KR20080007648A - Vaccine compositions and methods for the treatment of urinary incontinence - Google Patents

Abstract

The present invention relates to the use of compositions comprising an anti-LHRH vaccine for the control or treatment of urinary incontinence. The present invention further relates to methods of treating or controlling urinary incontinence in mammals, and in particular, in those that have been surgically neutered, e.g., spayed female dogs.

Description

Vaccine compositions and methods for treating urinary incontinence {VACCINE COMPOSITIONS AND METHODS FOR THE TREATMENT OF URINARY INCONTINENCE}

The present invention relates to the use of a composition comprising an anti-LHRH vaccine for the inhibition or treatment of urinary incontinence. The present invention also relates to a method for treating or controlling urinary incontinence in mammals, in particular surgically neutralized, such as female dogs that have had their ovaries removed.

Urinary incontinence is the inadvertent leakage or leakage of urine due to bladder or urethral sphincter dysfunction. During normal storage of urine, as the bladder is full, the urethral sphincter is in close contact and prevents leakage. Subsequently, when urine is to be excreted, the urethral sphincter relaxes, allowing urine to pass through the urethra. Urinary incontinence is often the result of loss of urethral sphincter tension, which reduces his tightness. In this case, urinary incontinence occurs in an unregulated manner whenever the pressure in the bladder exceeds the pressure required to close the urethra. This problem is sometimes more pronounced when the mammal is relaxed, such as during deep sleep, or when the pressure on the bladder is momentarily increased, such as when the mammal coughs or sneezes.

Those familiar with these conditions may find that urinary incontinence is widespread among adult women. Because of the social stigma associated with urinary incontinence, many suffering people do not report this problem to their health care providers. As a result, this medicinal problem is vastly undiagnosed and reported below.

Urinary incontinence is also fairly well known in dogs. Typically, this occurs in female dogs, most frequently in female dogs that have had their ovaries removed. Male dogs are also susceptible to this condition, but are less common than their corresponding females. Previously, the underlying mechanism of incontinence in ovariectomized or neutralized dogs has been described as being associated with low estrogen or testosterone levels after surgery after removal of the ovaries or testes. In female dogs, estrogen hormones are thought to play a role in providing tension to the urethral sphincter muscles. After ovarian removal, estrogen hormones are produced in significantly lower amounts than before ovarian removal. The adrenal gland produced a small amount of the estrogen hormone, while its produced levels were considered insufficient to provide sphincter tension. In this case, incontinence can occur, which requires some form of treatment to support sphincter tension. In a similar manner, male dogs may develop urinary incontinence after neutralization, in which the testosterone causing testosterone is removed.

Treatment of urinary incontinence has traditionally been aimed at improving urethral obstruction pressure. In response, drugs have been prescribed that increase the resistance of the bladder neck or urethra. In particular, these drugs include alpha-adrenergic agents such as ephedrine or phenylpropanolamine ("PPA"). Such treatments typically involve prolonged use of the drug and are known to cause severe cardiac electrophysiological side effects in humans. These side effects include, but are not limited to, ventricular fibrillation and cardiac arrhythmias. These potentially fatal cardiac side effects have led to the suspension of several human drugs on the market, including PPAs recently retrieved by the Food and Drug Administration ("FDA"). Alpha-adrenergic agents are contraindicated in glaucoma and progressive kidney disease

As an alternative therapy, alternative forms of estrogen or estradiol, such as diethylstilbestrol, (or testosterone in male dogs), may be used, which enhances the responsiveness of the catecholamine receptors of the urethra. However, estrogen therapy is widely considered to be less safe than the use of alpha-adrenergic agents, as it can lead to bone marrow suppression in dogs and can be fatal.

In recent years, increased serum concentrations of gonadotropins ("FSH") and luteinizing hormone ("LH"), which are lower than the levels of estrogen itself, are assumed to be the cause of incontinence in adult females and ovarian depleted female dogs. come. See Reichler, I.M. et al., The Effect of GnRH Analogs on Urinary Incontinence after Ablation of the Ovaries in Dogs, Theriogenology, 60: pp. 1207-1216 (2003); And US Patent Publication No. 2005/0043342 (hereinafter collectively referred to as "Reichler et al."). Reportedly, the pituitary gland in response to the production of luteinizing hormone secretion hormone ("LHRH", commonly referred to as gonadotropin-releasing hormone, "GnRH" or gonadotropin-releasing factor "GnRF") in the system LH and FSH Secrete. LHRH is secreted from the hypothalamus into the bloodstream in a pulsating manner in response to a range of factors, and its regulation is generally considered to be complex. Its secretion is actively affected by hypothalamic neurons but passively controlled by low levels of estrogen and other gonadoid steroids and hormones such as inhibin A and B. The secreted LHRH travels through the blood to the pituitary gland. Receptor cells in the anterior pituitary gland respond to elevations in LHRH by secreting gonadotropins FSH and LH. Then, the relative levels of FSH and LH that change over the course of the estrous cycle stimulate the production of estrogen by the ovaries. In female dogs from which the ovary has been removed, after removal of the ovary (eg, primary estrogen source), estrogen can only be measured at very low serum concentrations. Other hormones commonly derived from the gonad, such as inhibin secreted by the granules in the ovary, selectively inhibit pituitary FSH secretion. Thus, since it is no longer a feedback mechanism, FSH and LH are secreted undisturbed, resulting in an increase in the concentration of both gonadotropins. Lechler et al. Argue that the levels of FSH and LH are elevated above normal levels in both postmenopausal women suffering from urinary incontinence and female ovary-removed female dogs. Accordingly, Lechler et al. Have proposed the use of LHRH analogues, such as agents, antagonists or antibodies, in drugs that inhibit or treat urinary incontinence.

However, there are separate disadvantages with this approach. In particular, high doses of LHRH antagonists or agents are typically required to achieve the desired antibiotic stimulatory effect. In addition, the effect of using such analogs is immediate, but they are short-lived and require frequent administration. In addition, in some cases, large amounts of these analogs may lead to unwanted side effects or adverse effects. They are also very expensive since their use, and therefore their availability, is usually limited to the treatment of hormone dependent tumors. Likewise, LHRH antibodies, in addition to their significant cost, have the disadvantage that their half-life is short, resulting in potential requirements for short-term treatment and multiple administrations. Thus, the necessary frequent and continuous administration of LHRH antibodies using this approach is costly and inconvenient for dog owners. Therefore, LHRH analogs are not practical for such veterinary applications.

Thus, despite some reports of urinary incontinence treatments currently in use, complete responses to these treatments are rarely observed, or they involve serious side effects, lack long-lasting effects, are too expensive or inconvenient to maintain. . Therefore, there is a need for the development of new treatment regimens, particularly those that enable long-term inhibition and treatment of urinary incontinence in a cost-effective manner while reducing the potentially fatal side effects (eg, cardiac arrhythmias or bone marrow suppression) of current therapies. The present invention meets these needs.

Summary of the Invention

An alternative and / or additional approach to urinary incontinence therapy that has not been approved so far is to target the immune system through the use of vaccine compositions. A potential benefit of active immunotherapy is to provide improved efficacy by enhancing the subject's own immune response to the effective and long lasting regulation of gonadotropin, FSH and LH, with unlimited production and secretion.

Accordingly, the present invention relates to anti-LHRH vaccines and their use for the treatment or inhibition of urinary incontinence in mammals. In particular, the present invention provides a method of treating urinary incontinence in a mammal in need of such treatment, the method comprising administering to the mammal a therapeutically effective amount of an anti-LHRH vaccine. In one aspect of the invention, the mammal in need of treatment of urinary incontinence is a female, preferably a female with elevated luteinizing hormone or follicle stimulating hormone. In another aspect, the mammal in need of urinary incontinence treatment is a female dog. In a particularly preferred embodiment, the mammal is a female dog that has been neutralized by surgery, i.

In one aspect of the invention, the anti-LHRH vaccine comprises an immunogenic conjugate between the carrier and LHRH. Preferably, the carrier comprises a protein selected from a broad range of known carrier proteins including but not limited to bacterial toxins. Most preferably, the carrier comprises diphtheria toxoid. In another aspect, the carrier may be a synthetic peptide comprising a T-cell helper epitope. In either aspect, the effective form of the LHRH peptide is linked to the carrier, either through conjugation or during the synthesis of consecutive T-helper-LHRH peptides. In another aspect of the invention, the anti-LHRH vaccine further comprises an adjuvant. Preferably, the adjuvant is an ionic polysaccharide; Most preferably, diethyl aminoethyl dextran ("DEAE dextran") is included. In a particularly preferred embodiment, the adjuvant is DEAE dextran in combination with an immunostimulatory complex comprising saponin and cholesterol.

In another aspect, the invention relates to a method of treating or inhibiting urinary incontinence in a mammal comprising administering to the mammal a composition comprising an immunogenic conjugate between a therapeutically effective amount of a carrier and LHRH. For protein-LHRH peptide conjugate vaccines, the effective amount ranges from about 100 to about 300 ug and for T-helper-LHRH peptide based vaccines in the range of about 20 to about 200 ug.

The present invention relates to immunogenic compositions and vaccines suitable for eliciting an immune response against luteinizing hormone secretion hormone ("LHRH", or gonadotropin secreting hormone or "GnRH"). The present invention relates to the use of such anti-LHRH vaccines or physiochemically suitable compositions in a method of immunizing a mammal against LHRH to inhibit or treat urinary incontinence.

The term “anti-LHRH vaccine” includes any vaccine, composition, or pharmaceutical agent capable of eliciting an immune response to LHRH in a host. In particular, the anti-LHRH vaccine of the present invention comprises at least one LHRH antigen or immunogen of a pharmaceutically acceptable carrier useful for inducing an immune response in said host. By way of example only, anti-LHRH vaccines that can be used in connection with the present invention and methods for making them are described in International Patent Application No. PCT / AU99 / 01167 (International Publication No. WO 99/02180; Publication Date: January 21, 1999). ), International patent application PCT / AU98 / 00532 (International Publication No. WO 99/41720; published July 20, 2000) and US Pat. No. 6,685,947, but are not limited to these; These patent documents are incorporated herein by reference.

The term "LHRH" refers to all forms of LHRH and its derivatives, including but not limited to all peptides having LHRH immunogenicity. The term "derivative" includes fragments, portions, portions, chemical equivalents, variants, homologs and analogs from natural, synthetic or recombinant sources, including fusion proteins. LHRH can be obtained from any suitable source, and methods for preparing, isolating and purifying natural and synthetic LHRH are well known in the art. In a preferred embodiment, the LHRH comprises an LHRH C-terminal fragment of at least 5 amino acids.

Since LHRH itself is too small to become immunogenic, LHRH used in the context of the present invention needs to be in immunogenic form. It is well known to those skilled in the art that there are a variety of ways to produce immunogenic forms of materials that are not themselves immunogenic. One suitable method is to bind LHRH to an immunogenic carrier. Thus, in the context of the present invention, LHRH may be administered to a mammal after conjugation with an immunogenic carrier as an antigen that induces the formation of a host antibody against that LHRH. The carrier functions to stimulate T-helper cells required for an effective antibody response. Thus, the carrier may comprise small peptides of length having at least 9 amino acids that form T-cell helper epitopes. Such T-helper epitopes can be derived from Canine Distemper Virus (CDV), which is particularly suitable for application in dogs since most dogs are already vaccinated against CDV. Antibodies derived from such vaccines will then act on the LHRH of the body itself, thereby providing a long-term antigonadogenic effect. In a preferred embodiment, LHRH is administered to the mammal via a vaccine after conjugation to an immunogenic carrier to stimulate the immune system to produce an anti-LHRH antibody, which antibody reacts with LHRH to increase its concentration in the body. Reduce effectively. Thus, this effect due to the presence of LHRH, including high serum concentrations of FSH and LH that are believed to cause urinary incontinence, is reduced or eliminated.

The immunogenic carrier used for the preparation of the LHRH conjugate described above includes any substance that can increase the immunogenicity of LHRH. The immunogenic carrier used in the conjugates of the present invention may be natural or synthetic. In one aspect, the immunogenic carrier is a protein. Any of a variety of immunogenic carrier proteins that are well known to those skilled in the art can be used in the conjugate vaccines of the present invention. Suitable classes of proteins include hair, outer membrane proteins and secretory toxins of pathogens, nontoxic or "toxoid" forms of these secreting toxins, nontoxic proteins and other proteins antigenically similar to bacterial toxins. Non-limiting examples of bacterial toxins envisioned for use in the present invention include tetanus toxin / toxin, diphtheria toxin / toxin, detoxified P. aeruginosa toxin A, cholera toxin / toxin, pertussis toxin / toxin And Clostridium perfringens exotoxins / toxins. Toxic forms of these bacterial toxins are preferred. Most preferably, the immunogenic carrier used in the LHRH conjugate of the present invention is diphtheria toxin. Diphtheria toxins need to be understood to include all forms of diphtheria toxins and derivatives thereof. The term "derivative" has the same meaning as defined above. Derivatives include, for example, molecules comprising diphtheria toxic T cell epitopes or T cell epitopes in isolated form.

In another aspect of the invention, the carrier may be a synthetic peptide comprising at least one T-cell helper epitope. Epitopes may be singular or may be joined together to form a single polypeptide. It will be appreciated that when the epitopes are bound together in a single polypeptide, the epitopes may be contiguous or may be separated from each other by additional amino acids that are not part of the T helper cell epitope itself. In one embodiment, T-cell helper epitopes are included in peptide sequences selected from those described in US Pat. No. 6,685,947, which is incorporated herein by reference. In a preferred embodiment, the synthetic peptide vaccine further comprises at least one B cell epitope and / or at least one CTL epitope. In the most preferred embodiment, the synthetic peptide vaccine comprises about 3 to about 10 T-cell helper epitopes, each having 4 to 10 forms of LHRH in a linear sequence, with a total peptide content of about 80 μg of adjuvant. In combination with about 120 μg peptide / dose. Preferably, the adjuvant comprises an immune stimulating complex.

Suitable methods of binding LHRH to immunogenic carriers are well known to those skilled in the art. Without intending to limit the present invention to either method, LHRH is described, for example, in Ladd et al., American J. Reproductive Immunology 22: 56-63 (1990) or Bonneau et al., J. Animal Science 72: 14-20 (1994). The resulting conjugate can be treated to remove any remaining unbound peptide or other byproducts of the conjugate. Such treatment may be carried out by conventional dialysis or ultrafiltration.

The vaccine of the present invention may be used in combination with an adjuvant or may further comprise an adjuvant. Adjuvants are included or added to vaccines and other immunogenic formulations to increase the immune response and in some cases direct the immune response. Those skilled in the art will appreciate that any well known standard adjuvant may be used in the present invention. Suitable adjuvants for use in the vaccines of the present invention are insoluble ammonium salts, especially in hydroxide and phosphate forms (collectively "alum"), murabutide, Freund's incomplete adjuvant, Freund's complete adjuvant, ionic polysaccharides such as, but not limited to, diethyl-aminoethyl ("DEAE") groups, saponins, immune stimulating complexes, and combinations thereof. In a preferred embodiment, an adjuvant is used which has no side effects or only causes mild side effects. Other suitable adjuvants that can be used in connection with the present invention include International Patent Application No. PCT / AU99 / 01167 (Publication Number WO 99/02180, published January 21, 1999) and International Patent Application No. PCT / AU98 / 00532 (Publication Number: WO 99/41720, Publication Date: July 20, 2000), but are not limited to these, and these patent documents are incorporated herein by reference. . In a preferred embodiment, the adjuvant is DEAE-dextran. In a more preferred embodiment, the adjuvant is DEAE dextran in combination with an immune stimulating complex comprising saponin and cholesterol.

Without limiting the present invention to either theory or mode of operation, administration of an effective amount of an anti-LHRH vaccine of the present invention elicits an immune response to LHRH, thereby secreting hormones LH and FSH from the anterior pituitary gland. prevent. Preferably, the amount of anti-LHRH vaccine improves the side effects associated with ovarian resection or symptoms associated with genital aging, in particular urinary incontinence, by lowering serum concentrations circulating FSH and / or LH in female mammals to which the vaccine has been administered. Effective for Efficacy is a functional measure, and it is necessary to understand that the presence of circulating antibodies alone does not need to represent the dose of said circulating antibodies to inhibit urinary incontinence and is not defined based on anti-LHRH antibody titers. Thus, depending on all immunogenic compositions for eliciting antibodies, immunologically effective amounts of the vaccines of the present invention need to be determined experimentally. Factors to consider include adjuvant selection, immunogenic molecules to which LHRH binds, route of administration, and number of doses administered. Such factors are known in the vaccine art and one of ordinary skill in the art can make this determination without undue experimentation.

Thus, according to the method of the present invention, in female mammals such as female dogs with ovaries removed, side effects associated with ovarian ablation or symptoms related to genital aging, in particular, incontinence can be treated or suppressed. In a preferred embodiment, the present invention relates to a method of treating or inhibiting urinary incontinence in a mammal, the method comprising administering to said mammal an effective amount of an anti-LHRH vaccine. Preferably, the anti-LHRH vaccine is in the form of an LHRH-immunogenic carrier conjugate. In a more preferred embodiment, the conjugate comprises an LHRH-diphtheria toxin conjugate. In an alternate embodiment, the anti-LHRH vaccine is in the form of a synthetic peptide comprising a T-cell helper epitope. Preferably, the synthetic peptide vaccine comprises about 3 to about 10 T-cell helper epitopes, each having 4 to 10 forms of LHRH in a linear sequence, the total peptide content of the immune stimulating complex being about 80 Μg in combination is about 120 μg peptide / dose. Preferred mammals to be treated according to the vaccines of the invention are females, more preferably females with elevated serum levels of FSH or LH or both. In the most preferred embodiment, the mammal is a female dog, in particular a female dog that is surgically neutralized or ovarian removed. In another embodiment, the present invention provides a method for preventing the development of urinary incontinence in an ovarian depleted female dog comprising administering an effective amount of an anti-LHRH vaccine to the ovarian depleted female dog after removal of the ovary. to provide.

The vaccine of the present invention can be administered by any suitable method known in the art including, but not limited to, oral administration or injection. Pharmaceutical forms suitable for injectable use include sterile aqueous solutions (here water-soluble) or dispersants and sterile powders for the instant preparation of sterile injectable solutions or dispersants. They need to be stable both physically and chemically under the conditions of manufacture and storage and need to be protected against the contaminating action of microorganisms such as bacteria and fungi. The carrier may be, for example, a solvent or dispersion medium containing water, ethanol, polyols (eg, glycerol, propylene glycol and liquid polyethylene glycols, etc.), appropriate mixtures thereof, and vegetable oils. Proper fluidity can be maintained, for example, by the use of coatings such as lecithin, by the maintenance of the required particle size in the case of dispersants and by the use of surfactants. Prevention of the action of microorganisms can be obtained by various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like. In many cases, it will be preferable to include isotonic agents, such as sugars or sodium chloride. Prolonged absorption or delayed secretion of the injectable compositions can be obtained by the use in compositions of delayed absorption agents, such as aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by blending the required amount of the active compound in the appropriate various solvents with the various other ingredients described above, as required, followed by filtered sterilization. Generally, dispersants are prepared by incorporating the various sterilized active ingredients into a sterile vehicle which contains the basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, preferred methods of preparation are vacuum drying, freeze drying and spray drying techniques which previously obtain powders of any additional desired and active ingredients from their sterile filtered solutions.

If the active ingredients are adequately protected, they may be administered orally, for example with inert diluents or compatible edible carriers, or they may be enclosed in hard or soft shell gelatin capsules or compressed into tablets or directly with dietary foods. Can be combined. For oral administration, the active compound may be combined with excipients and used in the form of tablets for ingestion, oral tablets, oral tablets, capsules, elixirs, suspensions, syrups, oblates and the like. Such compositions and preparations need to contain at least 1% by weight of active compound. The percentage of compositions and formulations may, of course, vary and may conveniently be from about 5 to about 80% of the unit weight. The amount of active compound in such useful compositions ensures that an appropriate dosage is obtained. Preferred compositions or formulations according to the invention are prepared such that oral dosage unit forms contain from about 0.1 μg to 2000 μg of active compound.

Tablets, tablets, pills, capsules, and the like may also contain ingredients such as those listed below: binders such as gum, acacia, corn starch or gelatin; Excipients such as dicalcium phosphate; Disintegrants such as corn starch, potato starch, alginic acid; And lubricants such as magnesium stearate. If the dosage unit form is a capsule, it may contain a liquid carrier in addition to the materials of the aforementioned type. Various other materials may be present as coatings or may vary the physical form of the dosage unit. Syrups or elixirs may contain the active compounds, methylparabens and propylparabens as preservatives, and pigments. Of course, certain materials used to make any dosage unit form need to be veterinary pure and substantially non-toxic in their amount of use. In addition, the active compound (s) may be formulated in sustained release formulations and formulations.

Pharmaceutically acceptable carriers and / or diluents include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. The use of such media and agents for veterinary active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic compositions is contemplated. Additional active ingredients may also be formulated in the compositions.

It is particularly advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. For administration to livestock, it is particularly advantageous to use multi-dose containers connected to vaccination guns. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; Each unit contains a predetermined amount of active substance calculated to produce the desired therapeutic effect with respect to the required pharmaceutical carrier. The specification for the new dosage unit form of the present invention is intended for living subjects having (a) the unique features of the active substance and the specific therapeutic effect to be obtained and (b) the disease condition of which physical health is poor as described in detail herein. It is influenced and directly depends on the limitations inherent in the art of combining such active substances for the treatment of diseases.

The main active ingredient is conveniently formulated for effective administration in an effective amount with a suitable pharmaceutically acceptable carrier in dosage unit form as disclosed herein above. The unit dosage form may contain the main active compound, for example in an amount ranging from about 0.5 μg to about 2000 μg. The active compounds, expressed in proportions, are generally present in about 0.5 μg / ml to about 2000 μg / ml in the carrier. The relative amounts of the active ingredients, pharmaceutically acceptable carriers and any additional ingredients in the pharmaceutical compositions of the present invention will vary depending on the identity, size and condition of the subject to be treated, and further on the route through which the composition is administered.

The invention is explained in more detail with reference to the following experimental examples. These examples are provided for illustrative purposes only and are not intended to be limiting unless otherwise specified. Accordingly, the present invention should not be construed as limited to the following examples, but rather should be inferred to encompass any and all variations that become apparent as a result of the teachings provided herein.

While the invention has been described with reference to specific embodiments, it is clear that other embodiments and modifications of the invention can be invented by those skilled in the art without departing from the true spirit and scope of the invention. The appended claims need to be construed to include all such embodiments and equivalent modifications.

Example  One

Scope of clinical research:

Female dogs (females) that have been neutralized by surgery or have had their ovaries removed frequently suffer from incontinence. The incidence of this disease is reported to be as high as 20%. Current treatment relies on daily administration of drugs such as long-term synthetic estrogens, which is expensive and inconvenient. The use of long acting vaccines would be a significant improvement in the treatment of these common symptoms of female ovaries removed.

Review:

Formulations for treating incontinence in dogs include any effective anti-LHRH vaccine. Many such vaccines have been described in the prior art, but most of them have failed to provide sufficiently high safety and efficacy to be commercially viable. Very few examples of effective vaccines in dogs described in the literature. There are several examples of LHRH vaccines currently available, especially in Australia and New Zealand. Examples of these include IMPROVAC® for inhibiting boar infection in male pigs and EQUITY® for suppressing estrogenic-related behavior in mare and mare, all of which are Pfizer Animal Health. Commercially available from Animal Health. Since these formulations are intended for application to large animals, it can be expected to be unsafe in smaller animals, such as dogs, and is expected to require modifications for safe and effective use in dogs and other companion animals. Another formulation is conditionally registered by the United States Department of Agriculture (USDA) for the treatment of prostatic hypertrophy in grown male dogs. This vaccine has been demonstrated to be safe and effective in growing male dogs and can be used for inhibiting dog urinary incontinence (CUI) in the present invention as follows:

The study was conducted as a double-blind study for two groups receiving CUl vaccine or placebo, with the placebo consisting of adjuvant only. Neither the organizer nor the clinical volunteer knows the treatment given.

animal:

Bitches (up to 35) raised as pets as part of a community were selected from clinical registries such as, for example, registries at the University of Melbourne / Livestock Clinic. For this study, you have a clinical symptom of long-term incontinence, have already been treated for this condition (other accepted treatments such as stilbestrol, a synthetic form of estrogen), or alpha-adrenergics such as PPA. Only chicks whose ovaries had been removed more than two years old with stimulants were selected.

vaccine:

In one embodiment, the vaccine comprises conditionally approved from USDA for Pfizer Animal Health for the treatment of prostatic hyperplasia, including, but not limited to, International Patent Application PCT / AU99 / 01167 (Publication No. WO 99/02180; Conjugation of 2 to 10 forms of LHRH peptide conjugated to diphtheria toxin as a carrier with an adjuvant combination of 80 ug of immunostimulatory complex and 10 mg of DEAE dextran as described on Jan. 21, 1999) Contains 200 ug / dose. In another embodiment, the vaccine comprises synthetic peptide vaccines containing about 3 to about 10 T-cell helper epitopes, each having 4-10 forms of LHRH in a linear sequence, and the total peptide content is immune. 120 ug peptide / dose in combination with 80 ug of stimulatory complex.

The vaccine is administered to each dog at 28 day intervals in studies in at least two cases.

The following parameters, body temperature, mode of behavior including lethargy, pain at the injection site and swelling at the injection site were measured directly after each vaccination. Serum samples were also taken to measure the levels of LH and antibody response to LHRH. Starting two weeks before the first vaccination, individual owners were scored for incontinence in each bitch according to the usual scoring system. In this way, each individual dog acted as its own control for post-treatment changes.

Claims (33)

A method of treating urinary incontinence in a mammal comprising administering a therapeutically effective amount of an anti-LHRH vaccine to a mammal in need thereof. The method of claim 1, A method of treating urinary incontinence in a mammal, wherein said mammal is a female. The method of claim 1, The mammal is a female with elevated serum concentrations of gonadotropin, the method of treating urinary incontinence in mammals. The method of claim 3, wherein The gonadotropin is selected from the group consisting of luteinizing hormone and follicle stimulating hormone, method of treating urinary incontinence in mammals. The method of claim 1, A method for treating urinary incontinence in a mammal, wherein said mammal is neutralized by surgery. The method of claim 5, A method for treating urinary incontinence in a mammal, in which the mammal is ovary removed. The method of claim 1, Wherein said anti-LHRH vaccine comprises an immunogenic conjugate between a carrier and LHRH. The method of claim 7, wherein And said carrier comprises a protein. The method of claim 8, A method for treating urinary incontinence in mammals, wherein said protein comprises bacterial toxins. The method of claim 9, The bacterial toxin is a diphtheria toxin. The method of claim 7, wherein The carrier comprises a synthetic peptide comprising at least one T-cell helper epitope. The method of claim 1, The vaccine further comprises an adjuvant. The method of claim 12, The adjuvant comprises DEAE-dextran, the method of treating urinary incontinence in mammals. The method of claim 13, The adjuvant further comprises an immune stimulating complex consisting of saponin and cholesterol components, the method of treating urinary incontinence in mammals. The method of claim 7, wherein Wherein said immunogenic conjugate comprises an LHRH-diphtheria toxin conjugate. A method of treating or inhibiting urinary incontinence in a mammal comprising administering to a mammal the composition comprising a therapeutically effective amount of an immunogenic conjugate between the carrier and the LHRH. The method of claim 16, A method of treating or inhibiting urinary incontinence in a mammal, wherein said carrier comprises a protein. The method of claim 17, And said carrier comprises diphtheria toxic toxins. The method of claim 16, A method of treating or inhibiting urinary incontinence in a mammal, wherein said carrier comprises a T-cell epitope. The method of claim 16, The method of claim 1, wherein the composition further comprises an adjuvant. A vaccine for the treatment or inhibition of urinary incontinence in female mammals, comprising a therapeutically effective amount of LHRH. The method of claim 21, A vaccine further comprising an adjuvant. The method of claim 21, A vaccine in which LHRH is conjugated with an immunogenic carrier. The method of claim 23, The vaccine, wherein the immunogenic carrier comprises a protein. The method of claim 23, And wherein said immunogenic carrier comprises a T-cell helper epitope. The method of claim 24, A vaccine comprising the diphtheria toxin. The method of claim 21, And the female mammal comprises a dog whose ovary has been removed by surgery. A method of enhancing an immune response in a mammal comprising administering to said mammal suffering from incontinence due to unlimited production and release of gonadotropin, a therapeutically effective amount of LHRH. The method of claim 28, The LHRH is administered in the form of an antigen, immune response method of a mammal. The method of claim 28, The LHRH is administered in the form of a vaccine, a method for enhancing immune response in mammals. Administering a therapeutically effective amount of an anti-LHRH vaccine to a dog that has been neutralized by surgery or has had its ovary removed. The method of claim 31, wherein A method of treating dog incontinence, wherein said dog is a female. A method for preventing the development of urinary incontinence in an ovarian depleted female dog, comprising administering to the female ovary depleted female dog an effective amount of an anti-LHRH vaccine after removal of the ovary.