EP0527774A1 - Peptide apparente au gene de la calcitonine pour le traitement de testicules non descendus - Google Patents

Peptide apparente au gene de la calcitonine pour le traitement de testicules non descendus

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Publication number
EP0527774A1
EP0527774A1 EP91907817A EP91907817A EP0527774A1 EP 0527774 A1 EP0527774 A1 EP 0527774A1 EP 91907817 A EP91907817 A EP 91907817A EP 91907817 A EP91907817 A EP 91907817A EP 0527774 A1 EP0527774 A1 EP 0527774A1
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EP
European Patent Office
Prior art keywords
cgrp
scrotum
treatment
undescended
human
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EP91907817A
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German (de)
English (en)
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EP0527774A4 (en
Inventor
John Medwyn Hutson
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University of Melbourne
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University of Melbourne
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Application filed by University of Melbourne filed Critical University of Melbourne
Publication of EP0527774A1 publication Critical patent/EP0527774A1/fr
Publication of EP0527774A4 publication Critical patent/EP0527774A4/en
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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
    • A61K38/225—Calcitonin gene related peptide
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P13/00—Drugs for disorders of the urinary system
    • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
    • 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

Definitions

  • This invention relates to a method for the non- surgical treatment of undescended testicles in male animals.
  • the invention is also concerned with pharmaceutical compositions for the treatment of undescended testicles.
  • testes are two glandular organs which secrete semen, and are situated in the scrotum, being suspended by the spermatic cords.
  • testicular descent is controlled by male androgenic hormones (testosterone). Androgens were proposed to act on mesenchymal tissue in the groin known as the gubernaculum, which migrates across the pubic region from the groin to the scrotum during inguino-scrotal testicular descent. The gubernaculum was thought to guide the testes into the scrotum.
  • testes In humans, approximately 5% of male babies are born with undescended testicles. In 1 to 2% of males, the testes do not descend into the scrotum. In the remaining males, the testes may arrive in the scrotum a few weeks after birth as compared with the normal time of 30 to 36 weeks gestation. These "late descenders" are not quite normal and many have testicles which "re-ascend" out of the scrotum later in childhood.
  • the treatment for undescended testes is invasive surgery (orchidopexy) where the undescended testes are physically transferred to the scrotum. This is usually carried out at 1 to 3 years of age because the testes are known to develop progressive histological abnormality thereafter. This surgical procedure is traumatic for the individual and family involved, is costly, and has attendant risks associated with all forms of surgery requiring general anaesthesia. It has also been proposed to treat undescended testicles by administering the human androgen testosterone or by treatment with HCG (human chronic gonadotrophin) or luteinizing hormone releasing hormone (LHRH). These treatments have proved ineffective. A requirement accordingly exists for a non-surgical and convenient treatment for undescended testicles.
  • HCG human chronic gonadotrophin
  • LHRH luteinizing hormone releasing hormone
  • a method for the treatment of undescended testicles in male animals which comprises administering to a subject in need of such treatment calcitonin gene-related peptide (hereafter CGRP) or an analogue thereof optionally in association with a carrier and/or excipient, in an amount effective to cause testicular descent.
  • CGRP calcitonin gene-related peptide
  • the invention relates to a pharmaceutical composition for the treatment of undescended testicles in male animals, which comprises CGRP or an analogue thereof having CGRP activity in association with a pharmaceutically acceptable carrier and/or excipient.
  • this invention relates to the use of CGRP or an analogue thereof in the manufacture of a medicament for the treatment of undescended testicles.
  • CGRP is a 37 amino acid peptide produced by alternative splicing of calcitonin mRNA (Rosenfeld et al., Nature, Vol. 304, 1983).
  • CGRP is a neuropeptide and has been described in many sensory nerves but few motor nerves.
  • CGRP refers to CGRP from any animal species, such as human, horse, sheep, pig, rat, mouse, etc. Principally, but without limitation, CGRP refers to human CGRP.
  • the term CGRP extends to naturally occurring allelic variants of the CGRP peptide sequence.
  • Human CGRP has the following sequence: H-Ser-Cys-Asn-Thr-Ala-Thr-Cys-Val-Thr-His-Arg-Leu-Ala- Gly- eu-Leu-Ser-Arg-Ser-Gly-Gly-Val-Val- ys-Asp-Asn-Phe- Val-Pro-Thr-Asn-Val-Gly-Ser-Lys-Ala-Phe.
  • CGRP human is available commercially from a number of suppliers, such as Peninsular Laboratories. CGRP may be purified from tissues containing it according to well known procedures. Preferably, CGRP is produced by peptide synthetic techniques, such as solid phase peptide synthesis, or is produced by recombinant DNA methods.
  • Analogues of CGRP which comprise amino acid sequence variants fall into one or more of three classes: substitutional, insertional or deletional variants. Insertions include amino and/or carboxyl terminal fusions as well as intrasequence insertions of single or multiple amino acids. Generally, insertions within the mature - 4 - coding sequence of CGRP will be smaller than those with the amino or carboxyl terminal fusions, of the order of say 1 to 4 residues.
  • Insertional amino acid sequence variants of CGRP are those in which one or more amino acid residues are introduced into a predetermined site in the CGRP peptide.
  • Deletional variants are characterised by the removal of one or more amino acids from the CGRP peptide sequence. Typically, no more than about 2 to 6 residues are deleted at any one site within the CGRP molecule.
  • Amino acid substitutions are typically of single residues; insertions usually will be on the order of about 1 to 10 amino acid residues; and deletions will range from about 1 to 20 residues. Deletions or insertions preferably are made in adjacent pairs, i.e. a deletion of two residues or insertion of two residues.
  • substitutional variants are those in which at least one residue in the CGRP sequence has been removed and a different residue inserted in its place. Such substitutions generally are made in accordance with the following Table.
  • amino acids are replaced by other amino acids having like properties, such as hydrophobicity, hydropholicity, electronegativity, bulky side chains, etc.
  • amino acid variants of CGRP referred to above may readily be made using peptide synthetic techniques well known in the art, such as solid phase peptide synthesis (Merrifield, J. Am. Chem. Soc. , 85, p2149 (1964) and the like, or by recombinant DNA manipulations upon the gene encoding CGRP of any particular animal. Techniques for making substitution mutations at predetermined sites in DNA having a known sequence are well known, for example M13' mutagenesis. The manipulation of DNA sequences to produce variant proteins which manifest as substitutional, insertional or deletional variants are well known in the art and are described for example in Maniatis et al. (Molecular Cloning, A Laboratory Manual, Cold Spring Harbor Laboratory, 1982).
  • CGRP activity is defined as the ability to effect testes descent in animals having undescended testes.
  • a convenient in-vivo assay for CGRP activity utilises isolated male gubernaculum tissue.
  • CGRP bioactivity may also be tested in-vivo utilising male animals whose testes have not yet descended, such as new born rats, or animals which have congenitally undescended testes, such as the TS strain of rat. When compounds having CGRP activity are applied to the scrotum of such animals, for example by way of injection, testicular descent takes place. Again, in- vivo models such as described provide a ready means for assessing CGRP activity.
  • Examples of commercially available CGRP include chicken CGRP, human CGRP, biotinyl-CGRP (human), [Tyr']- CGRP (human), biotinyl-CGRPII (human), CGRP (rat and biotinyl-CGRP (rat).
  • CGRP should generally be administered under the guidance of a physician, and pharmaceutical compositions would usually contain an effective amount of the peptide or analogues thereof in conjunction with a conventional, pharmaceutically acceptable carrier.
  • the pharmaceutical carrier employed may be, for example, either a liquid or a solid.
  • liquid carriers include physiologically buffered saline, dextrose, sterile water, olive oil and the like.
  • the carrier may include time delay material well known to the art, such as glyceryl monostearate, ethyl cellulose, hydroxypropylmethyl cellulose, methylmethacrylate and the.like.
  • solid carriers are lactose, sucrose, talc, gelatin, agar, pectin, magnesium stearate, stearic acid and the like.
  • a wide variety of pharmaceutical forms can be employed.
  • I jectable forms of CGRP generally contain CGRP dissolved in a sterile vehicle such as water, saline, dextrose or the like.
  • Injectable solutions of CGRP may contain, for example, 50 ⁇ g to 500mg per ml. Injectable solutions may be provided in ampule or vial or non-aqueous liquid suspension.
  • the preparation can be tableted, placed in a hard gelatin capsule or admixed with a slow release polymer to form a dosage form.
  • the amount of solid carrier will vary widely but will preferably be from about O.lmg to lg.
  • CGRP may be formulated into a cream, ointment, paste, salve or the like for topical application to the scrotum.
  • Such forms may include skin penetrating agents to facilitate the passage of CGRP or analogues thereof into the scrotum.
  • Suitable skin penetrating agents include dimethylsulphoxide (DMSO) and the like.
  • Formulations for topical use containing CGRP or analogues thereof may be applied to the scrotum from 1 to 4 times per day for about 1 to 10 days.
  • the active ingredient may contain from about 0.001% to 10% w/w, e.g. from 1 to 2% by weight of the formulation.
  • Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetration through the skin to the site where treatment is required, such as: liniments, lotions, creams, ointments or pastes.
  • Lotions or liniments for application to the skin may also include an agent to hasten drying and to cool the skin, such as an alcohol or acetone and/or moisturiser such as glycerol or an oil such as caster oil.
  • Creams, ointments or pastes according to the present invention are semi-solid formulations of the active ingredients for external application. They may be made by mixing the active ingredient in finely-divided or powdered form, alone or in solution, or in suspension in an aqueous or non-aqueous fluid with the aid of suitable machinery, with a greasy or non-greasy base.
  • the base may comprise hydrocarbons such as hard, soft or liquid paraffin, glycerol, bees wax, a mucilage, an oil of natural origin such as almond, corn, caster or olive oils, wool fat or its derivatives or a fatty acid such as stearic acid together with an alcohol such as polypropylene glycol.
  • the formulation may incorporate any suitable surface active agent such as an ionic, cationic or non-ionic surfactant such as sorbitan esters or the like.
  • each parenteral dose of CGRP containing pharmaceutical forms will contain a reactive ingredient in an amount from about 0.05mg to about 500mg. If oral dosage units are employed, they will contain the active ingredient in an amount of from about 0.05mg to about 1.Omg.
  • CGRP or analogues thereof may be administered from an implantable or skin-adhesive sustained release article.
  • suitable systems include copolymers of L-glutamic acid and ⁇ -ethyl-L-glutamate (U. Sidman et al., 1983, "Biopolymers" 22, 1: 547-556), poly(2- hydroxyethyl- ethacrylate) (R. Langer et al., 1981, J. Biomed. Matter.
  • Such articles may, for example, be implanted sub-cutaneously in the scrotum or placed in contact with the skin of the - 10 - scrotum.
  • Animals which may be treated according to the present invention include humans, horses, and other domestic animals.
  • Medicaments or compositions may be prepared by admixing, dissolving, blending, grinding or the like, CGRP with a pharmaceutically acceptable carrier or excipient according to methods well known in the art.
  • CGRP is administered to such an animal in an effective amount.
  • effective amount refers to an amount effective to cause testicular descent. It will be recognised by one of skill in the art that the form and character of the pharmaceutically acceptable carrier or diluent is dictated by the amount of active ingredient with which it is to be combined, the route of administration and other well known variables.
  • the route of administration of CGRP or analogues thereof may be parenteral, topical, or oral.
  • parenteral as used herein includes intravenous, intramuscular or subcutaneous administration.
  • the subcutaneous form of parenteral administration to the scrotum is generally preferred.
  • CGRP or analogues thereof may be injected into the scrotum at birth or shortly after, on the side of the palpable but undescended testis as a way of stimulating normal migration of the gubernaculum into the scrotum.
  • the amount of CGRP or analogues thereof injected into the scrotum in a single bolus injection is from about 20 ⁇ g to lOOO ⁇ g per kilogram, and more preferably l ⁇ g per 5 to 10 g body weight.
  • a single scrotal injection may be provided for testis descent, or several injections over a limited time period such as from 1 to 10 days.
  • CGRP is slowly released to the scrotum to effect migration of gubernaculum and testis into the scrotum.
  • CGRP and analogues thereof will be determined by factors such as the route and site of administration, and the type and age of the particular animal being treated. Dosage and frequency of administration of CGRP or analogues thereof will often depend upon the judgement of a consulting physician or veterinarian in any particular case.
  • the optimal course of treatment that is, the number of doses of CGRP or analogues thereof given per day for a defined number of days, can be readily ascertained by those skilled in the art using conventional courses of treatment determination tests.
  • CGRP is the chemotactic signal released by the genitofemoral nerve within the scrotum.
  • FIG. 1 there is shown a displacement curve derived by computerized densitometry from serial gubernacular sections incubated with increasing concentrations of unlabelled human CGRP in the presence of radiolabelled human CGRP.
  • GNS Genitofemoral Nerve
  • the genitofemoral nerve is essential for inguino-scrotal testicular descent as transection of the nerve prevents gubernacular migration from groin to scrotum (Beasley, S.W. & Hutson, J.M. [1987] Aust. NZ J.Surg. 57, 49-51). It has been previously shown that cutting the GFN blocks the action of androgens without preventing testosterone secretion. This experiment examines the course of the GFN nerve in immature rats. Materials and Methods:
  • the animals were anaesthetised with oxygen and 2% halothane and a small transverse incision was made across the lower abdomen to open the peritoneum.
  • the bowel was pushed aside with cotton swabs, and the peritoneum was divided inferior to the renal vessels and medial to the ureter on one side.
  • Each genitofemoral nerve was picked up carefully with forceps, and divided.
  • Several crystals of either diamidinophenyl indole (DAPI - Sigma) or Fast Blue (Sigma) were applied to the distal end of the cut nerve, then the peritoneum and bowel were returned to the normal positions.
  • the wound was sutured with one layer of continuous 6/0 Ethicon silk.
  • the pelvis was embedded in OCT plastic medium cooled by liquid nitrogen and isopentane, then 20 ⁇ m frozen sections were cut in the sagittal plane onto 1% gelatin- coated slides. The fluorescence, and thus the course of the nerve, could then be followed by studying serial sections under an epifluorescence microscope (Leitz
  • Fluorescent tracing of the genital branch of the GFN showed that it runs through the inguinal canal posterolateral to the spermatic cord, and a small branch follows the cord to supply the head of the epididymis and tunica albuginea, but not the testis itself.
  • The' main trunk runs distally behind the testis on the surface of the cremaster muscle, supplying this, then turns - 14 - cranially to enter the gubernaculum from its distal attachment, where branches fan out supplying the gubernacular substance and the caudal epididymis.
  • a branch of the nerve continues past the testis to end in a network of fibres in the subcutaneous tissues and dermis of the region that will become the scrotum.
  • Fluorescent anterograde nerve labelling appeared to be a reliable and specific method of tracing the course of the GFN, as very little fluorescent straining was present outside the nerve itself, provided that the interval between labelling the nerve and sacrificing the animal did not exceed three days. After this time some leakage of dye out of the nerve occurred.
  • Example 2 hereafter provides evidence of a neuropeptide located within the GFN which is believed to act as a chemotactic signal drawing the gubernaculum and associated testis into the scrotum.
  • the GFN is analysed histochemically in this Example in an attempt to identify neuropeptides distributed therein which may act as possible transmitters mediating gubernacular and testis descent.
  • a transmitter is likely to be a peptide, as these are capable of exerting trophic and modulatory effects over extended periods of time, and often function in a neuro-endocrine manner.
  • the animals used in this Example were 12 male and 3 - 15 - female rats, aged 6 days at operation, and 5 male, 5 female and 5 Testicular Feminization Syndrome (TFM) mice aged 10 days old at operation.
  • TBM Testicular Feminization Syndrome
  • Antibodies against thirty different neuropeptides 5 were used in this example. These included antibodies against vasoactive intestinal peptide, 5- hydroxytryptamine, somatostatin 8, met-enkephalin, substance P, thyrotrophin releasing hormone, neuropeptide Y and calcitonin gene-relating peptide (CGRP). 0 Antibodies were obtained from Prof. J. Furness, Flinders Medical Centre, with the exception of calcitonin gene- related peptide rabbit anti-rat antibody which was purchased from Peninsula Laboratories, U.S.A. This latter antibody was used in a concentration 1:2000. 5 Animals were anaesthetised, surgically prepared, and tissues removed and fixed as for Example 1.
  • Tissues were embedded in OCT plastic medium, cooled with liquid nitrogen and isopentane, then 14 ⁇ m frozen serial sections were cut onto 1% gelatin-coated slides, 0 and left to dry. The spinal cords were cut transversely and the pelvises were cut in the sagittal plane. Slides were pre-incubated for 30 minutes with a covering to 10% normal sheep serum (NSS) to block non-specific staining. Excess serum was removed, then the slides were incubated overnight in a humidified box with various primary antibodies raised against different neuropeptides. The antibody mixture was diluted with PBS, and contained 10% NSS in the final solution.
  • NSS normal sheep serum
  • the location of immunohistochemical staining was compared with the retrograde labelling on the same 5 sections by simply changing the excitation filter, as different wave lengths are required in order for FITC (520nm) and DAPI (400nm) to emit light. To quantify cells consistently all sections were counted, and only those cells which stained brightly and in which the 0 nucleus could be clearly defined were counted as positive.
  • Negative controls include either PBS alone, or antiserum with excess synthetic rat CGRP, instead of the primary antibody.
  • the dorsal root ganglia served as a 5 positive control for CGRP.
  • CGRP is the transmitter which exerts its effect on migration of the gubernaculum into the scrotum.
  • This experiment examines gubernacula in-vitro and in-vivo and the effects of CGRP thereon.
  • Testicular descent was defined as descent of testis into the scrotum on compression of the abdomen.
  • CGRP 470 ⁇ mole/day for 14 days
  • Water injections at the same site acted as a control.
  • Two out of five right testes were undescended (40%) with water injections compared with four out of six (67%) when CGRP was injected nearby.
  • the binding of radioactive CGRP in the gubernaculum was examined in vitro, to determine whether the gubernaculum contained receptors for CGRP.
  • neonatal Sprague-Dawley male rats were sacrificed by decapitation at two days after birth.
  • each rat was placed in a supine position and its limbs taped on a metal tray on ice.
  • a Zeiss operating microscope Using a Zeiss operating microscope, a lower abdominal transverse incision about 2 mm below the umbilicus was made and, by excising flap-shaped inguinoscrotal skin, gubernacula were exposed to show a cone-shaped projection.
  • Increased abdominal pressure produced by pressing on the upper abdomen with cotton wool buds aided the resection of gubernaculum-.
  • Fine microsurgical scissors were used to divide the distal attachment of the caudal epididymis with the gubernaculum.
  • the excised gubernacula were rapidly removed and snap frozen by immersion in liquid nitrogen. Sagittal sections of 20- ⁇ m thickness were cut on a cryostat at -12°C and thaw mounted on 0.5% gelatin-coated slides. The sections were dried in a vacuum desiccator overnight at 4°C and stored at -70"C. Alternate sections were stained with haematoxylin and eosin in order to identify fine structures of the 125 I-labelled gubernacula at the time of quantitation.
  • Radiolabelled human CGRP ⁇ (2-[ 125 I)iodohistidyl 10 )- human CGRP ⁇ was obtained from Amersham International pic. (Buckinghamshire, England) (-2000 Cl/mmol).
  • the following synthetic peptides: human CGRP, rat CGRP (8- 37), rat CGRP (Tyr 27 , 28-37), salmon thyrocalcitonin, human vasoactive intestinal peptide (VIP), somatostatin, and substance P were purchased from Auspep Pty. Ltd. (Melbourne, Australia); human thyrocalcitonin and Serotonin (5-hydroxytryptamine) from Sigma Chemical Co. (St. Louis, MO).
  • the following buffer was used for incubating tissue sections with various ligands: 100 mM HEPES, containing 120 mM NaCl, 1.2 mM MgS0 4 , 2.5 mM KC1, 15 mM NaC 2 H 3 0 2 , 10 mM D-glucose, 1 mM EDTA, 0.5% BSA, and 0.35 mM Bacitoracin.
  • tissue sections were thawed and subsequently incubated for 24 hours at 4°C in the foregoing buffer at pH 8.0, containing 35 pM [ 125 I]-human CGRP. To determine non ⁇ specific binding, some sections were also incubated with 1 ⁇ M unlabelled human CGRP.
  • Displacement of [ 15 I]-human CGRP binding for characterisation of binding properties and specificity were derived by incubating a series of adjacent tissue sections with increasing concentrations of unlabelled CGRP or of the following peptide: rat CGRP (8-37), rat CGRP (Tyr 27 , 28-37), salmon thyrocalcitonin, human thyrocalcitonin, human VIP, somatostatin, serotonin, and substance P. After incubation, the sections were passed through four successive 30-seconds changes of buffer at pH 8.0 on ice, to remove nonspecifically-bound ligand.
  • Radioactive standards used autoradiographic [ 125 I] micro-scales (Amersham International pic, Buckinghamshire, England), which consist of 10 layers of [ 125 I]-incorporated polymer arranged in order of increasing specific activity separated by non-radioactive layers and were simultaneously exposed to the X-ray film.
  • the radioactivity standards were corrected for decay using the exponential equation, the decay constant and the elapsed time interval to the middle of the exposure period.
  • EyeCom Model 850 image processor Log E/Spatial Data systems, Springfield, Virginia, U.S.A.
  • incubated sections were processed for emulsion autoradiography as follows.
  • the sections were dipped for 1 to 2 seconds in Amersham LM-1 emulsion (Amersham International pic, Buckinghamshire, England) immersed in a waterbath at 43°C, dried for one hour at room temperature with a relative humidity of 60%, and stored in plastic slide box containing silica gel at 4°C for exposure for 4 weeks.
  • Amersham LM-1 emulsion Amersham International pic, Buckinghamshire, England
  • Binding saturation was achieved by 24 hours incubation at 4°C with the buffer pH at 8.0. Either longer incubation time, higher incubation temperature, or higher buffer pH than the optimal conditions resulted in higher non-specific binding. Definitive binding studies were, therefore, performed at 4°C for 24 hours, when specific binding was seen.
  • Displacement curves were derived by computerised densitometry from serial sections incubated with increasing concentrations of unlabelled human CGRP in the presence of radiolabelled human CGRP ( Figure 1). Computer-derived plots of these data also revealed a single class of binding sites.
  • Rat CGRP (8-37) was potent in competing with [ 125 I]- human CGRP with affinity constants similar to or greater than that of unlabelled human CGRP; salmon calcitonin competed at this site with a lower affinity than human CGRP, while the human calcitonin had only a very low potency; rat CGRP (Tyr 27 , 28-37) also competed with the binding sites but at a lower affinity than human CGRP. However the unrelated peptides, human VIP, somatostatin, serotonin, and substance P do not compete at all for concentrations of up to 1 X 10" 6 M.
  • mice were used in the study.
  • mice were injected with 25 ⁇ l of 10" 4 molar CGRP (2.5 n molar) within the first three days of life.
  • the mice were injected through the skin of the left iliac fossa and the developing left scrotum was entered obliquely. Once approximately half of the injected dose was administered, the scrotal sceptum was pierced to enter the right scrotum and the rest of the injection was continued.
  • CGRP (8-37) is a competitive inhibitor of CGRP and lacks the first seven amino acids which comprise the active site of CGRP.
  • mice were inspected weekly for four weeks for testicular descent. Any mice who demonstrated descended testis on inspection were not injected again on that side. Testicular descent was defined as descent of testis into the scrotum on compression of the abdomen. Descent of the gubernaculum was not considered significant. All rectractile testes were included under descended testes.
  • mice with undescended testis were sacrificed and subjected to histology to determine whether there were mechanical factors inhibiting testicular descent, i.e. fibrous tissue formation as a result of the injection.
  • the control group of mice were injected with phosphate buffered saline (PBS) at the same volume. Results were compared using non-parametric analysis.
  • the experimental group consisted of 138 mice while the control group consisted of 37 mice.
  • CGRP (8-37) binds to the receptors and competitively inhibits the activity of CGRP released from the genitofemoral nerve.
  • this inhibiting effect of CGRP (8-37) was not irreversible, probably because of diffusion and its biological half-life.
  • Sex cells also contain- containing CGRP ing

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Abstract

L'invention décrit des procédés servant au traitement des testicules non descendus chez les animaux mâles, comprenant l'administration à un sujet nécessitant un tel traitement, d'un peptide apparenté génétiquement à la calcitonine (CGRP) ou un de ses analogues possédant une activité CGRP, éventuellement associé à un agent porteur et/ou un excipient, en quantité efficace pour provoquer la descente des testicules. L'invention décrit également des compositions pharmaceutiques servant au traitement des testicules non descendus chez les animaux mâles, comprenant un peptide apparenté génétiquement à la calcitonine CGRP ou un de ses analogues possédant une activité CGRP, en association avec un agent porteur et/ou un excipient acceptable sur le plan pharmaceutique. L'utilisation d'un peptide apparenté génétiquement à la calcitonine CGRP dans le traitement des testicules non descendus représente une solution alternative efficace par rapport à la chirurgie interne.
EP19910907817 1990-04-10 1991-04-10 Calcitonin gene related peptide for the treatment of undescended testicles Withdrawn EP0527774A4 (en)

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AU9573/90 1990-04-10
AUPJ957390 1990-04-10

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EP0527774A4 EP0527774A4 (en) 1993-03-31

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US5567679A (en) * 1993-12-13 1996-10-22 Daly; Theodore J. Use of CGRP in treating alopecia

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1985002840A1 (fr) * 1983-12-23 1985-07-04 Evans Roland M Cgrp humain
WO1990012586A1 (fr) * 1989-04-27 1990-11-01 Georg Stief Medicaments (peptides apparentes au gene de calcitonine) pour soigner les dysfonctionnements erectiles

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BRAIN RESEARCH vol. 512, no. 2, 2 April 1990, pages 229 - 237 S. KAR ET AL. 'ORIGINS AND PROJECTIONS OF PEPTIDE-IMMUNOREACTIVE NERVES IN THE MALE RAT GENITOFEMORAL NERVE' *
JOURNAL OF PATHOLOGY vol. 154, no. 1-4, 1988, page 79A S. KAR ET AL. 'PRIMARY SENSORY NEURONES IMMUNOREACTIVE FOR CALCITONIN GENE-RELATED PEPTIDE (CGRP), GALANIN AND SUBSTANCE P AND CGRP-IMMUNOREACTIVE MOTORNEURONES CONSTITUTE THE MAJOR AFFERENT AND EFFERENT PEPTIDERGIC INPUT TO THE GENITOFEMORAL NERVE' *
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WO1991015246A1 (fr) 1991-10-17
JPH05506223A (ja) 1993-09-16
CA2080354A1 (fr) 1991-10-11
EP0527774A4 (en) 1993-03-31

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