2011 31 1 79-83 Acta Theriologica Sinica EP-1 1 2 3 2 1 1 1* 1 100193 2 100101 3 100081 EP 2 - -β- HPCD EP 4 EP-1 2 4 20% HPCD 4 mg /ml 0. 5 mg /ml 2 2 mg /kg EP-1 HPCD EP-1 EP-1 EP 2- -β- Q494 A 1000-1050 2011 01-0079 - 05 Preparation of hydroxypropyl-β-cyclodextrin inclusion with Levonorgestrel and Quinestrol EP-1 and its influence on the reproductive organs Brandt s voles Lasiopodomys brandtii WANG Dawei 1 2 LIU Qi 3 LIU Ming 2 LI Ning 1 HUANG Baohuan 1 LIU Xiaohui 1* 1 Key Laboratoryof Weed and Rodent Biology and Management Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100193 China 2 State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of Zoology Chinese Academy of Science Beijing 100101 China 3 Key Laboratory of Dryland Farming and Water-saving Agriculture MOA Institute of Environment and Sustainable Development in Agriculture Chinese Academy of Agricultural Sciences Beijing 100081 China Abstract EP series rodent contraception is mainly composed of levonorgestrel and quinestrol. However neither component do not readily dissolves in water which limits the precision of research and application. In the present study solubilization was measured in hydroxypropyl-β-cyclodextrin HPCD inclusion complexes of levonorgestrel and quinestrol respectively. Afterwards the effects of treatment times after 2 and 4 weeks and doses 0 1 2 4 mg /kg of EP-1 inclusion complexes on reproductive organs were investigated using a gastric gavage method. The results showed that the solubility of levonorgestrel and quinestrol reached 4 mg /ml and 0. 5 mg /ml in 20% aqueous HPCD solution respectively. For females there were no significant differences after 2 or 4 weeks between the control and treatment groups that received gavage. For males the average weight of reproductive organs of the group that received gavage for 2 weeks of 2 mg /kg group was less than that of the control group. Our results indicate that the solubility of EP-1 was significantly enhanced by forming inclusion complexes with HPCD the suppressive effects on reproduction in male Brandt s voles is more obvious than those in females. Moreover inclusion did not affect the anti-fertility effect of EP-1. Key words Antifertility control HPCD Inclusion complex Lasiopodomys brandtii Levonorgestrel Quinestrol 973 2007CB109104 ChineseIPM0804 2009YW15 1981 -. 2010-01 - 14 2010-09 - 27 * Corresponding author E-mail lxiaohui2000@ 163. com
80 31 120. 0 μg /ml 30 0. 25 ml 1. 00 ml Knipling 1959 1960 Davis 1961 1. 25 ml 2. 00 ml 3. 00 ml 4. 00 ml 6. 00 ml 25 ml Marsh 1988 1995 149. 0 μg /ml 4. 00 ml 5. 00 ml 6. 00 ml 7. 00 ml 10. 00 ml 15. 00 ml 18. 75 ml 1995 2001 25 ml 280 nm A C 1. 1. 2 HPCD EP 10 min EP-1 48 h 0. 45 μm 2 1 2004 2005 2006 2006 2007 ph = 7. 0 2005 1. 1. 3 0. 02 g /ml 0. 05 g /ml 0. 10 g /ml 0. 20 g /ml HPCD 2- -β- HPCD β- ph = 7. 0 β-cd 10 min 20 Szetjli 1998 2005 0. 45 μm 240 nm 280 nm 2006 Trapani et al. 2005 HPCD 1. 2 EP-1 EP-1 1. 2. 1 > 25 g 1 1. 1 2 4 1. 1. 1 0 mg /kg C HPCD 1. 0 mg /kg Ⅰ 2. 0 mg /kg Ⅱ UV-2550 HPCD 99% 4. 0 mg /kg Ⅲ 6 Wako 1. 2. 2 EP-1 2 1 λ max 240 nm 20% HPCD 3 280 nm HPCD 1. 0 2. 0 4. 0 mg /ml 240 nm A C 48 h 14L 10D 96 20% HPCD 4
1 EP-1 81 Δλ = 2. 0 nm 5 d 2 4 1. 2. 3 2. 1. 3 ph = 7. 0 g /100 g 4π 2 /3cm 3 HPCD 1 cm HPCD HPCD X 1. 3 SPSS 13. 0 Oneg /ml Y μg /ml Y = 3185. 1X + 6. 089 R 2 = Way ANOVA EP-1 0. 9995 Y = 20460X - 269. 15 R 2 = 0. 9985 P < 0. 05 1 20% HPCD 4 mg /ml 0. 5 mg 2 2. 1 HPCD 2. 1. 1 2. 39 ~ 28. 68 μg /ml A C A = 0. 0567C + 0. 0288 R 2 = 0. 9995 23. 84 ~ 111. 8 μg /ml A C A = 0. 0065C - 0. 0107 R 2 = 0. 9999 2. 1. 2 Fig. 1 λ max 1998 2004 1 ph = 7. 0 HPCD The solubility curve of levonorgestrel and quinestrol of HPCD solution ph = 7. 0 Table 1 1 EP-1 HPCD 2 4 ± The reproductive organs of male Brandt s voles 2 and 4 weeks after treated by HPCD inclusion with EP-1 Mean ± SE 2 Two weeks Testis g 1. 654 ± 0. 066 a 1. 551 ± 0. 273 a 1. 075 ± 0. 214 a 1. 664 ± 0. 152 a Epididymides g 0. 425 ± 0. 032 a 0. 395 ± 0. 049 a b 0. 279 ± 0. 045 b 0. 393 ± 0. 036 a b Seminal vesicle g 0. 949 ± 0. 100 a 0. 389 ± 0. 154 b 0. 335 ± 0. 151 b 0. 610 ± 0. 175 a b Volume of testicle cm 3 2. 581 ± 0. 117 a 1. 903 ± 0. 417 a b 1. 471 ± 0. 385 b 2. 069 ± 0. 332 a b 4 Four weeks Testis g 1. 730 ± 0. 325 a 1. 704 ± 0. 231 a 1. 686 ± 0. 171 a 1. 765 ± 0. 147 a Epididymides g 0. 381 ± 0. 093 a 0. 320 ± 0. 047 a 0. 395 ± 0. 034 a 0. 349 ± 0. 046 a Seminal vesicle g 0. 600 ± 0. 296 a b 0. 333 ± 0. 127 a 1. 035 ± 0. 207 b 0. 620 ± 0. 271 a b Volume of testicle cm 3 2. 282 ± 0. 599 a 2. 039 ± 0. 405 a 2. 682 ± 0. 210 a 2. 501 ± 0. 366 a P < 0. 05 C Ⅰ 1. 0 mg /kg Ⅱ 2. 0 mg /kg Ⅲ 4. 0 mg /kg Different superscripts in each row indicate significant differences P < 0. 05 C Control Ⅰ 1. 0 mg /kg Ⅱ 2. 0 mg /kg Ⅲ 4. 0 mg /kg
82 31 2. 2 EP-1 HPCD EP-1 HPCD 2. 2. 1 2 1. 0 mg /kg 2. 2. 2 2. 0 mg /kg 2 4 P = 0. 026 EP - 1 P = 0. 012 P = HPCD 0. 041 4. 0 mg /kg 4 Table 1 2 EP-1 HPCD 2 4 ± The reproductive organs of female Brandt s voles 2 and 4 weeks after treated by HPCD inclusion with EP-1 Mean ± SE 2 Two weeks Ovaries mg 22. 3 ± 1. 4 a 19. 6 ± 2. 0 a 19. 5 ± 1. 9 a 21. 5 ± 1. 6 a Uterus g 0. 159 ± 0. 0418 a 0. 2051 ± 0. 0664 a 0. 1513 ± 0. 0280 a 0. 1580 ± 0. 0321 a Length of Uterus cm 5. 97 ± 0. 22 a 5. 90 ± 0. 38 a 5. 78 ± 0. 40 a 5. 72 ± 0. 18 a 4 Four weeks Ovaries mg 22. 3 ± 1. 0 a 22. 6 ± 2. 1 a 23. 4 ± 3. 2 a 21. 8 ± 1. 3 a Uterus g 0. 0876 ± 0. 0203 a 0. 1497 ± 0. 0431 a 0. 0816 ± 0. 0227 a 0. 0834 ± 0. 0119 a Length of Uterus cm 4. 798 ± 0. 157 a 4. 549 ± 0. 367 a 4. 744 ± 0. 241 a 4. 725 ± 0. 084 a P < 0. 05 C Ⅰ 1. 0 mg /kg Ⅱ 2. 0 mg /kg Ⅲ 4. 0 mg /kg Different superscripts in each row indicate significant differences P < 0. 05 C Control Ⅰ 1. 0 mg /kg Ⅱ 2. 0 mg /kg Ⅲ 4. 0 mg /kg 3 2- -β- 2005 2006 2007 2005 EP-1 EP-1 HPCD Cricetulus migratorius Meriones meridianus Tscheskia triton Meriones unguiculatus Phodopus campbelli EP-1 HPCD 2004 2005 2006 2006 Szetjli 1998 2 mg /kg 2006 EP-1 HPCD Zhao 2007 EP-1 Zhao et al. 2007 EP-1 HPCD EP-1 - -
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