To assess the food effects and pharmacokinetic profile of oral prednisone (test preparation,5 mg) and prednisolone tablets (reference preparation,5 mg) using a randomized, two-period, two crossover, single-dose, fast and fed trial in 48 (24 in fast, 24 in fed) healthy Chinese adult subjects. In the trial, the plasma concentrations were determined at different time points up to 24 h and the pharmacokinetic parameters were analyzed according to the concentration data by non-compartmental analysis using WinNonlin software. All laboratory parameters, vital signs and adverse events (AEs) were monitored and recorded under the supervision of the clinicians throughout the whole process of the study. Prednisone and prednisolone undergo interconversion in liver. On average, the bioavailability of prednisolone after oral prednisone is approximately 80 http://www.chinadrugtrials.org.cn/ at 11 March 2020. • The food effects and pharmacokinetic evaluation of prednisone acetate and prednisolone were investigated in healthy Chinese subjects. • Prednisone and prednisolone undergo interconversion in liver. On average, the bioavailability of prednisolone after oral prednisone is approximately 80

Background EMLA cream is a local anesthetic. The pharmacokinetics and dermal effects of a topical anesthetic formulation has not been evaluated in healthy Chinese volunteers. Materials and methods The Pharmacokinetics of the lidocaine/prilocaine test (T) or reference (R, EMLA) cream were evaluated in a fasting, single-dose, two-period crossover bioequivalent study conducted in 40 healthy Chinese volunteers. Meanwhile, the dermal effects including blanching, erythema, temperature sensation, edema, and skin rash were also evaluated during the study. Results After applied 15 g of the cream for 4 h to a 100 cm 2 area under plastic occlusive film on the skin of the thigh of healthy volunteers, the results of the pharmacokinetic study showed that the active components absorbed in skin from topical products was relatively low compared with most system absorption drugs. After the removal of the residual anesthetic cream, there was a vascular biphasic response with initial transient blanching which reaches a peak at 4.5 h and later more persisting period erythema. The change of temperature sensory sensitivity reached the peak value at 4.5-6 h.There was no statistically significant difference of the changes after application the lidocaine/prilocaine T or R cream in subjects. In general, the lidocaine/prilocaine T or R cream was well tolerated. Conclusion The method described a model for investigations of pharmacokinetics and pharmacodynamics of topical lidocaine/prilocaine cream. Except the plasma drug level indicator, these pharmacodynamics data should also be evaluated in the anesthetic transdermal pharmacokinetics study. Clinical Trial Registration CTR20211544; registered in http://www.chinadrugtrials.org.cn/ at September 2021.

目的 探讨阿达帕林对人皮肤T细胞淋巴瘤(CTCL)Hut78细胞增殖、凋亡及周期的影响.方法 采用CCK-8法检测不同浓度(0、0.01、0.1、1、2.5、5、10、25、50和100μmol/L)阿达帕林对人CTCL Hut78、Myla和HH细胞的增殖抑制作用,并分别计算IC50.采用膜联蛋白(Annexin)V/碘化丙啶(PI)双染法和PI单染法分别检测阿达帕林0、2、4和6 μmol/L对Hut78细胞凋亡和周期分布的影响,Western blot法检测阿达帕林对Hut78细胞DNA损伤标志蛋白γ-H2AX表达的影响.结果 阿达帕林对Hut78、Myla和HH细胞增殖的抑制作用具有浓度依赖性,Hut78细胞的IC50为(1.45±0.47)μmol/L,HH 细胞的 IC50为(2.89±0.22)μmol/L,Myla 细胞的 IC50为(30.51±2.12)μmol/L.与阿达帕林0 μmol/L组相比,阿达帕林2、4和6 μmol/L组Hut78细胞的凋亡率升高[(10.82±0.46)％、(55.51±2.88)％、(73.26±2.21)％vs.(5.79±0.90)％](P＜0.01),S 期细胞比例升高[(72.51±1.89)％、(69.84±1.89)％、(91.68±10.84)％vs.(45.99±0.48)％](P＜0.01),阿达帕林4和6μmol/L组细胞中γ-H2AX表达增加(P＜0.05).结论 阿达帕林可选择性抑制CTCL中Hut78细胞的增殖,并诱导Hut78细胞凋亡和S期阻滞,其机制可能与阿达帕林诱导的DNA损伤有关.

Background and Objectives: Tissue engineering skin is a three-dimensional skin substitute cultured in the gas-liquid interface using the immortalized keratinocytes (HaCaT cells). In this study, the preliminary metabolism of betamethasone dipropionate by tissue engineering skin was studied and the pharmacokinetics methodology was established using betamethasone dipropionate gel as the target drug. Methods: The betamethasone dipropionate gel was applied on the tissue engineering skin after the skin was cultured. Then the medium (receiving liquid) and skin were taken on 0.25, 0.75, 1.75, 3, 5, 8, 12, 24, 36, 48 h time points. The betamethasone concentration in the medium and skin was determinated by the LC-MS method. Chromatographic analysis was conducted using isocratic elution on a C18 column (150 mm × 2.0 mm, 5 µm) in mobile phase consisting of methanol and water (70 : 30, v/v). The mobile phase was pumped at a flow rate of 0.2 mL/min. Results: This method exhibited linearity within the concentration range of 0. 1 to 50 µg /mL of betamethasone. The LLOQ was 0. 1 µg /mL. The intra- and inter-day precisions of betamethasone in the blank medium were all less than 10.69 % (RSD, %), while in the blank, skin homogenates were all less than 13.96 % (RSD, %). As a result, the betamethasone concentration in the medium and skin could both be detected, which suggested that betamethasone dipropionate could be metabolized to betamethasone through the tissue engineering skin. Conclusion: It was feasible to use tissue engineering skin as a model to study the dermatopharmacokinetics of topical betamethasone dipropionate gel. The research could build a foundation for the dermato-pharmacokinetic study approach.

Pharmacogenetics and pharmacometabolomics are the common methods for personalized medicine, either genetic or metabolic biomarkers have limited predictive power for drug response. In order to better predict drug response, the study attempted to integrate genetic and metabolic biomarkers for drug pharmacokinetics prediction. The study chose celecoxib as study object, the pharmacokinetic behavior of celecoxib was assessed in 48 healthy volunteers based on UPLC–MS/MS platform, and celecoxib related single nucleotide polymorphisms (SNPs) were also detected. Three mathematic models were constructed for celecoxib pharmacokinetics prediction, the first one was mainly based on celecoxib-related SNPs; the second was based on the metabolites selected from a pharmacometabolomic analysis by using GC–MS/MS method, the last model was based on the combination of the celecoxib-related SNPs and metabolites above. The result proved that the last model showed an improved prediction power, the integration model could explain 71.0% AUC variation and predict 62.3% AUC variation. To facilitate clinical application, ten potential celecoxib-related biomarkers were further screened, which could explain 68.3% and predict 54.6% AUC variation, the predicted AUC was well correlated with the measured values (r = 0.838). This study provides a new route for personalized medicine, the integration of genetic and metabolic biomarkers can predict drug response with a higher accuracy.

BackgroundThe treatment of Cutaneous T-cell lymphoma (CTCL) met huge challenges because of the heterogeneity and the scarcity of targeted drugs. ECPIRM derived from isotretinoin exhibited strong anti-proliferation effects in Hut78 and MJ cells rather than Myla cells. However, there was no data regarding the potential target of ECPIRM for its selective activity.ObjectivesTo investigate the potential target of ECPIRM for its selective anti-proliferation activity.MethodsWe evaluated the cell viability of CTCL cells after ECPIRM treatment, and detected the effects of ECPIRM on the biomarker genes of CTCL. Subsequently, the mRNA and protein level of Interleukin-2-inducible T-cell kinase (ITK) was determined. Then the induction of apoptosis triggered by ITK inhibitor BMS-509744 and ITK siRNAs were detected, and the docking of ECPIRM interacted with ITK and the effects of ECPIRM on ITK-mediated signaling pathway were analyzed. Finally, we evaluated the anti-growth activity of ECPIRM in Hut78-xenografted nude mice, and the relative expression of cleaved caspase-3, ITK, p-ERK and p-Akt were determined.ResultsITK was highly expressed in Hut78 and MJ cells rather than Myla cells, and targeted inhibition of ITK triggered cell apoptosis. ECPIRM efficiently bound the hydrophobic active pocket of ITK, and significantly inhibited ITK-mediated signaling pathway. In addition, ECPIRM suppressed tumor growth in Hut78-xenografted model, and upregulated the expression of cleaved caspase 3 and inhibited the expression of ITK, p-ERK and p-Akt in tumor tissues, which was consistent with in vitro study.ConclusionECPIRM might provide a novel strategy for CTCL by inhibiting ITK-mediated signaling pathway.

Malignant melanoma was the leading cause of mortality among the skin-associated cancer owing to its highly metastatic feature, increasing incidence and drug resistance requirement. Retinoids played important roles in the treatment of cancer via the activation of retinoid acid receptor (RAR) or retinoid X receptor (RXR). Our present study showed that the third-generation retinoid adapalene exhibited strong inhibitory effects on the proliferation of melanoma cells than other retinoids, such as all-trans-retinoic acid (ATRA), isotretinoin, acitretin and bexarotene, and adapalene exerted significant inhibitory effects on the colony formation of melanoma cells. Further study confirmed that adapalene treatment triggered dramatic S phase arrest and apoptosis, and S phase arrest was the potential mechanism of apoptosis induction. In addition, adapalene treatment dramatically regulated the expression of S phase-related protein, and increased the protein level of DNA damage marker，which were consistent with the results of the induction of the tail moment in comet assays. Meanwhile, DNA damage response was activated and the DNA repair pathway was simultaneously inhibited by adapalene treatment, which might furtherly potentiate S phase arrest and subsequent apoptosis. Taken together, these results showed that adapalene exhibited strong anti-cancer activity, and might be a candidate agent for the clinical treatment of melanoma.

Epidermal proliferative diseases consisted of a series of common skin diseases, most of which were recurrent chronic skin diseases, and had greatly negative influence on the life quality of patient. Retinoids exhibited vital roles in the treatment of many skin diseases. Our recent study demonstrated that adapalene significantly inhibited the growth of HaCat cells, and the inhibitory activity was stronger than other retinoids, such as all‐trans‐retinoic acid, acitretin, isotretinoin, tazarotene, and bexarotene. Further study showed that adapalene suppressed the colony formation of HaCat cells, and it dramatically triggered S‐phase arrest and apoptosis, rather than G1 phase arrest which was reported in other retinoids in several studies. Additionally, adapalene treatment greatly upregulated the protein expression of DNA damage marker γ‐H2AX, which was in accord with the results of the elongation of tail moment by comet electrophoresis analysis. Moreover, DNA damage was triggered and DNA repair was suppressed synchronously with adapalene treatment, which accounted for the mechanism of S‐phase arrest induced by adapalene. In summary, our recent work demonstrated that adapalene showed strong anti‐proliferation activity in HaCat cells and could be an alternative agent for the epidermal proliferative disease.

Individual variation in pharmacokinetics of faropenem is significant. We attempted to predict drug response of faropenem using a pharmacometabonomic approach. Metabolic profiling was performed on predose plasma samples from 36 healthy volunteers by gas chromatography-mass spectrometry (GC/MS), with 204 endogenous metabolites detected. Plasma concentration was measured after drug administration, using high pressure liquid chromatography-tandem mass spectroscopy (LC/MS/MS), and the pharmacokinetic parameters were calculated. Then a two-stage partial least squares strategy was employed to screen potential biomarkers and predict the pharmacokinetic parameters of faropenem. The results showed a good prediction of AUC and C-max with the screened biomarkers, and metabolites such as valine, proline, aspartic acid, gluconic acid, glucuronic acid, and 2-ketoisocaproic acid were indicated as candidate biomarkers. Finally, we explored the mechanism of individual variation by pathway enrichment analysis, and it suggested that organic anion transporter 1 (OAT1) and 3 (OAT3) might be responsible for individual variation of faropenem, and this hypothesis was verified by an experiment in rats.

Background: Retinoids are important agents for the treatment of cutaneous T-cell lymphomas (CTCL). But side effects and drug resistance caused by activation of RAR/RXR limited their clinical application. Therefore, it is urgent to develop new agents to fight against CTCL. ECPIRM, a 13-cis retinoic acid derivative, was reported that it inhibited proliferation and induced apoptosis of SCL-1 cells. Objective: The aim of this study was to evaluate the biological activities and mechanisms of ECPIEM. Methods: The effect of ECPIRM on cell proliferation was determined by MTT assay and Trypan blue exclusion assay while FACS analysis was used to detect changes in cell cycle and apoptosis in HUT78 cells. The influence of ECPIRM on RAR/RXR and JAK/STAT signaling was evaluated by western blot analysis. Results: ECPIRM, better than other agents (all-trans retinoic acid,13-cis-retinoic acid or bexarotene), inhibited proliferation and induced apoptosis significantly in HUT78 cells, but with little cytotoxicity on normal lymphocytes. Then ECPIRM induced G0/G1 phase arrest by decreasing the expression of cyclinD1, cyclinE, CDK2 and CDK4 while increasing p21. Furthermore, the unaffected expression of RAR and RXR members suggested that ECPIRM acted independently of RAR/RXR pathway in HUT78 cells. But decreased phosphorylation of JAK1, STAT3, STAT5 and downregulated Bcl-xL, Cyclin D1 and c-Myc indicated that ECPIRM inhibited the activation of JAK/STAT signaling. Conclusion: ECPIRM inhibited proliferation, induced apoptosis and G0/G1 phase arrest in HUT78 cells through inhibiting JAK/STAT pathway but not RAR/RXR pathway, which presented ECPIRM as a promising candidate for the treatment of CTCL patients.

A simple, rapid and highly sensitive LC-MS/MS method was developed and validated for the determination of ketoconazole in human plasma. Sample preparation was accomplished through a single step liquid-liquid extraction by ethyl acetate. The chromatography separation was carried out on a Hedera CN (150 mm x 2.1 mm, 5 mu m) column with isocratic elution using acetonitrile and 10 mM ammonium acetate containing 0.1% formic acid (45:55, v/v) as the mobile phase. The flow rate was 0.5 mL/min. Detection was performed in the positive ion electrospray ionization mode using multiple reaction monitoring of the transitions of 531.2 -> 489.3 and 286.1 -> 217.1 for ketoconazole and letrozole (the internal standard), respectively. The method exhibited good linearity over the concentration range of 0.01-12 ng/mL for ketoconazole. The intra- and inter-batch precision and accuracy of ketoconazole were all within the acceptable criteria. The method was successfully applied to a clinical study of the exposure to ketoconazole in Chinese seborrheic dermatitis patients after topical administration of two ketoconazole formulations of foam and lotion, respectively. The study results showed that there was little systemic absorption of ketoconazole in patients for the two formulations, and the ketoconazole foam and lotion are safe therapeutic drugs for seborrheic dermatitis patients. (C) 2016 Published by Elsevier B.V.

A high-performance liquid chromatography-positive electrospray ionization single quadrupole mass spectrometric (LC-ESI-MS) method for the determination of dexamethasone acetate in skin of nude mouse using triamcinolone acetonide acetate as the internal standard (I.S.) was developed and fully validated. Both compounds were precipitated from skin homogenate with methanol and were separated by HPLC on a Shimadzu Shim-pack VP-ODS C18 column (150mm×2.0mm, 5μm) with a mobile phase of methanol-water (80:20, v/v) at a flow rate of 0.2mL/min. Calibration curves were linear over the range of 0.05-5μg/mL. The intra-run relative standard deviations were less than 9.59%. The inter-run relative standard deviations were less than 7.82%. The mean recovery was in the ranges of 89.95-95.97%, respectively. The method was successfully applied to determinate the concentration of dexamethasone acetate in skin and study the percutaneous absorption process in skin of nude mouse.

Goodyeroside A 1, a natural product with hepatoprotective activity, was efficiently synthesized in a total yield of 47% in five steps starting from (S)-malic acid. The spectral data of the target compound 1 is identical with those of the natural product.