Use of deslorelin in chemical castration of non-human mammals related to PK / PD interaction

Abstract

The present invention relates to deslorelin in a pharmaceutical or non-pharmaceutical product for use in chemical castration of non-human mammals within one month after administration, wherein when administered, the Cmax of deslorelin after administration (about 1 to 2 hours after administration) exceeds 7000 pg / mL and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL. The present invention also relates to the sustained-release drug delivery system for carrying out the above use and other embodiments. The present invention further relates to a method for chemically castrating a non-human animal, comprising the step of administering an effective amount of deslorelin so as to obtain plasma deslorelin having a Cmax concentration of 7000 pg / mL to 40000 pg / mL or an equivalent amount of its pharmaceutically acceptable salt within the first 24 hours after injection, and then the step of administering a maintenance dose so as to obtain plasma deslorelin having an average concentration of more than 10 pg / ml or an equivalent amount of its pharmaceutically acceptable salt.

Description

Technical Field

[0001] The present disclosure generally relates to pharmaceutical compositions of deslorelin for use in chemical castration of non-human mammals, preferably companion animals.

Background Art

[0002] Deslorelin is a gonadotropin-releasing hormone (GnRH) gonadotropin analogue that acts by suppressing the function of the gonadotropic pituitary axis when administered continuously at low doses.

[0003] GnRH is a hypothalamic decapeptide that acts at the top of the cascade regulating the function of the hypothalamic-pituitary-gonadal axis. GnRH binds to GnRH receptors (seven-transmembrane receptors), producing and releasing the two important gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), in the pituitary gland, which then control gonadal function. - LH (Luteinizing Hormone): In addition to its role in gametogenesis, it promotes the production of sex steroids (androgens, estrogens, and progesterone); - FSH (Follicle-Stimulating Hormone): Stimulates the production of gametes (sperm and eggs).

[0004] When GnRH binds to its receptor, it increases the synthesis of the receptor's mRNA and enhances the pituitary's sensitivity to GnRH. Continuous secretion of GnRH has the opposite effect (a breakdown in LH and FSH secretion is observed, which can be called desensitization) (Finch A.R. et al., Agonist-induced internalization and downregulation of gonadotropin-releasing hormone receptors. Am. J. Physiol. Cell Physiol. 2009 Sep., 297(3)).

[0005] Desensitization of the pituitary gland to GnRH is caused by internalization of the GnRH receptor (no longer present and not binding to the cell surface) and inactivation of the intracellular signaling cascade. As soon as desensitization of pituitary cells to GnRH begins, LH concentration drops to undetectable levels and thus cannot support testosterone and sperm production.

[0006] To solve the problem of contraception in animals, there are people who consider using GnRH agonists as contraceptives continuously over a long period of time, and thus the idea of designing a deslorelin implant that continuously diffuses this GnRH agonist (deslorelin) has emerged.

[0007] This type of implant is used in two different ways. - Fertility induction (short-term implant): Ovuplant - Induction of infertility by long-term action: Suprelorin

[0008] These agonist-based implants are known in the state of the art to have a two-stage action during implant insertion, releasing deslorelin and inducing a strong stimulation of the LH and FSH production cascades. This step is called the "flare-up". In male dogs, this induces the production of LH and FSH, leading to strong synthesis of testosterone. Subsequently, the male has strong fertility (improvement in sperm quality, motility).

[0009] After this "flare-up" period, a period of infertility with harmful effects on all aspects of sperm quality and testosterone concentration is established (Goericke-Pesch S., Long-term effects of GnRH agonists on fertility and behaviour. Reprod. Domest. Anim., 2017, 52 Suppl2: 336-347).

[0010] In addition to testing dogs, studies have been conducted in cats and prepubertal cats (Fontaine C., Long-term contraception in a small implant: A review of Suprelorin (deslorelin) studies in cats., J. Feline Med. Surg., 2015, 17(9), pages 766-771). This has shown the efficacy of the implant as a contraceptive in both males and females. During the reproductive suppression period, steroid hormone concentrations of less than 1 ng / ml progesterone and 10 pg / ml estradiol were measured in females, and plasma testosterone concentrations were at basal levels (<0.1 ng / ml) in male cats. In males, the duration of action using a 4.7 mg implant was 78.8 ± 12.9 weeks, varying from 61.7 weeks (about 15 months) to 100.7 weeks (about 25 months).

[0011] Other studies are being conducted in leopards, rats, and coyotes.

[0012] According to the prior art, deslorelin has a contraceptive effect in females, but the effect varies by species.

[0013] As far as is known in female dogs, deslorelin is an effective long-term contraceptive but causes estrus as an undesirable effect in the short term. Progesterone analysis was performed after the contraceptive effect, and luteal phase concentrations were above 24 ng / ml (Brandli Sp et al., Long-term effect of repeated deslorelin acetate treatment in bitches for reproduction control., Theriogenology. 2021 Oct 1, 173, pages 73-82).

[0014] To solve the problem of estrus induction, a test was conducted to implant this implant in prepubertal female dogs. The efficacy of the implant was measured by measuring 17 beta-estradiol (E2) and progesterone (P4) using a fluorescence-based enzyme assay. This implant reduced these hormones (above the basal level of 10 ng / ml), P4 was between 0.3 and 1.1 ng / ml (0.6 ± 0.2 ng / ml), and E2 varied between 8 and 138 pg / ml. Thus, deslorelin is known to be an effective early contraceptive in prepubertal female dogs, delaying puberty by at least 4.5 months, and its effect can be maintained by repeated treatment (G Marino et al., deslorelin implants in pre-pubertal female dogs: short- and long-term effects on the genital tract. Reprod. Domest. Anim., 2014), 49(2), pages 297-301).

[0015] It is also known to have a contraceptive effect on cats, and ovarian activity was evaluated by monitoring the E2 (estradiol) concentration in feces. In all cats, estrus behavior was suppressed for approximately 668 days (23 months) and estradiol secretion was reduced by the implant (average 128.48 ng / g; control: 283.26 ng / g) (T S F Toydemir et al., Effects of the GnRH analogue deslorelin implants on reproduction in female domestic cats. Theriogenology. 2012 Feb, 77(3), pages 662-674; and the above Fontaine C. 2015).

[0016] In ferrets, a 4.7 mg deslorelin implant also showed a contraceptive effect, and the side effect was always the same estrus. Therefore, ferrets showed very strong signs of estrus with vulvar swelling within 4 days of implant insertion. Two weeks after insertion, estrus stopped naturally. However, fecal progesterone concentrations in these animals remained low throughout the 8.5-month sampling period, indicating that they did not ovulate. The deslorelin implant had an inhibitory effect on ovarian function for 698 (±122) days (A Prohaczik et al., Comparison of four treatments to suppress ovarian activity in ferrets (Mustela putorius furo)., Vet.Rec., 2010 Jan 16, 166(3), pages 74 - 78).

[0017] In female tigers, which are large mammals, this implant is also being tested. In fact, this implant can reversibly regulate the reproduction of female tigers by implantation in some cases of some implants. The average time to conception was 50.7 months (with a 4.7 mg implant) and 51.9 months with a 9.4 mg implant (A Guthrie et al., The past, present and future of hormonal contraceptive use in managed captive female tiger populations with a focus on the current use of deslorelin acetate., Zoo Biol., 2021, 40(4) pages 306 - 319).

[0018] Theoretical basis: To date, the known data have not shown a PK / PD (pharmacokinetic / pharmacodynamic) relationship, and the efficacy of the product has been explained by a very large initial burst immediately after implantation and a very long-lasting GnRH receptor saturation. Current data in dogs indicate the presence of deslorelin for 2.5 months, but its effect is observed for at least 6 months (source: EMA website: https: / / www.ema.europa.eu / en / documents / product-information / suprelorin-epar-product-information_en.pdf).

[0019] Currently, new data are being generated, indicating that deslorelin can be measured if its effect is observed in male and female dogs and cats.

[0020] Consequently, it is now demonstrated that in previously non-chemically castrated non-human animals, a "burst" (hereinafter also referred to as "flare-up") effect is required to induce castration after a strong fertility period. It is also demonstrated that it is necessary to maintain a minimum concentration of deslorelin (higher than a specific threshold) for a long period to maintain the chemical castration effect. Contrary to the prior art and common understanding regarding the mode of action of deslorelin, the PK / PD relationship between the in vivo levels of deslorelin and its PD effect has been further clearly demonstrated, enabling us to specify the efficacy threshold.

[0021] As a result, in non-human animals that have already been chemically castrated, the "burst" (hereinafter also referred to as "flare-up") effect is ultimately not necessary, but it has now been demonstrated that it is necessary to maintain a long-term concentration of deslorelin above a certain threshold. In addition to this, it has been shown that maintaining the concentration range of deslorelin in the bloodstream is necessary to obtain and maintain the effects of castration. Contrary to the prior art and common knowledge regarding the mode of action of deslorelin, the PK / PD relationship between the in vivo level and the PD effect of deslorelin has been further clearly demonstrated, allowing us to identify the efficacy threshold.

Prior Art Documents

Non-Patent Documents

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Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Summary of the Invention

Means for Solving the Problems

[0023] The present invention relates to deslorelin in a pharmaceutical or non-pharmaceutical product for use in chemical castration of non-human mammals within 1 month after administration, which, when administered, results in a Cmax of deslorelin (preferably about 1 to 2 hours after administration) exceeding 7000 pg / mL and a deslorelin concentration in the plasma of the non-human mammal of at least 10 pg / mL. In a preferred embodiment, deslorelin is administered subcutaneously or intramuscularly to a non-human mammal in need of deslorelin in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / ml after 8 days of administration.

[0024] In a specific embodiment of use, the deslorelin plasma concentration is from 10 pg / mL to 400 pg / ml from 8 days to at least 6 months after administration.

[0025] In another embodiment, deslorelin is used to induce temporary infertility in non-castrated male animals for several months, preferably at least 6 months, and is administered subcutaneously or intramuscularly to a mammal in need thereof in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / ml after 8 days of administration. In a preferred embodiment, the deslorelin plasma concentration is less than 400 pg / ml from 8 days to 6 months after administration. Preferably, after administration, deslorelin is released substantially continuously at a plasma concentration level of less than 400 pg / mL from 8 days to at least 6 months after administration.

[0026] In another embodiment, the present invention relates to a sustained-release drug delivery system (DDS) containing deslorelin for use as described above and administered to non-human mammals via a subcutaneous or intramuscular route. The sustained-release DDS releases more than 15%, preferably more than 20%, preferably more than 25%, preferably more than 30%, preferably more than 35%, preferably more than 40%, preferably more than 45%, more preferably more than 50% of the deslorelin contained in the DDS within 2 days, and the remaining effective dose of deslorelin is released over 3 to 6 months or more. In a preferred embodiment, the DDS is a subcutaneous or intramuscular implant containing deslorelin and a biodegradable polymer for use in chemical castration of non-human mammals. The implant releases deslorelin with a Cmax of more than 7000 pg / mL (preferably about 1 or 2 hours after administration), and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL and is maintained for at least 6 months. In a preferred embodiment, deslorelin is administered to non-human mammals in need thereof in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / ml from 8 days after administration, preferably from 8 days after administration to at least 6 months after administration.

[0027] In another embodiment, the present invention relates to a subcutaneous or intramuscular implant containing deslorelin and a biodegradable polymer for inducing temporary infertility in non-castrated male animals for at least 6 months. It involves administering deslorelin subcutaneously or intramuscularly to mammals in need thereof in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / ml from 8 days after administration.

[0028] In another embodiment, the present invention relates to deslorelin for use in chemical castration of non-human mammals, which, when administered, reduces testosterone levels in the plasma of the non-human mammals to less than 0.4 ng / ml, and the deslorelin concentration in the plasma of the non-human mammals is at least 10 pg / mL, preferably more than 15 pg / mL.

[0029] According to another embodiment, the present invention provides a method for chemically castrating a non-human animal, (a) administering an effective amount of deslorelin such that a plasma deslorelin having a Cmax concentration of from 7000 pg / mL to 40000 pg / mL, or an equivalent amount of a pharmaceutically acceptable salt thereof, is obtained within the first 24 hours after injection; and then (b) administering a maintenance dose such that the plasma deslorelin, or an equivalent amount of a pharmaceutically acceptable salt thereof, has an average concentration of more than 10 pg / ml, comprising.

[0030] Preferably, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. In a specific embodiment, in step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). In a preferred embodiment, the sustained-release drug delivery system (DDS) of step (a) is a subcutaneous or intramuscular implant. Preferably, the implant is a biodegradable polymer.

[0031] Another embodiment of the invention relates to a method for inducing temporary infertility in healthy, non-castrated and sexually mature male animals, preferably dogs, (a) administering an effective amount of deslorelin such that a plasma deslorelin having a Cmax concentration of from 7000 pg / mL to 40000 pg / ml, or an equivalent amount of a pharmaceutically acceptable salt thereof, is obtained within the first 24 hours after injection; and then (b) administering a maintenance dose such that the plasma deslorelin, or an equivalent amount of a pharmaceutically acceptable salt thereof, has an average concentration of more than 10 pg / ml, comprising.

[0032] Preferably, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. In a specific embodiment, in step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). In a preferred embodiment, the sustained-release drug delivery system (DDS) of step (a) is a subcutaneous or intramuscular implant. Preferably, the implant is a biodegradable polymer.

[0033] Another embodiment of the invention relates to a method of inducing temporary infertility in order to delay the first signs of estrus and to prevent prepubertal pregnancy in healthy, non-castrated, sexually immature female animals (preferably dogs). (a) administering an effective amount of deslorelin such that a plasma deslorelin Cmax concentration of from 7000 pg / mL to 40000 pg / ml, or an equivalent amount of a pharmaceutically acceptable salt thereof, is obtained within the first 24 hours after injection; and then (b) administering a maintenance dose such that the average concentration is greater than 10 pg / ml of plasma deslorelin, or an equivalent amount of a pharmaceutically acceptable salt thereof, is included.

[0034] Preferably, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. In a specific embodiment, in step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). In a preferred embodiment, the sustained-release drug delivery system (DDS) of step (a) is a subcutaneous or intramuscular implant. Preferably, the implant is a biodegradable polymer.

[0035] Another embodiment of the invention relates to a method of inducing temporary infertility in non-castrated male animals (preferably cats), and suppressing sexual behaviors such as urine odor, sexual impulse, meowing, urine marking, and aggression. (a) administering an effective amount of deslorelin such that a Cmax concentration of plasma deslorelin of from 7000 pg / mL to 40000 pg / ml, or an equivalent amount of a pharmaceutically acceptable salt thereof, is obtained within the first 24 hours after injection; and thereafter (b) administering a maintenance dose such that the average concentration is greater than 10 pg / ml of plasma deslorelin, or an equivalent amount of a pharmaceutically acceptable salt thereof, comprising.

[0036] Preferably, in step (a) and / or step (b), a subcutaneous or intramuscular sustained release drug delivery system (DDS) is injected. In a specific embodiment, in step (b), a subcutaneous or intramuscular sustained release drug delivery system (DDS) is injected simultaneously with the implant of step (a). In a preferred embodiment, the sustained release drug delivery system (DDS) of step (a) is a subcutaneous or intramuscular implant. Preferably, the implant is a biodegradable polymer.

[0037] According to yet another embodiment, the present invention provides a method for diagnosing behavioral disorders (aggression) in non-human mammals treated with a deslorelin sustained release implant, a) collecting a blood sample from the non-human mammal treated with deslorelin; b) measuring the testosterone plasma level in the blood sample; c) determining the testosterone level in the blood plasma, wherein the testosterone plasma level in the blood sample is greater than 0.2 ng / mL, preferably 0.4 ng / mL; d) finally, determining the dosage of deslorelin to be further administered to the non-human mammal to resolve the behavioral disorder.

[0038] In another embodiment, the present invention relates to the use of deslorelin to induce temporary infertility in non-castrated male animals over at least 6 months, comprising administering deslorelin subcutaneously or intramuscularly to a mammalian subject in need thereof at a dosage effective to induce a deslorelin plasma concentration, wherein upon administration, the Cmax of deslorelin 1 hour after administration exceeds 7000 pg / mL and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL.

[0039] Another embodiment of the invention relates to a method of contraception in non-human mammals, comprising administering subcutaneously or intramuscularly to a non-human mammal an implant comprising deslorelin and a biodegradable polymer, wherein the implant releases deslorelin with a Cmax of deslorelin exceeding 7000 pg / mL 1 hour after administration, the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL, and is maintained for at least 6 months. According to this embodiment, deslorelin is administered subcutaneously or intramuscularly to a non-human animal in need thereof at a dosage effective to induce a deslorelin plasma concentration of from 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration, and a plasma concentration of less than 400 pg / mL from 8 days after administration. Preferably, the deslorelin plasma concentration is less than 400 pg / mL from 8 days after administration to at least 6 months after administration.

[0040] According to another embodiment, the present invention provides a maintenance therapy for inhibiting or reducing the fertility of an animal, (a) first, administering a therapeutically effective amount of a deslorelin product to an animal in need thereof to induce a deslorelin plasma concentration of from 4000 pg / mL to 40000 pg / mL for a first predetermined period; (b) then, administering to the animal in need thereof a system that continuously releases a therapeutically effective amount of a deslorelin dosage for a second predetermined period to obtain a deslorelin plasma concentration of at least 10 pg / ml.

[0041] In another embodiment, the present invention relates to deslorelin for use in preventing behavioral disorders (including aggression) in non-human mammals, wherein deslorelin is administered subcutaneously or intramuscularly using a sustained-release implant, and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL (for at least 6 months).

[0042] Another embodiment of the present invention relates to deslorelin for use in preventing disorders of FSH / LH levels in non-human mammals, wherein deslorelin is administered subcutaneously or intramuscularly using a sustained-release implant, and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL (for at least 6 months).

[0043] Finally, generally the present invention - inducing chemical castration of non-human mammals, or - inducing temporary infertility in non-castrated male animals, preferably dogs, for several months, or - inducing temporary infertility to delay the first signs of estrus and prevent pregnancy in young, healthy and sexually immature female animals (preferably dogs) that are not castrated, or - inducing temporary infertility, and suppressing the odor of urine, and suppressing sexual behaviors such as sexual impulse, whining, urine marking, aggression, etc. in non-castrated male animals (preferably cats), or - contraception in non-human mammals, or inhibiting or reducing fertility, or - preventing behavioral disorders (including aggression) in non-human mammals, or - preventing disorders of FSH / LH levels in non-human mammals, also relates to the use of deslorelin or a sustained-release drug delivery system containing deslorelin for manufacturing a medicament therefor.

[0044] According to the present invention, all embodiments described herein regarding various uses of deslorelin, as well as various methods and compositions related to the use of deslorelin, are also applicable to the use of deslorelin for manufacturing the above medicament.

[0045] In addition to the above typical aspects and embodiments, further aspects and embodiments of the present invention will become apparent by referring to the drawings and studying the following description.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0047] Deslorelin is used as deslorelin acetate in a commercially available implant named Suprelorin® 4.7 mg or 9.4 mg implant for dogs. It is prescribed to induce temporary infertility in healthy, non-castrated, sexually mature male dogs. Deslorelin is a GnRH agonist and acts by suppressing the function of the pituitary-gonadal axis when administered continuously at a low dose. This suppression causes a disorder in the synthesis and / or release of follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which are hormones involved in maintaining fertility, in the treated animals.

[0048] When deslorelin is administered continuously at a low dose, male genital function, sexual drive, and spermatogenesis decrease, and plasma testosterone levels become low, starting from 4 to 6 weeks after implantation. A short-term transient increase in plasma testosterone may be observed immediately after implantation. Measurement of testosterone plasma concentration has demonstrated a sustained pharmacological effect due to the continuous presence of deslorelin in the blood circulation for at least 12 months after administration of the veterinary drug.

[0049] As used herein, examples of "behavioral disorders in non-human mammals" include, but are not limited to, urine odor, sexual drive, vocalization, urine marking, and aggression in non-castrated male animals (preferably cats).

[0050] The term "chemical castration" as used herein refers to the use of a chemical substance or drug that stops sex hormone production in order to reduce or suppress reproductive and sexual activities. Chemical castration is sometimes referred to as medical castration or hormone therapy. Chemical castration is reversible, i.e., when the use of the chemical substance or drug is stopped, hormone production resumes. Unlike surgical castration, which removes the gonads by laparotomy, chemical castration does not remove organs and is not a form of contraceptive surgery.

[0051] The term "sexual activity" as used herein refers to and encompasses sexual behaviors such as sexual drive, vocalization, urine marking or scent-marking behavior, and aggression, particularly in non-castrated male or female animals. Courtship, mating, and scent-marking behavior are also examples of sexual activity in non-castrated male or female animals.

[0052] The term "Cmax" of deslorelin as used herein means the maximum plasma concentration of deslorelin after administration.

[0053] The term "healthy animal" as used herein means an animal that does not have a contraindication to temporary infertility induction, surgical castration, or chemical castration.

[0054] As used herein, the term "non-castrated animal" refers to an animal that has not been surgically castrated and is not currently undergoing a temporary infertility treatment.

[0055] As used herein, the term "non-human animal" can refer to any living organism in the animal kingdom, excluding humans (Homo sapiens). Examples of "animals" as the term is used herein include, but are not limited to, companion animals such as dogs, cats, horses; and livestock animals such as cows, goats, sheep, pigs.

[0056] As used herein, the term "pharmaceutically acceptable salt thereof" in relation to deslorelin means a pharmaceutically acceptable salt of deslorelin. An example of such a pharmaceutically acceptable salt is deslorelin acetate.

[0057] As used herein, the term "sexually mature female animal" means a female animal with seasonal estrus signs.

[0058] In contrast, a "sexually immature female animal" is a female animal that has not yet reached the estrus period and shows no signs of estrus.

[0059] As used herein, the term "<temporary>" when referring to infertility means that the infertility is for a certain period, i.e., the infertility is reversible.

[0060] As used herein, the term "temporary infertility" refers to the inability to become pregnant in relation to a chemical substance that can induce this. Some chemotherapy treatments, although undergoing this treatment, may cause infertility. However, fertility may return after the treatment ends. This varies not only depending on the drug but also on other factors such as age and whether the patient is male or female. In the context of the present invention, "temporary infertility" may thus be made to coincide with contraception or the result of a contraceptive method.

[0061] The term "Tmax", as used herein, means the time at which the maximum plasma concentration (Cmax) of deslorelin is achieved or observed after deslorelin administration.

[0062] Pharmaceutical composition One aspect of the present disclosure provides a composition for use in inducing chemical castration in a non-human animal in need thereof, which comprises a therapeutically effective amount of deslorelin and an excipient as set forth in the claims.

[0063] In a preferred embodiment of the present invention, deslorelin is formulated into a solid implant formulation.

[0064] In a preferred embodiment, the implant formulation comprises stearin (also known as hydrogenated palm oil) and lecithin.

[0065] According to this particular embodiment of the present invention, the solid implant formulation preferably comprises about 2-12% (w / w) deslorelin (active basis), about 0.5-2.5% (w / w) lecithin and about 85-97.5% (w / w) stearin.

[0066] More preferably, the solid implant formulation comprises about 4-10% (w / w) deslorelin (active basis), about 0.5-1.5% (w / w) lecithin and about 85-94% (w / w) stearin.

[0067] Particularly preferred solid implant formulations are selected from the group comprising formulations containing 85-89% (w / w) stearin, 9-10% (w / w) deslorelin (active basis) and 0.8-1.2% (w / w) lecithin.

[0068] In a further preferred embodiment, the solid implant formulation may further comprise anhydrous sodium acetate.

[0069] Stearin is partially hydrogenated palm oil. Its main fatty acids are C16:0 (45%) and C18:0 (53%). The melting point is about 55°C.

[0070] Lecithin is phosphatidylcholine. It is a mixture in which diglycerides composed of stearic acid, palmitic acid and oleic acid are bonded to choline esters of phosphoric acid.

[0071] In another preferred embodiment, deslorelin is prepared as a liquid composition.

[0072] Deslorelin can be administered by any suitable means, for example, preferably by biodegradable implants over time, by sustained release drug delivery systems (DDS), or by a biocompatible polymer-solvent system containing a biodegradable polymer compatible with subcutaneous, transdermal or intramuscular administration.

[0073] According to the present invention, "DDS" refers to any system capable of delivering a drug to a living body. Preferred DDS according to the present invention include any type of implant. In particular, this term encompasses liquid and solid implants, which have at least a polymer or non-polymer material.

[0074] According to the present invention, "sustained release" when referring to DDS means that deslorelin is not released as a single administration of the total dose of deslorelin. Rather, a sustained release DDS can deliver deslorelin over a period of time, as disclosed in the present application.

[0075] In some embodiments, the implant is preferably biodegradable over time. It is selected from the group consisting of biodegradable materials and bioerodible dosage forms.

[0076] As used herein, the term "biocompatible" can mean "harmless to living tissues". This term encompasses both biodegradable and bioerodible.

[0077] As used herein, "degradable" means that the polymer degrades or regresses in vivo to form smaller chemical species, and the degradation can be due to, for example, enzymes, chemical ( "biodegradable" polymers) and / or physical processes ( "in vivo degradable" polymers).

[0078] As used herein, the term "biodegradable" can refer to any water-insoluble material that can be converted into one or more water-soluble materials under physiological conditions, regardless of any specific degradation mechanism or process.

[0079] In some embodiments, the biodegradable material is a biodegradable polymer (or biopolymer), preferably polylactic acid, polyglycolic acid, polylactide, polyglycolide, polycaprolactone, polyanhydride, polyamide, polyurethane, polyesteramide, polyorthoester, polydioxanone, polyacetal, polyketal, polycarbonate, polyphosphazene, polyhydroxybutyric acid, polyhydroxyvaleric acid, polyalkylene oxalate, polyalkylene succinate, poly(malic acid), polyethylene glycol, hyaluronic acid, chitin and chitosan, copolymers thereof, terpolymers thereof, and any combination thereof.

[0080] In some embodiments, the biodegradable polymer comprises monomers selected from the group consisting of lactide, glycolide, caprolactone, p-dioxanone, trimethylene carbonate, 1,5-dioxepan-2-one, 1,4-dioxepan-2-one, ethylene oxide, propylene oxide, sebacic anhydride, diketene acetal / diol, lactic acid, and combinations thereof.

[0081] As used herein, the term "implant" can also be understood as a biodegradable dosage form, and devices such as microspheres; "liquid polymer depot" formulations (or in-situ forming implants); and solid polymer implants such as pellets or minipellets. This implant contains a biodegradable polymer as deslorelin and an excipient.

[0082] As used herein, the term "bioerodible" can refer to any water-insoluble material that can be mechanically degraded by a biological process that at least partially solubilizes this material.

[0083] Upon degradation, the biodegradable polymer releases deslorelin into body fluids or tissues including blood and lymph. Examples of biodegradable polymers according to the present invention include polyhydroxy acids such as poly(lactide), poly(glycolide), poly(lactide-co-glycolide), poly(lactic acid), poly(glycolic acid) and poly(lactic acid-co-glycolic acid), polyanhydrides, polyorthoesters, polyether esters, polyethylene glycol, poly(ε-caprolactone), polyesteramides, polyphosphazines, polycarbonates, polyamides and their copolymers and mixtures, fats with a melting point above 50 °C, waxes, and long-chain C 12 -C 22 triglyceride esters of fatty acids, and mixtures thereof, preferably glyceryl tristearate. Preferably, the polymer is selected from the group consisting of poly(ε-caprolactone) (PCL), poly(lactide), poly(glycolide) and their copolymers or mixtures such as poly(lactide-co-glycolide) and glyceryl tristearate.

[0084] The injectable implant may further contain an antioxidant. Antioxidants useful for injectable implants are known in the art. One preferred antioxidant is BHT (butylated hydroxytoluene). Such implants, microspheres and in situ forming implants can be manufactured without dissolving deslorelin during the process using methods well known in the art described, for example, in European Patent Application Publication No. 3349726, International Publication No. 90 / 03768, International Publication No. 2006 / 063794, International Publication No. 2009 / 091737, International Publication No. 98 / 07423, European Patent Application Publication No. 1197207, European Patent Application Publication No. 0525307, International Publication No. 98 / 08533, International Publication No. 2020 / 222399, International Publication No. 2020 / 130585 and International Publication No. 2013 / 082373. In a preferred embodiment, the present invention relates to an implant as defined above, comprising deslorelin, poly(ε-caprolactone) (PCL), poly(lactide), poly(glycolide), poly(lactide-co-glycolide), copolymers or mixtures thereof, and a biodegradable polymer selected from the group consisting of glyceryl tristearate, and optionally an antioxidant.

[0085] In some embodiments, deslorelin is present in a liquid form composition at an effective dose for more than 1 month, more preferably more than 2 months, and most preferably more than 3 months. In a preferred embodiment, deslorelin is present in the composition at an effective dose for more than 4 months, more preferably more than 5 months, and most preferably more than 6 months.

[0086] The term "liquid" as used herein can refer to the ability of a composition to deform under shear stress, regardless of the presence of a non-aqueous solvent.

[0087] The liquid compositions according to the present disclosure are in a liquid state at ambient temperature and body temperature and remain liquid in vivo, i.e., in an aqueous environment.

[0088] The deslorelin composition is prepared to be released continuously, preferentially uniformly, by a sustained delivery system. For this purpose, the viscosity of the deslorelin liquid composition at ambient temperature (i.e., about 20 °C) is greater than about 0.5 Pa·s, greater than about 1.0 Pa·s, greater than about 2.0 Pa·s, greater than about 3.0 Pa·s, greater than about 4.0 Pa·s, greater than about 5.0 Pa·s, greater than about 6.0 Pa·s, greater than about 7.0 Pa·s, greater than about 8.0 Pa·s, greater than about 9.0 Pa·s, greater than about 10.0 Pa·s, greater than about 11.0 Pa·s, greater than about 12.0 Pa·s, greater than about 13.0 Pa·s, greater than about 14.0 Pa·s, greater than about 15.0 Pa·s, greater than about 16.0 Pa·s, greater than about 17.0 Pa·s, greater than about 18.0 Pa·s, greater than about 19.0 Pa·s, or greater than about 20.0 Pa·s. In some further embodiments, the viscosity of the substantially homogeneous composition is greater than about 30.0 Pa·s, greater than about 40.0 Pa·s, greater than about 50.0 Pa·s, greater than about 60.0 Pa·s, greater than about 70.0 Pa·s, greater than about 80.0 Pa·s, greater than about 90.0 Pa·s, greater than about 100.0 Pa·s, greater than about 110.0 Pa·s, greater than about 120.0 Pa·s, greater than about 130.0 Pa·s, greater than about 140.0 Pa·s, greater than about 150.0 Pa·s, greater than about 160.0 Pa·s, greater than about 170.0 Pa·s, greater than about 180.0 Pa·s, greater than about 190.0 Pa·s, or greater than about 200.0 Pa·s. Alternatively, the viscosity of the substantially homogeneous composition may be greater than any integer from about 0.50 to about 200.0 Pa·s.

[0089] In a configuration compatible with the deslorelin liquid composition, the viscosity can be measured using a suitable viscometer. For example, without limitation, a viscometer such as a Brookfield viscometer or an Anton Paar Rheoplus viscometer having a suitable configuration can be used to measure the viscosity. When the viscosity value is referred to / defined in this document, it relates to the viscosity measured using a Brookfield viscometer at 20 °C.

[0090] The various excipients commonly used in formulations, preferably pharmaceutical formulations, can be selected based on several criteria such as, for example, the desired dosage form and the release characteristics of the dosage form. Non-limiting examples of suitable excipients include substances selected from the group consisting of binders, fillers, preservatives, diluents, lubricants, dispersants, pH adjusters, stabilizers, and combinations of any of these substances.

[0091] Method of using a deslorelin composition Another aspect of the present disclosure includes a method of inducing chemical castration in a non-human animal in need thereof. The method includes administering to the non-human animal a composition comprising an effective amount of deslorelin.

[0092] In one embodiment, the effective amount of deslorelin is the dosages of 4.7 mg and 9.4 mg of Suprelorin® implant.

[0093] A first object of the present invention is deslorelin for use in chemical castration of non-human mammals within one month after administration, which when administered, the Cmax of deslorelin after administration exceeds 7000 pg / mL, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL.

[0094] Administration of deslorelin to an animal achieves a Cmax of deslorelin from about 7000 pg / mL to about 200000 pg / mL, for example, a Cmax of deslorelin from about 7500 pg / mL to about 170000 pg / mL, at about 0.4 to about 3.4 hours (Tmax), depending on the animal species. In a specific example, the Cmax of deslorelin 1 hour after administration exceeds 7429 pg / mL.

[0095] In other embodiments, the Cmax of deslorelin can be achieved when the Tmax is from about 0.7 to about 2 hours, for example, the Tmax can also be from about 0.5 to about 1.7 hours. In some embodiments, since the Cmax can vary in various non-human animals, the Cmax can be even higher. For example, the Cmax can be about 7200 pg / mL, about 7400 pg / mL, about 8000 pg / mL, about 9000 pg / mL, about 10000 pg / mL, about 11000 pg / mL, about 12000 pg / mL, about 13000 pg / mL, about 15000 pg / mL, about 20000 pg / mL, about 25000 pg / mL, about 30000 pg / mL, about 35000 pg / mL, about 40000 pg / mL, about 45000 pg / mL, about 50000 pg / mL, about 55000 pg / mL, about 60000 pg / mL, about 65000 pg / mL, about 70000 pg / mL, about 75000 pg / mL, about 80000 pg / mL, about 85000 pg / mL, about 90000 pg / mL, about 95000 pg / mL, or about 100000 pg / mL, or about 110000 pg / mL, or about 120000 pg / mL, or about 130000 pg / mL, or about 140000 pg / mL or about 150000 pg / mL or about 160000 pg / mL, or about 170000 pg / mL. In typical embodiments, the Cmax of deslorelin can be from 375 ng / ml to 10000 ng / ml. In some embodiments, the Cmax of deslorelin can be greater than 500 pg / mL. In other embodiments, the Cmax of deslorelin can be less than 300000 pg / mL.

[0096] In other embodiments, Tmax appears in individual non-human animals at 30-minute, 1-hour or 2-hour intervals and reaches Tmax in about 30 minutes to 2 hours. Tmax can be about 0.4 hours, about 0.5 hours, about 0.6 hours, about 0.7 hours, about 0.8 hours, about 0.9 hours, 1.0 hour, about 1.1 hours, about 1.2 hours, about 1.3 hours, about 1.4 hours, about 1.5 hours, about 1.6 hours, about 1.7 hours, about 1.8 hours, about 1.9 hours, about 2.0 hours, about 2.2 hours, about 2.4 hours, about 2.6 hours, about 2.8 hours, about 3.0 hours, about 3.2 hours or about 3.4 hours. In one embodiment, Tmax can be from 0.4 to 4 hours. In other typical embodiments, Tmax can be from 0.5 to 1.0 hour. In some embodiments, Tmax can be greater than 0.4 hour. In other embodiments, Tmax can be less than 3.4 hours. In typical embodiments, Tmax can be from 0.03 days to 1.33 days.

[0097] The area under the curve (AUC) can be from about 10000 day·pg / mL to about 110000 day·pg / mL. In typical embodiments, AUC can be from about 11000 day·pg / mL to about 15000 day·pg / mL, from about 15000 day·pg / mL to about 20000 day·pg / mL, from about 20000 day·pg / mL to about 25000 day·pg / mL, from about 25000 day·pg / mL to about 30000 day·pg / mL, from about 30000 day·pg / mL to about 35000 day·pg / mL, from about 35000 day·pg / mL to about 40000 day·pg / mL, from about 40000 day·pg / mL to about 45000 day·pg / mL, from about 45000 day·pg / mL to about 50000 day·pg / mL, from about 50000 day·pg / mL to about 55000 day·pg / mL, from about 55000 day·pg / mL to about 60000 day·pg / mL.

[0098] According to a first object of the present invention, deslorelin is administered subcutaneously or intramuscularly to a non-human mammal in need of deslorelin in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL, preferably 7000 pg / mL to 40000 pg / mL, between 1 hour and 24 hours after administration, and a plasma concentration of less than 400 pg / ml after 8 days of administration. Preferably, the deslorelin plasma concentration is 10 pg / mL to 400 pg / ml from 8 days after administration until at least 6 months after administration.

[0099] According to a specific embodiment of the first object of the present invention, deslorelin is used to induce temporary infertility.

[0100] Therefore, the present invention also relates to deslorelin for use in inducing temporary infertility in non-castrated male animals over at least 6 months, in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL, preferably 7000 pg / mL to 40000 pg / mL, between 1 hour and 24 hours after administration, and a plasma concentration of less than 400 pg / ml after 8 days of administration, and administering deslorelin subcutaneously or intramuscularly to a mammal in need thereof. Preferably, the deslorelin plasma concentration is less than 400 pg / ml from 8 days after administration until 6 months after administration. According to this specific embodiment, after administration, deslorelin is substantially continuously released at a plasma concentration level of less than 400 pg / mL from 8 days after administration until at least 6 months after administration.

[0101] The present invention also relates to deslorelin for use in inducing temporary infertility to delay the first signs of estrus and prevent young, healthy, non-castrated female animals from becoming pregnant at a young age. (a) Administering an effective amount of deslorelin such that a Cmax concentration of deslorelin of 4000 pg / mL to 40000 pg / ml in plasma, preferably 7000 pg / mL to 40000 pg / ml in plasma within the first 24 hours after administration, or an equivalent amount of its pharmaceutically acceptable salt is obtained; and then (b) Administering a maintenance dose of deslorelin such that the mean concentration of deslorelin or an equivalent amount of its pharmaceutically acceptable salt is greater than 10 pg / ml in plasma, comprising.

[0102] Preferably, when deslorelin is used to induce chemical castration or temporary infertility according to the first object of the present invention, deslorelin is prepared in the form of a sustained-release drug delivery system (DDS).

[0103] Accordingly, a second object of the present invention is a sustained-release drug delivery system (DDS) containing deslorelin for use in chemical castration of non-human mammals according to the first object of the present invention, wherein the DDS releases deslorelin with a Cmax of deslorelin exceeding 7000 pg / mL after administration, the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL, and is maintained for at least 6 months. Preferably, the DDS is a subcutaneous or intramuscular implant containing deslorelin and a biodegradable material such as a biodegradable polymer.

[0104] According to a specific embodiment of this second object, upon administration, deslorelin reduces the testosterone level in the plasma of the non-human mammal to less than 0.4 ng / ml, and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL, preferably more than 15 pg / mL.

[0105] A third object of the present invention is a sustained-release drug delivery system (DDS) containing deslorelin for use in inducing temporary infertility in intact male animals for at least 6 months, having a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL, preferably 7000 pg / mL to 40000 pg / mL in plasma within the first 24 hours after injection, and less than 400 pg / mL in plasma 8 days after administration, from 1 hour to 24 hours after administration, and administering deslorelin subcutaneously or intramuscularly to mammals in need thereof in an effective dose.

[0106] A fourth object of the present invention is a method for chemically castrating non-human mammals, (a) administering an effective amount of deslorelin such that the Cmax concentration of deslorelin in plasma is at least 7000 pg / mL, preferably 7000 pg / mL to 40000 pg / mL in plasma, or an equivalent amount of its pharmaceutically acceptable salt, within the first 24 hours after administration; and then (b) administering a maintenance dose such that the average concentration of deslorelin or an equivalent amount of its pharmaceutically acceptable salt is greater than 10 pg / ml in plasma.

[0107] According to a particular embodiment of this fourth object of the present invention, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. The sustained-release DDS of step (a) is preferably a subcutaneous or intramuscular implant. According to a preferred method, in step (b), the subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). The implant preferably comprises a biodegradable material such as a biodegradable polymer.

[0108] A fifth object of the present invention is a method for inducing temporary infertility in healthy, non-castrated, sexually mature male animals, preferably dogs, (a) Administering an effective amount of deslorelin such that a Cmax concentration of deslorelin of 4000 pg / mL to 40000 pg / ml, preferably 7000 pg / mL to 40000 pg / ml in plasma, or an equivalent amount of its pharmaceutically acceptable salt, is obtained within the first 24 hours after administration; and then (b) Administering a maintenance dose of deslorelin such that the average concentration is deslorelin of more than 10 pg / ml in plasma, or an equivalent amount of its pharmaceutically acceptable salt, and comprising.

[0109] According to a specific embodiment of the fifth object of the present invention, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. In that case, the DDS preferably releases deslorelin with a Cmax of deslorelin exceeding 7000 pg / mL 1 hour after administration, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL and is maintained for at least 6 months. More preferably, deslorelin is administered in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL in the plasma from 1 hour to 24 hours after administration, and a plasma concentration of less than 400 pg / ml from 8 days after administration, preferably at least 6 months later. According to a preferred method, in step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). The implant preferably comprises a biodegradable material such as a biodegradable polymer.

[0110] The present invention also contemplates the use of deslorelin to induce temporary infertility in non-castrated male animals for at least 6 months, which, when administered, has a Cmax of deslorelin exceeding 7000 pg / mL 1 hour after administration and induces a deslorelin plasma concentration of at least 10 pg / mL in the plasma of non-human mammals, and administering deslorelin subcutaneously or intramuscularly to a mammal in need thereof in an effective dose, which is.

[0111] A sixth object of the present invention is a method of inducing temporary infertility in order to delay the first signs of estrus and to prevent young, non-castrated, healthy, sexually immature female animals (preferably dogs) from becoming pregnant, (a) administering an effective amount of deslorelin such that a Cmax concentration of deslorelin or an equivalent amount of its pharmaceutically acceptable salt is obtained in plasma of from 4000 pg / mL to 40000 pg / mL, preferably from 7000 pg / mL to 40000 pg / mL in plasma within the first 24 hours after injection; and then (b) administering a maintenance dose of deslorelin such that the average concentration of deslorelin or an equivalent amount of its pharmaceutically acceptable salt is greater than 10 pg / ml in plasma.

[0112] According to a particular embodiment of the sixth object of the present invention, in step (a) and / or step (b), a subcutaneous or intramuscular sustained release drug delivery system (DDS) is injected. In that case, the DDS should preferably be administered at 12 to 16 weeks of age. The DDS contains a biodegradable material such as deslorelin and a biodegradable polymer, and the DDS preferably releases deslorelin with a Cmax of deslorelin exceeding 7000 pg / mL 1 hour after administration, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL and is maintained for at least 6 months. Further, even more preferably, deslorelin is administered in an effective dose to induce a deslorelin plasma concentration of from 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration, and a plasma concentration of 400 pg / ml from 8 days after administration, preferably at least 6 months later. According to a preferred method, in step (b), a subcutaneous or intramuscular sustained release drug delivery system (DDS) is injected simultaneously with the implant of step (a). The implant preferably contains a biodegradable material such as a biodegradable polymer.

[0113] The seventh object of the present invention is a method for inducing temporary infertility and suppressing the smell of urine in non-castrated male animals (preferably cats), and suppressing sexual behaviors such as sexual impulse, meowing, urine marking, and aggressiveness. (a) administering an effective amount of deslorelin such that a Cmax concentration in plasma is from 4000 pg / mL to 40000 pg / ml within the first 24 hours after injection, or an equivalent amount of a pharmaceutically acceptable salt thereof, preferably deslorelin such that a Cmax concentration in plasma is from 7000 pg / mL to 40000 pg / ml within the first 24 hours after injection, or an equivalent amount of a pharmaceutically acceptable salt thereof; and then (b) administering a maintenance dose such that an average concentration is deslorelin in plasma greater than 10 pg / ml, or an equivalent amount of a pharmaceutically acceptable salt thereof.

[0114] According to this seventh object of the present invention, the non-castrated male animal is preferably a cat, more preferably a 3-month-old cat.

[0115] According to a specific embodiment of this seventh object of the present invention, in step (a) and / or step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected. In that case, the DDS preferably contains a biodegradable material such as deslorelin and a biodegradable polymer, and the DDS releases deslorelin such that the Cmax of deslorelin exceeds 7000 pg / mL 1 hour after administration, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL and is maintained for at least 6 months. More preferably, deslorelin is administered in an effective dose to induce a deslorelin plasma concentration of from 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration, and a plasma concentration of 400 pg / ml from 8 days after administration, preferably at least 6 months later. According to a preferred method, in step (b), a subcutaneous or intramuscular sustained-release drug delivery system (DDS) is injected simultaneously with the implant of step (a). The implant preferably contains a biodegradable material such as a biodegradable polymer.

[0116] An eighth object of the present invention is a method for diagnosing behavioral disorders (aggression) in non-human mammals treated with a deslorelin sustained-release implant, a) measuring the testosterone plasma level in a blood sample of the non-human mammal treated with the deslorelin sustained-release implant; b) determining the testosterone level in the blood plasma, wherein the testosterone plasma level in the blood sample is greater than 0.2 ng / mL, preferably 0.4 ng / mL; c) finally, determining the dosage of deslorelin to be further administered to the non-human mammal to resolve the behavioral disorder.

[0117] A ninth object of the present invention is a method of contraception in non-human mammals, comprising subcutaneously or intramuscularly administering to the non-human mammal an implant containing a biodegradable material such as deslorelin and a biodegradable polymer, wherein the implant releases deslorelin with a Cmax of deslorelin exceeding 7000 pg / mL 1 hour after administration, and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL and is maintained for at least 6 months.

[0118] According to this ninth object, deslorelin is administered to non-human mammals in need of deslorelin in an effective dosage to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL, preferably 7000 pg / mL to 40000 pg / mL, between 1 hour and 24 hours after administration, and a plasma concentration of less than 400 pg / ml after 8 days of administration. Preferably, the plasma concentration of deslorelin is less than 400 pg / ml from 8 days to at least 6 months after administration.

[0119] A tenth object of the present invention is a maintenance therapy for inhibiting or reducing the fertility of animals. (a) First, administer a therapeutically effective amount of a deslorelin product to an animal in need thereof to induce a plasma concentration of deslorelin or an equivalent amount of a pharmaceutically acceptable salt thereof that is from 4000 pg / mL to 40000 pg / mL, preferably from 7000 pg / mL to 40000 pg / mL, during a first predetermined period; (b) Next, administer to an animal in need thereof a system that continuously releases a therapeutically effective amount of a deslorelin dose during a second predetermined period to obtain a plasma concentration of deslorelin or an equivalent amount of a pharmaceutically acceptable salt thereof of at least 10 pg / ml. This includes:

[0120] An eleventh object of the present invention is deslorelin for use in preventing behavioral disorders in non-human mammals. Deslorelin is administered subcutaneously or intramuscularly using a sustained-release drug delivery system, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL (for at least 6 months).

[0121] One embodiment relates to the administration of deslorelin that induces fluctuations in the hormonal pathway. Indeed, new data generated now reveals that deslorelin affects LH concentration. In fact, Example 10 shows fluctuations in LH hormone by the results of the mean (95% CI) LH ratio before and after GnRH stimulation in treated dogs (n = 6) measured over approximately 16 months of the study, as shown in FIG. 6.

[0122] Also, in Example 11, a correlation is shown between the injected deslorelin concentration and the hormone concentrations of LH, FSH, and testosterone in male cats treated with Suprelorin®. This study showed an inverse correlation between deslorelin and testosterone / LH and FSH. In fact, as soon as the deslorelin concentration decreased (to 12 pg / mL), the other concentrations began to increase.

[0123] Accordingly, a twelfth object of the present invention is deslorelin for use in chemical castration of non-human mammals, which, when administered, reduces the testosterone level in the plasma of said non-human mammals to less than 0.4 ng / ml, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL, preferably more than 15 pg / mL.

[0124] A thirteenth object of the present invention is deslorelin for use in preventing follicle-stimulating hormone (FSH) and luteinizing hormone (LH) disorders in non-human animals, which is administered subcutaneously or intramuscularly using a sustained-release implant, and the deslorelin concentration in the plasma of non-human mammals is at least 10 pg / mL (for at least 6 months).

[0125] In fact, it has now been demonstrated that deslorelin affects the hormonal pathway. However, this correlation extends beyond the hormonal region. The hormonal pathway also affects the metabolic region when changing / improving or altering several metabolic pathways such as the following, for example. Sperm concentration: As shown in Example 3 and Figure 2, deslorelin has an impact on sperm concentration. Testicular weight: As shown in Example 6 and Figure 4, deslorelin has an impact on testicular weight. Indeed, it has been shown that the testicular volume decreases over time when there is a deslorelin effect by deslorelin implant injection. Fat and carbohydrate fluctuations: Low levels of testosterone induce weight gain. Improvement / reduction of urine odor: Urine marking is · A pheromone signal to other animals, especially intact male cats · A territorial signal · Signs of stress or sexual excitement related to the social or physical environment the normal and intentional deposition of urine as

[0126] In such situations, the urine odor is intense and it is desirable to remove or mask the odor. Some commercially available enzyme-based urine odor neutralizers can be used on the surface after cleaning the area with a mild detergent and can be used to remove the odor, but these neutralizers require great care. Castration or ovariectomy has been proven especially as a treatment for cats that mark as part of their reproductive activity.

[0127] One reason for surgical castration is to alleviate the strong and persistent urine odor in male animals, especially cats and ferrets. It is known that the urine odor improves several days after surgical castration.

[0128] One goal of the present application is to show that administration of deslorelin (also known as Suprelorin®) reduces the odor of urine and / or feces. Surprisingly, the inventors have shown that the odor reduction is brought about via at least two pathways. Indeed, one such pathway is the sex hormone pathway mediated by testosterone. The inventors have also demonstrated another pathway not related to sex hormones. Thus, according to the present invention, the urine odor in non-castrated male animals administered deslorelin is improved and reduced compared to the urine odor reduction that occurs in surgically castrated male animals, especially cats.

[0129] One aspect of the present invention is to alleviate the urine odor in animals such as domestic animals, especially cats, preferably non-castrated male cats.

[0130] One goal of the present application is to show that administration of deslorelin / Suprelorin® reduces the urine odor by reducing the levels of testosterone / ferinine and its metabolites.

[0131] Testosterone has been known from the prior art (Hendriks WH et al., Testosterone increases urinary free felinine, N-acetylfelinine and methylbutanolglutathione excretion in cats (Felis catus). J Anim Physiol Anim Nutr (Berl). 2008 Feb;92(1):53-62) to increase the secretion of free felinine, N-acetylfelinine and 3-methylbutanolglutathione in castrated adult male cats and intact female cats, while estradiol does not modulate this effect.

[0132] Coxinin is a carboxylesterase secreted as a major urinary component and regulates felinine production. Coxinin secretion is also known to be sex-dependent. In mature cats, coxinin secretion is higher in intact males than in castrated males or intact or ovariectomized females. Coxinin secretion per day decreased immediately after castration. Immunohistochemistry confirmed that coxinin expression in the renal proximal straight tubules is higher in intact males than in castrated males. These results indicate that 1) coxinin secretion is regulated by sex hormones such as testosterone, 2) coxinin functions as an esterase in urine rather than in renal cells, and 3) the degradation products by coxinin are secreted in a species-, sex- and age-dependent manner like coxinin itself.

[0133] Castration immediately reduces corisin secretion, and it has been confirmed that the cause is the decrease in the expression level of corisin in the proximal straight tubule. Plasma testosterone concentrations in intact male cats have been reported to increase with age to 1.5, 6.8, and 12.6 pmol / mL at 6, 12, and 16 months after birth, respectively, and decrease to zero after castration (Miyazaki et al, Chemical Signals in Vertebrates, 1998, 11, pp51 - 60). Thus, it is hypothesized that the transcriptional activity of corisin is regulated by sex hormones such as testosterone.

[0134] Corisin regulates felinine production. In vitro enzyme assays have shown that corisin hydrolyzes 3 - methylbutanol cysteinylglycine, the felinine precursor, into felinine and glycine.

[0135] Therefore, chemical castration induced by deslorelin release affects corisin and felinine production, which also cause the odor of urine. Since felinine and N - acetylfelinine are detected in cat bile and fecal samples, it has been shown that felinine is secreted from the liver through bile into feces (LC - MS / MS quantification of felinine metabolites in tissues, fluids, and excretions from the domestic cat (Felis catus), Futsuta et al, J Chromatogr B Analyt Technol Biomed Life Sci., 2018 Jan 1;1072:94 - 99).

[0136] In addition to the sex hormone pathway mediated by testosterone that causes the odor of urine and / or feces, the inventor has discovered another pathway that is not related to sex-dependent hormones. In fact, as shown in the examples, the inventor has identified new molecules as the molecules that cause the odor of urine and / or feces. These molecules are secreted outside the sex hormone pathway. In fact, those molecules do not originate from the testosterone cascade.

[0137] Accordingly, one aspect of the present invention is the use of deslorelin for increasing or decreasing or down-regulating or up-regulating at least one molecular level selected from the list of crotonic acid; indole; p-cresol; cadaverine and its derivatives such as acetylcadaverine, glutathionylaminopropylcadaverine; putrescine and its derivatives such as N-acetylputrescine, p-coumaroylputrescine; mercapto derivatives such as 2-mercapto-3-butanone, 2-mercaptoethanol, 3-mercaptohexyl butyrate, mercaptopyruvic acid, 3-mercaptopropionic acid, 3-mercapto-3-methylbutan-1-ol, 7-mercaptoheptanoyl threonine, 1alpha,5alpha-dimercaptoandrostane-3alpha,17beta-diol, mercaptopyruvic acid; taurine; isovaline and isobutyine.

[0138] Accordingly, one object of the present invention is to reduce the odor of urine and / or feces by administering deslorelin to a non-human animal.

[0139] In another aspect of the present invention, deslorelin administration affects fertility on the one hand and animal behavior such as sexual and marking behaviors on the other hand. Indeed, as shown in Example 13, deslorelin administration affects penile spines and behavioral recordings as shown in FIGS. 15C and 15D, and for penile spines, the results are statistically significant from 7 weeks to 67 weeks after injection. For sexual behavior, the results were statistically significant at 10, 12, 34, 36, 38, 46, 52, 58, and 66 weeks after injection. Finally, with regard to reproductive behavior, no statistical difference was found between the two groups. Sexual impulses were affected by the individual personality characteristics of the cats. A tendency for a decrease in meowing, aggressiveness, urine marking, and the typical odor of male urine was also observed.

[0140] The present invention provides several advantages including, but not limited to, the following. First, it has an effect on two pathways that synthesize the odor of urine or feces. The results regarding odor reduction are clearly superior to the sole effect based on the synthesis of urine / feces odor by the sex hormone pathway, being at least the combined results of these two pathways.

[0141] It is now shown that in animals chemically castrated by deslorelin administration, molecules responsible for the odor of urine or feces are quantitatively and qualitatively reduced or decreased compared to surgically castrated animals.

[0142] Also, according to the present disclosure, the method of using deslorelin as described herein is a reversible, non-surgical, reliable castration measure, and it has been demonstrated to promote the health of the pet family while leaving options.

[0143] The effect on the testosterone pathway, particularly when reducing its blood level, can contribute to the prevention of kidney diseases in said animals. This is because these types of diseases are related to the accumulation of molecules such as trimethylamine.

[0144] Furthermore, it should also be mentioned that for the safety study of cats, high-dose deslorelin administration has not revealed any serious adverse events or life-threatening events. Also, according to several tolerance studies on male and female species, especially deslorelin administered as an implant (i.e., Suprelorin®) has been concluded to show good tolerance.

[0145] Another object of the present invention is the use of deslorelin to induce temporary infertility in non-castrated female animals, preferably prepubertal animals such as female cats or female dogs, for at least 6 months, including administering deslorelin subcutaneously or intramuscularly to the animals in need. Indeed, as shown in Example 14, deslorelin is effectively administered to prepubertal female dogs, and by using at least one deslorelin implant, the onset of puberty is delayed.

Example

[0146] Example 1: Determination of Deslorelin Plasma Concentration over Time in Cats The intention of this study was to determine the plasma concentration of deslorelin over time in the plasma of non-castrated male cats after a single subcutaneous (SC) administration of a 4.7 mg Suprelorin® implant containing hydrogenated palm oil, lecithin, and anhydrous sodium acetate as excipients.

[0147] 1.1 Materials and Methods: · Study period: D0 - 560 (18 months) · n = 20 adult, sexually mature non-castrated male cats · Blinded, randomized · Suprelorin® group: n = 16 ⇒ 4.7 mg Suprelorin® SC, neck · Control group: n = 4 ⇒ 0.5 ml saline SC, neck

[0148] Blood samples were collected from 16 healthy, non-castrated male cats administered with deslorelin implants. After solid-phase extraction using Oasis® WCX μElution plates, the samples were analyzed using a valid liquid chromatography combined with tandem mass spectrometry (LC-MS / MS) method. The calibration range was 4 pg / mL ± 2000 pg / mL. When the measured concentration was below the lower limit of quantification (LLOQ) of 4 pg / mL, it was reported as BLQ (below the limit of quantification).

[0149] Phoenix® software (WinNonlin 8.0, NLME 1.6, Satara, Farsite, St. Louis, Missouri, USA) was used to perform non-compartmental pharmacokinetic analysis of individual deslorelin concentrations in feline plasma to evaluate the pharmacokinetic parameters of deslorelin.

[0150] 1.2 Results: A series of data showing plasma deslorelin concentrations (pg / ml) measured over time in cats treated with Sprelorelin® are presented in Table 1 below.

[0151] [Table 1]

[0152] From this example, it was found that deslorelin was present in the blood at a very high concentration at D0 and then decreased continuously over time. In some cats, deslorelin was present in the blood for up to 560 days.

[0153] Example 2: Determination of Plasma Testosterone Concentration in Cats The intention of this study was to determine the plasma testosterone concentration to monitor the effectiveness of deslorelin implants.

[0154] 2.1 Materials and Methods: Samples were taken from 16 healthy, non-castrated male cats administered deslorelin implants (Suprelorin® 4.7 mg). Four healthy, non-castrated male cats in the control group were administered saline. Testosterone was analyzed using the previously described established and valid radioimmunoassay (RIA) (Basal testosterone concentrations after the application of a slow-release GnRH agonist implant are associated with a loss of response to buserelin, a short-term GnRH agonist, in the tom cat, Goericke-Pesch et al., Theriogenology. 2013 Jul 1;80(1):65-9). The lower limit of detection was 0.05 ng / mL. The coefficients of variation within and between assays were 3.7 and 7.6%, respectively.

[0155] 2.2 Results: The results are shown in Figure 1. The geometric mean and its 95% confidence interval (95% CI) of the testosterone ratio (Post / Pre) before and after GnRH stimulation are represented according to time (days).

[0156] At baseline, the ratios (Post / Pre) before and after GnRH stimulation were 9.39 in the control group and 5.65 in the treatment group. There was no significant difference between the two groups. The treatment / control ratio and its 95% confidence interval were 0.60 [0.18;1.96] and p = 0.3785.

[0157] On day 14, the ratio (Post / Pre) before and after GnRH stimulation was significantly higher in the control group, and the treatment / control ratio was equal to 0.07 (p < 0.0001). These differences persisted until day 252 as the treatment / control ratio was 1.37 and p = 0.7004 on day 280. At time points from day 308 to day 364, the control group exceeded again (p < 0.01). No statistically significant differences were observed after day 392.

[0158] Conclusion: This example shows that the testosterone concentration increases during deslorelin implantation (flare-up phase) and then decreases from 2 weeks to 252 weeks. After this date, the testosterone concentration slowly increases and returns to physiological concentrations.

[0159] Example 3: Effect of deslorelin on sperm concentration in male cats The purpose of this study was to examine whether deslorelin has an effect on sperm concentration in male cats.

[0160] 3.1 Materials and Methods: Tests were performed on the treatment group and control group of male cats in Example 2 (see 2.1 of Example 2).

[0161] 1 μL of semen was added to 199 μL of formalin citrate, the sperm concentration in the sample was counted, and when a sufficient amount was available, the sperm morphology was evaluated. The sperm concentration was determined according to standard operating procedures. The dilution ratio was 1:200.

[0162] 3.2 Results: The results are shown in Table 2 and Figure 2. The sperm concentration ( / μL) and its interquartile range are represented according to time (days).

[0163] Despite observing a low sperm concentration 56 days later in the treatment group, the median reached 0 from 112 days to 364 days, and the distribution of the change from the baseline between the two treatments was not statistically significant except at 252 days (p = 0.0262). Measure the sperm concentration ( / μL) over time in cats treated with (Suprelorin®). Details are shown in Table 2 below.

[0164]

Table 2

[0165] Example 4: Determination of the positive correlation between the injected deslorelin concentration and testosterone concentration The objective of this study was to examine whether there was a positive correlation between the injected deslorelin concentration and the testosterone concentration.

[0166] 4.1 Materials and Methods: From the testosterone and deslorelin concentrations obtained from 16 healthy male cats treated with 4.7 mg of Suprelorin® (registered trademark), the clear relationship between the plasma deslorelin concentration over time and the PD parameters was evaluated graphically.

[0167] The serum concentration of testosterone was measured according to the protocol detailed in Example 2, 2.1.

[0168] The plasma concentration of deslorelin was measured according to the protocol detailed in Example 1, 1.1.

[0169] 4.2 Results: The results of the mean deslorelin plasma concentration and mean testosterone (before and after GnRH stimulation) serum concentration over time are shown in Figure 3 (n = 16 cats treated). In this graph, the mean deslorelin plasma concentration (black circles) (pg / mL), mean pre-GnRH testosterone serum concentration (black squares) (pg / mL), and mean post-GnRH testosterone serum concentration (black triangles) (pg / mL) are represented according to time (days).

[0170] Conclusion: This study showed an inverse correlation between deslorelin and testosterone. In fact, as soon as the deslorelin concentration decreased (below 12 pg / mL), the testosterone concentration began to increase.

[0171] Example 5: Determination of the deslorelin plasma concentration over time and validation of the LC-MS / MS method This objective was to determine the deslorelin plasma concentration over time and to validate the LC-MS / MS method for determining deslorelin in the plasma of intact male cats after single subcutaneous administration of a 4.7 implant of Suprelorin® (registered trademark).

[0172] 5.1 Materials and Methods: Samples were collected from 10 intact male cats treated with Suprelorin® 4.7 implant. After solid-phase extraction using Oasis® WCXμElution plates, samples (treated with K2-EDTA as anticoagulant, based on a plasma volume of 150 μL) were analyzed by LC-MS / MS method. The compound (13C,15N)-Leu(7)-deslorelin was used as a standard. The calibration range was 4 pg / mL to 2000 pg / mL.

[0173] 5.2 Results: The measurement results of the plasma deslorelin concentration over time in cats treated with deslorelin implant (i.e., Suprelorin® 4.7 mg) are shown in Table 3 below.

[0174]

Table 3

[0175] Conclusion: This example shows that deslorelin is present in the blood at a very high concentration at D0 and then this concentration decreases over time. In some cats, deslorelin remains in the blood for up to 497 days.

[0176] Example 6: Determination of serum testosterone concentration using radioimmunoassay (RIA) method 6.1 Objectives: Serum testosterone concentrations were collected from intact male cats after single SC administration of Suprelorin® 4.7 implant, providing information on the baseline testosterone levels and the effect of this implant on testosterone production ability after stimulation.

[0177] 6.2 Materials and Methods: Samples were collected from 10 intact male cats treated with Suprelorin®. Effective extraction was performed, and the samples were analyzed using a radioimmunoassay (RIA) method with a lower limit of quantitation (LLOQ) of 0.01 ng / mL. The samples were analyzed twice at 350 μL each for duplicate analysis. Only results with a coefficient of variation (CV%) of 20% or less between 0.01 ng / mL and 2 ng / mL were reported.

[0178] 6.3 Results: The measurement results of serum testosterone concentration over time after a single subcutaneous (SC) administration of 4.7 implant of Suprelorin® to intact male cats are shown in Table 4 below.

[0179]

Table 4

[0180] Example 7: Effect of deslorelin on testis size 7.1 Objectives: The purpose was to confirm whether deslorelin affects the size of the testis.

[0181] 7.2 Materials and Methods: To conduct this study, two groups of cats were set up. - Control group (5 intact male cats) - Group treated with 4.7 mg of Suprelorin® (10 intact male cats).

[0182] The size of the testes of the cats (length, width, and depth of each testis, unit: mm) was measured using a caliper and recorded as a number rounded to the nearest 0.1 mm.

[0183] 7.3 Results: The time-course curve of the results of average testis volume (n = 5 cats in the control group, n = 10 cats in the treatment group) is shown in Figure 4.

[0184] At baseline, the mean testicular volume was similar between treatments, 323.5 and 340.8 mm for cats in the control and treatment groups, respectively 3 and the medians were 318 and 324.5 mm 3 respectively. Over time, the mean values increased dramatically in the control cats, reaching 3661.5 mm at 71 weeks / 18 months 3 (a 10-fold increase from baseline), while the mean values in the treatment group fluctuated gently and the final value was 1394.8 mm 3 (a 3-fold increase from baseline).

[0185] At 4 weeks / 1 month, the mean testicular volumes were significantly different from each other, with mean values (mm 3 ) and 95% CIs of 664.0 [465.7; 862.3] in the control cats and 466.4 [388.3; 544.5] in the treatment cats. The difference observed between the treatment and control cats, and the 95% CI, was -197.3 mm 3 [-347.7; -47.5]. This difference between treatments remained significant until the end of the study (-2266.7 mm 3 [-3313.7; -1219.6]), and at 71 weeks / 18 months, the mean values (mm 3 ) and 95% CIs were 3661.5 [3006.4; 4316.4] in the control cats and 1394.8 [676.8; 2112.7] in the treatment cats. Comparing with the testosterone ratios before and after GnRH stimulation where there was no significant difference between groups from D364 / 53 weeks (round 1 year), this indicates that the reversibility of the effect on testicular volume occurs later.

[0186] Conclusion: This study demonstrated that deslorelin induced a decrease in testicular volume in treated cats.

[0187] Example 8: Analysis of the pharmacokinetic parameters of deslorelin in dogs The intent of this study was a descriptive analysis of the pharmacokinetic parameters of deslorelin in dogs.

[0188] 8.1 Materials and Methods: This study was conducted in 6 intact male dogs implanted with deslorelin (4.7 mg), and blood was collected over the duration of the study (about 15 months). The objective of this biological analysis study was to validate an LC-MS / MS method for the determination of deslorelin in dog plasma (based on a plasma volume of 150 μL. K2-EDTA was used as the anticoagulant). The lower limit of quantification (LLOQ) was established at 4 pg / mL in dog plasma.

[0189] 8.2 Results: The results of the plasma concentration (pg / ml) of deslorelin measured over time in dogs treated with deslorelin are shown in Table 5 below.

[0190] [Table 5]

[0191] Conclusion: This example found that deslorelin was present in the blood at very high concentrations in the first few hours after administration and then decreased over time. Plasma deslorelin concentrations could be quantified in some dogs until the end of the study (D441), and the average concentration was 2 pg / mL.

[0192] Example 9: Effect of deslorelin on testosterone concentration 9.1 Objectives: The objective of this study was to confirm whether deslorelin implants affect testosterone concentrations.

[0193] 9.2 Materials and Methods: This study was conducted in 6 intact male dogs implanted with deslorelin (4.7 mg), and blood was collected over the duration of the study (about 15 months).

[0194] Before the test, testosterone was extracted from the serum. Serum samples were thawed at room temperature, vortexed, and processed for steroid extraction. Briefly, 500 μL of serum was transferred to a clean glass tube, and 2.5 mL of ethyl acetate was added. The mixture was vortexed for 3 minutes, separated into layers, and the upper phase was transferred to a clean test tube. This procedure was repeated twice. Subsequently, the combined extracts were evaporated by heating at 30 °C under a gentle stream of compressed air. The dried extracts were stored at -20 °C for 18 hours before analysis.

[0195] Testosterone levels in canine plasma were measured using a Testosterone ELISA Kit from Cayman Chemical (see 582701, Ann Arbor, USA).

[0196] 9.3 Results: Results of the mean ratio of testosterone (and its 95% CI) before and after GnRH stimulation, measured over approximately 16 months of the study, in treated dogs (n = 6) are shown in Figure 5.

[0197] Conclusion: This study demonstrated that implant injection (D0) was followed by an initial stimulatory phase of testosterone (from D0 to D28), and subsequently an inhibitory phase from 28 to 252 days. Thus, implant injection induced a long-term decrease in testosterone.

[0198] Example 10: Effect of deslorelin on LH concentration 10.1 Materials and Methods: This study was conducted in six intact male dogs implanted with Suprelorin® (4.7 mg), and blood was collected over the duration of the study (approximately 15 months).

[0199] LH was measured using the ELISA kit "LH Detect®" (ReproPharm Vet, France). Serum was diluted 1:5 and incubated with the coated capture antibody. After washing, the second antibody was incubated. After washing again, the third conjugated antibody was finally incubated. After washing once more, tetramethylbenzidine (TMB) was used as the substrate to detect the conjugated antibody that had bound. The reaction was stopped with an acidic solution. Absorbance was read at 450 nm, and the absorbance value of the sample was compared with the calibration curve values (0 - 8 ng / mL) to determine the sample concentration. LH values were expressed in ng / mL.

[0200] 10.2 Results: In the treated dogs (n = 6), the results of the mean LH ratio (95% CI) of the LH ratio before and after GnRH stimulation measured over approximately 15 months of the study are shown in Figure 6.

[0201] At baseline, the mean ratio before and after GnRH stimulation was 15.8. From D28 to D168, the mean ratio was equal to 1. At D252, this ratio began to increase and reached an average of 5.8. From there until the end of the study, the value of the mean ratio was approximately 10.

[0202] From D28 to D168, in all dogs, the ratio was less than 2. At D196 and D224, in only one dog, the ratio was 2 or more. After D252, in all dogs, the value of the ratio was ≥2.

[0203] Example 11: Correlation between the injected deslorelin concentration and the concentrations of the hormones LH, FSH, and testosterone in male cats treated with Suprelorin® 11.1 Objectives: The aim of this study was to examine whether there was a positive correlation between the injected deslorelin concentration and the concentrations of the hormones LH, FSH, and testosterone in male cats treated with Suprelorin® (4.7 mg).

[0204] 11.2 Materials and Methods: This study was conducted in six intact male dogs implanted with Suprelorin® (4.7 mg), and blood was collected during the duration of the study (approximately 15 months). From the testosterone / LH / FSH and deslorelin concentrations obtained in six healthy male cats treated with 4.7 mg of Suprelorin®, the clear relationship between plasma deslorelin concentration over time and PD parameters was evaluated graphically.

[0205] 11.3 Results: The results are reported in Figure 7. Figure 7 shows an initial spike, followed by an average LH concentration, then a dramatic decrease in testosterone concentration (slight for FSH), and a gradual decrease in deslorelin plasma concentration over time relative to GnRH pre-testosterone, LH, and FSH concentrations, with all hormone levels increasing at D224 where the deslorelin concentration is 12 ± 10.4 pg / mL.

[0206] Conclusion: This study demonstrated an inverse correlation between deslorelin and testosterone / LH and FSH. In fact, as soon as the deslorelin concentration decreased (below 12 pg / mL), the other concentrations began to increase.

[0207] Example 12: Field investigation of cats The intention of this study was to demonstrate the suppression of fertility and the persistence of infertility in male cats.

[0208] 12.1 Materials and Methods This example provides data by combining two studies (double-blind placebo-controlled randomized) conducted consecutively according to the following schedule. Study 1: Study 1 was conducted by dividing a group of 205 male cats into two groups, one control group receiving 1 mL of saline (n = 51, subcutaneous administration), and one treatment group receiving an implant of 4.7 mg of Suprelorin® (subcutaneous implant, deslorelin acetate) (n = 154). Study 2: At the end of the first period of 12 months, the Suprelorin® implants were removed from all male cats in the treatment group. Thereafter, 12 male cats in the treatment group of Study 1 were administered physiological saline (1 mL subcutaneously) (physiological saline group), and 22 cats in the treatment group of Study 1 were re-implanted with a new 4.7 mg Suprelorin® implant during the second period of 12 months (treatment twice group). The effectiveness and safety of the re-implantation were followed up for these additional 12 months, that is, for a total of 24 months.

[0209] The observation schedule was as follows. Study 1: V1: from -14 to -7 days, V2: day 0, V3: 1.5 months ± 3 days, V4: 3 months ± 5 days, V5: 6 months ± 5 days, V6: 9 months ± 5 days, V7: 12 months ± 5 days; Study 2: V8: 12 months ± 3 days, V9: 13.5 months ± 3 days, V10: 15 months ± 5 days, V11: 18 months ± 5 days, V12: 21 months ± 5 days, V13: 24 months ± 5 days.

[0210] Regarding the inclusion criteria, in both studies, the criteria were as follows. - Domestic short-haired cat - Age > 3 months or > 15 months in Study 2 - Intact indoor male - Cats owned by the applicant (including cats in a protective facility / cats taken in by foster families) The additional criteria for Study 2 were success in Study 1.

[0211] In these studies, the following primary evaluation items were measured. - Study 1: Suppression of fertility (*testosterone ≤ 0.10 ng / mL for 3 to 12 months from V4 to V7*) - Study 2: Suppression of fertility (*testosterone ≤ 0.10 ng / mL from V8 to V13 (12 to 24 months)) and time to effect regression.

[0212] In addition to the safety parameters (hematology and blood biochemistry, urine test, local reaction at the implantation site, body weight change, appetite, abnormal testicular evaluation, adverse events), the following secondary evaluation items are also reported. - Total sexual behavior score - Aggressiveness - Chirping - Urine marking - Urine odor / smell - Penile spine - Testicular volume

[0213] 12.2 Results: - Test Sterone: The results of the mean effectiveness are shown in Figure 8A. Overall, the success rates of both studies were very similar for 1 or 2 implants, and the effectiveness was >87% at all time points in Study 2. The detailed levels of testosterone are shown in Figure 8B for Study 1 and Figure 8C for Study 2, respectively.

[0214] - Reversibility: The reversibility of the deslorelin effect was investigated. Indeed, among the deslorelin-treated groups in Study 1, as described in 12.1 above, n = 12 subjects were recorded in the saline group of Study 2. The results showed that 100% of the cats that were in the treatment group in Study 1 and the saline group in Study 2 reversed at D279, as recorded in Study 2. The median time for reversibility based on serum testosterone concentration measurements was D379 ± 178.

[0215] - Secondary Evaluation Items: The effectiveness results of the secondary evaluation items are reported as follows. · The mean values of the total sexual behavior scores are shown in Figure 9A for Study 1 and Figure 9B for Study 2, respectively. · The mean chirping variation is shown in Figure 10A for Study 1 and 10B for Study 2. · The % change in urine marking from baseline is shown in Figure 11A for Study 1 and Figure 11B for Study 2, respectively. · The variation / decrease in urine smell measured as a change from baseline is shown in Figure 12A for Study 1 and Figure 12B for Study 2, respectively. ·The percentage change in testicular volume is shown in Figure 13A for Study 1 and in Figure 13B for Study 2, respectively. ·The percentage change in penile spines that appeared is shown in Figure 14A for Study 1 and in Figure 14B for Study 2, respectively.

[0216] During the course of the study, there were no safety concerns, and the deslorelin implant showed good tolerance.

[0217] In short, the combination of these studies has demonstrated the following. Deslorelin was effective for at least 12 months when chemically castrating the subjects. This effect has been proven to be reversible when the treatment process of the action ends. These studies have also demonstrated that sexual behavior decreases due to the rapid onset of the effectiveness of reduced behavior (about 7 - 31 days). Moreover, there were no safety concerns regarding single or repeated administrations.

[0218] Therefore, it can be concluded that deslorelin administered by sustained release, especially by implant, is an excellent alternative to surgical castration and a safe non - surgical method for controlling the reproduction of male cats.

[0219] The method of using deslorelin as described herein is a reversible, non - surgical, reliable castration measure that promotes the health of the pet family while leaving options.

[0220] Example 13: PK / PD pre - clinical study in intact male cats (intact male cats) The intention of this study was to demonstrate suppression of fertility, measure deslorelin levels along with PK data, evaluate reversibility by PD analysis, and record systemic and local tolerance.

[0221] This example provides data collected from a PK / PD pre - clinical study in intact male cats according to the following study design. - Product: Suprelorin® 4.7 mg vs placebo (NaCl solution) - Subjects: Cats treated at 15 weeks of age = 15 + 3 intact male cats (15 weeks old) were divided into two groups, one treated with a 4.7 mg Suprelorin® implant (n = 13, Group 2), and the other treated with saline (placebo) (n = 5, Group 1). At D14, 3 preliminary cats were excluded from the study. This study only included 5 + 3 female cats (Group 3) that were in puberty at the start of the study to evaluate the reproductive behavior of male cats. These cats were not treated with implants. - Duration: 18 months of study after injection (PI) (D502 / 72 weeks, i.e., 16.5 months)

[0222] Table 6 below shows the details of the study.

[0223]

Table 6

[0224] In this table, BW means body weight.

[0225] Research Results: - The PK study results are shown in Figure 15A. Overall, it should be noted that Tmax is at 2 hours, with an average decrease of 92% in the first 24 hours after injection (PI), followed by a constant slow decrease. In the first cats, at 1 year after PI (52 weeks after PI), the concentration was not detected in 5 / 10 male cats at the end of the study. - Testosterone levels are recorded in Figure 15B, showing that testosterone was low in treated cats until 1 year after PI (53 weeks after PI), and then increased (except for 3 / 10 cats). At the end of the study, the average testosterone was still low in treated cats (GnRH before: 1.59 vs 2.16 ng / mL, GnRH after: 2.34 vs 6.44 ng / mL in the treated group / control group, respectively).

[0226] Note that the ratio before and after is statistically significant from 2 weeks to 1 year after PI (p = 0.0001). - The penile spines and behavioral records are shown in FIGS. 15C and 15D. For penile spines, the results are statistically significant from 7 to 67 weeks post-injection. For sexual behavior, the results were statistically significant at 10, 12, 34, 36, 38, 46, 52, 58, and 66 weeks post-injection. Finally, for reproductive behavior, there was no statistical difference, and it was observed that one control male mated 35 weeks post-injection (1 year old), and one treated cat mated 65 weeks post-injection. Sexual impulse was affected by the individual personality characteristics of the cats. As shown in FIG. 15D, a tendency for a decrease in meowing, aggression, urine marking, and the typical male urine odor was also observed. It is clear from the graph that the control cats (animals C1 to C5) exhibited many sexual activities or reproductive behaviors, including meowing, aggression, urine marking, and the typical male urine odor, compared to the treated animals (T1 to T10) using deslorelin implants. - Puberty appeared at 5.5 months of age in 5 non-castrated control cats. The onset of puberty was delayed and lasted at least 1 year in deslorelin-treated cats.

[0227] A complete reversal of male and hormonal parameters was effective at the end of the study (72 weeks of PI) in 5 / 10 treated cats.

[0228] Example 14: Use of deslorelin in prepubertal female dogs The intention of this study was to evaluate the delay in the onset of puberty in prepubertal female dogs. As is known, puberty begins depending on the setting of several parameters. - Increased receptor numbers in the central nervous system (CNS) and gonads - Desensitization of the hypothalamus / pituitary to estrogen - Maturation of the gonads and secondary genitalia - Follicular maturation, spermatogenesis

[0229] The results of this study indicate that it is possible to delay puberty. Use of 1 implant: There was no impairment of body development, the size of the vulva was normal, but epiphyseal closure was delayed. There was a slight possibility of flare-up (swelling of the vulva). Use of 3 implants every 4.5 months: The vulva remained immature and the effect was found to be comparable to pre-pubertal castration.

[0230] Note that the deslorelin concentration is not related to the persistence of efficacy and the implant should not be removed.

[0231] This study determines the criteria for deslorelin administration to prepubertal female dogs. - The age at implant insertion is 4 - 5 months old - Exclude the onset of puberty determined by clinical examination, vaginal examination, vaginal cytology, E2, DHEA measurement (sexual behavior).

Claims

**Claim 1** Deslorelin for use in chemical castration of non-human mammals within one month after administration, wherein when administered, the Cmax of deslorelin after administration exceeds 7000 pg / mL and the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL. **Claim 2** The deslorelin according to claim 1, wherein the deslorelin is administered subcutaneously or intramuscularly to a non-human mammal in need of deslorelin in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / mL after 8 days of administration. **Claim 3** The deslorelin according to claim 2, wherein the deslorelin plasma concentration is from 10 pg / mL to 400 pg / mL from 8 days after administration to at least 6 months after administration. **Claim 4** Deslorelin for use in inducing temporary infertility in non-castrated male animals for at least 6 months, comprising administering deslorelin subcutaneously or intramuscularly to a mammal in need of deslorelin in an effective dose to induce a deslorelin plasma concentration of 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / mL after 8 days of administration. **Claim 5** The deslorelin according to claim 4, wherein after administration, the deslorelin is released substantially continuously at a plasma concentration level of less than 400 pg / mL from 8 days after administration to at least 6 months after administration. **Claim 6** A sustained-release drug delivery system (DDS) containing deslorelin for use in chemical castration of non-human mammals, wherein the DDS releases deslorelin with a Cmax of deslorelin after administration exceeding 7000 pg / mL, the deslorelin concentration in the plasma of the non-human mammal is at least 10 pg / mL, and is maintained for at least 6 months. **Claim 7** The sustained-release drug delivery system (DDS) according to claim 6, wherein the DDS is a subcutaneous or intramuscular implant containing deslorelin and a biodegradable material. **Claim 8** Deslorelin for use in chemical castration of non-human mammals, which, when administered, reduces the testosterone level in the plasma of said non-human mammals to less than 0.4 ng / ml and the deslorelin concentration in the plasma of said non-human mammals is at least 10 pg / mL, preferably more than 15 pg / mL.

9. A method of contraception in non-human mammals, comprising subcutaneously or intramuscularly administering to a non-human mammal an implant containing deslorelin and a biodegradable material such as a biodegradable polymer, wherein the implant releases deslorelin in a state where the Cmax of deslorelin after 1 hour of administration exceeds 7000 pg / mL, and the deslorelin concentration in the plasma of said non-human mammals is at least 10 pg / mL and is maintained for at least 6 months.

10. The method of contraception according to claim 9, wherein the deslorelin is administered to a non-human mammal in need thereof in an effective dose to induce a deslorelin plasma concentration of from 4000 pg / mL to 40000 pg / mL between 1 hour and 24 hours after administration and a plasma concentration of less than 400 pg / ml after 8 days of administration.

11. The method of contraception according to claim 9 or 10, wherein the plasma concentration of deslorelin is less than 400 pg / ml from 8 days after administration to at least 6 months after administration.

12. Deslorelin for use in preventing behavioral disorders in non-human mammals, wherein the deslorelin is administered subcutaneously or intramuscularly using a sustained-release drug delivery system, and the deslorelin concentration in the plasma of said non-human mammals is at least 10 pg / mL.

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