG is a growth and differentiation hormone and promotes blood flow to the uterus. It also has immunosuppressant, antiviral and antibacterial effects. In addition, eCG keeps the smooth muscles of the uterus quiet.
  • Equine chorionic gonadotropin is an immune protection hormone as well as a barrier protection and anti-infective hormone that acts especially during the second half of the cycle and throughout the gestation period.
  • Chorionic gonadotropin plays an essential role in reproductive physiology. Although the amino acid sequence of equine chorionic gonadotropin is significantly different from human chorionic gonadotropin, surprisingly similar effects are obtained. Disturbances of the endometrial eCG production lead to infections of the uterus and the fruit, which not infrequently induces fetal abortion and death. Lack of eCG formation, especially in early pregnancy, causes growth retardation and, in later pregnancy, placental insufficiency.
  • equine chorionic gonadotropin binds as a drug at equine eLH / eCG receptors in the pituitary gland (a), in the Girdle cells of the chorio-villous cup structure of the equine concept (b) and probably already in the normal luteal endometrium non-pregnant mare (c).
  • el_H and eCG bind with different affinity to the receptors.
  • This eCG antibody allows post-ovulatory monitoring and pregnancy monitoring by ELISA testing.
  • the equine chorionic gonadotropin secreted with the pregnancy of the mare in the Girdle cells of the chorio-villous cup structure is preferably determined in the peripheral blood with a peak from the 39th to the 130th but also to the 200th day of gestation.
  • the equine chorionic gonadotropin has a paracrine meaning in addition to the endocrine.
  • This effect of the endometrial eCG can already be used in the normal luteal phase of the non-pregnant uterus of the mare and continues to take further protective effect on the course of gestation with the chorio-villous eCG of the conceptus implantation.
  • the equine chorionic gonadotropin formed in the maternal endometrium of the uterus and the well-known chorio-villous eCG of Girdle cells of the conceptus are of importance throughout pregnancy (pregnancy). During the first 150 days of pregnancy, an abortion often goes unnoticed. Early pregnancy losses are associated with a lack of early eCG secretion in both the maternal endometrium and fetal chorio-villous Girdle cells of the mare. This lack of eCG is often not recognized or recognized too late.
  • the eCG's barrier protection and infection protection function can also be used outside the pregnancy in horses for the treatment of diseases of the bronchial tract (chronic obstructive inflammation), the gastrointestinal tract (colic) and the joints (osteoarthritides).
  • ECG administered daily at low doses of 3 to 6 ⁇ g / kg body weight may advantageously stimulate endogenous eCG production.
  • the equine chorionic gonadotropin used according to the invention is preferably produced recombinantly.
  • the recombinant production is preferably carried out in a eukaryotic expression system.
  • the synthesis preferably takes place as prehormonal subunits pre- ⁇ -eCG and pre- ⁇ -eCG.
  • the cleavage of the N-termini and thus the generation of the mature subunits ⁇ -eCG and ⁇ -eCG preferably takes place by proteases or during the synthesis of the proteins.
  • the equine chorionic gonadotropin preferably contains both the alpha ( ⁇ -eCG) and beta subunits ( ⁇ -eCG) of the eCG.
  • the alpha subunit has SEQ ID no. 3 ( ⁇ -eCG) or SEQ ID NO. 4.
  • the beta subunit ( ⁇ -eCG) has SEQ ID no. 1 or SEQ ID NO. Second
  • the sequence SEQ ID no. 1 and SEQ ID NO. 3 are each the amino acid sequences of the pre-hormonal subunits (pre- ⁇ -eCG or pre- ⁇ -eCG).
  • the mature ⁇ -eCG subunit (SEQ ID No 4) is formed by cleavage of the first 24 amino acids from pre- ⁇ -eCG (SEQ ID No. 3).
  • the mature ⁇ -eCG subunit (SEQ ID No 2) is formed by cleavage of the first 20 amino acids from pre- ⁇ -eCG (SEQ ID No. 1). The cleavage of the amino acid sequences in the prehormonal subunits to obtain the mature subunits occurs either before administration as a drug or after administration in the organism to be treated.
  • An alternative pre- ⁇ -eCG or ⁇ -eCG sequence is described in SEQ ID no. 10 or SEQ ID no. 11 indicated. Included here are in each case sequences which belong to one of these sequences SEQ ID no. 1 to 4 and 10 or 11 have a homology of at least 90%, preferably of at least 95%.
  • the alpha subunit ( ⁇ -eCG) is preferably at the amino acids selected from Asp-56, Asp-82 glycanome the specified amino acid positions refer to the sequence of the mature ⁇ -eCG, for the pre- ⁇ -eCG shift the positions corresponding to +24 amino acids (Asp-80, Asp-106).
  • the beta subunit ( ⁇ -eCG) is preferably selected from the amino acids selected from Asp-13 Ser-123, Thr-127, Ser-128, Thr-129, Ser-130, Thr-131, Thr-133, Ser-137 , Ser-140, Ser-141, Thr-148, Ser-149 glycated (the indicated amino acid positions refer to the sequence of the mature ß-eCG, for the pre-ß-eCG positions shift accordingly by +20 amino acids, Asp-33, Ser-143, .).
  • the glycan chains each contain preferably 2 to 15, more preferably 2 to 7 sugar residues.
  • Preferred sugar residues are N-acetylglucosamine (GlcNac), galactose, manose, fucose, sialic acid (N-acetylneuraminic acid).
  • the glycan side chains of the eCG beta subunit of the luteal eCG in the normal endometrium of the non-pregnant mare may differ slightly from the above-described glycan chains of the chorio-villous eCG.
  • the alpha subunit contains 2 to 5 disulfide bridges between the cysteine pairs selected from Cys-11 with Cys 36, Cys 14 with Cys-35, Cys-63 or Cys-64 with Cys-91, Cys-86 Cys-88 and Cys-32 with Cys-63 or Cys-64 (the indicated amino acid positions refer to the sequence of the mature ⁇ -eCG, whereas for the pre- ⁇ -eCG the positions shift accordingly by +24 amino acids).
  • the beta subunit contains 2 to 6 disulfide bridges between the cysteine pairs selected from.
  • the indicated amino acid positions refer to the sequence of the mature ß-eCG, for the pre-ß-eCG the positions shift accordingly by +20 amino acids).
  • Preferred nucleic acid sequences coding for the alpha subunit ( ⁇ -eCG) are SEQ ID NO. 7 and SEQ ID NO. 8, wherein SEQ ID no. 7 encoded for the prehomonal alpha subunit (pre- ⁇ -eCG).
  • Preferred nucleic acid sequences coding for the beta subunit ( ⁇ -eCG) are SEQ ID NO. 5 and SEQ ID NO. 6, wherein SEQ ID no. 5 encoded for the prehomonal beta subunit (pre- ⁇ -eCG). Included are also sequences which belong to one of these sequences SEQ ID no. 5 to 8 have a homology of at least 90%.
  • SEQ ID no. 1 to 11 or fragments thereof for the treatment of pregnancy disorders, in particular for the treatment of fertility disorders, implantation disorders, early pregnancy loss, early and habitual abortion and premature birth and growth retardation, in veterinary medicine, especially in Perissodactyla, especially in Equinae, such as horses.
  • the invention also encompasses the use of SEQ ID NO. 1 to 11 or fragments thereof for the treatment of infectious and immunological diseases, in particular of the intestine and the lungs and the joints, in veterinary medicine, especially in Perissodactyla, in particular in Equinae, such as horses.
  • Another object of the invention is the use of SEQ ID NO. 1 to 11 or fragments thereof to aid in transplantation, d. H. for the prevention of rejection reactions, in particular in the transplantation of the cornea and the skin, in veterinary medicine, in particular in perissodactyla, in particular in Equinae, such as horses.
  • the invention also provides the use of SEQ ID no. 1 to 1 1 or fragments thereof as a contraceptive in veterinary medicine, especially in Perissodactyla, especially in Equinae, such as horses.
  • the fragments of the alpha unit preferably have a length of 10 to 95 amino acids. Fragments of the beta unit preferably have a length of 20 to 148 amino acids. Particular preference is given in each case to fragments having a length of from 25 to 50 amino acids.
  • the eCG has hitherto only been produced in insect cells. In the method according to the invention, however, the production takes place in a culture of mammalian cells, such as.
  • CHO Chinese hamster ovary
  • the equine chorionic gonadotropin is administered intramuscularly, intravenously, intraamnically, orally, intra-articularly or in the form of aerosols, preferably 3-6 ⁇ g / kg body weight daily, preferably distributed over several single doses.
  • the serum chorionic gonadotropin levels are determined during pregnancy. In case of suspected fetomaternal functional unit therapy should be started. In case of impaired early pregnancy injections should i. m. be applied. After the serum eCG levels have dropped, diagnosis can be made by determining the chorionic gonadotropin in the amniotic fluid and substituted accordingly. Depending on the level of the amnial eCG, treatment should be limited to the completion of the 10. Pregnancy month.
  • nucleic acid sequence coding for the equine chorionic gonadotropin takes place according to the known methods of gene therapy, e.g. B. by means of adenoviral vectors.
  • the invention also provides a method for the treatment of pregnancy disorders, in particular for the treatment of fertility disorders, implantation disorders, early pregnancy losses, imminent and habitual abortions, as well as premature birth and growth retardation, treatment of infectious and immunological-related diseases, in particular of the intestine and the lungs and the Joints, arthrides and ischaemias and to assist transplantation, in particular of the cornea and the skin, in veterinary medicine, in particular in horses by administering equine chorionic gonadotropin or an equine chorionic gonadotropin encoding nucleic acid sequence (ecg) or fragments of the protein eCG or fragments of the for the equine chorionic gonadotropin-encoding nucleic acid sequence.
  • ecg equine chorionic gonadotropin encoding nucleic acid sequence
  • the agents according to the invention for the first time the implementation of a causal therapy of pregnancy disorders in Perissodactyla, especially in Equinae, such as horses allows.
  • the loss of decidual eCG which causes the pregnancy disorders is substituted by the agents according to the invention.
  • the delivered eCG stimulates the formation of eCG in the endometrium, which in turn calms the uterine musculature and improves circulation to the placenta.
  • the causal treatment of pregnancy disorders and premature birth in terms of premature birth are possible.
  • the medicament according to the invention is used in particular for the treatment of pregnancy disorders.
  • Pregnancy disorders include fertility disorders, implantation disorders, early pregnancy loss, abortion and habitual abortion, as well as premature birth and growth retardation.
  • a fertility disorder is a disorder characterized by the absence of pregnancy despite regular insemination or treatment by in vitro fertilization.
  • An abortion is a so-called miscarriage, which is characterized by an early death or resorption of a fruit.
  • a habitual abortion is present when a mare had a miscarriage three or more times in succession.
  • An intrauterine growth retardation of a fetus occurs when the fetus is too small and in growth for its gestation period.
  • the deviation of the estimated weight by two standard deviations is below the normal value. This variation in size growth is determined by measuring the fetus with ultrasound and then comparing it to growth curves.
  • the agents according to the invention also make it possible to treat infectious and immunologically-related diseases, in particular of the intestine, the lungs and the joints. Furthermore, the agents according to the invention are also suitable for inducing immune tolerance in transplants, in particular the cornea and the skin.
  • Immune tolerance refers to the absence of an immune response after administration of a particular antigen.
  • the autoimmune disease is an umbrella term for diseases whose cause is an excessive reaction of the immune system against the body's own tissue. The body's own tissue is mistakenly perceived as a foreign body to be controlled by the immune system. This leads to severe systemic or local inflammatory reactions, which lead to damage to the affected organs.
  • the medicament of the invention is administered by injection, for example.
  • a particularly preferred embodiment of the medicament is adapted to allow parenteral administration of the drug.
  • parenteral administration of the medicament according to the invention for example, 250 micrograms of the mature eCG are dissolved in 0.5 ml of an injection solution and transferred to a pre-filled syringe.
  • the drug is e.g. administered intramuscularly, intra-amnially, intravenously, orally, intra-articularly or in the form of aerosols. Under emergency conditions, intravenous administration is preferred.
  • administration of the drug is preferably by subcutaneous injection.
  • the drug is adapted so that the amount of equine chorionic gonadotropin administered is 3 to 6 ⁇ g per kg of body weight per day.
  • Injections are prescribed at the onset of regular labor in the event of premature birth. After the contractions have subsided, the injection with the medicament according to the invention takes place at intervals of 2 to 4 days.
  • intravenous infusion e.g. 1000 ⁇ g of the mature eCG dissolved in 500 ml of an infusion solution and administered over a period of four hours.
  • the injection is intraamnial.
  • mononuclear cells are removed from the animal, incubated with the abovementioned eCG forms in vitro and then again intramuscularly, intravenously, orally or in the form of aerosols to be treated Animal back given.
  • the mononuclear cells especially monocytes, NK cells and T cells
  • the mononuclear cells are changed in their properties so that they cause an immune tolerance.
  • the invention also includes a method for cycle synchronization of mares but also for the prevention of pregnancy (contraception) by administration of at least one of SEQ ID NO. 1 to 11 or fragments thereof as, especially in Perissodactyla, especially in Equinae, such as horses.
  • the eCG is preferably injected intramuscularly in the form of depot sticks or inserted intravaginally monthly in ring form.
  • the invention further relates to a method for the production of equine eCG with the subunits ⁇ -chorionic gonadotropin ( ⁇ -eCG) and ⁇ -chorionic gonadotropin ( ⁇ -eCG) in isolated endometrial epithelial cells, decidual cells or cells of the membranes (chorion or amnion).
  • ⁇ -eCG ⁇ -chorionic gonadotropin
  • ⁇ -eCG ⁇ -chorionic gonadotropin
  • the isolated endometrial epithelial cells are preferably cell lines of equine origin.
  • the epithelial cells are preferably obtained from native endometrial tissue.
  • the eCG expressed in these cells has the above-mentioned preferred glycation pattern and the above-mentioned disulfide bridges.
  • the invention also relates to the use of eCG or its fragments in medical diagnostics in particular for the diagnosis of said pregnancy disorders or implantation disorders or infectious and immunological-related diseases, arthrides and ischaemias.
  • the invention further relates to the differentiated diagnosis of the el_H, as well as the eCG of the endometrium and eCG of the placenta, preferably from the serum. Endometrial eCG and placental eCG make a distinction through the sugar side chains.
  • the invention further relates to the use of eCG or its fragments as a contraceptive. Preference is given thereto that eCG or its fragments are administered intramuscularly or intraperitoneally in implants.
  • the subject matter of the invention is also an antibody which specifically recognizes eCG.
  • This is preferably not cross-reactive with human CG (hCG) and preferably recognizes an epitope on the peptide according to SEQ ID no. 9th
  • FIG. 1 shows that eCG is also formed in the endometrium in the sexually mature mare in the second half of the cycle.
  • the eCG was detected by classical immunohistochemistry staining with rabbit anti-eCG (10 ⁇ resolution).
  • FIG. 2 shows that maternal eCG is already formed in the sexually mature mare from the early proliferative (estrus) phase until the second half of the cycle (secretory, diestrus phase) in the gland epithelium of the endometrium.
  • A inactive endometrium, an oestrus or early oestrus phase (proliferative phase);
  • B periovulatory oestrus phase;
  • C oestrus phase, beginning diestrus phase (secretory phase);
  • Fig. 3 shows that fetal eCG is released in pregnant mares in the Girdle cells of the chorio-villous cup structure (placenta, upper arrow) of the conceptus and, in addition, maternal eCG in the gland epithelium of the endometrium (decidua, lower arrows) at gestation (magnification 1: 100).
  • A human placenta as negative or positive control.
  • B to (D) different sections of equine chorio-villous cup structure and endometrium.
  • Left column polyclonal antibody to equine CG (anti-eCG).
  • Right column polyclonal antibody against humans CG (anti-hCG).
  • Embodiment 1 Genetic engineering of recombinant hCG (based on Loumaye et al., 1995)
  • the cDNA sequences coding for the mature ⁇ -eCG and ⁇ -eCG are respectively cloned into the expression vector with the pGEM-T vector system (Promega) according to the manufacturer's instructions.
  • the dehydrofolic acid reductase (DHFR) DNA sequence is cloned into the expression vector.
  • the thus-obtained ⁇ -eCG and the ⁇ -eCG expression vectors are cotransfected into the well-characterized DHFR-deficient CHO (Chinese hamster ovary) cell line, cultured and selected individual clones.
  • the single cell clones are screened for their ability to produce eCG, their eCG biological activity and their genetic stability.
  • the best clones are then cultured in a bioreactor for the production of the recombinant eCG.
  • the secreted into the culture medium eCG is collected and purified by ultrafiltration and conventional chromatography methods and sterile filtered.
  • Embodiment 2 is a diagrammatic representation of Embodiment 1:
  • the CNBr-activated Sepharose 4B column is coupled with the eCG peptide ligand (SEQ ID No. 9) and buffer washed.
  • eCG peptide ligand SEQ ID No. 9
  • buffer washed After capacity calculation of the AK binding, up to 50 ml of eCG antibody serum are applied, prepared and washed after buffer washing with an eluate buffer from the column.
  • the affinity-purified monospecific IgG buffer solutions of the eCG antibody are used for WB and ICH in the dilution 1: 200 to 1: 5000.
  • Example 4 Detection and Diagnostic Recording of the eCG Expression: Detection and Diagnostic Recording of the eCG Release in the Equine Endometrium / Decidua of the Non-Pregnant and Pregnant Mare Using Western Blot and Immunohistochemistry (ICH) for Therapy Control of the eCG Application with the eCG Described -specific antibodies according to SEQ ID No 9 (FIG. 1, FIG. 2, FIG. 3 and FIG. 4)
  • the recombinant eCG produced as in Example 1 or 2 is used for therapy in the following examples:
  • Embodiment 5 Sterility If sterility is suspected, eCG is given intramuscularly every 4 days to induce ovulation and to support the luteal phase at a dose of 3 - 6 ⁇ g / kg body weight until a sufficient increase in eCG is achieved.
  • Embodiment 6 Osteoarthritides
  • 300 ⁇ g / ml eCG is injected into the joint space of the affected joints every 3 to 4 days for a period of 4 weeks.
  • Embodiment 7 Treatment of Growth Retardation
  • Exemplary embodiment 8 Treatment of imminent abortion
  • Embodiment 9 Treatment of fertility disorders, implantation disorders and early pregnancy loss
  • the animals are injected with 500-1000 ⁇ g eCG intramuscularly on the 24th cycle day and further every 4 days.
  • Embodiment 10 Treatment of habitual abortion
  • the animals are given intramuscular injection of 1000 ⁇ g eCG once a week after the diagnosis of pregnancy until the 36th week of pregnancy.
  • Embodiment 11 Treatment for Induction of Contraception
  • 500-1000 ⁇ g eCG is intramuscularly injected in the incipient proliferative (oestrus) cycle phase of the endometrium.
  • administration is by an intraperitoneal eCG implant.
  • Embodiment 12 Treatment of immunological diseases and induction of immunological tolerance in transplantations
  • the eCG is prescribed for the treatment of immunological diseases of the lungs, intestines and joints.
  • the eCG in a dosage of 3 to 5 ug / ml in aerosol form is prescribed to the animals.
  • the eCG in capsule form is administered at a concentration of 3000 ⁇ g per capsule daily for a period of 4 weeks.
  • the animals are injected intraarticularly with 1000 ⁇ g for 3 weeks.
  • the grafts are transported in a solution of 3 ⁇ g / ml eCG to avoid a graft-versus-host reaction.
  • an eCG medium is created at the recipient sites by eCG rinsing in a concentration of 3-6 ⁇ g / ml, which is continued for 3 weeks by injecting the transplant in the same concentration.
  • biomembranes are applied for 4 to 6 weeks, which release eCG.
  • Embodiment 13 Ovulation timing and contraception of the mare
  • eCG depot sticks of 5000 ⁇ g are injected intramuscularly every quarter or intravaginally administered monthly in ring form.
  • Embodiment 14 Treatment of Acute Viral and Bacterial Inflammations
  • eCG is injected intravenously at a daily dose of three times 500 to 1000 ⁇ g.
  • Embodiment 15 Treatment of severe tissue ischemia
  • SEQ ID No. 1 to 4 the positions to which the respective protein carries N-glycosidic side chains are emphasized by simple underlining, positions with O-glycoside side chains are underlined twice.
  • SEQ ID no. 5 to 8 are corresponding nucleic acid sequences.

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Abstract

La présente invention concerne des agents utilisés pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques (maladies auto-immunes) et pour des transplantations, ainsi que des procédés de fabrication de ces agents pour une application en médecine vétérinaire, en particulier chez les chevaux. Selon cette invention, une composante protéique de la gonadotrophine chorionique équine (eCG) ou une séquence d'acides nucléiques (ecg) codant une composante protéique de la gonadotrophine chorionique équine est utilisée pour traiter ou diagnostiquer des troubles de la gravidité, notamment des troubles de la fertilité, des troubles d'implantation, des fausses couches précoces, des avortements imminents et habituels, des naissances prématurées et des retards de croissance, et pour traiter ou diagnostiquer des maladies infectieuses et immunologiques, des arthrites et des ischémies, pour favoriser une transplantation, pour synchroniser des cycles et pour assurer une contraception dans le cadre de la médecine vétérinaire, avant tout chez les périssodactyles, et en particulier chez les équidés.
EP08759974A 2007-05-23 2008-05-23 Médicament utilisé pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques et pour une transplantation, appliqué à la médecine vétérinaire, notamment chez les chevaux, et procédé de fabrication et de contrôle thé Withdrawn EP2148694A2 (fr)

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DE102007025107A DE102007025107A1 (de) 2007-05-23 2007-05-23 Arzneimittel zur Behandlung von Fertilitäts- und Schwangerschaftsstörungen und immunologisch-bedingten Erkrankungen und Transplantationen zur Anwendung in der Tiermedizin, insbesondere bei Pferden sowie Verfahren zur Herstellung
PCT/EP2008/056374 WO2008142162A2 (fr) 2007-05-23 2008-05-23 Médicament utilisé pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques et pour une transplantation, appliqué à la médecine vétérinaire, notamment chez les chevaux, et procédé de fabrication et de contrôle thé

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EP2148694A2 true EP2148694A2 (fr) 2010-02-03

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EP08759974A Withdrawn EP2148694A2 (fr) 2007-05-23 2008-05-23 Médicament utilisé pour traiter des troubles de la fertilité et de la gravidité et des maladies immunologiques et pour une transplantation, appliqué à la médecine vétérinaire, notamment chez les chevaux, et procédé de fabrication et de contrôle thé

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BR102015032660B1 (pt) 2015-12-28 2019-05-28 Ouro Fino Saúde Animal Participações S.A. PROCESSO DE PRODUÇÃO DE UMA GONADOTROFINA CORIÔNICA EQUINA RECOMBINANTE (reCG): COMPOSIÇÃO VETERINÁRIA E USO

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WO1990002757A1 (fr) * 1988-09-02 1990-03-22 Integrated Genetics, Inc. Proteine heteropolymerique
AU4964197A (en) * 1996-11-12 1998-06-03 Teikoku Hormone Mfg. Co., Ltd. Recombinant single-stranded equine chorionic gonadotropin
EP1541168B1 (fr) * 2003-12-09 2010-11-24 Intervet International BV Methode de synchronisation de l'ovulation chez le bétail

Non-Patent Citations (1)

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Title
MAUREL M-C ET AL: "Immunochemical study of equine chorionic gonadotropin (eCG/PMSG): Antigenic determinants on alpha- and beta-subunits", BIOCHIMICA ET BIOPHYSICA ACTA, ELSEVIER, NL, vol. 1159, no. 1, 1 January 1992 (1992-01-01), pages 74 - 80, XP009030538, ISSN: 0006-3002 *

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WO2008142162A2 (fr) 2008-11-27
WO2008142162A3 (fr) 2009-02-12

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