【摘要】：目的 制備符合眼部應用要求的托吡卡胺（TR）溫敏凝膠。運用流變學方法評價并篩選處方，對優(yōu)化處方所制備凝膠的質量、凝膠溶蝕、藥物釋放、眼部滯留時間和刺激性進行全面考察。 方法 1.星點設計-效應面法優(yōu)化TR眼用溫敏凝膠處方 通過單因素試驗，以P407和P188作為溫敏材料基質，獲取了對相變溫度影響較大的因素；采用星點設計-效應面法設計并進行處方篩選試驗，星點設計試驗結果以SPSS13.0統(tǒng)計學軟件進行方程擬合，建立了模擬淚液稀釋前、后凝膠的相變溫度（T1、T2）對P407、P188質量分數(shù)的線性模型；運用Matlab2011b統(tǒng)計學軟件繪制以上兩方程的三維效應面、二維等高線圖，由兩等高線的疊加獲得兩溫敏材料質量分數(shù)的優(yōu)化區(qū)域；代表性處方從上述優(yōu)化區(qū)域中選出后，對所擬合方程進行預測效果的驗證。根據2010版《中國藥典》的要求考察抑菌劑、滲透壓調節(jié)劑和（或）粘附性材料對T1、T2的影響，根據結果調整溫敏材料的含量即獲得符合眼部應用要求的處方。 2. TR眼用溫敏凝膠流變學性質的考察 采用Kinexus Pro旋轉流變儀，在動態(tài)振蕩模式下進行處方的流變學評價試驗。分別應用應力掃描選取處方的線性粘彈區(qū)，溫度掃描獲得相變溫度，頻率掃描考察粘度隨頻率的變化情況，時間掃描對TR溫敏凝膠由液態(tài)轉變?yōu)槟z狀態(tài)的時間進行考察。 3.質量控制方法考察 根據2010版《中國藥典》二部對眼用制劑的相關規(guī)定，對TR溫敏凝膠的含量測定方法、制劑的含量、性狀、pH、初步穩(wěn)定性、無菌檢查等進行全面考察，建立TR溫敏凝膠的質量控制標準。 4. TR眼用溫敏凝膠的凝膠溶蝕、藥物釋放行為的考察 根據TR眼用溫敏凝膠的含量測定方法，運用無膜溶出法考察TR溫敏凝膠的凝膠溶蝕和藥物的釋放行為。 5. TR溫敏凝膠兔眼滯留時間和刺激性考察 以熒光素鈉標記已制備好的TR溫敏凝膠和普通滴眼液制劑，交叉給藥，用紫外燈觀察熒光消失時間即代表各制劑的滯留時間。 采用自身對照法、Draize評分法對TR溫敏凝膠單次給藥、多次給藥的兔眼局部刺激性進行評價。 結果 1.由單因素試驗結果可知兩溫敏材料的質量分數(shù)均顯著影響處方的相變溫度，且P407質量分數(shù)與相變溫度呈負相關，P188質量分數(shù)與相變溫度呈正相關；星點設計試驗結果以SPSS13.0擬合的方程為：T1=74.270-2.578X+0.934Y (r=0.9187，P 0.01)；T2=89.264-3.018X+1.191Y (r=0.8899，P 0.01)。Matlab2011b繪出優(yōu)化的溫敏材料質量分數(shù)區(qū)域并從該區(qū)域中選取了4個代表性處方，對擬合方程進行預測能力的驗證，結果可知T1、T2偏差均5%，表明其預測能力良好。在上述代表處方中加入滲透壓調節(jié)劑、抑菌劑和（或）HPMC K4M或HPMC E50后，發(fā)現(xiàn)處方的相變溫度降低；由相變溫度的改變調整P407和P188的質量分數(shù)，獲得了相變溫度適宜的符合眼部應用的A~G七個處方，分別為A：20%P407-4%P188；B：20%P407-5%P188-0.5%HPMCK4M；C：21%P407-8%P188-0.5%HPMC K4M；D：22%P407-10%P188；E：20%P407-5%P188-1%HPMC E50；F：21%P407-6%P188；G：21%P407-9%P188-1%HPMCE50，各處方均含有0.25%TR、0.03%尼泊金乙酯和0.9%氯化鈉（至全量）。 2.應力掃描結果顯示模擬淚液稀釋前、后的線性粘彈區(qū)范圍分別為5Pa~13Pa、1Pa~13Pa，因此剪切應力分別選擇為10Pa、5Pa。TR眼用溫敏凝膠的彈性模量（G’）和粘性模量（G”）隨溫度升高呈S形趨勢變化；溫度低于相變溫度時呈自由流動的液體狀態(tài)，達相變溫度時發(fā)生相變，轉變?yōu)榘牍虘B(tài)；同時，模擬淚液稀釋后溫敏凝膠的G’和G”相比之前均有所下降，，說明其凝膠強度降低；隨著溫度的升高，相角δ逐漸降低。同樣地，隨著剪切頻率的升高，粘度η’也有所降低；且模擬淚液稀釋后仍呈現(xiàn)較好的粘度；TR溫敏凝膠不同處方的相變時間隨著溫敏材料含量的增加而縮短。 通過對比處方A~G各流變學參數(shù)，處方D：0.25%TR-22%P407-10%P188-0.03%尼泊金乙酯-0.9%氯化鈉（至全量）相變溫度適宜，粘度隨剪切頻率升高下降最為緩慢，而且具有最短的相變時間，因此確定其為最優(yōu)處方。 3. HPLC測定結果顯示，TR的保留時間為7min；輔料對其的含量測定無干擾；峰面積（Y）對TR濃度（C）進行線性回歸并繪制標準曲線，得標準曲線為，Y=16.163C+52.731(R2=0.9997)，表明TR濃度在19.9~199μg/mL范圍內與峰面積呈現(xiàn)良好的線性關系；24h溶液穩(wěn)定性試驗和精密度試驗中，峰面積的RSD分別為0.35%(n=6)、0.19%(n=6)；低、中、高濃度的平均回收率分別為99.01%、98.94%、100.21%，對應的RSD分別為0.97%、0.14%、0.12%(n=6)；質量研究顯示TR溫敏凝膠低溫時呈透明、均勻的可流動液體，溫度高于相變溫度即變?yōu)橥该骶鶆虻哪z；各處方pH值均位于6.6~6.7之間，符合眼部用藥的要求；無菌檢查結果符合要求；初步穩(wěn)定性結果顯示制劑6個月的穩(wěn)定性良好。 4. TR溫敏凝膠的凝膠溶蝕和藥物釋放試驗顯示TR呈緩慢釋放；凝膠溶蝕量與TR釋放量呈正比，說明藥物釋放是由凝膠溶蝕控制的釋放行為。 5.測得熒光標記的TR溫敏凝膠與市售TR滴眼液的眼部滯留時間分別為20min、5min，溫敏凝膠滯留時間更長；且其單次給藥、多次給藥對兔眼均無刺激性。 結論 本課題采用星點設計-效應面法優(yōu)化了TR眼用溫敏凝膠的處方，并運用旋轉流變儀對處方的流變學進行評價，表征了其相變過程，獲得了符合眼部應用要求的最佳處方；采用HPLC法建立了TR溫敏凝膠的含量測定方法；考察了其質量控制方法、凝膠溶蝕和TR的釋放；考察了TR溫敏凝膠的兔眼滯留時間和局部刺激性。所得制劑不僅給藥劑量準確、減少患者用藥次數(shù)，提高患者的順應性，而且與普通滴眼液相比延長了藥物在眼內的滯留時間，提高藥物局部作用效果，具有良好的臨床應用前景。
[Abstract]:objective
To prepare topicamide (TR) thermosensitive gel which meets the requirements of ophthalmic application, rheological method was used to evaluate and screen the prescription, and the gel quality, gel dissolution, drug release, eye retention time and irritation were investigated.
Method
Optimization of TR ophthalmic thermosensitive gel formulation by central composite design response surface methodology 1.
Through single factor experiment, using P407 and P188 as the matrix of temperature-sensitive materials, the factors which have great influence on the phase change temperature were obtained; Star design-response surface methodology was used to design and carry out the prescription screening test; Star design test results were fitted by SPSS 13.0 statistical software, and the phase change temperature of the gel before and after the dilution of the simulated tear was established. T1, T2) Linear model of mass fraction of P407 and P188; 3-D effect surface and 2-D contour plot of the above two equations were drawn by using MATLAB 2011B statistical software, and the optimal region of mass fraction of two thermosensitive materials was obtained by the superposition of two contours; representative prescriptions were selected from the above optimized region, and the predicted results of the fitting equation were obtained. The effects of antimicrobial agents, osmotic pressure regulators and/or adhesive materials on T1 and T2 were investigated according to the requirements of the Chinese Pharmacopoeia (2010 edition).
Investigation on rheological properties of 2. TR ophthalmic thermosensitive gel
The Kinexus Pro rotary rheometer was used to evaluate the rheology of the prescription under dynamic oscillation mode. The linear viscoelastic region of the prescription was selected by stress scanning, and the phase transition temperature was obtained by temperature scanning. The change of viscosity with frequency was investigated by frequency scanning, and the time of time scanning for the transition of TR thermosensitive gel from liquid to gel state was investigated by time scanning. Investigate.
3. quality control method investigation
According to the related regulations of ophthalmic preparations in the second part of Chinese Pharmacopoeia 2010 edition, the content determination method, properties, pH, initial stability and sterility test of TR thermosensitive gel were investigated comprehensively, and the quality control standard of TR thermosensitive gel was established.
Investigation of gel dissolution and drug release behavior of 4. TR ophthalmic thermosensitive gel
According to the content determination method of TR ophthalmic thermosensitive gel, the gel dissolution and drug release behavior of TR thermosensitive gel were investigated by non-membrane dissolution method.
Retention time and irritation of 5. TR thermosensitive gel in rabbit eyes
The prepared TR thermosensitive gel and common eye drops were labeled with sodium fluorescein, and the fluorescence disappearance time was observed by ultraviolet lamp.
The local irritation of TR thermosensitive gel was evaluated by Draize score and self-control method.
Result
1. The results of single factor experiment showed that the mass fraction of the two thermosensitive materials significantly affected the phase transition temperature of the prescription, and the mass fraction of P407 was negatively correlated with the phase transition temperature, and the mass fraction of P188 was positively correlated with the phase transition temperature. The equation fitted by SPSS 13.0 was T1 = 74.270-2.578X + 0.934Y (r = 0.9187, P 0.01); T2 = 89.2. 64-3.018X+1.191Y (r=0.8899, P 0.01). The optimized mass fraction region of temperature-sensitive materials was plotted by MATLAB 2011b. Four representative prescriptions were selected from the region. The predictive ability of the fitting equation was verified. The results showed that the deviations of T1 and T2 were all 5%, indicating good predictive ability. (or) After HPMC K4M or HPMC E50, it was found that the phase transition temperature of the prescription decreased; P407 and P188 mass fractions were adjusted by the change of phase transition temperature, and seven prescriptions of A~G with suitable phase transition temperature were obtained, which were A: 20% P407-4% P188; B: 20% P407-5% P188-0.5% HPMCK4M; C: 21% P407-8% P188-0.5% HPMCK4M; D: 22% P407-10% P188-0.5% HPMCK4M; D: 22% P407-10% P407-10% P188-0.5% MCK % P188; E: 20% P407-5% P188-1% HPMC E50; F: 21% P407-6% P188; G: 21% P407-9% P188-1% HPMC E 50. Each prescription contains 0.25% TR, 0.03% ethyl Nipagin ester and 0.9% sodium chloride (to the full amount).
2. The results of stress scanning show that the linear viscoelastic region of the simulated tear before and after dilution is 5Pa~13Pa and 1Pa~13Pa respectively, so the shear stress is 10Pa and 5Pa. At the same time, the G'and G'of the thermosensitive gel decreased after dilution, indicating that the gel strength decreased; with the increase of temperature, the phase angle delta decreased gradually. Similarly, with the increase of shear frequency, the viscosity_'also decreased; and the simulated tear dilution decreased. The phase transition time of TR thermosensitive gel was shortened with the increase of the content of thermosensitive materials.
By comparing the rheological parameters of prescription A~G, prescription D: 0.25% TR-22% P407-10% P188-0.03% ethyl Nipagin ester-0.9% sodium chloride (to the full amount) has the best phase transition temperature, viscosity decreases most slowly with the increase of shear frequency and has the shortest phase transition time, so it is the best prescription.
3. The results of HPLC showed that the retention time of TR was 7 minutes; the excipients had no interference in the determination of TR content; the peak area (Y) was linearly regressive to the concentration of TR (C) and the standard curve was drawn. The standard curve was Y=16.163 C+52.731 (R2=0.9997), indicating that the TR concentration had a good linear relationship with the peak area in the range of 19.9-199 ug/mL; the solution was stable in 24 hours. In qualitative test and precision test, the RSD of peak area was 0.35% (n=6) and 0.19% (n=6), respectively; the average recoveries of low, medium and high concentrations were 99.01%, 98.94%, 100.21%, and the corresponding RSDs were 0.97%, 0.14%, 0.12% (n=6), respectively; the quality study showed that TR thermosensitive gel was a transparent and uniform flowable liquid at low temperature, and the temperature was higher than the phase transition temperature. The pH value of each prescription ranged from 6.6 to 6.7, which met the requirements of ophthalmic medication. The results of sterility test met the requirements. The preliminary stability results showed that the preparation had good stability for 6 months.
4. The gel dissolution and drug release test of TR thermosensitive gel showed that TR was released slowly, and the gel dissolution was proportional to the TR release, indicating that the drug release was controlled by the gel dissolution.
5. The retention time of fluorescence-labeled TR thermosensitive gel and commercial TR eye drops were 20 minutes and 5 minutes respectively, and the retention time of thermosensitive gel was longer than that of the commercial TR eye drops.
conclusion
In this study, the prescription of TR ophthalmic thermosensitive gel was optimized by star design-response surface methodology, and the rheology of the prescription was evaluated by rotational rheometer, and the phase transformation process was characterized, and the best prescription was obtained, and the content determination method of TR ophthalmic thermosensitive gel was established by HPLC. Methods: The gel dissolution and release of TR were studied. The retention time and local irritation of TR thermosensitive gel in rabbit eyes were investigated. The prospect of clinical application.
【學位授予單位】：第三軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R943

【參考文獻】

相關期刊論文 前10條

1 刁雨輝;周建平;胡雄林;;眼用溫敏凝膠釋放度及其影響因素研究[J];廣東藥學院學報;2008年06期

2 張利;畢宏生;解孝鋒;郭俊國;;眼用原位凝膠的藥學研究進展[J];國際眼科雜志;2011年10期

3 吳偉,崔光華;星點設計-效應面優(yōu)化法及其在藥學中的應用[J];國外醫(yī)學.藥學分冊;2000年05期

4 邱永鋒;李曼;沙先誼;方曉玲;;促透劑對氫溴酸高烏甲素凝膠體外透皮吸收的影響[J];貴陽中醫(yī)學院學報;2012年04期

5 郜琪臻;劉華麗;魏靜;丁平田;;鹽酸米諾環(huán)素原位凝膠的流變學研究[J];化學與生物工程;2012年01期

6 黃蔚茹;;美多麗與阿托品眼膏在兒童弱視驗光作用的比較探討[J];醫(yī)藥論壇雜志;2011年06期

7 沈雪;常笛;姜同英;王思玲;;硫普羅寧眼用原位凝膠的研制[J];沈陽藥科大學學報;2009年02期

8 林友文;莊小慧;李光文;康建軍;;殼聚糖-羧甲基殼聚糖凝膠的溫敏性及體外藥物緩釋試驗[J];中國現(xiàn)代應用藥學;2010年07期

9 仇海鎮(zhèn);李娟;;原位凝膠的研究進展及其在藥劑學中的應用[J];醫(yī)學信息;2010年05期

10 吳麗萌;朱國嬋;陳光龍;;加替沙星熱敏型凝膠滴眼液的制備與質量控制[J];中國藥業(yè);2009年23期

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