In vivo Pharmacokinetic Analysis of Shenbai Nanosuspension in Rats Based on Overall Composition of Chinese Materia Medica / 中国实验方剂学杂志

ObjectiveTo clarify the scientific validity of in vivo pharmacokinetic determination of the whole drug composition in Shenbai nanosuspension in rats， and to provide methodological guidance and theoretical basis for the in vivo study of multi-component complex system of traditional Chinese medicine（TCM） preparations. MethodThe concentration of the overall components， mainly total saponins and total polysaccharides in Shenbai decoction and Shenbai nanosuspension， was determined in rat plasma at different times by area under the absorbance-wavelength curve method（AUAWC）， and the concentration of individual ginsenoside Rg1 was determined by high performance liquid chromatography（HPLC）， and the methodology was verified. The pharmacokinetic parameters of the whole component were compared with those of ginsenoside Rg1 to evaluate the in vivo operational characteristics of the two preparations. ResultThe methodological investigations of AUAWC and HPLC were in accordance with the requirements. AUAWC analysis showed that the overall components in both the decoction group and the nanosuspension group showed a one-compartment model， with half-life（t1/2） of 2.43 h and 2.04 h， respectively. The relative bioavailability of Shenbai nanosuspension was 138.99%. HPLC assay showed that ginsenoside Rg1 in the decoction group and the nanosuspension group showed a two-compartment model， with distribution half-life（t1/2α） of 0.13 h and 2.55 h， and elimination half-life（t1/2β） were 14.28 h and 3.85 h， respectively. The relative bioavailability of Shenbai nanosuspension was 127.49%. Compared with Shenbai decoction， the time to peak（tmax）， peak concentration（Cmax） and area under the drug-time curve（AUC） of the overall components and ginsenoside Rg1 in Shenbai nanosuspension were increased. ConclusionThe established AUAWC can be used for the pharmacokinetic study of the overall components of TCM preparations， which is complementary to the results of individual components measured by HPLC， and can provide useful reference for the in vivo study of new dosage forms of TCM.

Research progress on the preparation and application of flavonoid nanocrystals / 浙江大学学报·医学版

Flavonoids have been reported to possess significant pharmacological activities,such as antioxidant, anti-inflammatory and anticancer effects. However, the low solubility and low bioavailability limits their clinical application. Nanocrystal technology can solve the delivery problems of flavonoids by reducing particle size, increasing the solubility of insoluble drugs and improving their bioavailability. This article summaries nanosuspension preparation methods and the stabilizers for flavonoid nanocrystals, and reviews the drug delivery routes including oral, Injection and transdermal of flavonoid nanocrystals, to provide information for further research on nanocrystal delivery system of flavonoids.

Optimization of formulation of paclitaxel nanosuspension encapsulated by erythrocyte membrane based on Box-Behnken method / 中国中药杂志

In view of the longevity and innate immune escape of red blood cells, this study designed the red blood cell membrane-coated paclitaxel nanosuspension [RBC-(PTX)NS] and investigated its physicochemical properties and antitumor effect in vitro. Paclitaxel nanosuspension [(PTX)NS] was prepared by ultrasonic precipitation and then RBC-(PTX)NS by ultrasonic coating. The formulation of(PTX)NS was optimized with Box-Behnken method and indexes of particle diameter, zeta potential, and stability. The morphology, particle diameter, stability, in vitro dissolution, and antitumor effect of(PTX)NS and RBC-(PTX)NS were characterized. The results showed that the particle diameter and zeta potential were(129.38±0.92) nm and(-22.41±0.48) mV, respectively, for the optimized(PTX)NS, while(142.5±0.68) nm and(-29.85±0.53) mV, respectively, for RBC-(PTX)NS. Under the transmission electron microscope,(PTX)NS was spherical and RBC-(PTX)NS had obvious core-shell structure. RBC-(PTX)NS remained stable for 5 days at 4 ℃. The in vitro dissolution test demonstrated that the cumulative release rate of RBC-(PTX)NS reached 79% within 20 min, which was significantly higher than that(25%) of(PTX)NS(P<0.05). As evidenced by MTT assay, RBC-(PTX)NS highly inhibited the proliferation of HepG2 cells in a dose-dependent manner. The cell membrane-coated nano-preparation preparation method is simple and reproducible. It improves the solubility of PTX and endows RBC-(PTX)NS with higher stability and stronger cytotoxicity. Thus, it is a new method for the delivery of PTX via nanocrystallization.

Effect of solubilizing strategies on oral absorption of felodipine / 中国药科大学学报

@#As a typical BCS Ⅱ drug, felodipine exhibits low solubility and high permeability. We herein investigated the effects of different solubilization strategies on the oral absorption of felodipine. Firstly felodipine tablets based on 200 μm, 150 μm and 25 μm particle size of bulk drug were prepared. Meanwhile, felodipine solid dispersion and felodipine nanosuspension with average particle size of (168.90 ± 6.22) nm, PDI of 0.11 ± 0.06 were prepared. The absorption rate, apparent permeability coefficient (Papp), absorption quality in duodenum, jejunum, ileum and colon of rats and in vivo pharmacokinetics of the above different felodipine preparations were investigated. The results of rat single-pass intestinal perfusion showed that the absorption of felodipine preparations in duodenum, jejunum and ileum was better than in colon. Felodipine had a wide absorption window in the small intestine, with the best absorption site in the small intestine. Papp of different felodipine preparations was greater than 2.0 × 10-5 cm/s. Thus, the low solubility was the main factor limiting the absorption. In vivo pharmacokinetic experiments demonstrated the solubilization strategies significantly improved the bioavailability. The bioavailabilities of felodipine tablets with particle sizes of 150 and 25 μm, as well as nanosuspension, and solid dispersion were 138.75%, 173.01%, 208.65% and 314.53% that of the tablets with particle size of 200 μm, respectively. Solubilization strategies can significantly improve the gastrointestinal absorption rate and absorption quality of felodipine, and thus improve its bioavailability, which provides some reference for the research on the improvement of oral absorption of BCS II drugs.

Nanosuspension enhances dissolution rate and oral bioavailability of Terminalia arjuna bark extract in vivo and in vitro

Objective: To enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract by formulating its nanosuspension. Methods: Nanoprecipitation approach was used for the formulation of nanosuspension using polysorbate-80 as a stabilizer. The formulated nanosuspension was assessed for particle size, polydispersity index, zeta potential value and for in vitro dissolution study. Oral bioavailability studies were carried out in Wistar male albino rats by administering a single dose (50 mg/kg. b. wt) of the formulated nanosuspension and coarse suspension. The storage stability of the formulated nanosuspension was determined after three months of storage at room temperature and under the refrigerated condition. Mutagenicity assay was carried out to evaluate the toxicity of the formulated nanosuspension using two mutant strains (Salmonella typhimurium TA100 and Salmonella typhimurium TA98). Results: The mean particle size of the formulated nanosuspension was 90.53 nm with polydispersity index and zeta potential values of 0.175 and-15.7 mV, respectively. Terminalia arjuna nanosuspension showed improved dissolution rate and 1.33 fold higher oral bioavailability than its coarse suspension. The formulated nanosuspension also showed better stability under the refrigerated condition and was non-mutagenic against both strains. Conclusions: Our study demonstrates that nanosuspension technology can effectively enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract. Zafar Fatiqa 1 Department of Chemistry, University of Okara, Okara Jahan Nazish 2 Department of Chemistry, University of Agriculture, Faisalabad Khalil-Ur-Rahman 3 Department of Biochemistry, University of Agriculture, Faisalabad Asi Muhammad 4 Food Toxicology Lab, Plant Protection Division, Nuclear Institute for Agriculture and Biology, Faisalabad Zafar Waseeq-Ul-Islam 5 Department of Computer Science, COMSATS University of Information and Technology, Islamabad Pawar SS, Dahifale BR, Nagargoje SP, Shendge RS. Nanosuspension technologies for delivery of drugs. Nanosci Nanotech Res 2017; 4(2): 5966. Kilor V, Sapkal N, Daud A, Humne S, Gupta T. Development of stable nanosuspension loaded oral films of glimepiride with improved bioavailability. Int J Appl Pharm 2017; 9(2): 28-33. He J, Han Y, Xu G, Yin L, Neubi MN, Zhou J, et al. Preparation and evaluation of celecoxib nanosuspensions for bioavailability enhancement. RSC Adv 2017; 7: 13053-13064. Wang Y, Zheng Y, Zhang L, Wang Q, Zhang D. Stability of nanosuspensions in drug delivery. J Control Release 2013; 172(3): 11261141. ElShagea HN, ElKasabgy NA, Fahmy RH, Basalious EB. Freeze-dried self-nanoemulsifying self-nanosuspension (snesns): A new approach for the preparation of a highly drug-loaded dosage form. AAPS Pharm Sci Tech 2019; 20: 1-14. Gao L, Zhang D, Chen M, Duan C, Dai W, Jia L, et al. Studies on pharmacokinetics and tissue distribution of oridonin nanosuspensions. Int J Pharm 2008; 355(1-2): 321-327. Srivalli KMR, Mishra B. Drug nanocrystals: A way toward scale-up. Saudi Pharm J 2016; 24(4): 386-404. Geng T, Banerjee P, Lu Z, Zoghbi A, Li T, Wang B. Comparative study on stabilizing ability of food protein, non-ionic surfactant and anionic surfactant on BCS type Π drug carvedilol loaded nanosuspension: Physicochemical and pharmacokinetic investigation. Eur J Pharm Sci 2017; 109: 200-208. Jahan N, Rehman KU, Ali S, Bhatti IA. Antioxidant activity of gemmo therapeutically treated indigenous medicinal plants. Asian J Chem 2011; 23: 3461-3470. Zafar F, Jahan N, Rahman KU, Khan A, Akram W. Cardioprotective potential of polyphenolic rich green combination in catecholamine induced myocardial necrosis in rabbits. Evid Based Complement Alternat Med 2015; 2015: 734903. Ramesh R, Dhanaraj T. GC-MS analysis of bioactive compounds in Terminalia arjuna root. Int J Multidiscip Res Dev 2015; 2: 460-462. Shanbhag D, Khandagale A. Screening and standardization of Terminalia arjuna used as medicine in homeopathy using hptlc method. Int J Ana Bioana Chem 2011; 1: 57-60. Pooja S. Production of flavonoids from Terminalia arjuna (ROXB.) in vivo and in vitro tissue cultures. Int J ChemTech Res 2014; 6: 881-885. Gao L, Liu G, Wang X, Liu F, Xu Y, Ma J. Preparation of a chemically stable quercetin formulation using nanosuspension technology. Int J Pharm 2011; 404(1-2): 231-237. Arshad MS, Sohaib M, Nadeem M, Saeed F, Imran A, Javed A, et al. Status and trends of nutraceuticals from onion and onion by-products: A critical review. Cogent Food Agric 2017; 3: 1-14. Penalva R, Gonzalez-Navarro CJ, Gamazo C, Esparza I, Irache JM. Zein nanoparticles for oral delivery of quercetin: Pharmacokinetic studies and preventive anti-inflammatory effects in a mouse model of endotoxemia. Nanomedicine 2017; 13(1): 103-110. Kumar S, Pandey AK. Chemistry and biological activities of flavonoids: An overview. Sci World J2013; 2013: 162750. Thadkala K, Nanam PK, Rambabu B, Sailu C, Aukunuru J. Preparation and characterization of amorphous ezetimibe nanosuspensions intended for enhancement of oral bioavailability. Int J Pharm Investig 2014; 4(3): 131-137. Khan S, Iqbal T, Ahmed N, Jamil A. Antioxidant, hemolytic and mutagenic potential of Psoralea corylifolia. J Animal Plant Sci 2015; 25(5): 1451-1456. Gera S, Talluri S, Rangaraj N. Formulation and evaluation of naringenin nanosuspensions for bioavailability enhancement. AAPS Pharm Sci Tech 2017; 18(8): 3151-3162. Sun W, Mao S, Shi Y, Li LC, Fang L. Nanonization of itraconazole by high pressure homogenization: Stabilizer optimization and effect of particle size on oral absorption. J Pharm Sci 2010; 100(8): 3365-3373. Jahan N, Rehman KU, Ali S, Asi MR, Akhtar A. Cardioprotective potential of gemmomodified extract of Terminalia arjuna against chemically induced myocardial injury in rabbits. Pak Vet J 2012; 32: 255-259. Huang S, Chang WH. Advantages of nanotechnology-based chinese herb drugs on biological activities. Curr Drug Metab 2009; 10(8): 905-913. Dizaj SM, Vazifehasl Z, Salatin S, Adibkia K, Javadzadeh Y. Nanosizing of drugs: Effect on dissolution rate. Res Pharm Sci 2015; 10(2): 95-108. Abd-Elsalam WH, ElKasabgy NA. Mucoadhesive olaminosomes: A novel prolonged release nanocarrier of agomelatine for the treatment of ocular hypertension. Int J Pharm 2019; 560: 235-245. Rachmawati H, Shaal LA, Muller RH, Keck CM. Development of curcumin nanocrystal: Physical aspects. J Pharm Sci 2013; 102(1): 204214. Hong C, Dang Y, Lin G, Yao Y, Li G, Ji G, et al. Effects of stabilizing agents on the development of myricetin nanosuspension and its characterization: An in vitro and in vivo evaluation. Int J Pharm 2014; 477(1-2): 251-260. Karadag A, Ozcelik B, Huang Q. Quercetin nanosuspensions produced by high-pressure homogenization. J Agric Food Chem 2014; 62(8): 18521859. Papdiwal A, Pande V, Sagar K. Design and characterization of zaltoprofen nanosuspension by precipitation method. Der Pharma Chemica 2014; 6(3): 161-168. Sumathi R, Tamizharasi S, Gopinath K, Sivakumar T. Formulation, characterization and in vitro release study of silymarin nanosuspension. Indo Am J Pharm Sci 2017; 4: 85-94. [31]Thakkar HP, Patel BV, Thakkar SP. Development and characterization of nanosuspensions of olmesartan medoxomil for bioavailability enhancement. J Pharm Bioall Sci 2011; 3(3): 426-434. Mohd-Fuat AR, Kofi EA, Allan GG. Mutagenic and cytotoxic properties of three herbal plants from Southeast Asia. Trop Biomed 2007; 24(2): 4959. Ravichandran R. Studies on dissolution behaviour of nanoparticulate curcumin formulation. Adv Nanoparticles 2013; 2(1): 51-59. Hussain N, Jaitley V, Florence AT. Recent Advances in the understanding of uptake of microparticulates across the gastrointestinal lymphatics. Adv Drug Deliv Rev 2001; 50(1-2): 107-142. Yuan H, Chen J, Du YZ, Hu FQ, Zeng S, Zhao HL. Studies on oral absorption of stearic acid sln by a novel fluorometric method. Colloids Surf B Biointerfaces 2007; 58(2): 157-164. Gursoy RN, Benita S. Self-emulsifying drug delivery systems (sedds) for improved oral delivery of lipophilic drugs. Biomed Pharmacother 2004; 58(3): 173-182. Liu D, Pan H, He F, Wang X, Li J, Yang X, et al. Effect of particle size on oral absorption of carvedilol nanosuspensions: In vitro and in vivo evaluation. Int J Nanomed 2015; 10: 6425-6434. Wang Y, Zhang D, Liu Z, Liu G, Duan C, Jia L, et al. In vitro and in vivo evaluation of silybin nanosuspensions for oral and intravenous delivery. Nanotechnology 2010; 21(15): 1-12. Hao J, Gao Y, Zhao J, Zhang J, Li Q, Zhao Z, et al. Preparation and optimization of resveratrol nanosuspensions by antisolvent precipitation using box-behnken design. AAPS Pharm Sci Tech 2015; 16(1): 118-128.

Preparation of herpetolide A nanosuspension lyophilized powder and evaluation of its anti-hepatitis B virus activity / 中国中药杂志

To prepare the herpetolide A nanosuspension lyophilized powder(HPA-NS-LP), in order to investigate its anti-hepatitis B virus(HBV) activity and the dissolution in vitro. Herpetolide A nanosuspension(HPA-NS) was prepared by ultrasonic precipitation method. The formulation and process of HPA-NS were optimized by the single factor experiment. Lyophilized powder(HPA-NS-LP) was prepared by freeze-drying method. Scanning electron microscopy was used to observe morphology of HPA-NS-LP. Paddle method was used to determinate the dissolution of HPT-NS-LP in vitro. The anti-HBV activity of herpetolide A coarse suspension lyophilized powder(HPA-CS-LP) and HPA-NS-LP was evaluated by HepG2.2.15 cell model. The mean particle size of optimized HPA-NS was(173.46±4.36) nm, with a polydispersity index of 0.110±0.012. After redispersion, the mean particle size and the polydispersity index of HPA-NS-LP increased, with changes within a rational range. Scanning electron microscopy showed that HPA-NS-LP was spherical in shape. Cumulative dissolution rate of HPA-NS-LP was more than 90% in 2 hours, which was higher than that of HPA-CS-LP. Both HPA-CS-LP and HPA-NS-LP could effectively inhibit the secretion of HepG2.2.15 cell antigens(HBsAg and HBeAg), and the inhibitory effect of HPA-NS-LP was significantly higher than that of HPA CS-LP(P<0.05). HBV-DNA test showed that high, medium and low-dose HPA-NS-LP(50, 25, 12.5 mg·kg~(-1)) significantly decreased the level of HBV-DNA(P<0.05), and the effect was better than that of the same dose of HPA-CS-LP(P<0.05). The results revealed that HPA-NS-LP exhibited anti-HBV activity in vitro, and its effect was superior to that of HPA-CS-LP.

Preparation and quality evaluation of orodispersible film containing ginkgolide B novel nanosuspension lyophilized powder / 中国中药杂志

To prepare a new dosage form that can improve the drug loading of the film--ginkgolide B nanosuspension lyophilized powder orodispersible film(GB-NS-LP-ODF) and to evaluate its quality. Firstly, ginkgolide B nanosuspension(GB-NS) was prepared by media milling method, and then ginkgolide B nanosuspension lyophilized powder(GB-NS-LP) was prepared with freeze-drying method. The mannitol was used as lyoprotectant and its dosage was also investigated. GB-NS-LP-ODF was prepared by solvent casting method and its formulation was screened by single factor test method and optimized by orthogonal test. The appearance, mechanical properties, content uniformity and in vitro dissolution of the optimized GB-NS-LP-ODF were investigated. The particle size of prepared GB-NS was about 201 nm, and the optimal dosage of mannitol was 8%. According to the optimal formula, the GB-NS-LP-ODF was prepared with GB-NS-LP 35.6%, PVA 0588 49.4%, PEG 400 10.7% and CMS-Na 4.3%, and completely disintegrated in about 30 s, and the particle size of reconstituted GB nanoparticles from ODF was about 210 nm. The film with smooth appearance and good mechanical properties was stable within 30 days and the content uniformity(A+2.2 S<15) conformed to the regulations. Scanning electron microscope(SEM) showed that GB-NS-LP-ODFs were evenly distributed and the particle size was about 200 nm. X-rays diffraction(XRD) showed that its crystallinity was significantly lower than that of GB raw drug and GB-ODF. The results of in vitro release test showed that the drug film was completely dissoluted within 10 minutes. These results indicated that nanosuspension lyophilized powder was prepared by freeze drying of nanosuspensions, and then loaded into the orodispersible film to effectively increase the drug loading of the ODF and have broad application prospects.

Nanosuspension enhances dissolution rate and oral bioavailability of Terminalia arjuna bark extract in vivo and in vitro

Objective: To enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract by formulating its nanosuspension. Methods: Nanoprecipitation approach was used for the formulation of nanosuspension using polysorbate-80 as a stabilizer. The formulated nanosuspension was assessed for particle size, polydispersity index, zeta potential value and for in vitro dissolution study. Oral bioavailability studies were carried out in Wistar male albino rats by administering a single dose (50 mg/kg. b. wt) of the formulated nanosuspension and coarse suspension. The storage stability of the formulated nanosuspension was determined after three months of storage at room temperature and under the refrigerated condition. Mutagenicity assay was carried out to evaluate the toxicity of the formulated nanosuspension using two mutant strains (Salmonella typhimurium TA100 and Salmonella typhimurium TA98).Results: The mean particle size of the formulated nanosuspension was 90.53 nm with polydispersity index and zeta potential values of 0.175 and ?15.7 mV, respectively. Terminalia arjuna nanosuspension showed improved dissolution rate and 1.33-fold higher oral bioavailability than its coarse suspension. The formulated nanosuspension also showed better stability under the refrigerated condition and was non-mutagenic against both strains. Conclusions: Our study demonstrates that nanosuspension technology can effectively enhance the dissolution rate and oral bioavailability of Terminalia arjuna bark extract.

Preparation and in vitro release study of ligustrazine pamoate sustained-release nanosuspension / 中草药

Objective: To prepare ligustrazine pamoate (Lig-PAM) sustained-release nanosuspension (Lig-PAM-NSps) and determine its in vitro release characteristics. Methods: Lig-PAM was prepared by hydrophobic salt formation method and its physicochemical properties were characterized. Then, Lig-PAM-NSps was prepared by miniaturized medium grinding method. The prescription and preparation process of Lig-PAM-NSps were optimized by the single factor and orthogonal experiment with average particle size, polydispersity index (PDI) and stability coefficient (SI) as indicators. Lig-PAM-NSps was characterized, and its stability and in vitro release was also investigated. Results: The compound ratio of Lig-PAM prepared by Lig and PAM in the amount of 1:1 was (97.48 ± 0.04)%. Compared with Lig, the solubilities of Lig-PAM in water and simulated body fluids were decreased by 95.50% and 77.39%, respectively. Fourier transform infrared spectroscopy (FT-IR) and X-ray powder diffraction (XRD) showed that the Lig and PAM formed Lig-PAM. The optimum prescription size of Lig-PAM-NSps was (585 ± 5) nm, PDI was (0.328 ± 0.015) and SI was (0.928 ± 0.012). The scanning electron microscopy (SEM) showed that Lig-PAM-NSps was spherical with uniform size distribution, and the particle size was about 600 nm and its physical stability was good within 60 d. The results of in vitro release showed that Lig-PAM-NSps had obvious sustained-release effect compared with Lig solution within 48 h, and showed the first-order release characteristics [ln(1-Q) = 0.153 67 t + 80.458 14, r = 0.998 26]. Conclusion: The preparation progress of Lig-PAM-NSps is stable and can release Lig slowly in vitro.

Preparation of silymarin nanosuspension with glycyrrhizic acid as stabilizer and study of its stabilization mechanism / 中草药

Objective: To prepare silymarin nanosuspension (SM-NS) with glycyrrhizic acid as stabilizer, and investigate the in vitro release characteristics and charge stabilization mechanism. Methods: SM-NS was prepared by high-speed shear-high pressure homogenization method. SM-NS lyophilized powder were prepared by freeze-drying method and characterized by physical and chemical characterization and in vitro release. The stability mechanism of SM-NS was studied from the ionic strength and pH value. Results: The dosage of glycyrrhizic acid (GA) was 0.15%. The preparation process was shear rate of 19 000 r/min, shear time of 4 min, homogenization pressure of 100 MPa, homogenization times of 12 times, and lyoprotectant was mannitol 3%, the average particle size of SM-NS lyophilized powder was (516.4 ± 10.4) nm, PDI was (0.260 ± 0.046); The in vitro release results showed that the dissolution rate and solubility of SM-NS lyophilized powder were significantly higher than the physical mixture; The study of charge stability mechanism showed that licorice acid can provide good charge stabilization and strong resistance to environmental impact. Conclusion: SM-NS is a potential and new nano-drug with high safety, which is formed by the charge stability of GA to significantly improve the solubility and stability of silymarin.

Study on Tissue Distribution of Apigenin Nanosuspension in Mice / 中国药房

OBJECTIVE：To study the distri bution and targeting characteristics of Apigenin nanosuspension （AP-NPs）in mice. METHODS：AP-NPs was prepared with ultrasound microprecipitation. Kunming mice were randomly divided into apigenin （AP） solution group and AP-NPs suspension group ，with 45 mice in each group. The mice were given relevant medicine intragastrically （80 mg/kg）；blood sample of eyeball 500 μL were collected before medication（0 h）and 0.25，0.5，1，2，4，6，8，10 h after medication. After the last blood collection ，the mice were sacrificed and their heart ，liver，spleen，lung，kidney and brain tissues were taken. After protein precipitation with methanol ，HPLC method was adopted for determining plasma and tissues. The determination was performed on Shimadzu ODS-SP column with mobile phase consisted of methanol-water （70 ∶ 30，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 340 nm，and column temperature was 35 ℃. The sample size was 20 μL. The concentration of AP in different samples was calculated according to standard curve，main pharmacokinetic parameters （AUC，cmax）of AP and the ratio of peak concentration （ce），relative uptake rate （RUE），uptake ratio and its change value were calculated with DAS 2.0 software and Excel 2010 software；the tissue distribution and targeting characteristics of AP were analyzed. RESULTS：The linear range of AP in plasma and tissue s were 0.1-25.0 μg/mL（all r＞0.99）；the lower limits of quantification were 0.1 μg/mL. RSDs of intra-day and inter-day were all lower than 15％，and the accuracy were 94.37％-117.48％. The extraction recovery rates were all more than 80％. Compared with AP solution group ，the concentrations of AP in plasma sample （during 0.5-6 h），liver tissue （during 0.25-8 h），spleen tissue （during 0.25-8 h）and cerebral tissue （during 0.25-4 h）were increased significantly in AP-NPs suspension group （P＜0.05 or P＜0.01），and the highest in liver tissue. The concentrations of AP in heart tiusse （6 h），liver tissue （10 h），lung tissue （0.5 h），spleen tissue （during 0.25-10 h）were decreased significantly （P＜ 0.05 or P＜0.01）. There was statistical significance in AUC and cmax of AP in plasma and tissue samples between 2 groups（P＜ 0.05）. The ce，RUE，uptake ratio and its change value of liver tissue were the highest ，being 1.34±0.40，1.99±0.29，48.49％ and 15.71％ . CONCLUSIONS ：After AP is made into nanosus- pension，the distribution of drug tissue is changed ，especially targeting effect on liver tissue is improved.

Multiparticulate Drug Delivery System For Gastrointestinal Tuberculosis

Drug solubility poses numerous challenges in design of formulations for drugs with poor aqueous solubility. Ethionamide is an antitubercular drug belonging to biopharmaceutical classification system class II drug having less aqueous solubility. Nanosuspensions were prepared by using various solvents such as methanol, ethanol, acetone and chloroform and it was prepared using anti-solvent precipitation technique by using probe sonication. Various stabilizers such as tocopherolpolythytlene glycol succinate, polyvinylpyrrolidone and tween 80 singly or in combination were studied. A 32 factorial design was employed to study the effect of independent variables, concentration of stabilizers and stirring speed on particle size and cumulative percent drug release. The particle size of the optimized batch was 97.54 ± 8.47 nm with polydispersity index of 0.36 and zeta potential -10.1 ± 2.3 mV. The cumulative percent drug release of optimized batch was found to be 95.01 ± 1.16% in 60 min. Optimized batch was ultracentrifuged and evaluated for saturation solubility studies, stability and powder X-ray Diffraction studies. Optimized nanosuspension was loaded on Espheres by spraying in a coating pan and then coating of Eudragit controlled release polymers. The coated Espheres were evaluated for drug content, friability, scanning electron microscopy, ex-vivo permeation studies and drug release kinetics studies. The friability value for primary coated sphere was found to be 0.8 ± 0.12% and for secondary was 1% and the best fit model was found to be Korsmeyer-Peppas model which is indicative of diffusion controlled release. Ex vivo diffusion studies revealed a moderate increase in permeability.

Preparation and in vitro anti-hepatic fibrosis evaluation of herpetone nanosuspensions / 中国中药杂志

Herpetone( HPT) is a bioactive lignan extracted from Herpetospermum pedunculosum,which can protect liver,lower aminotransferase and inhibit hepatitis B virus. However,HPT has a poor oral bioavailability due to its poor water solubility. And there is no report about whether HPT has an anti-hepatic fibrosis activity. To improve the dissolution of HPT and study its anti-hepatic fibrosis activity and mechanism,the study group prepared herpetone nanosuspensions( HPT-NS) by the miniaturized media milling method. The formulation and process of HPT-NS were optimized by the single factor experiment. Scanning electron microscopy was used to observe morphology of HPT-NS. Dialysis method was used to study dissolution of HPT-NS in vitro. CCK8 method was used to assess the effect of HPT-NS on proliferation of the rat hepatic stellate cells( HSC-T6). Flow cytometry was used to assess the effect of HPT-NS on apoptosis and cell cycle of HSC-T6. The mean particle size of optimized HPT-NS was( 196±7) nm with a polydispersity index of 0.279±0.009.SEM showed that HPT-NS was in a regular rod shape. The cumulative dissolution rate of HPT-NS reached 93% in 18 h,and was higher than that of herpetone coarse suspensions( HPT-CS,28%). CCK8 experiment showed that the inhibition rate of HPT-NS on HSC-T6 was higher than that of HPT-CS. Flow cytometry showed that HPT-NS could block HSC-T6 cells in G2/M phase and induce apoptosis of HSC-T6 cells,with a significantly stronger effect than HPT-CS. The results revealed that HPT-NS significantly increased the in vitro dissolution of HPT,and enhanced the inhibitive effect on HSC-T6 cell proliferation by blocking cells in the G2/M phase and inducing late apoptosis.

Preparation of disulfiram naonosuspensions and their anti-tumor efficacy in vitro and in vivo / 药学学报

Disulfiram (DSF) is a traditional anti-alcohol drug, but it was recently found that DSF has strong inhibitory effect on the growth of a variety of cancer cells. However, its clinical application is greatly limited due to its poor solubility, instability in gastrointestinal tract and short plasma half-life. In this study, DSF is fabricated into nanosuspensions with the aim of trying to solve these problems. DSF nanosuspensions (DSF-NSps) were prepared by the anti-solvent precipitation method under ultrasonication, and the suitable stabilizer was screened according to the size, polydispersity index (PDI), and zeta potential of the resultant nanosuspensions, along with their particle size change during the storage at room temperature. The particle size, PDI, and zeta potential of DSF-NSps were determined using dynamic light scattering method, while the morphology of DSF-NSps was observed by transmission electronic microscope (TEM). The stability of DSF-NSps in media was examined according to their particle size change in different physiological media. The concentration of DSF was measured by HPLC assay. The in vitro drug release was evaluated on basis of dialysis. MTT assay was employed to evaluate the in vitro cytotoxicity of DSF-NSps against cancer cell lines. The 4T1 tumor-bearing mouse model was used to evaluate the in vivo therapeutic efficacy of DSF-NSps. All the animal experiments were acquired according to the Regulations for Animal Experiments and Guidelines for Ethical as defined by Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College. As a result, the combinational use of soyabean lecithin (SPC) and D-alpha tocopherol acid polyethyene glycol succinate (TPGS) was determined to best stabilize DSF-NSps when the ratio of DSF-SPC-TPGS was 24∶20∶4 (weight ratio), with small particle size and good storage stability. The resultant DSF-NSps showed a regular spherical morphology and drug loading content of (45.36 ± 2.09) %, with average particle size of 175.00 ± 0.75 nm, PDI of 0.24 ± 0.07 and zeta potential of -14.3 mV. DSF-NSps displayed good particle size stability in a variety of biological media including phosphate buffer saline, normal saline, 5% glucose, artificial gastric fluid, artificial intestinal fluid and plasma, which would meet the demand of both intravenous and oral administration. The in vitro study demonstrated that nano-encapsulation greatly increased the stability of DSF in aqueous media, DSF-NSps exhibited sustained release of the encapsulated drug and significantly inhibited 4T1 cells compared to free DSF (IC50, 1.07 vs 5.53 μg·mL-1, P<0.01). DSF-NSps showed a good dose-response relationship on the 4T1 tumor-bearing mice with the tumor inhibition rates at the three doses being 80.22%, 75.14% and 66.10%, all higher than that of paclitaxel injections (55.01%, P<0.05). The in vivo biodistribution study displayed that DSF-NSps were mainly distributed into liver, spleen and tumor. In sum, disulfiram nanoparticles could be expected to provide an effective anti-cancer drug for the treatment of breast cancer.

Beneficial effects of nanosuspensions of honokiol in mice on high fat diet through suppression of hepatic gluconeogenesis / 药学学报

To investigate the potential hypoglycemic effect of nanosuspensions of honokiol and explore the underlying mechanisms, a high fat diet (HFD) was studied in C57BL/6J mice divided into five groups: normal diet (ND), HFD, HFD/honokiol-sodium carboxymethyl cellulose (CMC-Na) (Hono-CMC, 100 mg·kg-1), HFD/honokiol- Nano (Hono-Nano, 80 mg·kg-1), HFD/metformin (HFD/Met, 200 mg·kg-1). Fasting blood glucose (FBG) and body weights (BW) of mice were measured every seven days. After 30-day treatment, an oral glucose tolerance test (OGTT) was performed, and blood and tissue samples were collected for analysis. All animal experiments were approved by the Research Animal Care Committee of Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine. The data showed Hono-Nano and metformin reduced FBG, BW, and markedly improved OGTT of mice compared to HFD group (P<0.05). Hono-CMC produced nonsignificant impact on FBG, BW of mice, while OGTT of mice was improved by Hono-CMC (P<0.05). Meanwhile, none of these treated groups showed significant effects on regulating serum insulin levels, but all of them exhibited decreased serum glucagon levels notably compared to the HFD group (P<0.05). Western blot analysis revealed that honokiol up-regulated levels of p-AMPK and p-FOXO1 in liver tissue of HFD mice (P<0.05), which resulted in activation of AMPK and inhibition of FOXO1. Moreover, the expression of PEPCK (a key enzyme of gluconeogenesis) was decreased by honokiol (P<0.05). Taken together, our findings demonstrate that nanosuspension of honokiol is more effective than CMC-Na-suspension of honokiol on blood glucose controlling in HFD mice. The hypoglycemic effects of honokiol might rely on suppressing hepatic gluconeogenesis via activating AMPK and inhibiting FOXO1.

Optimization of compounded nanosuspension of paclitaxel and betulinic acid by Box-Behnken design-response surface method / 中草药

Objective: To prepare compounded nanosuspensions of paclitaxel and betulinic acid nanosuspension and optimize the preparation process. Methods: Nanosuspension was prepared by high pressure homogenization with poloxamer 188 and lecithin as stabilizer. Stabilizer concentration, stabilizer ratio, homogeneous pressure, and cycle times were selected as investigation factors and the particle size and PDI of nanosuspension were selected as evaluation index to optimize the prescription by Box-Behnken design-response surface method and the in vitro evaluation was determined. Results: The optimal formulation was as follows: stabilizer concentration 0.6 mg/mL, poloxamer 188-lecithin (2∶1), homogenous pressure 100 MPa, cycle times 20 times. The average particle size was (282.54 ± 5.40) nm, PDI was 0.242 ± 0.020. The nano-particles in the paclitaxel-betulinic acid compounded nanosuspension were rod-shaped. The nanosuspension had perfect redispersibility and satisfactory short-term stability. Paclitaxel and betulinic acid in nano lyophilized powders all existed in amorphous form. After being prepared into nanosuspension formulation, the solubility of paclitaxel in water was increased by about 90 times while that of betulinic acid was increased by about 100 times. In 2 h, the cumulative dissolution rate of paclitaxel and betulinic acid in nanosuspension of paclitaxel and betulinic acid were all up to 95%. Conclusion: Optimizing the preparation process of nano-suspension with Box-Behnken design-response surface method is effective and feasible and nanosuspension formulation can improve the dissolution of paclitaxel and betulinic acid observably.

In-vitro study of formulation and evaluation of nanosuspension of tamoxifen

Background: Nanosuspension technology has been developed as a promising candidate for efficient delivery of hydrophobic drugs. It could maintain the required crystalline state of the drug with reduced particle size, leading to an increased reporting on dissolution rate and therefore improved bioavailability.Methods: In this paper, we report on the preparation of Tamoxifen nanosuspension by high-pressure homogenization (HPH). The aim is to obtain a stable nanosuspension with an increased drug saturation solubility and dissolution velocity. The morphology and particle size distribution of the modified nanosuspensions were characterized by the means of several analyses that included: transmission electron microscopy (TEM), polarized light microscopy (PLM), scanning electron microscopy, differential scanning calorimetry (DSC) and powder X- ray diffractometry (XRD).Results: HPH was employed to produce aqueous drug nanosuspensions with fine solubility and dissolution properties, which render the produced particles stable up to one month. In addition, the prepared nanosuspensions possessed a high drug-loading efficiency (10%). The recoded zeta potential values (? -27 mV) indicated that the prepared nanosuspensions possess a higher degree of long-term stability. TEM data showed narrow size distribution with average size 322.7 nm. Morphologically, as indicated from results, the produced nanosuspensions have a homogenous distribution even after redispersion, indicating the stability of the product.Conclusions: It was possible to obtain Tamoxifen nanosuspensions with fine solubility and dissolution properties. Nanosuspensions possessed a high drug- loading (10%), which could reduce the dosage administration and gastrointestinal side effects. HPH can be employed to produce aqueous drug nanosuspensions that are stable up to one month. Aqueous nanosuspension can be converted to dry nanocrystals by lyophilization which offer superior physicochemical properties.

Preparation and in vitro Dissolution of Febuxostat Nanosuspension Sustained-release Pellets / 中国药师

Objective:To prepare febuxostat nanosuspension and prepare sustained-release pellets,and investigate the in vitro dis-solution.Methods: Febuxostat nanosuspension was prepared by a high pressure homogenization method. Febuxostat nanosuspension pellets were prepared by fluidized bed coating technique. Eudragit RL30D and Eudragit RS30D were used to prepare the sustained-re-lease pellets. The dissolution mechanism of febuxostat nanosuspension sustained-release pellets was evaluated. Results: The average particle size of the prepared febuxostat nanosuspension was (212.5 ± 36.3) nm, PdI was (0.193 ± 0.018), zeta potential was ( -12.4 ± 0.3) mV,and the scanning electron microscopy showed that the particle size distribution of febuxostat nanosuspension was narrow. The in vitro dissolution of febuxostat nanosuspension sustained-release pellets was more stable and conformed to the first-order release model. Conclusion: The in vitro dissolution of febuxostat nanosuspension sustained-release pellets is more stable, and the preparation provides a new choice for febuxostat clinical application.

Effects of three preparation technologies on the oral bioavailability of berberine α-hydroxy β-decanoylethyl sulfonate / 中成药

AIM To compare the effects of three preparation technologies on the oral bioavailability of HB (berberine α-hydroxy β-decanoylethyl sulfonate,houttuyn berberine).METHODS Solid dispersions,HP-β-CD inclusion complexes and nanosuspension freeze-dried powders were prepared.The suspensions of crude drug and these three preparations were intragastrically administered to SD rats,respectively.HPLC-MS/MS was adopted in the content determination of HB in plasma.then pharmacokinetics parameters were calculated.RESULTS Compared with the crude drug,three preparation technologies could significantly increase the Cmax value of this component (P ＜ 0.05),especially for HP-β-CD inclusion complexes (P ＜ 0.01).And HP-β-CD inclusion complexes demonstrated much higher AUC0-6h than the crude drug and the other two preparation technologies (P ＜ 0.05).CONCLUSION HP-β-CD inclusion complexes can effectively increase the oral bioavailability of HB.

Preparation of astilbin amorphous nanosuspension and its evaluation / 中国中药杂志

Astilbil nanosuspension (AT-NS) was prepared by an antisolvent precipitation method. The formula and process of AT-NS were optimized by the single factor experiment. AT-NS was prepared under the optimal conditions, and its morphology and crystallinity were characterized. In vitro release of AT-NS was also determined. The particle size of AT-NS stabilized by PVP K30 was (149±3) nm, and the polydispersity index (PDI) and stability index (SI) were 0.137±0.014 and 0.940±0.012, respectively. The results of SEM showed that AT-NS was spherical. Both XRD and DSC showed that AT was amorphous in nanosuspension. In the release test, AT-NS showed a significantly increased dissolution. This simple low-cost approach could prepare AT-NS successfully. AT-NS could significantly improve the dissolution of AT and provide the reference to break the limitation on the clinical application of AT.