In vitro and in vivo skin anti-aging evaluation of gel containing niosomes loaded with a semi-purified fraction containing gallic acid from Terminalia chebula galls

Context: The galls of Terminalia chebula Retz. (Combretaceae) frequently appear in many Thai Lanna medicinal plant recipes for promotion of longevity.

Objective: The objective of this study was to evaluate the skin anti-aging of gel containing niosomes loaded with a semi-purified fraction containing gallic acid from T. chebula galls.

Method: The semi-purified fraction containing phenolic compounds including gallic acid isolated from T. chebula galls loaded in non-elastic or elastic niosomes, and its developed gel, were evaluated for rabbit skin irritation by the closed patch test and skin anti-aging in human volunteers by measuring skin elasticity and roughness.

Results: Gel containing the fraction unloaded (SS) or loaded in non-elastic (SN) or elastic (SE) niosomes and gallic acid loaded in non-elastic (GN) or elastic (GE) niosomes showed no skin irritation, whereas the unloaded gallic acid (GS) gave the irritation in rabbit’s skin by the closed patch test. The % parameter changes of skin elastic recovery and skin elastic extension when applied with SN and SE gels were +28.73 and +32.57; ?21.25 and ?22.63%, respectively. SN and SE gel also showed a significant decrease of the maximum and average roughness values with the parameter changes of ?29.43 and ?32.38; ?39.47 and ?35.28%, respectively.

Conclusion: The semi-purified fraction loaded in niosomes indicated not only higher chemical stability of gallic acid containing in the fraction, but also more in vivo anti-aging activities than the unloaded fraction when incorporated in gel.     

Anti-inflammatory activity of gel containing novel elastic niosomes entrapped with diclofenac diethylammonium

The objective of this study was to develop a novel elastic bilayer vesicle entrapped with the non-steroidal anti-inflammatory drug (NSAID), diclofenac diethylammonium (DCFD) for topical use. Eighteen bilayer vesicular formulations composing of DPPC or Tween 61 or Span 60 mixed with cholesterol (at 1:1, 3:7 and 1:1 molar ratios, respectively) and ethanol at 0-25% (v/v), by chloroform film method with sonication were developed. The elastic Tween 61 niosomes which gave no sedimentation, no layer separation, unchanged particle sizes (about 200 nm) were selected to entrap DCFD. The entrapment efficiency of the drug in the conventional and elastic Tween 61 niosomes was 65 and 93%, respectively. At least 87% of DCFD determined by HPLC remained in elastic Tween 61 niosomes when kept at 4, 27 and 45 degrees C for 3 months. The deformability index values of the elastic niosomes were 13.76 and 3.44 times higher than the conventional niosomes entrapped and not entrapped with the drug, respectively, indicating the higher flexibility of the elastic vesicle especially, when entrapped with the drug. Transdermal absorption through excised rat skin was performed by vertical Franz diffusion cell at 32+/-2 degrees C for 6h. Gel containing elastic niosomes exhibited fluxes of DCFD in the stratum corneum (SC), deeper skin layer (viable epidermis and dermis, VED) and receiver chamber at 191.27+/-9.52, 16.97+/-2.77 and 3.76+/-0.54 microg/(cm2 h), whereas the commercial emulgel, containing an equivalent DCFD, gave 60.84+/-13.63, 7.33+/-1.70 and 0.14+/-0.01 microg/(cm2 h), respectively. The in vivo anti-inflammatory activity was evaluated by ethyl phenylpropiolate (EPP)-induced rat ear edema (n=3). DCFD entrapped in the developed elastic niosomes and incorporated in gel gave the same ear edema inhibition percentages of 23.81% at 30 min, but 2 and 9 times more inhibition percentages at 45 and 60 min than the commercial emulgel, respectively. This result has not only demonstrated the enhancement of transdermal absorption through rat skin, but also the in vivo anti-inflammatory effect of DCFD when entrapped in the developed novel elastic Tween 61 niosomes, as well.     

Triterpenoids from the barks of Terminalia chebula

Two new triterpenoids, termichebuloside A (1), an unusual dimeric triterpenoid saponin, and termichebulolide (2), an oleanolic acid-type lactone, along with 11 known triterpenoids, were isolated from MeOH extract of the barks of Terminalia chebula. The structures of 1 and 2 were elucidated to be arjunglucoside I-(3-O-19′,23-O-19′)-18,19-seco-19-hydroxyarjunglucoside I (1) and 2α,3β,23-trihydroxyolean-11,13(18)-dien-28,19β-olide (2), respectively, on the basis of spectroscopic evidences and biogenetic consideration.     

HPLC测定没食子中没食子酸的含量

目的　建立没食子中没食子酸的含量测定方法。方法　采用HPLC法测定没食子酸的含量 ,色谱柱为WatersSymmetryR○C18柱 (5 μm ,4 .6mm× 15 0mm) ;流动相为甲醇 - 0 .0 5 %磷酸溶液 (5∶95 ) ;流速 0 .8ml·min-1;检测波长 2 73nm。结果　没食子酸进样量在 0 .0 5 06～ 0 .4 0 6 0 μg范围内线性关系良好 ,平均加样回收率为 98.99% ,RSD =1.5 %。 结论　所用方法简便、快速、准确。     

Transdermal delivery enhancement of haloperidol from gel formulations by 1,8-cineole

The feasibility of using 10% 1,8-cineole as an enhancer for transdermal delivery of haloperidol has been examined. In-vitro transdermal delivery across full-thickness human, rabbit and hairless mouse skins was measured from three polymer gel systems, hypromellose (hydroxypropylmethylcellulose), Carbomer (Carbopol) 940 and macrogol (polyethylene glycol) using Franz cells. Values for the permeability coefficient kp, calculated as the product (Kh)x(D/h2) where these two factors were obtained from curve fitting of the non-steady-state equation over 24 h, were similar from the three formulations. The value of kp from hypromellose was significantly enhanced by cineole by factors of 6.2 (4.6-8.1), 5.6 (5.0-6.2) and 3.0 (2.6-3.4) for human, rabbit and mouse, respectively (mean and 95% confidence intervals). Enhancement ratios for K: 13.3 (8.3-20), 3.1 (2.5-3.9) and 2.0 (1.5-2.6), were higher than those for D: 0.47 (0.41-0.55), 1.8 (1.6-2.1) and 1.5 (1.3-1.8). This suggested that the barrier function of the skin lipids was marginally affected and the main effect was to increase the thermodynamic activity of the drug in the barrier. The enhancement achieved in human skin suggested that delivery could be safely enhanced by terpenoids.     

Transfollicular enhancement of methyl myristate loaded in cationic niosomes incorporated in hair lotion

Hair lotion containing methyl myristate loaded in cationic niosomes (HL-MMnio) composed of Brij72/cholesterol/DDAB at 7:3:0.65 molar ratio was developed. The remaining percentages of MM loaded in cationic niosomes in hair lotion were higher than free MM in hair lotion of about 1.2 times. The cumulative amounts in porcine skin and the receiver compartment of MM loaded in cationic niosomes incorporated in hair lotion were higher than those of free MM in hair lotion of 1.45 and 1.32 times, respectively. HL-MMnio showed very slightly irritation on rabbit skin, which was disappeared after 4 d. For melanogenesis induction in C57BL/6 mice with aged-induced grey body coat hairs, the highest pigmentation scores of HL-MMnio applied on the dorsal area were observed after 21 days, while hair lotion containing the free MM indicated after 35 days. This study has suggested that HL-MMnio was the high potential formulation for canities treatment.     

Transdermal delivery of drugs and enhancement of percutaneous absorption

This paper describes 1) the drug delivery through the skin to produce systemic effects, 2) the enhancement of percutaneous absorption by absorption enhancers, heating and complex formation, 3) the mechanism for the enhancement effect by enhancers, 4) the percutaneous absorption of peptides, and 5) the pharmacokinetic analysis for percutaneous absorption. 1,3-Dinitroglycerin, indomethacin (IND) and many drugs were efficiently absorbed via rat and rabbit skins in the presence of some enhancers, and using a microporous membrane therapeutic plasma concentrations were maintained for a long time. Enhancement of percutaneous absorption by the complex formation with fatty acid was observed for propranolol (PL) in vitro and in vivo. Heating at 42-45 degrees C also enhanced the percutaneous absorption dramatically, with decreased activation energies. The following mechanisms for the enhancement effect by enhancers were found: a) an increase in the fluidity of the stratum corneum lipids and reduction in the diffusional resistance to permeants, b) the removal of intercellular lipids and dilation between adherent cornified cells, c) an increase in the thermodynamic activity of drugs in vehicles, d) the exfoliation of stratum corneum cell membranes, the dissociation of adherent cornified cells and elimination of the barrier function. Peptides such as enkephalin, elcatonin and insulin were effectively absorbed through the skin in the presence of some enhancers and specific inhibitors, with no proteolytic degradation. The pharmacokinetic model with two parallel absorption processes, lipidic and aqueous pore transport pathways, in skin could adequately describe the percutaneous absorption of IND, PL and valproic acid. With peptides, a kinetic model including zero-order input rate, first-order permeation rate and first-order degradation rate was able to describe well the steady-state flux of peptides.     

Enhancement of gene expression and melanin production of human tyrosinase gene loaded in elastic cationic niosomes

Objectives Disturbance in the synthesis of tyrosinase might be one of the major causes of vitiligo. The enhancement of tyrosinase gene expression and melanin production by loading the plasmid in elastic cationic niosomes was investigated in tyrosinase gene knocked out human melanoma (M5) cells and in tyrosine‐producing mouse melanoma (B₁₆F₁₀) cells. Methods Niosomes composed of Tween 61/cholesterol/dimethyl dioctadecyl ammonium bromide at 1 : 1 : 0.5 molar ratio were prepared by the freeze‐dried empty liposomes method. The thin lipid film was redissolved in distilled water or 25% ethanol to obtain the non‐elastic or elastic cationic niosomes, respectively. Key findings The maximum loading of the plasmid in non‐elastic and elastic niosomes was 130 and 100 µg per 16 mg of the niosomal contents, respectively. The plasmid‐loaded elastic cationic niosomes exhibited high specific tyrosinase activity of 1.66 and 1.50 fold in M5 cells and 6.81 and 4.37 fold in B₁₆F₁₀ cells compared with the free plasmid and the plasmid‐loaded non‐elastic cationic niosomes, respectively. Conclusions This study has demonstrated not only the enhancement of the expression of human tyrosinase gene by loading in elastic cationic niosomes, but also the potential application of this gene delivery system for the further development of vitiligo gene therapy.     

水杨酸及其衍生物透皮吸收的研究

目的研究水杨酸及其衍生物的透皮吸收.方法用丙二醇作促透剂,用改良的Franz扩散池进行离体大鼠皮体外释放试验,紫外法测定累积释放量,计算平均透皮速率.结果含丙二醇乳膏中各药物的透皮速率分别为水杨酸38.722 5μg·cm2·h-1,赖氨匹林16.882 1μg·cm2·h-1,阿司匹林10.121 5 μg·cm2·h;不含丙二醇乳膏各药物透皮速率分别为水杨酸29.765 7μg·cm2·h-1,赖氨匹林10.690 0μg·cm2·h-1,阿司匹林6.965 3 μg·cm2·h-1.结论两种乳膏水杨酸、赖氨匹林、阿司匹林透皮速率差异显著;丙二醇对赖氨匹林和阿司匹林均有显著的促透作用.     

Effects of different processing methods in Tibet on content of chebulinic acid in Terminalia chebula Retz.

To study the change of the content of chebulinic acid in Terminalia chebula Retz. by different processing methods in Tibet. We taking chebulinic acid as an indicator, the contents in Terminalia chebula Retz. and its processed products were analyzed by using HPLC. Also, the change rule was explored. Compared with Terminalia chebula Retz., the contents of chebulinic acid have decreased after processing, declining 50.18% for the Rubia cordifolia L. processed product, while 11.2% reduce for the Euphorbia fischeriana Steud processed product. After the specific preparation process in Tibet, the contents of chebulinic acid have reduced significantly, which may lead to different pharmacological effects.     

Transdermal delivery of low molecular weight heparin loaded in flexible liposomes with bioavailability enhancement: comparison with ethosomes

Low molecular weight heparin (LMWH)-loaded flexible liposomes (flexosomes) were formulated for transdermal delivery, and their physicochemical and pharmacokinetic parameters were compared with LMWH-loaded ethosomes. Flexosomes had similar particle size compared with ethosomes, but their deformability was higher than that of ethosomes (76.7% vs. 46.8%). In vitro, flexosomes demonstrated 2.6-fold higher permeability coefficient than ethosomes. In comparison to LMWH aqueous solution, skin deposition of flexosome increased 3.2-fold, while that of ethosome increased only 2.0-fold. In vivo, after the topical application of flexosome to hairless mouse, [anti-Xa](max) was 1.11?IU/mL, while ethosomes showed only 0.32?IU/mL. Moreover, AUC(0-24?h) of flexosomes was 2.5-fold higher than ethosomes. In conclusion, the enhanced skin permeation and bioavailability of LMWH can be achieved with flexosomes in comparison with ethosomes. The LMWH transdermal delivery via flexosomes has the potential to replace the parenteral dosage forms for the treatment of venous thromboembolism, pulmonary embolism and cardiovascular events.     

Transdermal delivery of low molecular weight heparin loaded in flexible liposomes with bioavailability enhancement: comparison with ethosomes

Low molecular weight heparin (LMWH)-loaded flexible liposomes (flexosomes) were formulated for transdermal delivery, and their physicochemical and pharmacokinetic parameters were compared with LMWH-loaded ethosomes. Flexosomes had similar particle size compared with ethosomes, but their deformability was higher than that of ethosomes (76.7% vs. 46.8%). In vitro, flexosomes demonstrated 2.6-fold higher permeability coefficient than ethosomes. In comparison to LMWH aqueous solution, skin deposition of flexosome increased 3.2-fold, while that of ethosome increased only 2.0-fold. In vivo, after the topical application of flexosome to hairless mouse, [anti-Xa]_max was 1.11?IU/mL, while ethosomes showed only 0.32?IU/mL. Moreover, AUC_0–24?h of flexosomes was 2.5-fold higher than ethosomes. In conclusion, the enhanced skin permeation and bioavailability of LMWH can be achieved with flexosomes in comparison with ethosomes. The LMWH transdermal delivery via flexosomes has the potential to replace the parenteral dosage forms for the treatment of venous thromboembolism, pulmonary embolism and cardiovascular events.     

Transdermal absorption enhancing effect of the essential oil of Rosmarinus officinalis on percutaneous absorption of Na diclofenac from topical gel

Abstract

Context: Rosemary essential oil has been used topically for several purposes (analgesic, anti acne, and anti-inflammatory) in Iranian traditional medicine.

Objectives: This investigation aimed to study the effect of essential oil of Rosmarinus officinalis L. (Lamiaceae) on the transdermal absorption of Na diclofenac from topical gel.

Material and methods: Diclofenac sodium topical gel was prepared with HPMC K4M and Carbopol 934P as a gelling agent, and several vehicles. The most stable gel was chosen and enhancing effects of the essential oil with different concentrations (0.1, 0.5, and 1.0% w/w) on the permeation of diclofenac were evaluated. The anti-nociceptive effect of preparations was evaluated based on the formalin and tail flick tests in mice.

Results: The major constituents of the essential oil were 1,8-cineol (15.96%), α-pinene (13.38%), camphor (7.87%), bornyl acetate (6.54%), verbenone (5.82%), borneol (5.23%), camphene (4.96%), and (E)-caryophyllene (3.8%). Topical diclofenac containing 0.5% essential oil showed more analgesic effect after 25, 30, and 35?min (p?<?0.001) than the reference drug in the tail flick test. The analgesic effect of preparation containing 1% essential oil was more than reference gel after 15?min (p?<?0.05). This difference was observed after 20, 25, 30, 35, and 40?min (p?<?0.001) too. Rosemary essential oil 1% promoted analgesic effect of drug in comparison with diclofenac gel in the formalin early phase (p?<?0.05). The enhancing effect of rosemary was observed in 0.5 and 1% concentration (p?<?0.05 and p?<?0.001, respectively) in the late phase.

Conclusion: This study proved the enhancing effect of 0.5 and 1% of rosemary essential oil on diclofenac percutaneous absorption.     

Inhibition of HIV-1 integrase by galloyl glucoses from Terminalia chebula and flavonol glycoside gallates from Euphorbia pekinensis

The bioassay-directed isolation of Terminalia chebula fruits afforded four human immunodeficiency virus type 1 (HIV-1) integrase inhibitors, gallic acid ( 1) and three galloyl glucoses ( 2 - 4). In addition, four flavonol glycoside gallates ( 5 - 8) from Euphorbia pekinensis containing the galloyl moiety also showed the inhibitory activity at a level comparable to those of 2 - 4. By comparison with the activities of the compounds not bearing this moiety, it is proposed that the galloyl moiety plays a major role for inhibition against the 3'-processing of HIV-1 integrase of these compounds.     

In vitro evaluation on the antioxidant capacity of triethylchebulate, an aglycone from Terminalia chebula Retz fruit

Objectives:

To evaluate the antioxidant and free-radical scavenging activities of triethylchebulate (TCL), an aglycone isolated from the fruit of Terminalia chebula Retz.

Materials and Methods:

Microsomes, mitochondria and red blood cells (RBCs) were isolated from rat liver. The antioxidant capacities were evaluated by determining the inhibitory effects of TCL on lipid peroxidation, hydrogen peroxide (H₂O₂)-induced RBCs hemolysis and RBCs autoxidative hemolysis. The free-radical scavenging activities were evaluated by 1,1-diphenyl-2-picrylhydrazyl (DPPH) method and 2´,7´-dichlorodihydrofluorescin diacetate (DCFH₂-DA) assay.

Result:

TCL significantly inhibited FeSO₄/Cys-induced microsomes lipid peroxidation and protected both H₂O_2--induced RBCs hemolysis and RBCs auto-hemolysis in a dose-dependent manner. Furthermore, TCL demonstrated potent DPPH free-radical scavenging ability with IC₅₀ at 2.4×10^-5 M. In addition, TCL also moderately suppressed azide-induced mitochondria ROS formation.

Conclusion:

These results demonstrated that TCL was a strong antioxidant and free-radical scavenger, which might contribute to the anti-oxidative ability of Terminalia chebula Retz.     

Potent enhancement of transdermal absorption and stability of human tyrosinase plasmid (pAH7/Tyr) by Tat peptide and an entrapment in elastic cationic niosomes

Abstract

Enhancement of transdermal absorption through rat skin and stability of the human tyrosinase plasmid (P) using Tat (T) and an entrapment in elastic cationic niosomes (E) were described. E (Tween61:cholesterol:DDAB at 1:1:0.5 molar ratio) were prepared by the freeze-dried empty liposomes (FDELs) method using 25% ethanol. TP was prepared by a simple mixing method. TPE was prepared by loading T and P in E at the T:P:E ratio of 0.5:1:160 w/w/w. For gel formulations, P, TP, PE and TPE were incorporated into Carbopol 980 gel (30?µg of plasmid per 1?g of gel). For the transdermal absorption studies, the highest cumulative amounts and fluxes of the plasmid in viable epidermis and dermis (VED) were observed from the TPE of 0.31?±?0.04?µg/cm² and 1.86?±?0.24?µg/cm²/h (TPE solution); and 4.29?±?0.40?µg/cm² and 25.73?±?2.40?µg/cm²/h (TPE gel), respectively. Only plasmid from the PE and TPE could be found in the receiving solution with the cumulative amounts and fluxes at 6?h of 0.07?±?0.01?µg/cm² and 0.40?±?0.08?µg/cm²/h (PE solution); 0.10?±?0.01?µg/cm² and 0.60?±?0.06?µg/cm²/h (TPE solution); 0.88?±?0.03?µg/cm² and 5.30?±?0.15?µg/cm²/h (PE gel); and 1.02?±?0.05?µg/cm² and 6.13?±?0.28?µg/cm²/h (TPE gel), respectively. In stability studies, the plasmid still remained at 4?±?2?°C and 25?±?2?°C of about 48.00–65.20% and 27.40–51.10% (solution); and 12.34–38.31% and 8.63–36.10% (gel), respectively, whereas at 45?±?2?°C, almost all the plasmid was degraded. These studies indicated the high potential application of Tat and an entrapment in elastic cationic niosomes for the development of transdermal gene delivery system.     

Oral administration of ethyl acetate-soluble portion of Terminalia chebula conferring protection from streptozotocin-induced diabetic mellitus and its complications

Terminalia chebula has been widely used in India as a folk medicine. This study investigated the in vivo anti-hyperglycemia and anti-diabetic complication effects of the EtOAc-soluble portion of ethanolic extract of T. chebula fruit (EETC) containing 29.4% chebulic acid. Rats were divided into non-diabetic, untreated diabetic and diabetic groups. Streptozotocin (40 mg/kg body weight (BW))-induced diabetic rats were orally administered the aminoguanidine (100 mg/kg BW), high dose (500 mg/kg BW; HEETC) and low dose (100 mg/kg BW; LEETC) for 13 weeks. HEETC administration reduced the levels of blood glucose and serum lipids, decreased malondialdehyde concentrations of serum and thoracic aorta in diabetic rats, and significantly improved serum biochemical values and the pathomorphological changes of the liver and kidney in diabetic rats. Also, HEETC decreased the advanced glycation end products (AGEs) distribution in testis seminiferous tubules. Therefore, HEETC has a merit to be a potent candidate to control glycemic and diabetic complications.     

Transdermal delivery system of triamcinolone acetonide from a gel using phonophoresis

Triamcinolone acetonide (TA) is a corticosteroid that is used in the systemic and topical treatment of many inflammatory diseases. In this study, a phonophoretic drug delivery system was designed to enhance the TA permeability and the influence of ultrasound was examined. In order to establish the transdermal delivery system for TA, a hydrophilic carbopol gel containing TA was prepared after adopting phonophoresis. A permeation study through mouse skin was performed at 37 degrees C using a Franz diffusion cell, and the ultrasound treatment was carried out for 10 h. The level of TA permeation through the skin was evaluated under various ultrasound conditions including the frequency (1.0, 3.0 MHz), intensity (1.0, 2.5 W/cm2), and duty cycle (continuous, pulse mode) using a 0.5% TA gel. The highest permeation was observed under the ultrasound treatment conditions of low frequency, high intensity, and in continuous mode.     

诃子制草乌模拟炮制品在人工胃液与肠液中的水解行为研究

目的:研究诃子制草乌模拟炮制品在人工胃液与肠液中的水解行为,明确二者相互作用机制,为诃子制草乌的炮制减毒提供依据。方法:采用高相液相色谱(HPLC)法测定诃子制草乌在人工胃肠液中孵育不同时间(胃液中孵育2 h,肠液中孵育5、6、7、8 h)后其降解产物乌头碱、苯甲酰乌头原碱和苯甲酸的含量。结果:诃子制草乌中乌头碱含量在人工胃液中减少,在人工肠液中明显回升,在人工肠液孵育6~8 h时含量稳定;诃子制草乌中苯甲酰乌头原碱在人工胃液中未测得,在人工肠液中测得;诃子制草乌中苯甲酸在人工胃肠液中含量变化呈先递增再递减的趋势,且在人工肠液中孵育6 h时达最大值,7~8 h时维持稳定。结论:诃子制草乌炮制过程中鞣质与乌头碱可能结合生成难溶性物质,胃肠液p H对游离乌头碱的释放和水解有一定影响,人工胃液酸性环境对其水解有抑制作用,而人工肠液碱性环境对其水解有促进作用。