Methods and compositions for synchronizing insemination time

Abstract

Compositions and methods for synchronizing the timing of insemination in pigs are described. More specifically, methods for synchronizing the timing of insemination by administering a composition comprising a hormone are described, wherein the pigs are inseminated only once after administration of the hormone, and wherein there is no estrus detection.

Description

[0001] This application is a divisional application of an invention patent application filed on April 23, 2010, with application number 201080029042.8 (PCT / US2010 / 032258), and invention name “Methods and compositions for synchronizing insemination time”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority under 35 USC §119(e) to U.S. Provisional Application No. 61 / 172,009, filed April 23, 2009, which is expressly incorporated herein by reference. Field of the Invention

[0004] The present invention relates to methods and compositions for synchronizing the timing of insemination in animals. More particularly, the invention relates to methods and compositions for synchronizing the timing of insemination in pigs without heat detection. Background of the Invention

[0005] Gonadotropin-releasing hormone is a 10 amino acid peptide also known as luteinizing hormone-releasing hormone (LHRH). Gonadotropin-releasing hormone is produced in the hypothalamus and is responsible for the release of follicle-stimulating hormone and luteinizing hormone from the pituitary gland. Gonadotropin-releasing hormone is released from neurons in the hypothalamus and plays a role in the complex regulation of follicle-stimulating hormone and luteinizing hormone release. Follicle-stimulating hormone and luteinizing hormone, in combination, regulate the function of the gonads, the production of testosterone in the testes and progesterone and estrogen in the ovaries, and regulate the production and maturation of gametes. For example, follicle-stimulating hormone stimulates the growth and recruitment of immature follicles in the ovaries, and luteinizing hormone triggers ovulation.

[0006] There are differences in the secretion of gonadotropin-releasing hormone in females and males. In males, gonadotropin-releasing hormone is secreted in pulses of constant frequency, but in females, the pulse frequency varies during the estrous cycle, and there is a large surge of gonadotropin-releasing hormone just before ovulation. The secretion of gonadotropin-releasing hormone is pulsatile in all vertebrates and is essential for proper reproductive function. Thus, gonadotropin-releasing hormone controls the complex process of follicular growth, ovulation and maintenance of the corpus luteum in females and spermatogenesis in males.

[0007] Gonadotropin-releasing hormone has been isolated and characterized as a decapeptide. Synthetic forms of gonadotropin-releasing hormone are available, and modifications to the decapeptide structure of gonadotropin-releasing hormone have resulted in a variety of gonadotropin-releasing hormone analogs that stimulate (e.g., gonadotropin-releasing hormone agonists) or inhibit (e.g., gonadotropin-releasing hormone antagonists) the release of gonadotropins such as luteinizing hormone and follicle-stimulating hormone.

[0008] For commercial pig production, it is important to maximize the efficiency of reproduction to make pig production more profitable. Daily estrus detection of individual female pigs has been heavily relied upon, and the associated labor costs are used for manual detection of estrus in female pigs, based on daily inspections of gilts or sows, to achieve the best results for, for example, artificial insemination. Estrus detection with labor-intensive methods (such as daily inspections) increases the likelihood of successful artificial insemination. Therefore, it is necessary to invest time, manual labor and material costs in daily inspections of estrus detection, because it is difficult to predict the time of estrus (i.e., the best time for insemination) without using a method that requires a daily plan to monitor estrus detection. Therefore, there is a need for a method for optimizing the success of insemination of animals, which does not have estrus detection, to reduce labor costs and increase the profitability of pig production. Brief description of the invention

[0009] Applicants have made the unexpected discovery that controlling the timing of ovulation by administering hormones can eliminate mating based on estrus detection and allow pigs to be inseminated only once to optimize fertility and optimize cost performance. In one embodiment, a method of synchronizing the timing of insemination of pigs without estrus detection is described. The method includes the steps of administering a dose of hormone to the pig on the fourth day after weaning, the hormone selected from gonadotropin releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof, wherein the pig is inseminated only once about 15 to about 24 hours after the administration of the hormone, and wherein there is no estrus detection.

[0010] In the above described embodiments, the following features or any combination thereof apply. In the above embodiments, 1) the pig can be a postpartum sow; 2) the pig can be a gilt; 3) the insemination can be artificial insemination; 4) the insemination can be by natural mating; 5) ovulation can be synchronized; 6) the hormone can be administered in an amount effective to stimulate ovulation of the follicle; 7) the dose of hormone can be administered using a method selected from the following: placement of a catheter (deposition catheter), artificial administration and injection; 8) the pig can be inseminated about 15 to about 18 hours after the hormone is administered; 9) the pig can be inseminated about 18 hours after the hormone is administered; 10) the pregnancy rate of the pig can be increased relative to pigs not given the hormone; 11) the litter size of the pig can be increased relative to pigs not given the hormone; 12) the hormone can be administered to more than one pig; 13) the percentage of pigs that ovulate about 48 hours after the hormone is administered can be increased relative to pigs not given the hormone; 15) the hormone can be administered intravaginally; 16) the hormone can be administered to the anterior vagina; 17) the hormone can be gonadotropin-releasing hormone receptor agonist; 18) the hormone may be a luteinizing hormone receptor agonist; 19) the hormone may be a human chorionic gonadotropin receptor agonist; 20) the hormone may be triptorelin; 21) the hormone may be a triptorelin salt; 22) the hormone may be a synthetic substance; 23) the hormone may be in acetate form; 24) the hormone may be administered in a composition comprising a gel; 25) the gel may comprise a polysaccharide; 26) the polysaccharide may be selected from: cellulose, dextran and alginate; 27) the gel may comprise cellulose; and 28) the cellulose may be methylcellulose. Any combination of the embodiments of the above paragraph

[0007] with 1-28 or any combination thereof is contemplated.

[0011] In another embodiment, a kit comprising a dose of a hormone selected from the group consisting of gonadotropin-releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof is described. The kit further comprises instructions for use, wherein the hormone is in a composition comprising a gel, and the composition has a pH of about 5 to about 6.

[0012] In the embodiment described in the above paragraph

[0009] , the following features or any combination thereof apply. In the embodiment described in paragraph

[0009] , 1) the instructions may indicate that the hormone should be administered to the pig on about the fourth day after weaning, and the pig can be inseminated once about 15 to about 24 hours after the administration of the hormone; 2) the instructions may indicate that the insemination should be artificial insemination; 3) the instructions may indicate that the insemination should be through natural mating; 4) the kit may further include a catheter for placing, an applicator or a syringe for manual administration; 5) the instructions may indicate that the pig should be inseminated about 15 to about 18 hours after the administration of the hormone; 6) the instructions may indicate that the pig should be inseminated about 18 hours after the administration of the hormone; 7) the instructions may indicate that the hormone should be administered intravaginally; 8) the instructions may indicate that the hormone is to be administered to the anterior vagina; 9) the hormone may be a gonadotropin-releasing hormone receptor agonist; 10) the hormone may be a luteinizing hormone receptor agonist; 11) the hormone may be a human chorionic gonadotropin receptor agonist; the hormone may be triptorelin; 12) the hormone may be a triptorelin salt; 13) the hormone may be a synthetic substance; 14) the hormone may be in acetate form; 15) the gel may contain a carbohydrate; 16) the gel may contain a polysaccharide; 17) the polysaccharide may be selected from cellulose, dextran and alginate; 18) the gel may contain cellulose; and 19) the cellulose may be methylcellulose. Any combination of the embodiments of paragraph

[0009] above and 1-19 or any combination thereof is contemplated.

[0013] In another embodiment, a composition comprising a gonadotropin-releasing hormone receptor agonist and a gel is described, wherein the composition has a pH of about 5.0 to about 6.0.

[0014] In the embodiment described in the above paragraph

[0011] , the following features or any combination thereof apply. In the embodiment described in paragraph

[0011] , 1) the composition may further comprise a preservative; 2) the preservative is selected from methylparaben and propylparaben; 3) the composition may further comprise a stabilizer; 4) the stabilizer may be an L-amino acid; 5) the stabilizer may be L-methionine; 6) the gel may comprise a polysaccharide; 7) the polysaccharide may be selected from cellulose, dextran and alginate; 8) the gel may comprise cellulose; 9) the cellulose may be methylcellulose; 10) the composition may further comprise a tonicity agent; 11) the agonist may be triptorelin; 12) the composition may also be combined with instructions for use; 13) the instructions combined with the composition may indicate that the agonist should be administered to a pig and the pig should be inseminated once about 15 to about 24 hours after administration of the agonist; and 14) the instructions may indicate that the pig should be inseminated once about 18 hours after administration of the agonist. Any combination of the embodiment of the above paragraph

[0011] with 1-14 or any combination thereof is contemplated.

[0015] The following various implementations are provided.

[0016] 1. A method for synchronizing the timing of insemination of pigs without estrus detection is described. The method comprises the step of administering to the pigs a dose of a hormone selected from the group consisting of gonadotropin releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof, on the fourth day after weaning, wherein the pigs are inseminated only once, about 15 to about 24 hours after the administration of the hormone, and wherein there is no estrus detection.

[0017] 2. The method of clause 1, wherein the pig is a postpartum sow.

[0018] 3. The method of clause 1, wherein the pig is a gilt.

[0019] 4. The method of clauses 1 to 3, wherein the insemination is artificial insemination.

[0020] 5. The method of clauses 1 to 4, wherein the insemination is by natural breeding.

[0021] 6. The method of clauses 1 to 5, wherein ovulation is synchronized.

[0022] 7. The method of clauses 1 to 6, wherein the hormone is administered in an amount effective to stimulate ovulation of the ovarian follicle.

[0023] 8. The method of clauses 1 to 7, wherein the effective amount of the hormone is from about 10 μg to about 1000 μg.

[0024] 9. The method of clauses 1 to 7, wherein the effective amount of the hormone is from about 10 μg to about 500 μg.

[0025] 10. The method of clauses 1 to 9, wherein the dose of hormone is administered using a method selected from the group consisting of: use of a placed catheter, manual administration, and injection.

[0026] 11. The method of clauses 1 to 10, wherein the pig is inseminated about 15 to about 18 hours after administration of the hormone.

[0027] 12. The method of clauses 1 to 10, wherein the pig is inseminated about 16 to about 24 hours after administration of the hormone.

[0028] 13. The method of clauses 1 to 10, wherein the pig is inseminated about 18 to about 22 hours after administration of the hormone.

[0029] 14. The method of clauses 1 to 13, wherein the pig is inseminated about 18 hours after administration of the hormone.

[0030] 15. The method of clauses 1 to 14, wherein the pregnancy rate of the pig is increased relative to pigs not administered the hormone.

[0031] 16. The method of clauses 1 to 15, wherein the total number of healthy fetuses is increased relative to pigs not administered the hormone.

[0032] 17. The method of clauses 1 to 16, wherein the percentage of litters born is increased relative to pigs not administered the hormone.

[0033] 18. The method of clauses 1 to 17, wherein the total number of piglets born is increased relative to pigs not administered the hormone.

[0034] 19. The method of clauses 1 to 18, wherein the total number of piglets born per semen dose is increased relative to pigs not administered the hormone.

[0035] 20. The method of clauses 1 to 19, wherein the total number of piglets born per semen dose is increased relative to pigs inseminated following estrus detection.

[0036] 21. The method of clauses 1 to 20, wherein the piglet index is increased relative to pigs not administered the hormone.

[0037] 22. The method of clauses 1 to 21 wherein the piglet index is increased relative to pigs inseminated following estrus detection.

[0038] 23. The method of clauses 1 to 22, wherein the pig has an increased litter size relative to pigs not administered the hormone.

[0039] 24. The method of clauses 1 to 23, wherein the hormone is administered to more than one pig.

[0040] 25. The method of clauses 1 to 24, wherein the percentage of pigs that ovulate is increased about 48 hours after administration of the hormone relative to pigs not administered the hormone.

[0041] 26. The method of clauses 1 to 25, wherein the hormone is administered intravaginally.

[0042] 27. The method of clauses 1 to 26, wherein the hormone is administered to the anterior vagina.

[0043] 28. The method of clauses 1 to 27, wherein the hormone is a gonadotropin-releasing hormone receptor agonist.

[0044] 29. The method of clauses 1 to 28, wherein the hormone is a luteinizing hormone receptor agonist.

[0045] 30. The method of clauses 1 to 29, wherein the hormone is a human chorionic gonadotropin receptor agonist.

[0046] 31. The method of clauses 1 to 30, wherein the hormone is triptorelin.

[0047] 32. The method of clauses 1 to 31, wherein the hormone is a triptorelin salt.

[0048] 33. The method of clauses 1 to 32, wherein the gonadotropin-releasing hormone has the formula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH 2 .

[0049] 34. The method of clauses 1 to 33, wherein the gonadotropin-releasing hormone receptor agonist has the formula

[0050]

[0051] or a solvate, a hydrate or a pharmaceutically acceptable salt thereof,

[0052] in

[0053] R 1 and R 2 is independently at each occurrence hydrogen, or is independently selected from: alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl, each of which is optionally substituted, or R 1 and R 2 Form a carbocyclic or heterocyclic ring with the attached carbon;

[0054] X is hydrogen, or X is selected from: alkyl, cycloalkyl, heteroalkyl, optionally substituted alkylenecarboxamide; and

[0055] HNC(O)NR 3 R 4 , where R 3 and R 4Each occurrence is independently selected from hydrogen, alkyl, heteroalkyl, and haloalkyl.

[0056] 35. The method of clause 34, wherein R1 is methylene-heteroaryl, wherein heteroaryl is selected from the group consisting of pyridyl, thiazolyl, pyridazolyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl, pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and

[0057] R 2 is hydrogen or methyl.

[0058] 36. The method of clauses 34 to 35, wherein X is CH 2 C(O)NH 2 .

[0059] 37. The method of clauses 1 to 36, wherein the hormone is a synthetic substance.

[0060] 38. The method of clauses 1 to 37, wherein the hormone is in acetate form.

[0061] 39. The method of clauses 1 to 38, wherein the hormone is administered in a composition comprising a gel.

[0062] 40. The method of clause 39, wherein the gel comprises a polysaccharide.

[0063] 41. The method of clause 40, wherein the polysaccharide is selected from the group consisting of: cellulose, dextran and alginate.

[0064] 42. The method of clauses 39 to 41 wherein the gel comprises cellulose.

[0065] 43. The method of clause 42, wherein the cellulose is methyl cellulose.

[0066] 44. The method of clauses 39 to 43 wherein the gel comprises from about 0.25% to about 10% by weight methylcellulose.

[0067] 45. The method of clauses 39 to 43, wherein the gel comprises about 0.5 wt% to about 4.0 wt% methylcellulose.

[0068] 46. ​​The method of clauses 1 to 45, wherein the hormone is administered with a stabilizer, wherein the stabilizer is L-methionine.

[0069] 47. A kit comprising a dose of a hormone selected from the group consisting of gonadotropin-releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof, and instructions for use, wherein the hormone is in a composition comprising a gel, the composition having a pH of about 5 to about 6.

[0070] 48. The kit of clause 47, wherein the instructions indicate that the hormone should be administered to the pig on about the fourth day after weaning, and the pig should be inseminated about 15 to about 24 hours after administration of the hormone.

[0071] 49. The kit of clauses 47 to 48, wherein the instructions indicate that the insemination is to be artificial insemination.

[0072] 50. The kit of clauses 47 to 49, wherein the instructions indicate that the insemination is to be by natural breeding.

[0073] 51. The kit of clauses 47 to 50, wherein the kit further comprises a placement catheter, an applicator or a syringe for manual administration.

[0074] 52. The kit of clauses 47 to 51, wherein the instructions indicate that the pig should be inseminated about 15 to about 18 hours after administration of the hormone.

[0075] 53. The kit of clauses 47 to 51, wherein the instructions indicate that the pig should be inseminated about 16 to about 24 hours after administration of the hormone.

[0076] 54. The kit of clauses 47 to 51, wherein the instructions indicate that the pig should be inseminated about 18 to about 22 hours after administration of the hormone.

[0077] 55. The kit of clauses 47 to 54, wherein the instructions indicate that the pig should be inseminated approximately 18 hours after administration of the hormone.

[0078] 56. The kit of clauses 47 to 55, wherein the instructions indicate that the hormone is to be administered intravaginally.

[0079] 57. The kit of clauses 47 to 56, wherein the instructions indicate that the hormone is to be administered in an effective amount of about 10 μg to about 1000 μg.

[0080] 58. The kit of clauses 47 to 56, wherein the instructions indicate that the hormone should be administered in an effective amount of about 10 μg to about 500 μg.

[0081] 59. The kit of clauses 47 to 58, wherein the instructions indicate that the hormone is to be administered to the anterior vagina.

[0082] 60. The kit of clauses 47 to 59 wherein the hormone is a gonadotropin releasing hormone receptor agonist.

[0083] 61. The kit of clauses 47 to 60, wherein the hormone is a luteinizing hormone receptor agonist.

[0084] 62. The kit of clauses 47 to 61 wherein the hormone is a human chorionic gonadotropin receptor agonist.

[0085] 63. The kit of clauses 47 to 62, wherein the hormone is triptorelin.

[0086] 64. The kit of clauses 47 to 63, wherein the hormone is a triptorelin salt.

[0087] 65. The kit of clauses 47 to 64, wherein the gonadotropin-releasing hormone has the formula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH 2 .

[0088] 66. The kit of clauses 47 to 65, wherein the gonadotropin-releasing hormone receptor agonist has the formula

[0089]

[0090] or a solvate, a hydrate or a pharmaceutically acceptable salt thereof,

[0091] in

[0092] R 1 and R 2 is independently at each occurrence hydrogen, or is independently selected from: alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl, each of which is optionally substituted, or R 1 and R 2 Form a carbocyclic or heterocyclic ring with the attached carbon;

[0093] X is hydrogen, or X is selected from: alkyl, cycloalkyl, heteroalkyl, optionally substituted alkylenecarboxamide; and

[0094] HNC(O)NR 3 R 4 , where R 3 and R 4 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, heteroalkyl, and haloalkyl.

[0095] 67. The kit of clause 66, wherein R 1 is methylene-heteroaryl, wherein heteroaryl is selected from the group consisting of pyridinyl, thiazolyl, sulfopyridinyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl, pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and

[0096] R 2 is hydrogen or methyl.

[0097] 68. The kit of clauses 66 to 67, wherein X is CH 2C(O)NH 2 .

[0098] 69. The kit of clauses 47 to 68, wherein the hormone is a synthetic substance.

[0099] 70. The kit of clauses 47 to 69 wherein the hormone is in acetate form.

[0100] 71. The kit of clauses 47 to 70 wherein the gel comprises a sugar.

[0101] 72. The kit of clauses 47 to 71 wherein the gel comprises a polysaccharide.

[0102] 73. The kit of clause 72, wherein the polysaccharide is selected from cellulose, dextran and alginate.

[0103] 74. The kit of clauses 47 to 73 wherein the gel comprises cellulose.

[0104] 75. The kit of clause 74, wherein the cellulose is methylcellulose.

[0105] 76. The kit of clauses 47 to 75, wherein the gel comprises about 0.25% to about 10% by weight methylcellulose.

[0106] 77. The kit of clauses 47 to 75, wherein the gel comprises about 0.5% to about 4.0% by weight methylcellulose.

[0107] 78. The kit of clauses 47 to 77, further comprising a stabilizer, wherein the stabilizer is L-methionine.

[0108] 79. A composition comprising a gonadotropin-releasing hormone receptor agonist and a gel is described, wherein the composition has a pH of about 5.0 to about 6.0.

[0109] 80. The composition of clause 79 further comprising a preservative.

[0110] 81. The composition of clauses 79 to 80 wherein the preservative is selected from: methylparaben and propylparaben.

[0111] 82. The composition of clauses 79 to 81 further comprising a stabilizer.

[0112] 83. The composition of clause 82, wherein the stabilizer is an L-amino acid.

[0113] 84. The composition of clauses 82 to 83 wherein the stabilizer is L-methionine.

[0114] 85. The composition of clauses 79 to 84 wherein the gel comprises a polysaccharide.

[0115] 86. The composition of clause 85, wherein the polysaccharide is selected from the group consisting of: cellulose, dextran and alginate.

[0116] 87. The composition of clauses 79 to 86 wherein the gel comprises cellulose.

[0117] 88. The composition of clause 87 wherein the cellulose is methyl cellulose.

[0118] 89. The composition of clauses 79 to 88 wherein the gel comprises from about 0.25% to about 10% by weight methylcellulose.

[0119] 90. The composition of clauses 79 to 88 wherein the gel comprises from about 0.5 wt% to about 4.0 wt% methylcellulose.

[0120] 91. The composition of clauses 79 to 90 further comprising a tonicity agent.

[0121] 92. The composition of clauses 79 to 91 wherein the agonist is triptorelin.

[0122] 93. The composition of clauses 79 to 92 in combination with instructions for use.

[0123] 94. The composition of clause 93, wherein instructions associated with said composition indicate that said agonist should be administered to a pig, and that the pig should be inseminated once about 15 to about 24 hours after administration of said agonist.

[0124] 95. The composition of clause 93, wherein instructions associated with the composition indicate that the agonist should be administered to a pig, and the pig should be inseminated once about 16 to about 22 hours after administration of the agonist.

[0125] 96. The composition of clause 93, wherein instructions associated with said composition indicate that said agonist should be administered to a pig, and that the pig should be inseminated once about 18 to about 22 hours after administration of said agonist.

[0126] 97. The composition of clauses 93 to 96, wherein the instructions indicate that the pig should be inseminated once about 18 to about 22 hours after administration of the agonist.

[0127] 98. The composition of clauses 79 to 97, wherein the gonadotropin releasing hormone receptor agonist is gonadotropin releasing hormone, and wherein the gonadotropin releasing hormone has the formula pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH 2 .

[0128] 99. The composition of clauses 79 to 98, wherein the gonadotropin-releasing hormone receptor agonist has the formula

[0129]

[0130] or a solvate, a hydrate or a pharmaceutically acceptable salt thereof,

[0131] in

[0132] R 1 and R 2 is independently at each occurrence hydrogen, or is independently selected from: alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl, each of which is optionally substituted, or R 1 and R 2 Form a carbocyclic or heterocyclic ring with the attached carbon;

[0133] X is hydrogen, or X is selected from: alkyl, cycloalkyl, heteroalkyl, optionally substituted alkylenecarboxamide; and

[0134] HNC(O)NR 3 R 4 , where R 3 and R 4 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, heteroalkyl, and haloalkyl.

[0135] 100. The composition of clause 99, wherein R 1 is methylene-heteroaryl, and wherein heteroaryl is selected from the group consisting of pyridyl, thiazolyl, sulfapyridinyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl, pyrrolyl, indolyl, benzopyrazolyl, and benzimidazolyl; and

[0136] R 2 is hydrogen or methyl.

[0137] 101. The composition of clauses 99 to 100, wherein X is CH 2 C(O)NH 2 .

[0138] 102. The composition of clauses 79 to 101 wherein the gonadotropin releasing hormone receptor agonist is in an effective amount of about 10 μg to about 1000 μg.

[0139] 103. The composition of clauses 79 to 101 wherein the gonadotropin releasing hormone receptor agonist is in an effective amount of about 10 μg to about 500 μg.

[0140] 104. The composition of clauses 79 to 103 comprising methylparaben, propylparaben, sodium chloride, sodium citrate, L-methionine, citric acid, triptorelin and methylcellulose.

[0141] 105. The composition of clauses 79 to 104, wherein the composition comprises methylparaben in an amount of about 0.09% by weight per volume, propylparaben in an amount of about 0.01% by weight per volume, sodium chloride in an amount of about 0.91% by weight per volume, sodium citrate in an amount of about 0.186% by weight per volume, L-methionine in an amount of about 0.1% by weight per volume, citric acid in an amount of about 0.07% by weight per volume, triptorelin in an amount of about 0.01% by weight per volume, and methylcellulose in an amount of about 1.2% by weight per volume. BRIEF DESCRIPTION OF THE DRAWINGS

[0142] Figure 1 The percentage of sows that ovulated after treatment was given at 96 + / - 2 hours post-weaning (0-hours) is shown.

[0143] Figure 2 The stability of triptorelin after 13 days at 0-4°C (triangles), 30°C (squares) and 50°C (diamonds) is shown.

[0144] DETAILED DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0145] In one embodiment, a method of synchronizing the timing of insemination of pigs without estrus detection is described. The method comprises the step of administering to a pig a dose of a hormone selected from the group consisting of gonadotropin releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof, on day four after weaning, wherein the pig is inseminated only once, about 15 to about 24 hours after the administration of the hormone, and wherein estrus detection is not performed.

[0146] In one embodiment, a kit is described comprising a dose of a hormone selected from, for example, gonadotropin-releasing hormone, luteinizing hormone, human chorionic gonadotropin, and combinations thereof. In another embodiment, the kit further comprises instructions for use. In another embodiment, the hormone is in a composition comprising a gel as described herein. The composition typically has a pH of about 5 to about 6, but the pH may range from about 4 to about 9.

[0147] In one embodiment, a composition comprising a gonadotropin releasing hormone receptor agonist is described. In another embodiment, the composition further comprising a gel is described. The composition typically has a pH of about 5 to about 6, but the pH may range from about 4 to about 9.

[0148] All illustrative embodiments, variations thereof, and alternative forms thereof described below will apply to the embodiments described previously in paragraphs

[0015] to

[0017] in the Detailed Description and to the embodiments described in the Summary of the Invention.

[0149] A method for synchronizing the timing of pig insemination without estrus detection, comprising the step of administering a dose of a hormone to a pig, the hormone being, for example, gonadotropin-releasing hormone, luteinizing hormone, human chorionic gonadotropin; a derivative or analog of gonadotropin-releasing hormone, luteinizing hormone, or human chorionic gonadotropin; or a combination thereof. According to one embodiment, the hormone is administered to a pig. Any type of pig can be used for the methods described herein and for administering the compositions described herein, for example, gilts including adolescent gilts (i.e., female pigs before their first mating), and sows (including postpartum sows), or any other type of pig.

[0150] Illustratively, as described in the text, pigs are weaned on day 0. Four days after weaning (i.e., the fourth day after weaning), animals typically receive a single dose of hormones. Treated animals are typically inseminated a single time 15 hours (or 15 hours ± 2 hours), 16 hours (or 16 hours ± 2 hours), 17 hours (or 17 hours ± 2 hours), 18 hours (or 18 hours ± 2 hours), 19 hours (or 19 hours ± 2 hours), 20 hours (or 20 hours ± 2 hours), 21 hours (or 21 hours ± 2 hours), 22 hours (or 22 hours ± 2 hours), 23 hours (or 23 hours ± 2 hours), 24 hours (or 24 hours ± 2 hours), 27 hours (or 27 hours ± 2 hours) or 30 hours (or 30 hours ± 2 hours) after hormone administration.

[0151] Animals can be bred by any means of artificial insemination (AI) or by natural breeding. In any embodiment described herein, a second breeding or subsequent breeding can be performed. However, in another embodiment, the pigs are only inseminated once. In another illustrative aspect, there is no estrus detection. In another aspect, there is no estrus detection between the administration of hormones and 48 hours after ovulation.

[0152] In any embodiment described herein, the hormone is given at about 96 hours after weaning, i.e., on the fourth day after weaning. In various illustrative embodiments, the hormone can be given at about 80, about 82, about 84, about 86, about 88, about 90, about 91, about 92, about 93, about 94, about 95, about 96, about 97, about 98, about 99, about 100, about 102, about 104 or about 108 hours after weaning. More typically, the hormone is given at about 94-about 98 hours after weaning, i.e., about 94, about 95, about 96, about 97 or about 98 hours after weaning. In another embodiment, the hormone is given at about 92-about 106 hours after weaning.

[0153] In any of the embodiments described herein, for example, the pig is inseminated once about 15 to about 24 hours after the hormone is administered. In various further illustrative embodiments, the pig is inseminated about 15 to about 18 hours after the hormone is administered, about 13 to about 18 hours, about 15 to about 20 hours, or about 13 to about 20 hours after the hormone is administered. In other illustrative aspects, the pig is inseminated about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, or about 24 hours after the hormone is administered. Exemplarily, the pig is inseminated about 15 hours, about 18 hours, about 20 hours, about 22 hours, or about 24 hours after the hormone is administered.

[0154] In any embodiment described herein, relative to pigs not given hormones (e.g., given a placebo), the pregnancy rate of the pigs can be increased. For example, the pregnancy rate in hormone-treated animals can be about 75%-about 90%. In another embodiment, relative to pigs not given hormones (e.g., given a placebo), the litter size of the pigs increases. For example, the typical litter size in hormone-treated animals can be about 20-about 24 per pregnancy. In another embodiment, the litter size in hormone-treated animals can be about 14-about 18, about 14-about 24, or about 14-about 20 per pregnancy. However, in another embodiment, relative to pigs not given hormones (e.g., given a placebo), about 48 hours after hormone administration, the percentage of ovulated pigs can be increased. For example, about 48 hours after hormone administration, the percentage of ovulated animals can be about 65%-about 85%.

[0155] In other embodiments described herein, relative to pigs not given hormones, the total number of healthy fetuses can be increased. For example, the number of healthy fetuses of hormone-treated animals can be about 9-about 18, about 9-about 16, about 9-about 14, or about 9-about 12. In other embodiments described herein, when compared to pigs not given hormones, the litter percentage of hormone-treated animals can be increased. For example, the litter percentage of hormone-treated animals can be about 76%-about 90%, about 76%-about 85%, or about 76%-about 80%. In other embodiments described herein, relative to pigs not given hormones, the total number of piglets born of hormone-treated animals can be increased. For example, the number of piglets born can be about 13-about 18, about 13-about 17, about 13-about 16, about 13-about 15, or about 13-about 14. In any of the embodiments described herein, for hormone treated animals, the pregnancy rate of pigs, litter size of pigs, total number of healthy litters, litter percentage, and total number of piglets born can be similar to animals inseminated by estrus detection.

[0156] In any of the embodiments described herein, the total number of piglets born per semen dose may be increased relative to pigs not given hormones, and the total number of piglets born per semen dose may be increased relative to animals inseminated by estrus detection. For example, the total number of piglets born per semen dose may be about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 14, about 16, about 18, or about 20. In any of the embodiments described herein, the piglet index (survival piglets born / 100 sows allocated) may be increased relative to pigs not given hormones, and the piglet index may be increased relative to animals inseminated by estrus detection.

[0157] In any of the embodiments described herein, a composition for synchronizing the timing of porcine insemination without estrus detection comprises: a) a hormone; and b) a pharmaceutically acceptable pH buffer to provide a pH in the range of about pH 4 to about pH 9. The pH of the compositions described herein may be in the range of from about 4 to about 9. In other embodiments, the pH may be in the range of about 4 to about 8, about 4 to about 7, about 4.5 to about 6.5, about 4.5 to about 6, or about 5 to about 6.

[0158] In addition, the hormone composition can be produced by conventional methods according to the dosage form by appropriately mixing, diluting or dissolving additives, such as various excipients, disintegrants, binders, salts, lubricants, local anesthetics (e.g., lidocaine), diluents, preservatives, chelating agents, buffers, tonicity agents, antiseptic agents, wetting agents, emulsifiers, dispersants, stabilizers, solubilizers or combinations thereof.

[0159] Illustratively, the composition comprising the hormone may be in the form of a gel having a viscosity of about 10 (centipoise) cP to about 300,000 cP. In various illustrative embodiments, the viscosity of the composition may be about 100 cP to about 100,000 cP, about 250 cP to about 400 cP, about 300 cP to about 400 cP, about 500 cP to about 100,000 cP, about 700 cP to about 100,000 cP, about 200 cP to about 20,000 cP, about 200 cP to about 100,000 cP, about 1 ... 0,000 cP, about 200 cP to about 5,000 cP, about 200 to about 1,000 cP, about 200 cP to about 600 cP, about 100 cP to about 600 cP, about 100 cP to about 500 cP, about 200 cP to about 500 cP, about 200 cP to about 450 cP, or about 100,000 cP to about 250,000 cP. According to various embodiments described herein, the viscosity of the composition can be about 200 cP, about 250 cP, about 300 cP, about 400 cP, about 500 cP, about 1,000 cP, about 15,000 cP, about 20,000 cP, about 30,000 cP, about 40,000 cP, about 50,000 cP, about 75,000 cP, about 100,000 cP, about 200,000 cP or about 300,000 cP. The viscosity of the solution can be measured with a viscometer (such as a rheometer) based on known techniques in the art.

[0160] Typically, the gels described herein comprise: about 0.001-about 3.0% weight / weight (w / w) of the hormone or its salt, more typically about 0.5-5.0% (w / w) or about 0.1-5.0% (w / w) of the hormone or its salt, a preservative, a gelling agent (i.e., a viscosity adjusting agent), a buffer to maintain the pH between about 5 and about 6, and a tonicity agent to maintain the tonicity between about 200 and about 400 mOsm / kG.

[0161] According to any of the embodiments described herein, the composition has sufficient viscosity to allow the composition to remain adsorbed to the target tissue for a sufficient time to release an effective amount of the hormone. For example, typical viscosity depends on factors such as, for example, the permeability of the hormone applied and the amount of hormone. Suitable viscosity modifiers include, but are not limited to: ionic or non-ionic water-soluble polymers; cross-linked acrylic acid polymers; hydrophilic polymers such as polyethylene oxide, polyoxyethylene-polyoxypropylene copolymers and polyvinyl alcohol; cellulosic polymers and cellulosic polymer derivatives such as hydroxypropyl cellulose, hydroxyethyl cellulose, hydroxypropyl methylcellulose, hydroxypropyl methylcellulose phthalate, methylcellulose, carboxymethyl cellulose and etherified cellulose; gums such as tragacanth and xanthan gum; sodium alginate; gelatin, hyaluronic acid and its salts, chitosan, gellan or any combination thereof.

[0162] The viscosity modulator can be in the form of a gel, paste, cream, ointment, etc. In one embodiment, the composition comprises a hormone and a gel, the gel being used as a viscosity modifier, and the hormone is administered in a composition comprising the gel. In one embodiment, the gel is a hydrogel, a liposome gel, or a viscous solution. In another embodiment, the gel is a hydrogel. The gel can be prepared by any method known in the art, for example, such as those described in U.S. Patent Nos. 6,908,623 and 7,456,207, which are incorporated herein by reference.

[0163] In any embodiment described herein, the gel (i.e., viscosity modifier) ​​comprises a polysaccharide. According to the methods and compositions described herein, the polysaccharide may include, for example, alginate and glucose, such as glycogen, precipitate (e.g., amylose and amylopectin), cellulose, and dextran. The polysaccharide may be, for example, methyl, ethyl, or propyl cellulose ester, ether, hydroxy ether, hydroxy alkyl, or hydroxy ester. To achieve the required viscosity, one or more polysaccharides in sufficient amounts may be used. Typically, about 0.25-about 10% by weight of polysaccharide (based on the gross weight of the composition) is appropriate. In another embodiment, the weight % of the polysaccharide is about 0.25% by weight-about 3.0% by weight, about 1.0% by weight-about 7% by weight, about 1.0% by weight-about 4.0% by weight, or about 1.0% by weight-about 2.0% by weight. In other embodiments, the weight % of the polysaccharide is about 0.1%, about 0.5%, about 0.75%, about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.4%, about 1.8%, about 2.0%, about 5%, about 8%, or about 10% (all by weight / weight). To increase the viscosity of the composition, the polysaccharide may be used with one or more non-polysaccharide viscosity enhancers known in the art. Examples of possible non-polysaccharide viscosity enhancers that can be used with one or more polysaccharides include xanthan gum, alginic acid and its salts, magnesium aluminum silicate, dextrins, sucrose and its derivatives and mixtures thereof. The amount of non-polysaccharide viscosity enhancer (if present) may be about 0.1 weight % to about 10 weight %, depending on the desired viscosity.

[0164] In any embodiment described herein, the gel comprises cellulose. The cellulose of the illustrative embodiments described herein includes methylcellulose, ethylcellulose, hydroxypropylcellulose, carboxymethyl (carbomethyl) cellulose, hydroxypropylmethylcellulose and hydroxyethylmethylcellulose. The cellulose can be a cellulose derivative, preferably a nonionic cellulose ester, ether, hydroxyether or hydroxyester, or a nonionic starch derivative. Typically, suitable cellulose is about 0.25 wt %-about 10 wt % (based on the gross weight of the composition). In another embodiment, the weight % of cellulose is about 0.25 wt %-about 3.0 wt %, about 0.5 wt %-about 3.0 wt %, about 0.5 wt %-about 4.0 wt %, about 1.0 wt %-about 7 wt %, about 1.0 wt %-about 4.0 wt % or about 1.0 wt %-about 2.0 wt %. In other embodiments, the weight % of cellulose is about 0.1%, about 0.5%, about 0.75%, about 0.8%, about 0.9%, about 1.0%, about 1.1%, about 1.2%, about 1.4%, about 1.8%, about 2.0%, about 5%, about 8%, or about 10% (all on a weight / weight basis). If a uniform gel is desired, a dispersant such as an alcohol, sorbitol, or glycerol may be added, or the gelling agent may be dispersed by tituration, mechanical mixing or stirring, or a combination thereof.

[0165] Acceptable stabilizers for the described methods and compositions include L-amino acids and L-methionine. In other embodiments, stabilizers that can be used include, but are not limited to, polysaccharides (such as gum arabic, agar, alginic acid, guar gum, and tragacanth), gelatin, and synthetic and semi-synthetic polymers (such as carbomer resins, cellulose ethers, and carboxymethyl chitin). The amount of the stabilizer is typically about 0.05-about 10%, about 0.05-about 5%, about 0.05-about 2.0%, about 0.05-about 1.0%, about 0.05-about 0.5%, about 0.05-about 0.2%, about 0.1-about 5%, about 0.1-about 10%, about 0.1-about 20%, about 1-about 5%, about 1-about 10%, about 1-about 20% (all by weight / volume). In one embodiment, in the presence of a stabilizer described herein, the composition can have a shelf life of at least 12 months, at least 18 months, or at least 24 months. In another embodiment, the composition can be stored at a temperature ranging from about 2°C to about 8°C. Inert carriers such as lactose, starch, dextrin, dicalcium phosphate, and calcium sulfate may also be included. In one embodiment including a stabilizer, the composition is chemically stable and maintains at least 99% purity, at least 99.5% purity, or at least 99.7% purity for at least three months.

[0166] The tonicity agent can be nonionic or ionic. Illustratively, acceptable tonicity agents for use in the described methods and compositions include, for example, ionic agents such as sodium chloride, potassium chloride, or balanced salt solutions. According to one embodiment, the tonicity agent is present in an amount to achieve a tonicity of about 200-400 mOsm / kG, about 220-380 mOsm / kG, or about 250-340 mOsm / kG. Nonionic tonicity agents include glycols such as glycerol, mannitol, erythritol, polyethylene glycol, propylene glycol; and sugars such as sucrose and dextrose. The amount of tonicity agent is typically from about 0.05 to about 10%, about 0.05 to about 5%, about 0.05 to about 2.0%, about 0.05 to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about 0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, 0.5 to about 2.0%, about 0.6 to about 2.0%, about 0.5 to about 1.8%, about 0.6 to about 1.8%, about 1.0 to about 5.0%, about 1.0 to about 10%, or about 1.0 to about 20% (all on a weight / volume basis).

[0167] In any embodiment described herein, the pH buffers used in the compositions and methods described herein are those known to the skilled person as pH buffers or compositions, and include, for example, acetate, borate, carbonate, citrate and phosphate buffers, and various biological buffers, for example, TAPS, N,N-bicine (Bicine), Tris, N-tris(hydroxymethyl)methylglycine (Tricine), HEPES, TES, MOPS, PIPES, cacodylate and MES. Other pH buffers include hydrochloric acid, sodium hydroxide, magnesium oxide, potassium dihydrogen phosphate, bicarbonate, ammonia, carbonic acid, sodium citrate, citric acid, acetic acid, disodium hydrogen phosphate, borax, boric acid, etc. The amount of buffer is typically about 0.01 to about 10%, about 0.02 to about 10%, about 0.02 to about 5%, about 0.02 to about 2.0%, about 0.02 to about 1.0%, about 0.02 to about 0.5%, about 0.05 to about 10.0%, about 0.05 to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about 0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1 to about 5%, about 1 to about 10%, about 1 to about 20% (all on a weight / volume basis).

[0168] The buffer used for the formulations described herein can be used in any concentration required to obtain the required pH range. For example, the buffer can be used in the following concentrations: about 0.001M-about 1M, about 0.001M-about 2M, about 0.001M-about 5M, about 0.05M-about 0.1M, about 0.05M-about 0.2M, about 0.05M-about 1M, 0.05M-about 2M, about 0.05-about 5M, about 0.1M-about 1M, about 0.1M-about 2M, about 0.1M-about 5M. Any amount of the buffer required to obtain the required pH range can be used for the formulations described herein. Typically, pharmaceutically acceptable pH buffers can be used to provide a pH range of about pH 4-about pH 9. The pH range of the compositions described herein can be about 3-about 10 or about 4-about 9. In any of the embodiments described herein, the pH range can be about 4 to about 8, about 4 to about 7, about 4.5 to about 6.5, about 4.5 to about 6, about 5 to about 6, about 5 to about 5.5, about 4 to about 6, or about 4.5 to about 5.5.

[0169] In any of the embodiments described herein, the compositions described herein comprise one or more pharmaceutically acceptable preservatives. As used herein, the term "preservative" includes an agent or combination of agents that helps stabilize the composition, inhibit microbial growth, or both. Examples of suitable preservatives include parabens (e.g., methyl, ethyl, propyl and butyl esters of parabens), propyl gallate, sorbic acid and its sodium and potassium salts, propionic acid and its calcium and sodium salts, "dioxane" (6-acetoxy-2,4-dimethyl-m-dioxane), "bronopol" (2-bromo-2-nitropropane-1,3-diol) and salicylanilides (e.g., dibromosalicylanilide, tribromosalicylanilide), "Cinaryl" 100 and 200 or "Dowicil" 100 and 200 (the cis isomer of 1-(3-chloroallyl-3,5,7-triaza-1-azanidadamantane chloride) , hexachlorophene, sodium benzoate, citric acid, ethylenediaminetetraacetic acid and its alkali metal and alkaline earth metal salts, butylated hydroxyanisole, butylated hydroxytoluene, phenolic compounds (such as chloro- and bromocresols and chloro- and bromo-oxylenol), quaternary ammonium compounds (such as benzalkonium chloride), aromatic alcohols (such as phenylethyl alcohol, benzyl alcohol, etc.), chlorobutanol, quinoline derivatives (such as iodochlorohydroxyquinoline), etc. When the preservative is present, the total amount is about 0.005% to about 2% by weight, about 0.001% to 1.0% by weight, about 0.005% to about 0.25% by weight, or about 0.05% to about 0.2% by weight, typically about 0.01% to about 0.1% by weight (all on a weight / weight basis).

[0170] In any embodiment described herein, the pharmaceutical composition contains a chelating agent, such as those known to those skilled in the art, for example, ethylenediaminetetraacetate (EDTA), diethylenetriaminepentaacetic acid (DTPA) and N,N-di(carboxymethyl)glycine (NTA) or salts thereof. The composition may contain about 0.003 wt% to about 1.0 wt%, about 0.02 wt% to about 0.2 wt%, about 0.01 wt% to about 1.0 wt%, or about 0.02 wt% to about 0.5 wt% (all on a weight / volume basis) of the chelating agent.

[0171] In any embodiment described herein, an antimicrobial agent may be included in the composition described herein. Such agents may include, but are not limited to, 5-chloro-2-(2,4-dichlorophenoxy)-phenol, 8-hydroxyquinoline, copper II compounds, phthalic acid, chlorhexidine, Alexidine, hexetidine, sanguinarine, benzalkonium chloride, salicylanilide, domiphen bromide, cetylpyridinium chloride, tetradecylpyridinium chloride, N-tetradecyl-4-ethylpyridinium chloride, octenidine, iodine, sulfonamides, bisbiguanides, phenolic compounds, delmopinol, octacyclol and other piperidinyl derivatives, and nicotinic acid preparations, any applicable antibiotics such as levotetin, amoxicillin, tetracycline, doxycycline, minocycline, metronidazole, neomycin, kanamycin and clindamycin, and any salts of any of these compounds applicable, and any combination of these compounds. However, in another embodiment, an antifungal compound may be included, either alone or in combination with any of the above antimicrobial agents. Antifungal agents suitable for use in the compositions described herein include, but are not limited to, nystatin, miconazole, econazole nitrate, clotrimazole, and flucytosine. The amount of antimicrobial or antifungal agent that can be added to the formulations described herein is about 0.01 to about 10%, about 0.01 to about 5%, about 0.01 to about 2.0%, about 0.01 to about 1.0%, about 0.01 to about 0.5%, about 0.01 to about 0.2%, 0.05 to about 10%, about 0.05 to about 5%, about 0.05 to about 2.0%, about 0.05 to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about 0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1 to about 5%, about 1 to about 10%, about 1 to about 20% (all on a weight / volume basis).

[0172] In any embodiment described herein, an antioxidant may also be added. For example, the antioxidants used herein include beta-carotene, vitamin E, vitamin C, vitamin A, tocopherol, butylated hydroxytoluene, butylated hydroxyanisole, tert-butylhydroquinone, propyl gallate, ascorbic acid, sodium metabisulfite, uric acid, carotenoids, flavonoids, melatonin and ethoxyquin. Antioxidants can be added to the formulations described herein in amounts of about 0.01 to about 10%, about 0.01 to about 5%, about 0.01 to about 2.0%, about 0.01 to about 1.0%, about 0.01 to about 0.5%, about 0.01 to about 0.2%, 0.05 to about 10%, about 0.05 to about 5%, about 0.05 to about 2.0%, about 0.05 to about 1.0%, about 0.05 to about 0.5%, about 0.05 to about 0.2%, about 0.1 to about 5%, about 0.1 to about 10%, about 0.1 to about 20%, about 1 to about 5%, about 1 to about 10%, about 1 to about 20% (all on a weight / volume basis).

[0173] As described herein, the composition contains a hormone selected from, for example, gonadotropin-releasing hormone, luteinizing hormone, human chorionic gonadotropin, their derivatives or analogs, and combinations thereof, which, when used in the methods described herein, are used in an amount effective to synchronize the time of pig insemination without estrus detection. Other examples of acceptable hormones for the methods and compositions described herein include: prostaglandins, progestins, progesterone, androgens, testosterone, estrogens, estradiol, gonadotropins, their derivatives or analogs, their combinations, and the like. The hormone can be in the form of acetate. In addition, the hormone can be a gonadotropin-releasing hormone, luteinizing hormone, or human chorionic gonadotropin agonist, or a gonadotropin-releasing hormone, luteinizing hormone, or human chorionic gonadotropin antagonist. As used herein, "gonadotropin-releasing hormone" refers to any gonadotropin-releasing hormone, including gonadotropin-releasing hormone analogs and derivatives, and gonadotropin-releasing hormone agonists and antagonists. In one embodiment, the gonadotropin-releasing hormone can be a synthetic substance. In another embodiment, the gonadotropin-releasing hormone may be GnRH (pGlu-His-Trp-Ser-Tyr-Gly-Leu-Arg-Pro-GlyNH 2) (see, e.g., U.S. Pat. No. 5,688,506, which is incorporated herein by reference) or triptorelin. As used herein, "luteinizing hormone" refers to any luteinizing hormone, including luteinizing hormone analogs and derivatives and luteinizing hormone agonists and antagonists. In one embodiment, the luteinizing hormone may be a synthetic substance. In another embodiment, the luteinizing hormone may be LH (see, e.g., U.S. Pat. No. 5,444,167, which is incorporated herein by reference). As used herein, "human chorionic gonadotropin" refers to any human chorionic gonadotropin, including human chorionic gonadotropin analogs and derivatives, and human chorionic gonadotropin agonists and antagonists. In one embodiment, the human chorionic gonadotropin may be a synthetic substance. In another embodiment, the human chorionic gonadotropin can be hCG (see, eg, US Patent Nos. 6,469,139, 4,400,316, and 4,804,626, which are incorporated herein by reference).

[0174] Examples of gonadotropin-releasing hormone agonists useful herein include, but are not limited to, leuprolide, nafarelin, buserelin, [DAla 6 , go to Gly-NH 2 10 ]GnRH, [DLys 6 ]GnRH,[DAla 6 ]GnRH, [2-Me-Ala 6 ]GnRH, [D-α-aminobutyryl 6 , de-GlyNH 2 10 ] GnRH, triptorelin, luteolin, goserelin, deslorelin and histrelin. In general, gonadotropin-releasing hormone agonists mimic the natural gonadotropin-releasing hormone decapeptide, typically shaped by specific amino acid substitutions at positions 6 and 10. Triptorelin is an example of a gonadotropin-releasing hormone agonist that has only a single substitution at the 6-position.

[0175] Examples of gonadotropin-releasing hormone antagonists include anti-peptides (with the formula D-Ac-D-2-Nal 1 -DpClPhe 2 -D-3-Pal 3 -Ser4-NiLys 5 -D-NicLys 6 -Leu 7 -ILys 8 -Pro 9 -D-Ala 10 The decapeptide represented by [Ac-D4ClDPhe 1 ,D4ClDPhe2 ,DTrp 3 ,DArg 6 ,DAla 10 ]GnRH, [Ac-4ClDPhe 2 ,D 3 Pal 3 ,Arg 5 ,D 2 Nal 6 ,DAla 10 ]GnRH, [Ac-D2-Na l 1,4ClDPhe 2 ,DTrp 3 ,DArg 6 ,DAla 10 ]GnRH, [Ac-D2Nal 1 ,4FDPhe 2 ,DTrp 3 ,DArg 6 ]GnRH, [Ac-D2Nal 1 ,4ClDPhe2,DTrp 3 ,DhArg(Et 2 ) 6 ,DAla 10 ]GnRH and [Ac-Na l 1,DME4ClPhe 2 ,DPal 3 ,Ser 4 ,Tyr 5 ,DArg 6 ,Le 7 ,ILys 8 ,Pro 9 ,DAla 10 ]GnRH.

[0176] In any of the embodiments described herein, a gonadotropin-releasing hormone agonist of the formula

[0177]

[0178] or its solvate, hydrate or pharmaceutically acceptable salt,

[0179] in

[0180] R 1 and R 2 is independently at each occurrence hydrogen, or is independently selected from: alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, haloalkyl, aryl, heteroaryl, arylalkyl and heteroarylalkyl, each of which is optionally substituted, or R 1 and R2 forms a carbocyclic or heterocyclic ring with the attached carbon; and

[0181] X is hydrogen, or X is selected from: alkyl, cycloalkyl, heteroalkyl, optionally substituted alkylene-carboxamide, and HNC(O)NR 3 R 4 , where R 3 and R 4 Each occurrence is independently selected from the group consisting of: hydrogen, alkyl, heteroalkyl, and haloalkyl.

[0182] In another embodiment, the above use is described, wherein R 1 is methylene-heteroaryl, wherein heteroaryl is selected from the group consisting of pyridyl, thiazolyl, sulfopyridinyl, pyrimidinyl, quinolinyl, pyrazolyl, imidazolyl, pyrrolyl, indolyl, benzopyrazolyl and benzimidazolyl; and R 2 is hydrogen or methyl.

[0183] In yet another embodiment, any of the uses described above is described, wherein X is CH 2 C(O)NH 2 .

[0184] The gonadotropin-releasing hormone agonists and antagonists used herein and their analogs, such as the analogs described above, can be administered in the form of pharmaceutically acceptable non-toxic salts or complexes. The salts include acid addition salts, such as hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, oxalates, fumarates, gluconates, tannates, maleates, acetates, benzoates, succinates, alginates, malates, ascorbates, tartrates, etc. The complexes can contain metals, such as zinc, barium, calcium, magnesium, aluminum, etc.

[0185] The effective amount of a hormone used in accordance with the methods and compositions described herein depends on a number of parameters, including the molecular weight of the hormone, its route of administration, and whether it is in its native form. In the description herein, an "effective amount" of a hormone is an amount sufficient to synchronize the time of insemination of pigs without estrus detection using the methods described herein. An effective amount of a hormone administered to a pig can range from about 10 μg to about 2000 μg, about 10 μg to about 1000 μg, about 10 μg to about 500 μg, about 10 μg to about 100 μg, about 10 μg to about 50 μg, about 50 μg to about 2000 μg, about 50 μg to about 1000 μg, about 50 μg to about 500 μg, about 50 μg to about 300 μg, about 50 μg to about 200 μg, about 100 μg to about 200 μg, about 100 μg to about 300 μg, about 100 μg to about 500 μg, about 100 μg to about 1000 μg, about 200 μg to about 2000 μg, or about 0.05 mg to about 50 mg. In various exemplary aspects, the dose of hormone administered to the pig is about 20 μg, about 50 μg, about 75 μg, about 100 μg, about 150 μg, about 180 μg, about 200 μg, about 225 μg, about 250 μg, about 300 μg, about 400 μg, about 500 μg, about 750 μg, about 1000 μg, about 1500 μg or about 2000 μg of hormone. The hormone can be administered in one or more doses.

[0186] The hormone in the compositions described herein can be administered at a concentration of about 50 μg / mL to about 500 μg / mL, about 50 μg / mL to about 400 μg / mL, about 50 μg / mL to about 300 μg / mL, about 50 μg / mL to about 200 μg / mL, about 50 μg / mL to about 150 μg / mL, about 50 μg / mL to about 250 μg / mL, or about 100 μg / mL. The compositions can be administered in various volumes, for example, including a dosage volume of 1 mL, 2 mL, 3 mL, 4 mL, or 5 mL.

[0187] In any of the embodiments described herein, the hormone is administered in an amount effective to stimulate ovulation of the ovarian follicle and to effectively synchronize ovulation according to the methods described herein. The dose of hormone can be administered using a method selected from the following: 1) use of a catheter, 2) manual administration, 3) injection, or any other means known in the art for administering a pharmaceutical composition, for example, any other means known in the art for vaginal administration of a pharmaceutical composition (such as a composition comprising a hormone). In one embodiment, the hormone can be administered to more than one pig.

[0188] Examples of methods for effective hormone administration, in addition to vaginal administration, include parenteral administration to animals, for example, subcutaneous, intramuscular, intraperitoneal, intrathecal or intravenous administration, or in combination with an acceptable carrier. Suitable methods for parenteral administration include needle (including microneedle) injection, needle-free injection and infusion techniques. The parenteral composition used in the present invention can be a reconstituted lyophilized form containing one or more doses of hormone composition. Examples of parenteral dosage forms include aqueous solutions of the composition in known acceptable liquid carriers, such as liquid alcohols, glycols (e.g., polyethylene glycol), glucose solutions (e.g., 5%), esters, amides, sterile water, buffered saline (including buffers such as phosphates or acetates; e.g., isotonic saline).

[0189] In any embodiment described herein, the composition can be administered locally to an animal. Examples of topical administration methods used herein include: topical, intravaginal, and intrarectal administration. Examples of dosage forms used in this embodiment include creams, ointments, gels, pastes, powders, lotions, transdermal patches, intrauterine devices, vaginal rings, and vaginal tablets. In an illustrative embodiment, the composition is administered to the anterior vagina of an animal. The compound can also be formulated into vaginal or rectal compositions such as suppositories, for example, containing general suppository bases, such as cocoa butter, carbowax, polyethylene glycols, or other glycerides, all of which melt at body temperature and solidify at room temperature.

[0190] The hormones may be administered to the animal by any useful procedure and in any effective dose and suitable dosage form, including oral dosage forms known in the art, such as pills, tablets or capsules, and the effective dose may be administered in a standard dosage form or a sustained release dosage form. Sustained release dosage form formulations include delayed, sustained, pulsatile, controlled, targeted and programmed release formulations.

[0191] The composition may also contain a suitable solid or gel phase carrier or excipient. Examples of such carriers or excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin, and polymers (such as polyethylene glycols).

[0192] In another illustrative aspect of the invention, a kit is provided. The kit comprises one or more doses of a hormone as described herein. In this embodiment, the kit may also comprise an applicator for manual administration, a catheter for placement, and / or a syringe for administering the hormone composition to an animal. In another embodiment, the hormone is in a composition comprising a gel as described herein. In an illustrative embodiment, the kit may comprise a separate hormone and gel for mixing before administration to an animal. In another embodiment, the kit comprises a hormone and gel mixed in a container for direct administration.

[0193] In another embodiment, the kit contains instructions for use. The instructions may indicate that the insemination should be by natural mating; the insemination should be by artificial insemination; the pig should be inseminated about 15-about 18, about 18-about 22, or about 15-about 24 hours after the hormone is administered, or the pig should be inseminated about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, or about 30 hours after the hormone is administered. Other applicable kit components include: excipients, disintegrants, binders, salts, lubricants, local anesthetics (e.g., lidocaine), diluents, preservatives, chelating agents, buffers, tonicity agents, antibacterial agents, wetting agents, emulsifiers, dispersants, stabilizers, etc. These ingredients can be available separately or mixed with hormones and / or gels when necessary. Any hormone embodiment and any composition embodiment described herein can be used to prepare a kit.

[0194] In another embodiment, the article of manufacture is provided. The article of manufacture may include any composition described herein. The composition may be in a primary container, such as a glass vial (such as a brown glass vial with a rubber stopper and / or an aluminum seal). In another embodiment, the primary container may be plastic or aluminum, and the primary container may be sealed. In another embodiment, the primary container may be contained in a secondary container to further protect the composition from light. The secondary container may be, for example, cardboard. Any of these embodiments may also be used in the above-mentioned kit embodiment, and any of the hormone and composition embodiments described herein may be used in the article of manufacture. Example

[0195] Example 1

[0196] Study Design and Treatment Groups

[0197] About 120 sows with parity from one to seven (30 sows per treatment group) were used to obtain ovulation and reproductive tract data. All sows were of the same genotype (PIC C22). On days 16-24 of lactation, sows were divided according to the length of lactation and parity and randomly assigned to one of four treatment groups: 1) a placebo group that received a placebo gel 96 ± 2 hours after weaning and a single AI 24 hours thereafter, 2) a triptorelin gel 96 ± 2 hours after weaning and an AI 18-20 hours thereafter. 3) a triptorelin gel 96 ± 2 hours after weaning and an AI 24-26 hours thereafter, and 4) a triptorelin gel 96 ± 2 hours after weaning and an AI 30-32 hours thereafter. The placebo formulation was identical to the active formulation but without the presence of hormones. All treatments were placed within 2 cm of the anterior vaginal cervix. Follicular development and ovulation were monitored by real-time ultrasound imaging at 24, 32, 40, 48 and 56 hours after treatment. Thirty (30) days after insemination, all pigs were slaughtered and the reproductive tracts were obtained. Pregnancy rate and embryo survival were determined.

[0198] Example 2

[0199] Test substance

[0200] The active ingredient is triptorelin (pGlu-His-Trp-Ser-Tyr-D-Trp-Leu-Arg-Pro-Gly-NH 2 ), which was provided as the acetate salt (MW: 1371.6) from Bachem, Torrance, CA (Catalog No. H-4075 CGMP grade). Triptorelin gel (200 μg / 2 mL), batch 054023833-1, was prepared by Chem Laboratories Ltd (Auckland, New Zealand) in a 1.2% Methocel in sodium citrate, pH 5.5. TM Premium A4000 (Dow Chemicals) in a gel consisting of methyl and propyl parabens, NaCL, EDTA and L-methionine. The formulation vehicle, batch 054006645-1, was formulated at Chem Laboratories Ltd. and consisted of 1.2% Methocel in sodium citrate, pH 5.5. TMPremium A4000 (Dow Chemicals) was composed with NaCl and methyl and propyl parabens. Fifty milliliters of triptorelin gel (200 μg triptorelin acetate / mL) or formulation vehicle were packaged in 50 mL brown borosilicate glass serum bottles (610206-50) with gray butyl rubber pharmaceutical serum bottle stoppers (73828A-SS) and standard aluminum closures (SAS20NAT). The test materials were stored frozen (approximately 5°C) and transported in insulated containers with appropriate ice packs.

[0201] Example 3

[0202] Example Formulation

[0203] Example formulations for the compositions described herein are shown in Tables 1 and 2.

[0204] Table 1

[0205] Element Function Weight (%w / v) Methylparaben, sodium salt (USNF) Antimicrobial preservatives 0.0900 Propylparaben, sodium salt (USNF) Antimicrobial preservatives 0.0100 Sodium Chloride, Laboratory Reagent Tonicity Agents 0.910 Sodium citrate, dihydrate Buffer 0.186 L-Methionine, laboratory reagent Stabilizer 0.100 Citric Acid, Anhydrous Buffer 0.0700 Triptorelin acetate Active Pharmaceutical Ingredients (API) 0.0100 Water(USNF) Dissolving solvent 98.4 Methylcellulose (A4MPremium) (USP) Thickener 1.20

[0206] Table 2.

[0207]

[0208] *Nominal weight

[0209] **Tested to compendial standard

[0210] Example 4

[0211] Estrus observation

[0212] Pigs were housed in gestation crates after weaning (day 0). Boars were housed in separate rooms and / or at least 12 m away and downwind. To determine the onset and duration of estrus, sows were observed daily for estrus, (1) from day 3 until the end of estrus was confirmed, or (2) until day 6, whichever came first. To elicit estrus signals, a mature boar was allowed to walk slowly through a passage in front of the sow crate, and each test sow was exposed to visual, auditory, and olfactory cues from the boar for up to 5 minutes. In keeping with standard practice on commercial farms, estrus was tested by an experienced person applying back pressure to the midsection of the sow in combination with lateral rubbing as the boar approached the front of the sow crate. Estrus was confirmed when the sow showed signs of standing upright in response to back pressure, no vocalization, and some ear reflexes.

[0213] Example 5

[0214] Drug administration and insemination

[0215] Triptorelin was placed about 1 to about 2 cm posterior to the cervix using a modified artificial insemination catheter as a single 2 mL dose at 96 hours (± 2 hours) after weaning. The pigs were weaned between 10 a.m. and noon on day (0). The insemination schedule for each group of sows was given as indicated in the experimental design. Those sows scheduled to be inseminated 18 hours after the administration of triptorelin were generally inseminated between 6:00 a.m. and 8:00 a.m. on the following day, those sows scheduled to be inseminated 24 hours after the administration of triptorelin were generally inseminated between noon and 2:00 p.m. on the following day, and those sows scheduled to be inseminated 30 hours after the administration of triptorelin were generally inseminated between 6:00 p.m. and 8:00 p.m. on the following day. The single insemination of each sow was performed as they were routinely performed on the farm.

[0216] Example 6

[0217] Ovulation and estrus determination

[0218] All sows were observed by transrectal ultrasound scanning. Ultrasound scanning was performed 24 hours (± 1.5 hours) after treatment on the 5th day and again at 8:00 p.m. (± 1.5 hours) on the 5th day. Transrectal ultrasound scanning was performed at 4:00 a.m. (± 1 hour), noon (± 1 hour) and 8:00 p.m. (± 1 hour) on the 6th day until ovulation was completed or until the ultrasound scanning at 8:00 p.m. on the 6th day (56 hours from the 96-hour animal treatment after weaning), whichever came first. Aloka 500 ultrasound instrument was used for this purpose, with a 7.5MHz linear array transducer connected to a fixed-angle PVC stabilizing rod for easy insertion into the rectum. The transducer and PVC rod were coated with gynecological lubricant and gently inserted into the rectum until the ovary was visible. The diameter of the three largest follicles (to the nearest 0.1mm) was recorded for each scan. When the number of large follicles (≥ 6.5mm) dropped to less than 3, the sow was said to have ovulated.

[0219] On day 5 post-weaning, the average size of the largest follicle was 7.3 mm and was not affected by treatment. The rate of estrus expression within 7 days of weaning averaged 79% and did not differ significantly among treatments. Treatment also had no effect on the interval from weaning to estrus (average 112 hours).

[0220] The percentage of sows that had ovulated at 24, 32 or 40 hours after treatment was not significantly affected by treatment (see Tables 3 and Figure 1). However, the percentage of sows ovulating 48 hours after treatment was increased in all triptorelin treatment groups [74.0% (at 18 hours AI), 77.0% (at 24 hours AI), and 76.8% (at 30 hours AI)] compared to vehicle treated sows (42.6%) (p=0.0054). Treatment did not affect the percentage of sows ovulating 56 hours after treatment (p=0.10), however, sows treated with triptorelin had a higher percentage of sows ovulating (average 80.4%) compared to sows treated with vehicle treatment (58.3%). There was no difference in the mean weaning-to-ovulation interval between triptorelin-treated and vehicle-treated animals, but more synchronous ovulation was observed in triptorelin-treated sows, with 70-77% of sows ovulating during their most synchronous 24-hour cycle (120-144 hours after weaning) compared to only 39.9% of vehicle-treated sows ovulating during their most synchronous cycle (128-156 hours after weaning).

[0221] Example 7

[0222] Pregnancy rate and litter size

[0223] Pregnancy rates, calculated as the percentage of animals assigned to each treatment group that were pregnant at slaughter, averaged 66.6% and did not differ significantly among treatment groups, although pregnancy rates were numerically highest in triptorelin-treated sows inseminated 18 hours post-treatment (74.4%), followed by triptorelin-treated sows inseminated 24 hours post-treatment (68.1%), triptorelin-treated sows inseminated 30 hours post-treatment (62.3%), and placebo-treated sows inseminated 24 hours post-treatment (61.8%). There was a trend (p=0.09) for triptorelin-treated sows inseminated 18 hours post-treatment (12.4 parities) and 24 hours post-treatment (11.8 parities) to have a higher number of healthy fetuses than triptorelin-treated sows inseminated 30 hours post-treatment (8.6 parities) and vehicle-treated sows inseminated 24 hours post-treatment (8.6 parities). There were no significant differences in healthy fetal weights among the treatment groups.

[0224] Table 3. Least Squares (LS) Means for Response Variables

[0225]

[0226] Means with different superscripts in a row are significantly different at p = 0.05

[0227] The abbreviations are as follows: N (number of sows), estrus (estrus during the cycle from day 3 to day 6), OV24 (percentage of sows ovulating 24 hours after treatment), and pregnancy (pregnant sows among all allocated sows).

[0228] AIOVh standard error: 1.8379 1.5804 1.5483 1.5700

[0229] #Standard error of healthy fetus: 1.3966 1.3966 1.3968 1.4105

[0230] Example 8

[0231] Study 1 - Combined Feeding Research Farm

[0232] Three study replicates were conducted on a research farm of 800 sows. All replicates included data on estrus and ovulation, and two replicates included results for AI, pregnancy, and farrowing. At weaning, mixed parity sows (n=32 / treatment) received vehicle 96 h after weaning and a single AI 24 h thereafter (placebo), test substance OVUGEL 96 h after weaning, and placebo 24 h thereafter. TM and a single AI 18 hours later (OG18), and OVUGEL 96 hours after weaning TM and a single AI 24 hours thereafter (OG24) or OVUGEL 96 hours after weaning TM and a single AI 30 hours later (OG30). TM It was given to 2 cm behind the cervix. Estrus detection was performed daily from day 3 (d3) until day 7 (d7) after weaning. Ultrasound was performed every 8 h from d 5 to d 6. On d30 of pregnancy, the pigs were sacrificed and the reproductive tract was examined.

[0233] Treatment had no effect on the number of sows presenting estrus within 7 days of weaning (79%) or on the interval from weaning to estrus (4.7 days). The percentage of sows ovulating 48 h after treatment was greater in OG18, OG24, and OG30 compared with placebo treatment (P = 0.005) (Table 4). The interval from AI to ovulation differed among treatment groups (P < 0.001, Table 4), with the interval furthest from ovulation occurring in OG18, intermediate in placebo and OG24, and closest in OG30. Pregnancy rate was not affected by treatment (P = 0.60, Table 4), but was affected by the interval from AI to ovulation (P = 0.02). Treatment had a significant effect on the number of healthy fetuses at day 30 (P = 0.003), with OG18 and OG24 having more healthy fetuses than OG30 and placebo treatments (Table 4). Treatment had no effect on the number of CLs (P = 0.30), but showed a trend towards an effect on embryonic survival (P = 0.09) (Table 4).

[0234] The results showed that OVUGEL TMEffectively synchronize ovulation without affecting pregnancy rates, and OVUGEL TM Compared with OVUGEL after 18 or 24 hours of single AI TM Litter size was larger at gestational day 30 after 30 hours of AI or after a single insemination compared with placebo treatment.

[0235] Table 4. Unadjusted Means for Treatment Response

[0236] Placebo OG18 OG24 OG30 SE Ovulation within 48h, % <![CDATA[40.6 a ]]> <![CDATA[71.9 b ]]> <![CDATA[75.0 b ]]> <![CDATA[75.0 b ]]> 4.2 AI* to ovulation (h) <![CDATA[20.3 a ]]> <![CDATA[26.4 b ]]> <![CDATA[19.1 a ]]> <![CDATA[15.5 c ]]> 1.0 Pregnant(%) 68.2 86.4 81.8 68.2 4.6 fetal <![CDATA[12.9 a ]]> <![CDATA[16.5 b ]]> <![CDATA[17.4 b ]]> <![CDATA[12.6 a ]]> 0.6 CL quantity 18.5 22.1 23.0 17.6 1.2 ES(%) 53.2 65.9 67.3 57.0 2.4

[0237] AI = Artificial Insemination

[0238] CL = Corpus Luteum

[0239] ES = embryonic survival

[0240] OG=OVUGEL TM

[0241] Example 9

[0242] Study 2 - Bache Farm Study

[0243] Sows were weaned (0 days) and screened for inclusion in the study, with body condition recorded and assigned to treatment groups. Postpartum sows from parity 1-10 were divided into zones based on postpartum length, parity and body condition score. Sows with body condition scores below 2.5 or above 3.5 were not used. The genotypes used were those typically used commercially in the United States and Canada. The treatment groups were housed in separate areas so that those sows that were mated later in the day would not receive multiple encounters with boars. Each treated sow received a single dose of triptorelin combination (OVUGEL TM ). Control sows were not treated.

[0244] Estrus determination was performed on day 4 and continued daily until 4 days after treatment (day 8) or until the sows no longer showed estrus. Control sows (untreated) were inseminated during the intervals after estrus detection as routinely practiced on the farm. OVUGEL was inseminated with a single insemination at 15 hours (6:00 am + / - 0.5 hours), 18 hours (9:00 am + / - 0.5 hours), 21 hours (12:00 pm + / - 0.5 hours), 24 hours (3:00 pm + / - 0.5 hours) and 27 hours (6:00 pm + / - 0.5 hours) after treatment. TM Treated sows were inseminated.

[0245] Estrus determination was performed daily between 18-24 days after insemination to determine if the sows were recirculating. Sows that had returned to estrus were recorded and then re-bred according to standard farm standard operation (SOP) (these animals left the study and received a withdrawal period). Pregnancy in those sows that did not return to estrus was confirmed by transabdominal ultrasound (once) 28-32 days after insemination. The withdrawal period was confirmed to be 51 days after treatment. Data on farrowing date, number of live piglets born 24 hours after birth, and number of stillborn piglets were recorded. Piglets were raised according to usual farm protocols.

[0246] At least 90 sows were assigned to each of the following groups. Control sows were not treated. All other sows were given OVUGEL containing 200 μg of triptorelin as acetate. TM .

[0247] 1) Control: untreated and inseminated at intervals after estrus detection as routinely performed at each site;

[0248] 2) Triptorelin: insemination once 15 hours (+ / - 0.5 hours) after triptorelin treatment;

[0249] 3) Triptorelin: insemination once 18 hours (+ / - 0.5 hours) after triptorelin treatment;

[0250] 4) Triptorelin: insemination once 21 hours (+ / - 0.5 hours) after triptorelin treatment;

[0251] 5) Triptorelin: insemination once 24 hours (+ / - 0.5 hours) after triptorelin treatment;

[0252] 6) Triptorelin: Insemination was performed once 27 hours (+ / - 0.5 hours) after triptorelin treatment.

[0253] Table 5. Insemination time after administration of OvuGel

[0254]

[0255] Data were collected from 366 sows (61 sows per treatment group).

[0256] LS means were calculated using the PROC GLM procedure of SAS, assuming a model with treatment and group.

[0257] There were no treatment by group interactions for any of the variables tested.

[0258] LS means with different superscripts are different at P < 0.05.

[0259] Example 10

[0260] Hormone stability

[0261] Figure 2 The stability of triptorelin (250 μg / mL -1 The effect of pH was analyzed at 0-4°C, 30°C and 50°C (Mcllvaine buffer). Figure 2 It was shown that the optimal pH range for hormone stability is about 5 to about 6.

[0262] Embodiment 11

[0263] Stability analysis

[0264] The stability of formulations containing various stabilizers was studied at the 3 month time point. The data summarized in Table 6 indicate that L-methionine provided the most stable formulation.

[0265] Table 6. Summary of stability of formulations containing stabilizers

[0266]

[0267] P-80 is polysorbate 80

[0268] Example 12

[0269] Method for preparing hormone composition

[0270] In one embodiment of the method of preparation, the hormone is dissolved in a container containing citric acid and water but separate from the other ingredients. The other ingredients are then added to the hormone composition, and the water is added before the methylcellulose component is added. The methylcellulose component is added slowly under high shear mixing to ensure uniformity of the composition and to avoid agglomeration.

[0271] Embodiment 13

[0272] Synchronous insemination of postpartum sows following intravaginal GnRH agonist treatment

[0273] This study shows the effect of a single, timed insemination following administration of GnRH on subsequent farrowing rate and litter size compared to control sows bred daily at the onset of estrus and during estrus (Tables 7 and 8). One hundred and ninety-nine weaned sows (PIC) were stratified and assigned to one of two treatments based on parity (parity 1 to 6; mean parity 2.9), previous lactation length (range 13-23 d; mean duration 19 d), and body condition score (range 2.0 to 4.0; mean score 3.1). Control sows were observed behaviorally for estrus 7 d after weaning and were inseminated on the day they were observed to be in estrus and at 24 h intervals throughout the duration of estrus. The GnRH agonist formulation (OvuGel TM ) GnRH-treated sows (GnRH-treated sows) were given intravaginal administration and in OvuGel TM GnRH sows were also observed for behavioral signs of estrus 7 days after weaning, but insemination was not considered to be ongoing.

[0274] For 99 control sows, 90.9% (90) were bred 7d after weaning compared to 100% (100) breds 7d after weaning (P<0.01). Control sows had an average of 2.1 inseminations per sow, while all GnRH sows had 1 insemination per sow (P<0.01). There was no difference between control and GnRH sows in the number of sows farrowing (P>0.75), which were 79.2% (79 sows) and 80.7% (81 sows), respectively. There was no effect of treatment on the number of stillbirths (P>0.45) and mummified fetuses (P>0.15). GnRH sows farrowed an average of 11.2 live piglets born compared to 11.4 live piglets born for control sows (P>0.65). The number of piglets born per semen dose in control and GnRH sows was 5.6 and 9.6, respectively (P<0.01). These data indicate that the use of OvuGel TM Treated sows inseminated once after treatment achieved farrowing rates and litter sizes comparable to sows inseminated multiple times during behavioral estrus, as well as OvuGel TM The number of piglets born per semen dose was higher in treated animals.

[0275] Table 7. Response to treatment and comparison between control and GnRH sows 1

[0276] Comparison GnRH Number of sows allocated 99 100 Time interval from weaning to estrus, d 4.4 4.3 Insemination during estrus, % 100 90 Number of inseminations at 7 days after weaning 90 100 Semen dosage per sow inseminated 2.1 1.0 The response rate of inseminated sows at 21 days, % 6.7 8 Number of replies in 21 days 6 8 The allocated sows were pregnant at 30 days, % 81.8 85 Number of pregnancies at 30 days 81 85 Number of sows farrowing 79 81 Farrowing sows among allocated sows, % 79.8 81.0 Total births per litter 12.1 11.8 Total births per semen dose 5.3 9.6 Survival per litter 11.4 11.2 Stillbirths per litter 0.7 0.6 Mummies per litter 0.8 0.6 Piglet index (born alive piglets / 100 allocated sows) 906 909

[0277] 1GnRH sows were treated between 99 and 102 hours after weaning, and inseminated between 21 and 22 hours after treatment. Control sows were not treated and bred normally. The numbers in the tables are unadjusted means and are raw data.

[0278] Table 8. Response to treatment (LS mean ± SEM) and comparison between control and GnRH sows 1

[0279] Comparison GnRH SEM Comparison, P< Sows assigned to treatment 99 100 * * Sows inseminated during the 7-day period after weaning 90 100 * * Sows in estrus AI (%) 100 90.0 2.28 0.005 Time interval from weaning to estrus (d) 4.4 4.3 0.06 0.33 Number of inseminations / sow 2.1 1.0 0.02 0.0001

[0280] Pregnant sows among allocated sows (%) 81.4 84.8 3.74 0.52 Number of sows farrowing 79 81 * * Farrowing sows among allocated sows (%) 79.2 80.7 3.94 0.78 Total piglets born / semen dose 5.2 9.6 0.45 0.0001 Total piglets born alive (all litters) 901 910 * * Total piglets born / litter 12.1 11.8 0.36 0.56 Number of piglets born alive / litter 11.4 11.2 0.34 0.67

[0281] 1 Data were collected from 199 sows (99 control, 100 GnRH). Control sows were bred according to normal farm SOP. GnRH sows were treated on day 4 after weaning and inseminated once 21-22 hours thereafter.

[0282] LS means were calculated using PROC GLM procedure of SAS and assuming a model with treatment and group. There were no treatment by group interactions for any of the tested variables.

[0283] Embodiment 14

[0284] Timed insemination after administration of GNRH agonist to weaned pigs

[0285] This study shows the effect of a single, fixed-time insemination after GnRH administration on subsequent farrowing rate and litter size (Tables 9 and 10). Three hundred weaned sows (PIC) were zoned and assigned to one of two treatments according to parity (parity 1 to 6; mean parity 2.8), previous lactation length (range 17-25d; mean duration 21d), and body condition score (range 2.5 to 3.5; mean score 2.8). Control sows were observed for behavioral estrus 7d after weaning and were inseminated on the day they were observed to be in estrus and at 24h intervals throughout the duration of estrus. Sows in the GnRH-treated group (GnRH sows) were also observed for behavioral signs of estrus 7d after weaning, but were inseminated with the GnRH agonist formulation described in Examples 2 and 3 (OvuGel TM ) intravaginal treatment followed by OvuGel TM Insemination was performed once 24±2h after treatment, regardless of whether they showed persistent signs of estrus. 80.7% (121) of the 150 control sows were bred 7d after weaning, compared to 100% (150) of the GnRH sows bred 7d after weaning (P<0.01).

[0286] Control sows had an average of 2.3 inseminations per sow, while all GnRH sows had 1 insemination per sow (P<0.01). There was no difference in the number of sows farrowing between control and GnRH sows (P>0.40), 72.7% (109 sows) and 76.7% (115 sows), respectively. There was no effect of treatment on the number of stillbirths (P>0.30) and mummified fetuses (P>0.45). GnRH sows farrowed an average of 11.3 live piglets, compared to 10.9 live piglets born in control sows (P<0.37). The number of piglets born per semen dose was 5.6 and 9.6 for control and GnRH sows, respectively (P<0.01). These data indicate that treatment with the GnRH agonist OvuGel TM Treatment and timing of GnRH treatment in sows treated with single insemination achieved farrowing rates and litter sizes comparable to sows inseminated multiple times during behavioral estrus. These results also suggest that OvuGel TM The number of piglets born per semen dose was higher in treated animals than in control animals.

[0287] Table 9. Response to treatment and comparison between control and GnRH sows 1

[0288] Comparison GnRH Number of sows allocated 150 150 Time interval from weaning to estrus, d 4.4 4.4 Insemination during estrus, % 100 84 Number of inseminations at 7 days after weaning 121 150 Number of inseminations 8-15 days after weaning 5 . Semen dosage per sow inseminated 2.2 1.0 % of sows that responded to insemination at 21 days 3.3 8.7 Number of replies in 21 days 4 13 The allocated sows were pregnant at 30 days, % 74.0 80.0 Number of pregnancies at 30 days 111 120 Number of sows farrowing 109 115 Farrowing sows among allocated sows, % 72.7 76.7 Total births per litter 12.2 12.6 Total births per semen dose 5.6 9.6 Birth survival per litter 10.9 11.3 Stillbirths per litter 1.0 0.9 Mummies per litter 0.3 0.4 Piglet index (born alive piglets / 100 allocated sows) 794 867

[0289] 1 Sows were treated 96 hours after weaning and inseminated 24±2 hours thereafter on day 5 after weaning (120-122 hours after weaning). Control sows were not treated and bred normally. The numbers in the tables are unadjusted means and are raw data.

[0290] Table 10. Response to treatment (LS mean ± SEM) and comparison between control and GnRH sows 1

[0291] Comparison GnRH SEM Comparison, P< Sows assigned to treatment 150 150 * * Sows inseminated during the 7-day period after weaning 121 150 * * Sows in estrus AI (%) 100 84.0 2.34 0.0001 Time interval from weaning to estrus (d) 4.4 4.4 0.05 0.75 Number of inseminations / sow 2.3 1.0 0.04 0.0001 Pregnant sows among allocated sows (%) 74.0 80.0 3.44 0.22 Number of farrowing sows 109 115 * * Farrowing sows among allocated sows (%) 72.7 76.7 3.56 0.43 Total piglets born / semen dose 5.6 9.6 0.43 0.0001 Total piglets born alive (all litters) 1191 1300 * * Total piglets born / litter 12.2 12.6 0.31 0.41 Number of piglets born alive / litter 10.9 11.3 0.29 0.37

[0292] 1 Data were collected from 300 sows (150 control, 150 GnRH). Control sows were bred according to normal farm SOP. GnRH sows were treated on day 4 post weaning and inseminated once 24+ / -2 hours thereafter.

[0293] LS means were calculated using PROC GLM procedure of SAS, assuming a model with treatment and group. There were no treatment by group interactions for any of the variables tested.

[0294] Embodiment 15

[0295] Statistical analysis

[0296] Data were analyzed using Proc MIXED and general linear model procedures of SAS (Cary, NC) for continuous and discrete variables. For all analyses, models contained fixed effects of treatment (placebo control insemination 24 hours after treatment, triptorelin-treated sows insemination 18 hours after treatment, triptorelin-treated sows insemination 24 hours after treatment, and triptorelin-treated sows insemination 30 hours after treatment) and replicates. Records of sow parity and lactation length were included as covariates. First-order interactions of treatment and replicates were tested and removed when not significant. Differences between treatments were tested for least squares mean estimates using T-tests at P < 0.05.

Claims

1. A method for synchronizing pig ovulation, the method comprising the following steps: A dose of hormone is administered to pigs on the fourth day after weaning, wherein the hormone is triptorelin or a solvate, a hydrate or a pharmaceutically acceptable salt thereof, and wherein the pigs are inseminated only once 15-24 hours after the administration of the hormone.

2. The method of claim 1, wherein the hormone is triptorelin acetate.

3. The method of any one of claims 1-2, wherein the hormone is administered to the pig using a placed catheter.

4. The method of any one of claims 1-2, wherein the hormone is administered by injection.

5. The method of any one of claims 1-2, wherein the hormone is administered in an effective amount, the effective amount of the hormone being 10 μg to 500 μg.

6. The method of any one of claims 1-2, wherein a single hormone is administered to the pig.

7. The method of any one of claims 1-2, wherein the hormone is administered in a composition comprising a gel, and wherein the composition has a pH of 5.0-6.

0.

8. The method of any one of claims 1-2, wherein the pig is inseminated 18-22 hours after administration of the hormone.

9. The method of any one of claims 1-2, wherein the pig is inseminated 22 hours after administration of the hormone.

10. The method of any one of claims 1-2, wherein the method is performed in the absence of estrus detection.

11. The method of any one of claims 1-2, wherein the hormone is administered 96 hours after weaning.

12. A kit for synchronizing pig ovulation, comprising: a placement catheter for intravaginal administration, and A composition comprising 0.09% by weight per volume of methylparaben, 0.01% by weight per volume of propylparaben, 0.91% by weight per volume of sodium chloride, 0.186% by weight per volume of sodium citrate, 0.1% by weight per volume of L-methionine, 0.07% by weight per volume of citric acid, 0.01% by weight per volume of triptorelin or a solvate, hydrate or pharmaceutically acceptable salt thereof, and 1.2% by weight per volume of methylcellulose, wherein the composition has a pH of 5.0-6.

0.

13. The kit of claim 12, wherein the triptorelin is triptorelin acetate.

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