Experimental acute anti-inflammatory activity of preparations with complexed cannabidiol in carriers

Abstract Cannabidiol (CBD) is a bioactive compound with promising anti-inflammatory results but has low aqueous solubility. Complexation of drugs with this characteristic in carriers is an alternative to improve their efficiency. This study aimed to prepare and characterize CBD complexes in different carriers, and to evaluate the anti-inflammatory effect of such preparations using an experimental model of edema induction in rat paws. The results were compared to a reference drug, ibuprofen (IBU). The carriers evaluated were beta cyclodextrin (bCD) and activated charcoal (AC). Quantification of the drugs in the complexes was determined, and different qualitative analyses were also performed. Oral treatments in single doses with CBD showed inhibitory effects similar to that of IBU, potentiating its bioactivity without significant adverse effects. CBD*bCD doses at 4.375, 8.75, 17.5, and 35 mg/kg significantly reduced the intensity of edema compared to equivalent doses of pure bioactive. In contrast, CBD*AC did not generate benefits. There was no significant inhibitory effect on myeloperoxidase activity, requiring more specific analyses to assess this parameter. The results suggest that the CBD*bCD complexation is perfectly feasible, increasing its anti-edematogenic efficacy in the experimental model used.

Study on the inclusion technic of hydroxypropyl-β-cyclodextrin in tea tree oil and evaluation of its included compounds / 国际中医中药杂志

Objective@#This study takes hydroxypropyl-β-cyclodextrin (HP-β-CD) as the inclusion materials to optimize the preparation technic of tea tree oil (TTO) and evaluate its pharmaceutical performance.@*Methods@#Take the production rate of HP-β-CD tea tree oil inclusion and entrapment rate as the evaluation index, taking the orthogonal test method to optimize the production technic of tea tree oil (HP-β-CD inclusion and using infrared (IR), differential thermal scanning (DSC) method to characterize the inclusion compound to analyze the stability of TTO-HP-β-CD.@*Results@#The best technic to produce HP-β-CD tea tree oil is as follow: the ratio of TTO and HP-β-CD should be equal to 1/10, at 40 ℃, within 1 h. The average drug loading shoud be 9.25% ± 3.25%. The IR, DSC characterization results showed that the characteristic peak of tea tree oil disappeared after the microspheres, which indicated the HP-β-CD encapsulated the tea tree oil with good compatibility. In 80 ℃ water bath, the TTO-HP-β-CD was stable with the retention rate 40% after 8 h, the retention rate was 4.32 times than that of the unwrapped tea tree oil.@*Conclusions@#The HP-β-CD tea tree oil obviously has higher rate of inclusion and stability. Therefore, it’s worth to promoting and being used in the pharmacy preparations and cosmetics field.

Extraction Technology of Volatile Oil from Qingjie Granules and Optimization of Inclusion Technology by Box-Behnken / 中国中医药信息杂志

Objective To optimize the extraction of volatile oil from Qingjie Granules and process of inclusion compound of beta-cyclodextrin. Methods Water Distillation was used for extraction. Extraction time, grinding degree, and the amount of water were set as inspection factors, and volatile oil volume was set as the evaluation index to inspect extraction process of volatile oil from Qingjie Granules. With inclusion rate as the evaluation index, the single factor test and the Box-Behnken combined with the response surface method were used to choose the optimum inclusion process. Results The optimum extraction process for Schizonepetae Herba, Saposhnikoviae Radix and Forsythiae Fructus coarse powder should be with 10 times amount of water, extracting 3 h. Inclusion method should be saturated water solution method, and the inclusion process of volatile oil as feed and beta-cyclodextrin inclusion ratio was 1:12; the temperature was 40 ℃; inclusion time was 3.5 h. By means of TLC, UV and IR spectra, the formation of the inclusion compound of volatile oil in clear solution particles was preliminarily proved. Conclusion The optimum extraction and inclusion process of volatile oil from Qingjie Granules are stable and feasible, which can be used in industrial production.

Lipid Raft Integrity Is Required for Survival of Triple Negative Breast Cancer Cells / 한국유방암학회지

PURPOSE: Lipid rafts are cholesterol enriched microdomains that colocalize signaling pathways involved in cell proliferation, metastasis, and angiogenesis. We examined the effect of methyl-β-cyclodextrin (MβCD)-mediated cholesterol extraction on the proliferation, adhesion, invasion, and angiogenesis of triple negative breast cancer (TNBC) cells. METHODS: We measured cholesterol and estimated cell toxicity. Detergent resistant membrane (DRM) and non-DRM fractions were separated using the OptiPrep gradient method. Cell cycles stages were analyzed by flow cytometry, apoptosis was assessed using the TdT-mediated dUTP nick end-labeling assay, and metastasis was determined using a Matrigel invasion assay. Neo-vessel pattern and levels of angiogenic modulators were determined using an in vitro angiogenesis assay and an angiogenesis array, respectively. RESULTS: The present study found that the cholesterol-depleting agent MβCD, efficiently depleted membrane cholesterol and caused concentration dependent (0.1–0.5 mM) cytotoxicity compared to nystatin and filipin III in TNBC cell lines, MDA-MB 231 and MDA-MB 468. A reduced proportion of caveolin-1 found in DRM fractions indicated a cholesterol extraction-induced disruption of lipid raft integrity. MβCD inhibited 52% of MDA-MB 231 cell adhesion on fibronectin and 56% of MDA-MB 468 cell adhesion on vitronectin, while invasiveness of these cells was decreased by 48% and 52% respectively, following MβCD treatment (48 hours). MβCD also caused cell cycle arrest at the G2M phase and apoptosis in MDA-MB 231 cells (25% and 58% cells, respectively) and in MDA-MB 468 cells (30% and 38% cells, respectively). We found that MβCD treated cells caused a 52% and 58% depletion of neovessel formation in both MDA-MB 231 and MDA-MB 468 cell lines, respectively. This study also demonstrated that MβCD treatment caused a respective 2.6- and 2.5-fold depletion of tyrosine protein kinase receptor (TEK) receptor tyrosine kinase levels in both TNBC cell lines. CONCLUSION: MβCD-induced cholesterol removal enhances alterations in lipid raft integrity, which reduces TNBC cell survival.

Preparation and characterization of nifedipine skeleton frame sustained release tablets with hydroxypropyl-beta-cyclodextrin and polyethylene glycolas as matrix / 重庆医学

Objective To prepare nifedipine (NP)skeleton frame sustained release tablets by adopting hydroxypropyl-beta-cy-clodextrin (HPC)and polyethylene glycol (PEG)as the matrix and to investigate its sustained release effect on NP.Methods NP skeleton frame sustained release tablets were prepared by the direct compression method after forming the inclusion complex with NP and HPC on a 1∶1 ratio and mixing with PEG.The in vitro drug release curves and the in vivo concentration-time curves of NP skeleton frame sustained release tablets were detected with the NP-PEG tablet without inclusion complex,NP-HPC tablet and NP powder capsule (NP capsule)as the control.Results The 5 h cumulative release rate of NP skeleton frame sustained release tablet in the simulated gastric fluid test(pH 1.2)was 36.4%,its sustained release effect was significantly stronger than that of the NP-HPC tablet group (80%,P<0.01)and the NP-PEG tablet group (100%,P<0.01).The in vivo test showed that AUC0-12 [(6 413±436)h/(ng·mL)],Cmax[(983±192)ng/mL]and tmax[(5.7±1.1)h]in the NP skeleton frame sustained release group were significantly higher than those in the NP-HPC tablet group,the NP-PEG tablet group and the NP Capsule group (all P<0.01 or 0.05).Conclusion NP skeleton frame sustained release tablets exhibits the sustained release effect on NP,can extend the NP in vitro release time,and improve the NP′s bioavailability,which is a novel NP sustained release tablet with excellent properties.

An Investigation on Enhancement of Solubility of 5 Fluorouracil by Applying Complexation Technique- Characterization, Dissolution and Molecular-Modeling Studies.

The drug 5 fluorouracil is sparingly soluble in water. The aqueous solubility and dissolution rate of 5-fluorouracil can be increased by inclusion complexation with β-cyclodextrin. Molecular-modeling studies support the formation of stable molecular inclusion complexation of 5-fluorouracil with β-cyclodextrin monomer (1:1). Complexes were prepared by physical mixture, kneading, co evaporation and freeze drying methods. Two ratios 1:1 and 1:2 were formulated. These eight complexes were subjected to Phase-solubility study, molecular modeling and dissolution study. The complexes formed were confirmed by DSC studies. Phase solubility profile indicated that the solubility of 5-fluorouracil increased in the presence of β-cyclodextrin monomer. Results obtained by different characterization techniques clearly indicate that the freeze-drying method leads to formation of solid state complexes between 5-fluorouracil and β-cyclodextrin. The complexation of 5-fluorouracil with β- cyclodextrin lends an ample credence for better therapeutic efficacy.

Comparative Study of Beta Cyclodextrin and Calix-8-Arene as Ionophores in Potentiometric Ion-Selective Electrodes for Sitagliptin Phosphate.

Two novel Sitagliptin (STG) selective electrodes were investigated with di-octyl phthalate as a plasticizer in a polymeric matrix of polyvinyl chloride (PVC). Sensor 1 was fabricated using β-cyclodextrin, while sensor 2 was constructed using calix-8-arene as ionophores. Linear responses of STG within the concentration ranges of 10−7 to 10−2, and 10−8 to 10−2 mol L−1 were obtained using sensors 1 and 2, respectively. Nernstian slopes of 58.57 and 59.88 mV/decade over the pH range of 5-6.5 were observed. The selectivity coefficients of the developed sensors indicated excellent selectivity for STG. The proposed sensors displayed useful analytical characteristics for the determination of STG in bulk powder, different pharmaceutical formulations, and biological fluids (plasma and urine).The two novel electrodes offer the advantage of determination of STG in biological fluids without pretreatment which is convenient for monitoring STG levels in clinical studies.

Synergistic activity of ofloxacin and ornidazole on biomedical fabrics against nosocomial pathogens.

Nosocomial infections in the hospitals disseminated from the cotton fabrics of health care professionals and patients leads to severe complications like respiratory, gastrointestinal and urinary tract infections. Since the hospital based textile materials like nylon and polyester has good surface properties, it can harbour large number of microorganisms. Hence in this study, two different antibacterial drugs showing synergistic properties were attached to different fabricsusing tocopherol acetate as a cross-linker with the aim that, treated fabric could act as barriers against transmission of challenge organisms. Inorder to decrease the drug resistant property of the nosocomial pathogens, a fluoroquinolone and a nitroimidazole compounds were mixed at suitable composition based on their synergistic behaviour. Both the compounds were modified to act as reactive dyes and were covalently bonded to the surface of nylon and polyester in order to impart antibacterial properties. The assay used for measuring antibacterial properties was based on the AATCC Test Method-100. The treated fabric was also subjected to multiple washings to determine its durability based on the AATCC Test Method-124. To determine the mode of action of these drugs, DNA of the drug exposed and unexposed challenge organisms were extracted and analysed by agarose gel electrophoresis. The difference in the number of viable bacteria after ‘0’ contact time and 18 hours contact time with treated fabrics were statistically calculated with P<0.05 considered significant.

Validation of quantitative analysis method for triamcinolone in ternary complexes by UV-Vis spectrophotometry / Validação de método de análise quantitativa para a triancinolona a partir de complexo ternário por espectrofotometria de UV-Vis

Triamcinolone (TRI), a drug widely used in the treatment of ocular inflammatory diseases, is practically insoluble in water, which limits its use in eye drops. Cyclodextrins (CDs) have been used to increase the solubility or dissolution rate of drugs. The purpose of the present study was to validate a UV-Vis spectrophotometric method for quantitative analysis of TRI in inclusion complexes with beta-cyclodextrin (B-CD) associated with triethanolamine (TEA) (ternary complex). The proposed analytical method was validated with respect to the parameters established by the Brazilian regulatory National Agency of Sanitary Monitoring ANVISA). The analytical measurements of absorbance were made at 242nm, at room temperature, in a 1-cm path-length cuvette. The precision and accuracy studies were performed at five concentration levels (4, 8, 12, 18 and 20µg.mL-1. The B-CD associated with TEA did not provoke any alteration in the photochemical behavior of TRI. The results for the measured analytical parameters showed the success of the method. The standard curve was linear (r2 > 0.999) in the concentration range from 2 to 24 µg.mL-1. The method achieved good precision levels in the inter-day (relative standard deviation-RSD <3.4%) and reproducibility (RSD <3.8%) tests. The accuracy was about 80% and the pH changes introduced in the robustness study did not reveal any relevant interference at any of the studied concentrations. The experimental results demonstrate a simple, rapid and affordable UV-Vis spectrophotometric method that could be applied to the quantitation of TRI in this ternary complex.

A triancinolona (TRI) é um fármaco amplamente utilizado no tratamento de doenças inflamatórias do globo ocular e é praticamente insolúvel em água, o que limita sua utilização na forma de colírio. As ciclodextrinas (CDs) têm sido utilizadas com sucesso para aumentar a solubilidade ou velocidade de dissolução de fármacos. O presente estudo teve como objetivo a validação de uma metodologia analítica para a TRI a partir de complexos de inclusão com beta-ciclodextrina (B-CD) associada com a trietanolamina (TEA) (complexo ternário) por espectrofotometria de UV-Vis. A validação do método proposto foi realizada de acordo com os parâmetros analíticos estabelecidos pela Agência Nacional de Vigilância Sanitária (ANVISA). As análises quantitativas foram realizadas a 242nm a temperatura ambiente, utilizando cubeta de quartzo de 1cm. Os estudos de precisão e exatidão foram realizados para cinco níveis de concentração (4, 8, 12, 18 e 20µg.mL-1). A B-CD associada a TEA não alterou o comportamento fotoquímico da TRI. Os resultados da avaliação dos parâmetros analíticos demonstraram o sucesso da metodologia. A curva padrão apresentou linearidade (r2 > 0.999) na faixa de concentração de 2 a 24µg.mL-1. A metodologia apresentou bons níveis de precisão para o estudo inter dia (desvio padrão relativo-DPR <3.4%) e reprodutibilidade (DPR<3.8%). A exatidão ficou em torno de 80% e a variação de pH inserida no estudo de robustez não revelou uma interferência significativa em todas as concentrações estudadas. Os resultados experimentais demonstraram um simples, rápido e viável método de espectrofotometria de UV-Vis com aplicabilidade para a análise quantitativa da TRI a partir do complexo ternário.

Cholesterol, a Major Component of Caveolae, Down-regulates Matrix Metalloproteinase-1 Expression through ERK/JNK Pathway in Cultured Human Dermal Fibroblasts

BACKGROUND: Cholesterol is a major component of specialized membrane microdomains known as lipid rafts or caveolae, which modulate the fluidity of biological membranes. Membrane cholesterol therefore plays an important role in cell signaling and vesicular transport. OBJECTIVE: In this study, we investigated the effects of cholesterol on matrix metalloproteinase-1 (MMP-1) expression in human dermal fibroblasts. METHODS: MMP-1 mRNA and protein expression were determined by RT-PCR and Western blotting, respectively. AP-1 DNA binding activity was detected by electrophoretic mobility shift assays. The amount of cholesterol was analyzed by cholesterol assay kit. RESULTS: We observed that MMP-1 mRNA and protein expression was dose-dependently decreased by cholesterol treatment. In contrast, cholesterol depletion by a cholesterol depletion agent, methyl-beta-cyclodextrin (M beta CD) in human dermal fibroblasts, increased MMP-1 mRNA and protein expression in a dose-dependent manner. Also, we investigated the regulatory mechanism of M beta CD-induced MMP-1 expression: cholesterol depletion by M beta CD, activated ERK1/2 and JNK, but not p38 MAPK cascade, and it also significantly increased c-Jun phosphorylation, c-Fos expression and activator protein-1 binding activity. Furthermore, the inhibition of ERK or JNK with specific chemical inhibitors prevented M beta CD-induced MMP-1 expression, which indicates that ERK and JNK play an important role in cholesterol depletion-mediated MMP-1 induction. In addition, M beta CD-induced phosphorylation of ERK and JNK and MMP-1 expression were suppressed by cholesterol repletion. CONCLUSION: Our results suggest that cholesterol regulates MMP-1 expression through the control of ERK and JNK activity in human dermal fibroblasts.

GD3 Accumulation in Cell Surface Lipid Rafts Prior to Mitochondrial Targeting Contributes to Amyloid-beta-induced Apoptosis

Neuronal apoptosis induced by amyloid beta-peptide (A beta) plays an important role in the pathophysiology of Alzheimer's disease (AD). However, the molecular mechanism underlying A beta-induced apoptosis remains undetermined. The disialoganglioside GD3 involves ceramide-, Fas- and TNF-alpha-mediated apoptosis in lymphoid cells and hepatocytes. Although the implication of GD3 has been suggested, the precise role of GD3 in A beta-induced apoptosis is still unclear. Here, we investsigated the changes of GD3 metabolism and characterized the distribution and trafficking of GD3 during A beta-induced apoptosis using human brain-derived TE671 cells. Extracellular A beta induced apoptosis in a mitochondrial-dependent manner. GD3 level was negligible in the basal condition. However, in response to extracellular A beta, both the expression of GD3 synthase mRNA and the intracellular GD3 level were dramatically increased. Neosynthesized GD3 rapidly accumulated in cell surface lipid microdomains, and was then translocated to mitochondria to execute the apoptosis. Disruption of membrane lipid microdomains with methyl-beta-cyclodextrin significantly prevented both GD3 accumulation in cell surface and A beta-induced apoptosis. Our data suggest that rapidly accumulated GD3 in plasma membrane lipid microdomains prior to mitochondrial translocation is one of the key events in A beta-induced apoptosis.

Encapsulation of lycopene using spray-drying and molecular inclusion processes

This study aimed to obtain encapsulated lycopene in a powder form, using either spray-drying or molecular inclusion with beta -cyclodextrin ( beta -CD) followed by freeze-drying. The encapsulation efficiency using spray-drying ranged from 94 to 96 percent, with an average yield of 51 percent, with microcapsules showing superficial indentations and lack of cracks and breakages. Lycopene- beta -CD complexes were only formed at a molar ratio of 1:4, and irregular structures of different sizes that eventually formed aggregates, similar to those of beta -CD, were observed after freeze-drying. About 50 percent of the initial lycopene did not form complexes with beta -CD. Lycopene purity increased from 96.4 to 98.1 percent after spray-drying, whereas lycopene purity decreased from 97.7 to 91.3 percent after complex formation and freeze-drying. Both the drying processes yielded pale-pink, dry, free-flowing powders.

Técnicas de encapsulamento, como "spray-drying" e formação de complexos por inclusão com ciclodextrinas, vêm sendo avaliadas para viabilizar a adição de carotenóides em sistemas hidrofílicos e aumentar a sua estabilidade durante o processamento e estocagem. Portanto, o objetivo do presente trabalho foi obter licopeno encapsulado na forma de pó, utilizando processos de "spray-drying" ou de inclusão molecular com beta -ciclodextrina (CD) seguido de liofilização. A eficiência do encapsulamento utilizando "spray-drying" variou de 94 a 96 por cento e o rendimento médio foi de 51 por cento, com as microcápsulas apresentando indentações superficiais, porém sem falhas ou aberturas na superfície. A formação de complexo licopeno- beta -CD ocorreu apenas quando utilizada razão molar de 1:4, e estruturas irregulares de diferentes tamanhos que eventualmente formaram agregados, similares às da beta -CD, foram observadas após liofilização. O licopeno não complexado neste processo ficou em torno de 50 por cento. A pureza do licopeno ( por cento área do all-trans-licopeno) aumentou de 96,4 para 98,1 por cento após o encapsulamento, enquanto que a pureza do licopeno diminuiu de 97,7 para 91,3 por cento após complexação e liofilização. Os dois processos de secagem resultaram em pós rosa claro, secos e com bom fluxo.

Methyl-beta-cyclodextrin inhibits cell growth and cell cycle arrest via a prostaglandin E(2) independent pathway

Methyl-beta-cyclodextrin, a cyclic oligosaccharide known for its interaction with the plasma membrane induces several events in cells including cell growth and anti-tumor activity. In this study, we have investigated the possible role of cyclooxygenase 2 (COX-2) in cell growth arrest induced by methyl-beta-cyclodextrin in Raw264.7 macrophage cells. Methyl-beta-cyclodextrin inhibited cell growth and arrested the cell cycle, and this cell cycle arrest reduced the population of cells in the S phase, and concomitantly reduced cyclin A and D expressions. Methyl-beta-cyclodextrin in a dose- and time-dependent manner, also induced COX-2 expression, prostaglandin E(2) (PGE(2)) synthesis, and COX-2 promoter activity. Pretreatment of cells with NS398, a COX-2 specific inhibitor completely blocked PGE(2) synthesis induced by methyl-beta-cyclodextrin, however inhibition on cell proliferation and cell cycle arrest was not effected, suggesting non-association of COX-2 in the cell cycle arrest. These results suggest that methyl-beta-cyclodextrin induced cell growth inhibition and cell cycle arrest in Raw264.7 cells may be mediated by cyclin A and D1 expression.

Current and Emerging Antifungal Agents in Japan / 대한의진균학회지

Currently available antifungal agents for the treatment of systemic fungal infections in Japan are smaller in number than those in the United States and Europe and probably in Korea. Therefore, the development of novel antifungal drugs for clinical use, including new formulations of approved agents, with advantages over and/or complimentary to existing agents is particularly needed in Japan. In this review, I have described historical perspectives of existing systemic antifungal agents and provided a brief overview of the current status of clinical development of several different categories of new drugs as follows: (1) liposomal amphotericin B; (2) itraconazole-hydroxypropyl-beta-cyclodextrin complexes; (3) phosphatyl fluconazole (phosfluconazole) ; (4) voriconazole; and (5) micafungin (FK463). At this moment, micafungin, a member of echinocandins attracts the greatest attention of Japanese medical mycologists because it has just been introduced into the clinic and has unique chemical and biological characteristics distinct from any other existing class of antifungal drugs. Micafungin, as well as other new drugs under clinical development, should constitute effective new options for the management of a variety of systemic fungal infections.