臺灣博碩士論文加值系統

Indenopyridine衍生物是一種有效之抑制造精作用的藥物。本論文實驗乃藉由觀察大鼠及犬的睪丸病變病程、分析公犬精液性狀變化及檢測血中睪固酮濃度來了解同屬indenopyridine衍生物的Indenopyridine Hydrochloride (IH)對造精作用抑制的情形。公犬及公大鼠口服30 mg IH /kg BW單一劑量之後，發現睪丸生精上皮細胞發生變性及脫落病變，同時在穿透式電子顯微鏡觀察下，大鼠的生殖細胞在IH投藥後第1天便開始顯現細胞凋亡(apoptosis)傾向。公大鼠連續兩次隔週投藥30 mg IH/kg BW抑制造精作用的病程與單一劑量並無明顯差異。公犬口服60 mg IH/kg BW單一劑量後二至八週會有寡精及甚至無精的現象，在投藥後三個月，其每次射精的精子總數又恢復至投藥前的正常數量，；而服用IH後的兩個月內其精漿鹼性磷酸酶(Alkaline Phosphatase)活性顯著上升(p<0.05)，表示睪丸的造精作用已有受到破壞。內泌素分析方面，發現公犬的血漿中睪固酮濃度並不受影響，此外公犬在服藥後依然具有正常的交配能力。另在毒物學試驗方面IH除抑制造精作用外，並無肝腎毒性，對腸道及骨髓等增殖分裂快速的細胞亦無毒性，且經Salmonella/microsome 檢側確定無致變性。IH會造成可逆的造精作用抑制，控制公犬授精能力達三個月之久，經毒物學測試無一般毒性，經進一步的試驗，可開發成理想之雄性避孕藥劑。

Indenopyridine derivatives are known to be effective antispermatogenic agents. An indenopyridine derivatives, indenopyridine hydrochloride (IH), was examined in this study to understand its extent of antispermatogenic effect. Examinations including testicular histopathology, semen characteristics, and sera testosterone profiles, were conducted. Either male dogs and rats treated with single oral dosage of 30 mg IH/kg BW, resulted in germ cells degeneration, that sloughed into the lumen of the seminiferous tubule. When rats were administrated by 30 mg IH/kg BW for 24 hours, the germ cells showed heterochromatin margination, a character of cell apoptosis observed under the transmissible electron microscopy. From two weeks onwards to eight weeks after IH administration, the dogs exhibited oligospermia or azoospermia. Three months later, the sperm counts per ejaculate recovered. Alkaline phosphatase in seminal plasma elevated significantly (p<0.05) during experimental periods. This suggests that process of spermatogenesis was interrupted. When rats were administrated with double dosage of 30 mg IH/kg in one week apart, the antispermatogenic effect was not different from those treat with single dose. Testosterone profiles were normal in all dog cases examined. Sexual behaviors including, libido, erection capability, and ejaculation were not impaired by IH. IH treatment showed no toxicity to hepatic- renal- functions. Similarly, this compound didn’t affect bone marrow and intestinal epithelium, nor mutagenic potential. Conclusively, IH exclusively inhibits spermatogenesis in a reversible manner. This might result in an episode of male infertility. IH possesses the clinical potential in development of male chemical castration medicine.

目錄 I
表次 II
圖次 III
中文摘要 IV
英文摘要 V
第一章、緒言 1
第二章、文獻探討
第一節、睪丸組織學構造及生理學 2
第二節、睪丸病變的觀察 5
第三節、睪丸傷害的特徵 7
第四節、公犬精液收集及分析 9
第五節、抑制造精作用藥物的種類 11
第六節、Indenopyridine衍生物 13
第三章、材料與方法
第一節、藥品與器材 18
第二節、重要儀器 19
第三節、實驗方法 20
第四章、結果
第一節、 Indenopyridine Hydrochloride對Sprague-Dawley大鼠造精作用的影響 27
第二節、 Indenopyridine Hydrochloride對公犬造精作用的影響 30
第三節、 Indenopyridine Hydrochloride之毒物學試驗 35
第五章、討論 40
參考文獻 45
附錄 50

1. 沈永紹。獸醫實驗診斷學提要。華香園，台北，195-236，1991。
2. Amann RP, Schanbacher BD. Physiology of male reproduction. J Anim Sci 57 (suppl 2): 380-403, 1983.
3. Amann RP. Reproductive physiology and endocrinology of the dog. In: Morrow DA, ed. Current therapy in theriogenology: treatment, diagnosis, and prevention of reproductive diseases in small and large animals, 2nd ed. Philadelphia, WB Saunders Co, 532-538, 1986.
4. Ames BN, Mccann J, Yamasaki E. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian microsome mutagenicity test. Mutat Res 31: 347-364, 1975.
5. Augstein J, Ham AL, Leeming PR. Relationship between antihistamine and antidepressantactivity in hexahydroindenopyridines. J Med Chem 15: 466-469, 1972.
6. Bauck L, Bihun C. Basic anatomy, physiology, husbandry, and clinical techniques in Small rodents. In: Hillyer EV, ed. Ferrets, rabbits, and rodents clinical medicine and surgery. Philadelphia, WB Saunders Co, 291-306, 1997.
7. Bergh A. Development of stage-specific paracrine regulation of leydig cells by the seminiferous tubules. Int J Androl 8: 80-85, 1985.
8. Bjordahl JA, Jost LK, Fail PA, Cook CE, Evenson DP. Flow cytometric analysis of the antispermatogenic effects of an indenopyridine derivative. Biol Reprod 54(suppl 1): 212(abstract 518), 1997.
9. Blackburn DM, Gray AJ, Lloyd SC, Sheard CM, Foster PM. A comparison of the effects of the three isomers of dinitrobenzene on the testis in the rat. Toxicol Appl Pharmacol 92: 54-64, 1988.
10. Blom E. Pathological conditions in genital organs and sperm as a cause for the rejection of breeding bulls for import into and export from Denmark (an andrologic retrospective, 1958-1982). Nord Vet Med 35: 105-130, 1983.
11. Bruce N, Ames JM, Edith Y. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian-microsome mutagenicity test. Mutat Res 31: 347-364,1975.
12. Chang LT, Fung HP, Lin YF, Kuo CY, Chien CW. Indenopyridine hydrochloride testicular spermatogenesis failure with high seminal alkaline phosphatase levels in male dog. Biological & Pharmaceutical Bulletin.(in press)
13. Clermont Y. Kinetics of spermatogenesis in mammals: seminiferous epithelium cycle and spermatogonial renewal. Physiol Rev 52: 198-236, 1972.
14. Cook CE, Jump JM, Zhang P, Stephens JR, Lee YW, Fail PA, Anderson SA. Exceptionally potent antispermatogenic compounds from 8-halogenation of (4aRS,5SR,9bRS)- hexahydroindeno[1,2-c]pyridines. J Med Chem 40: 2111-2112, 1997.
15. Cook CE, Wani MC, Jump JM, Lee YW, Fail PA, Anderson SA, Gu YQ, Petrow V. Structure-activity studies of 2,3,4,4a,5,9b-hexahydroindeno[1,2-c]pyridines as antispermatogenic agents for male contraception. J Med Chem 38: 753-763, 1995.
16. Creasy DM, Flynn JC, Gray TJ, Butler WH. Aquantitative study of stage-specific spermatocyte damage following administration of ethylene glycol monomethyl ether in the rat. Exp Mol Pathol 43:321-336, 1985.
17. Creasy DM, Ford GR, Gray TJ. The morphogenesis of cyclohexylamine-induced testicular strophy in the rat: in vivo and in vitro studies. Exp Mol Pathol 52: 155-169, 1990.
18. Creasy DM, Foster PM. The morphological development of di-N-pentyl phthalate induced testicular atrophy in the rat. J Pathol 139: 309-321, 1983.
19. Creasy DM, Foster PM. The morphological development of glycol ether-induced testicular strophy in the rat. Exp Mol Pathol 40: 169-176, 1984.
20. Creasy DM. Evaluation of testicular toxicity in safety evaluation studies: the appropriate use of spermatogenic staging. Toxicol Pathol 25: 119-131, 1997.
21. Dube JY, Frenette G, Chapdelaine P, Paquin R, Tremblay RR. Biochemical characteristics of the proteins secreted by dog prostate, a review. Exp Biol 43: 149-159, 1985.
22. Fail PA, Anderson SA, Cook CE. 28-day toxicology test: indenopyridine RTI 4587-056 in male Sprague-Dawley rats. Reprod Toxicol 14: 265-274, 2000.
23. Frenette G, Dube JY, Tremblay RR. Origin of alkaline phosphatase of canine seminal plasma. Arch Androl 16: 235-241, 1986.
24. Gill HP, Kaufman CF, Foote RH, Kirk RW. Artificial insemination of beagle bitches with freshly collected, liquid-stored, and frozen-stored semen. Am J Vet Res 31: 1807-1813. 1970.
25. Gürtler J, Donatsch P. Effect of two structurally different antispermatogenic compounds on the synthesis of steroids in rat testes. Arch Toxicol. Supplement 2: 381-385, 1979.
26. Hair WM, Ketteridge K, Wu FCW. A new male contraceptive pill/patch combination: oral desogestrel and transdermal testosterone in suppression of spermatogenesis in men. J Androl 20: 30, 1999.
27. Heindel JJ, Treinen KA. Physiology of the male reproductive system: endocrine, paracrine and autocrine regulation. Toxicol Pathol 17: 411-445, 1989.
28. Hess RA, Chen P. Computer tracking of germ cells in the cycle of the seminiferous epithelium and prediction of changes in cycle duration in animals commonly used in reproductive biology and toxicology. J Androl 13: 185-190, 1992.
29. Hess RA, Linder RE, Strader LF, Perreault SD. Acute effects and long-term sequelae of 1,3-dinitrobenzene on male reproduction in the rat. II. Quantitative and qualitative histopathology of the testis. J Androl 9: 327-342, 1988.11
30. Hess RA. Quantitative and qualitative characteristics of the stages and transitions in the cycle of the rat seminiferous epithelium: light microscopic observations of perfusion-fixed and plastic-embedded testes. Biol Reprod 43: 525-542, 1990.
31. Hild SA, Meistrich ML, Blye RP, Reel JR. Lupron Depot prevention of antispermatogenic/antifertility activity of the indenopyridine, CDB-4022, in the rat. Biol reprod 65: 165-172, 2001A.
32. Hild SA, Reel JR, Attardi BJ, Blye RP. Age-dependent effects of CDB-4022 on testicular function and fertility in male rats. Biol reprod 62 (Suppl 1): 337(abstract 588), 2000.
33. Hild SA, Reel JR, Larner JM, Blye RP. Disruption of spermatogenesis and function by the indenopyridine CDB-4022 in rats. Biol Reprod 65: 1771-1779, 2001B.
34. Hild-Petito S, Reel JR, Larner JM, Blye RP. Mechanism of action of a male antifertility drug, CDB-4022([4αRS,5SR,9βRS]-2-ethyl-2,3,4,4α,5,9β-hexahydro-8-iodo-7-methyl-5-[4-carbomethoxyphenyl]-1H-Indeno[1,2-c]pyridine-hydrochloride). In: 31st Annual meeting of the society for the study of reproduction: Abstract 312, 1998.
35. Hodel C, Suter K. Reversible inhibition of spermatogenesis with an indenopyridine (20-438). Arch Toxicol.(Suppl 1): 323-326, 1978.
36. Ibach B, Weissbach L, Hilscher B. Stages of the cycle of the seminiferous epithelium in the dog. Andrologia 8: 297-307, 1976.
37. Johnston SD, Kustritz MVR, Olson PNS. Sexual differentiation and normal anatomy of the dog. In: Johnston SD et al., ed. Canine and feline theriogenology 1st ed. Philadelphia, WB Saunders Co, 275-286, 2001.
38. Johnston SD. Performing a complete canine semen evaluation in a small animal hospital. Vet Clin North Am Small Anim Pract 21: 545-551, 1991.
39. Kerr JB. Macro, micro, and molecular research on spermatogenesis: the quest to understand its control. Microsc Res Tech 32: 364-384, 1995.
40. Ku WW, Chapin RE, Wine RN, Gladen BC. Testicular toxicity of boric acid (BA): relationship of dose to lesion development and recovery in the F344 rat. Reprod Toxicol 7: 295-196, 1993.
41. Lewis-Jones DI, Johnson PM, Desmond AD, McLaughlin PJ. Germ cell alkaline phosphatase in human seminal plasma following vasectomy. Br J Urol 69: 418-420, 1992.
42. Linder RE, Strader LF, Slott VL, Suarez JD. Endpoints of spermatotoxicity in the rat after short duration exposures to 14 reproductive toxicants. Reprod Toxicol 6: 491-505, 1992.
43. Matter BE, Jaeger I, Suter W, Tsuchimoto T, Deyssenroth H. Action of an antispermatogenic, but non-mutagenic, indenopyridine derivative in mice and Salmonela typhimurium. Mutat Res 66: 113-127, 1979.
44. McLaughlin PJ, Lewis-Jones I, Hutchinson GE, Johnson PM. Placental-type alkaline phosphatase in human seminal plasma from fertile and infertile men. Fertil Steril 46: 934-937, 1986.
45. Millan JL, Manes T. Seminoma-derived Nagao isozyme is encoded by a germ-cell alkaline phosphatase gene. Proc Natl Acad Sci USA 85: 3024-3028, 1988.
46. Olson PN. Clinical approach for evaluating dogs with azoospermia or aspermia. Vet Clin North Am Small Anim Pract 21: 591-607, 1991.
47. Parkinson T. Fertility and infertility in male animals. In: Noakes DE et al., ed. Arthur’s veterinary reproduction and obstertrics 8th ed. London, WB Saunders Co, 695-750, 2001A.
48. Parkinson T. Normal reproduction in male animals. In: Noakes DE et al., ed. Arthur’s veterinary reproduction and obstertrics 8th ed. London, WB Saunders Co, 673-694, 2001B.
49. Parvinen M, Vihko KK, Toparri J. Cell interactions during the seminiferous epithelial cycle. Int Rev Cytol 104: 115-151, 1986.
50. Parvinen M. Regulation of the seminiferous epithelium. Endocrine Rev 3: 404-417, 1982.
51. Paulou SN, Brewer K, Farley MC. Combined administration of a gonadotropin-releasing hormone antagonist and testosterone in men induces reversible azoospermia without loss of libido. J Clin Endocrinol Metab 73: 1360-1368, 1991.
52. Pineda MH, Faulkner LC. The biology of sex. In: McDonald LE, ed. Veterinary endocrinology and reproduction 3rd ed. Philadelphia, LEA & FEBIGER Co, 208-234, 1980.
53. Post K. Clinical examination of the reproductive system: dogs and cats. In: Radostits OM et al., ed. Veterinary clinical examination and diagnosis 1st ed. Philadelphia, WB Saunders Co, 663-674, 2000.
54. Russell LD, Ettlin RA, Giesel LO. Quantitative studies on spermatogenesis in the albino rat. Am J Anat 87: 1-22, 1950.
55. Sharpe RM, Maguire SM, Saunders PT, Millar MR, Russell LD, Ganten D, Bachmann S, Mullins L, Mullins JJ. Infertility in a transgenic rat due impairment of cytoplasmic elimination and sperm release from the Sertoli cells. Biol Reprod 53: 214-226, 1995.
56. Silvestrini B, Burberi S, Catanese B, Cioli V, Coulston F, Lisciani R, Barcellona PS. Antispermatogenic activity of 1-p-chlorobenzyl-1H-indazol-3-carboxylic acid (AF 1312/TS) in rats. Exp Mol Pathol 23: 288-307, 1975.
57. Sirivaidyapong S, Bevers MM, Gadella BM, Colenbrander B. induction of the acroaome reaction in dog sperm cell is dependent on epididymal maturation: the generation of a functional progesterone receptor is involved. Mol Reprod Dev 58: 451-459, 2001.
58. Stockner PK, Bardwick C. The relationship of semen parameters to fertility in the dog. Canine Pract 16: 15-23, 1991.
59. Suter KE, Hodel C, Gadient F. Antispermatogenic activity of an indenopyidine derivative. Experientia 33: 810, 1977.
60. Takayama S, Akaike M, Kawashima K, Takahashi M, Kurokawa Y. A collaborative study in japan on optimal treatment period and parameters for detection of male fertility disorders induced by drugs in rats. J Am Coll Toxicol 14:266-292, 1995.
61. Ulbrich B, Palmer AK. Detection of effects on male reproduction─A literature survey. J Am Coll Toxicol 14: 2293-3327, 1995.
62. Whitsett JM, Noden PF, Cherry J, Lawton AD. Effert of transitional photoperiods on testicular development and puberty in male deer mice (peromyscus maniculatus). J Reprod Fertil 72: 277-286, 1984.
63. Zschauer A, Hodel C. Drug-induced histological changes in rat seminiferous tubular epithelium. Arch Toxicol. Supplement 4: 466-470, 1980.