Synthesis and antitumor activity evaluation of glycoconjugates derived from natural product harmine / 中国药科大学学报

@#Based on our previous work, the study herein designed and synthesized eight glycoconjugates of natural product harmine (14a-14h)by introducing a cyclohexylmethyloxyl group at its C7 position and coupling a methyl-2-amino-β-D-glucopyranoside to the N9 position through different lengths of alkyl chains.In vitro anti-tumor activity screening and structure-activity relationship studies showed that the antitumor activity of the conjugates increased with the lengthening of the alkyl chain in the linker.Compound 14h exhibited significantly better proliferative inhibitory activity against MDA-MB-231 breast cancer cells than harmine.As compared to harmine, the introduction of the carbohydrate moiety improved the water solubility of compound 14h and enhanced its tumor cell selectivity through the Warburg effect.Mechanism of action studies revealed that compound 14h induced apoptosis and G0/G1 cell cycle arrest in MDA-MB-231 cells, and inhibited tumor cell migration by interfering with epithelial-mesenchymal transition process.This study provides a new approach for the development of antitumor drugs based on harmine.

Preparation of Legumain enzyme and mitochondrial double-stage targeted harmine liposome and evaluation of its in vitro characterization / 中国药房

OBJECTIVE To prep are Legumain enzyme and mitochondrial double-stage targeted harmine （HM） liposome （KA@HM-LPS）and preliminary evaluate its pharmaceutical properties ，in vitro antitumor effect and biocompatibility. METHODS Firstly，the preparation and homogenization methods of KA@HM-LPS was screened ，and prepared liposomes were characterized. Secondly，the serum stability ，in vitro release rate ，hemolysis percentage of KA@HM-LPS and cell survival rate under KA@BLPS were determines respectively. Finally ，the cell surivival rate ，mitochondrial targeting and inhibitory effects on cell migration and invasion of KA@HM-LPS were determined. RESULTS KA@HM-LPS was prepared by the thin-film dispersion method ，with encapsulation efficiency of （90.50 ± 0.62）％ . The extrusion moulding method was selected as homogenization method of KA@HM-LPS. The particle size ，polydispersity index ，and Zeta potential of KA@HM-LPS were （211.40±11.67）nm，0.316± 0.014 and（－14.20±0.49）mV，respectively. In 37 ℃，10％ FBS，the particle size of KA@HM-LPS kept stable after 12 h. In vitro release curve of KA@HM-LPS in 20％ plasma conformed to Weibull distribution and had the property of sustained release. When HM concentration was 160 μg/mL，the hemolysis percentage of KA@HM-LPS was （4.23±0.19）％，which was much lower than that of free HM ，with safety. When the mass concentration of KA@BLPS reaches 400 μg/mL，the survival rate of LO 2 cells was （94.40 ± 6.12）％ ，and the biocompatibility was good. Cell test results in vitro showed that ，inhibitory effect of KA@HM-LPS on liver cancer cells with overexpression of Legumain enzyme （LGMN -SK-Hep-1） was significantly higher than that of normal liver cancer cells SK-Hep- 1； compared with SK-Hep-1，LGMN +-SK-Hep-1 cells had a higher uptake efficiency of the liposome ；KA@HM-LPS could significantly inhibit the migration and invasion of LGMN +- SK-Hep-1 cells. CONCLUSIONS KA@HM-LPS is prepared successfully ，which can effectively inhibit the migration and invasion of liver cancer cells with Legumain enzyme overexpression ，and improve the blood compatibility of HM.

Heme peroxidases are responsible for the dehydrogenation and oxidation metabolism of harmaline into harmine / 中国天然药物

Harmaline and harmine are β-carboline alkaloids with effective pharmacological effects. Harmaline can be transformed into harmine after oral administration. However, enzymes involved in the metabolic pathway remain unclear. In this study, harmaline was incubated with rat liver microsomes (RLM), rat brain microsomes (RBM), blood, plasma, broken blood cells, and heme peroxidases including horseradish peroxidase (HRP), lactoperoxidase (LPO), and myeloperoxidase (MPO). The production of harmine was determined by a validated UPLC-ESI-MS/MS method. Results showed that heme peroxidases catalyzed the oxidative dehydrogenation of harmaline. All the reactions were in accordance with the Hill equation. The reaction was inhibited by ascorbic acid and excess H2O2. The transformation of harmaline to harmine was confirmed after incubation with blood, plasma, and broken blood cells, rather than RLM and RBM. Harmaline was incubated with blood, plasma, and broken cells liquid for 3 h, and the formation of harmine became stable. Results indicated an integrated metabolic pathway of harmaline, which will lay foundation for the oxidation reaction of dihydro-β-carboline. Moreover, the metabolic stability of harmaline in blood should not be ignored when the pharmacokinetics study of harmaline is carried out.

Effect of harmine on osteosarcoma cell proliferation and apoptosis by down regulating COX-2 expression / 中国医师杂志

Objective To investigate the inhibitory effect of harmine on osteosarcoma cell proliferation and apoptosis by down regulating cyclooxygenase-2 (COX-2) expression.Methods Human osteosarcoma cell line U2OS was cultured in vitro and randomly divided into control group,study group 1,study group 2 and study group 3.The cells were cultured in 0,5 μmol/L,10 μmol/L and 20 μmol/L concentration of harmine for 48 hours.Cell counting kit-8 (CCK-8) method and flow cytometry were used to detect cell viability and apoptosis.The expression level of COX-2,proliferation and apoptosis related proteins and mRNA were detected by Western blot and real-time quantitative polymerase chain reaction (RT-qPCR),respectively.Results After cultured with different concentrations of harmine for 12,24 hours and 48 hours,the cell viability of the three study groups were significantly lower than that of the control group (P ＜ 0.05),while that of the study groups 2 and 3 were significantly lower than that of the study group 1 (P ＜ 0.05).The apoptosis rate of the three study groups were significantly higher than that of the control group (P ＜0.05),while that of the two groups were significantly higher than that of the study group 1 (P ＜ 0.05).After 48 hours of culture,the levels of COX-2,cyclin D1,proliferating cell nuclear antigen (PCNA),B-cell lymphoma-2 (Bcl-2) protein and mRNA expression in study group 2 were significantly lower than those in control group,while the expression levels of cleaved caspase-3 and BCL2-Associated X (Bax) in study group 2 were significantly higher than those in control group (P ＜ 0.05).Conclusions Harmine can inhibit the proliferation and promote the apoptosis of osteosarcoma cells by inhibiting the expression of COX-2 and regulating the expression of cell cycle and apoptosis related protein.

Study on Quality Standard for Kazakhstan Medicine Peganum harmala / 中国药房

OBJECTIVE： To establish the quality standard for Kazakhstan medicine Peganum harmala. METHODS： Ten batches of P. harmala collected in Xinjiang Kazakh region were selected as research objects to investigate their characteristics. Qualitative identification of harmaline and harmine was conducted by TLC. The contents of water， total ash， acid-insoluble ash and ethanol extract were tested according to Chinese Pharmacopoeia（2015 edition）. The contents of harmaline and harmine were determined by HPLC. The determination was performed on X-bridge C18 column（250 mm×4.6 mm，5 μm） with mobile phase consisted of acetonitrile-ammonium acetate buffer （adjusted to 6 with glacial acetic acid， gradient elution） at the flow rate of 1 mL/min. The detection wavelength was set at 267 nm， and column temperature was 25 ℃. The sample size was 5 μL. RESULTS： TLC identification results showed that 10 batches of medicinal material showed clear spots at the same position as harmaline and harmine reference substances. Water， total ash， acid-insoluble ash should not be more than 12％，22％，2％， respectively； ethanol extract must not be less than 16％. HPLC results showed that the linear ranges of harmaline and harmine were 15.22-301.40，15.09-301.80 μg/mL； RSDs of precision， reproducibility and stability tests were all lower than 4％； average recoveries were 100.22％ and 100.94％（all RSD＜2％）. The determination results showed that the content of total alkaloids （harmaline and harmine） should not be less than 6.5 mg/g. CONCLUSIONS： Based on the original standard， test items are added in this study. TLC method is established to identify harmaline and harmine. HPLC method is established to determine their contents. Established quality standard can be used for comprehensive quality control of P. harmala from Xinjiang Kazakh region.

Determination of Plasma Concentration of Harmine Derivative DH- 330 by UPLC-MS and Its Pharmacokinetics Ev aluation in Rats / 中国药房

OBJECTIVE： To establish a method for the determination of harmine derivative DH-330 in rat plasma and to use it for pharmacokinetic behavior evaluation of DH-330 in rats after intragastric administration. METHODS： Using tinidazole as internal standard， after pre-treatment of acetonitrile precipitated protein， UPLC-MS method was adopted to determine the plasma concentration of DH-330. UPLC analysis was performed on Waters ACQUITY BEH C18 column （50 mm×2.1 mm，1.7 μm） with mobile phase consisted of acetonitrile-methanol-0.5％ formic acid aqueous solution（15 ∶ 55 ∶ 30， V/V/V） at flow rate of 0.4 mL/min， while the column temperature was 30 ℃， and sample size was 5 μL. MS analysis was conducted by electrospray ionization source， positive ion scanning， ion source temperature at 124 ℃， DH-330 detection of mass to charge ratio （m/z） of 335.8→334.8， and internal standard m/z of 247.0→81.0. Six Wistar rats were given DH-330 suspension（50 mg/kg） intragastrically. Blood samples were collected from fundus venous plexus capillary before administration （0 h） and 0.25，0.5，1，2，4，6，8，12，24 h after administration. Plasma concentration of DH-330 was determined and plasma concentration-time curves were drawn. Pharmacokinetic parameters were calculated by using Kinetica 5.0 software. RESULTS： The linear ranges of DH-330 were 25.05-2 004 ng/mL（r＝0.999 8），and the limits of quantitation was 25.05 ng/mL. RSDs of intra-day and inter-day were all less than 10％. The accuracy RE was －9.76％ to 4.55％. The extraction recovery was higher than 85％（RSD＜5％）. Stability RE was －2.53％ to 2.29％. They were not affected by matrix effect or residual effect of injection. The pharmacokinetic parameters of DH-330 in rats after intragastric administration included that cmax was （1 162.43±241.72）ng/mL，AUC0-∞ was （3 242.93±652.31）ng·h/mL，t1/2 was （1.93±0.61）h， MRT was （3.23±0.30）h，CL was （16.80±5.30）L/h·kg， Vss was （54.78±19.64）L/kg. CONCLUSIONS： The established method is simple， specific， sensitive， precise and recovery， which can be used for the plasma concentration determination of DH-330 in rats. DH-330 has short half-life， rapid absorption and large apparent distribution volume after intragastric administration in rats， which indicates that it has high lipophilicity and may be mainly distributed in tissues.

Harmine inhibits COX-2 expression in gastric cancer cells through PTEN/Akt/MDM2 signaling pathway / 中国药理学通报

Aim To further analyze the effects of PTEN/Akt/MDM2 signaling pathway in the harmine (HM)-mediated inhibition of COX-2 expression in gastric cancer cells. Methods PTEN-siRNA, Akt-siR-NA, MDM2-siRNA were constructed and respectively transfected into SGC-7901 and MKN45 cells,and then added or not added HM for 24 h. The expression of PTEN, Akt and phosphorylated Akt ( p-Akt), MDM2 and phosphorylated MDM2 ( p-MDM2), as well as COX-2 expression was detected by Western blot. Results HM increased PTEN expression, but inhibited p-Akt,p-MDM2 and COX-2 expression in SGC-7901 and MKN45 cells. Knockdown of PTEN blocked HM-induced inhibition of Akt and MDM2 phosphorylation, as well as down-regulation of COX-2 expression. Knockdown of Akt and treatment with HM synergisti-cally inhibited p-MDM2 and COX-2 expression. Knockdown of MDM2 and treatment with HM synergis-tically inhibited COX-2 protein expression. Conclusions HM down-regulates the expression of COX-2 protein in gastric cancer cells via PTEN/Akt/MDM2 signaling pathway.

Effect and Underlying Mechanism of Harmine on Proliferation and Apoptosis of Gastric Cancer Cells / 胃肠病学

Background:Previous study has found that harmine inhibited the proliferation of gastric cancer cells by down-regulating cyclooxygenase-2(COX-2)expression. However,its molecular mechanism is not fully clear. Aims:To investigate the effect of harmine on proliferation and apoptosis of gastric cancer cells,and explore the role of PTEN/Akt/MDM2 signaling pathway in this process. Methods:Human gastric adenocarcinoma cell line SGC-7901 and MKN-45 were treated with harmine at different concentrations(2,4,8,16,32 μg/mL)for 24,48,and 72 hours. The cell proliferation and apoptosis were detected by MTT assay and Hoechst staining,respectively. The expressions of PTEN,COX-2, phosphorylated Akt(p-Akt)and p-MDM2 were measured by Western blotting. Results:Harmine dose- and time-dependently inhibited proliferation and induced apoptosis of SGC-7901 and MKN-45 cells. Also,harmine dose-dependently increased PTEN expression,and inhibited p-Akt,p-MDM2 and COX-2 expressions in SGC-7901 and MKN-45 cells. Conclusions:Harmine may inhibit proliferation and induce apoptosis of gastric cancer cells via down-regulating COX-2 expression through PTEN/Akt/MDM2 signaling pathway.

Preparation of Transferrin Modified Harmine Magnetoliposomes and Evaluation of Its Antitumor Effect / 中国药学杂志

OBJECTIVE: To prepare the transferrin modified harmine magnetoliposomes and investigate its in vitro properties and antitumor effect. METHODS: Transferrin modified harmine magnetoliposomes was prepared by reverse phase evaporation method. The size, Zeta potential, entrapment efficiency, drug loading efficiency, release rate and plasma stability of liposomes were investigated. Furthermore, cytotoxicity to liver cancer and glioma were investigated too. RESULTS: The transferrin modified harmine magnetoliposomes were successfully prepared. The average size of liposomes was (184.7±8.08)nm.The average Zeta potential was (-12.4±0.896) mV and average entrapment efficiency was (84.67±6.26)%. Drug loading efficiency was (9.14±4.54)%. Transferrin modified harmine magnetoliposomes released more slowly and were more stable in rat plasma than harmine solution. The results of cytotoxicity test showed that the cell inhibitory effect of transferrin modified harmine magnetoliposomes was enhanced compared with other groups, and the cell inhibitory effect of this preparation was more obvious underwhen cells were put under magnetic field compared with those under the non-magnetic field(P<0.05). CONCLUSION: This method can be successfully used to prepared transferrin modified harmine magnetoliposomes, and liposomes can inhibit the cell visbility of liver cancer and human glioma cells significantly.

Effects of harmine on the epithelial mesenchymal transition of melanoma A375 cells / 天津医药

Objective To explore the inhibitory effect of harmine on melanoma A375 cells and its mechanism thereof.Methods (1) Melanoma A375 cells were treated with harmine at 0,0.5,1,2,5,10,20,50 and 100 mg/L for 48 h in vitro.CCK-8 method was used to detect the cell viability and confirm the experimental concentrations.(2) After the cells were treated with 0,1,2 mg/L harmine,the scratch and transwell assays were used to detect the cell migration and invasion ability.Western blot assay was used to detect the expression levels of epithelial mesenchymal transition (EMT)-related protein E-cadherin,N-cadherin,Snail and p53.(3) Three groups of ceils were set up.The control group was transfected with empty vector ordy.The empty vector group was transfected with empty vector after treated with 2 mg/L harmine for 24 h.The Snail transfection group was transfected with Snail cDNA after treated with 2 mg/L harmine for 24 h.The cell migration and invasion ability were detected after the transfection.Results (1) When the concentration of harmine was above 2 mg/L,the survival rate of A375 cells was significantly lower than that of the control group with the increase of harmine concentrations (P ＜ 0.05).Then,the concentrations of 0,1 and 2 mg/L of harmine were used in the following experiments.(2) With the increase of the harmine concentrations,the number of cells in the scratched area and the number of trans-membrane cells in each group were significantly decreased.The migration and invasion ability of the ceils were decreased gradually.The expression levels of E-cadherin and p53 were increased,while the expression levels of N-cadherin and Snail were decreased.(3) Cell transfection experiments showed that the migration and invasion ability of the cells were increased compared with those of empty vector group after transfection with Snail.Conclusion Harmine can inhibit the proliferation of A375 cells and decrease the abilities of metastasis and invasion,which may be achieved by decreasing the expression of Snail after activating the p53,thereby increasing E-cadherin and down-regulating N-cadherin to inhibit the EMT process.

Antidepressive and anxiolytic effects of ayahuasca: a systematic literature review of animal and human studies

Objective: To conduct a systematic literature review of animal and human studies reporting anxiolytic or antidepressive effects of ayahuasca or some of its isolated alkaloids (dimethyltryptamine, harmine, tetrahydroharmine, and harmaline). Methods: Papers published until 3 April 2015 were retrieved from the PubMed, LILACS and SciELO databases following a comprehensive search strategy and using a predetermined set of criteria for article selection. Results: Five hundred and fourteen studies were identified, of which 21 met the established criteria. Studies in animals have shown anxiolytic and antidepressive effects of ayahuasca, harmine, and harmaline, and experimental studies in humans and mental health assessments of experienced ayahuasca consumers also suggest that ayahuasca is associated with reductions in anxiety and depressive symptoms. A pilot study reported rapid antidepressive effects of a single ayahuasca dose in six patients with recurrent depression. Conclusion: Considering the need for new drugs that produce fewer adverse effects and are more effective in reducing anxiety and depression symptomatology, the described effects of ayahuasca and its alkaloids should be further investigated.

Antidepressant effects of a single dose of ayahuasca in patients with recurrent depression: a preliminary report

Objectives: Ayahuasca (AYA), a natural psychedelic brew prepared from Amazonian plants and rich in dimethyltryptamine (DMT) and harmine, causes effects of subjective well-being and may therefore have antidepressant actions. This study sought to evaluate the effects of a single dose of AYA in six volunteers with a current depressive episode. Methods: Open-label trial conducted in an inpatient psychiatric unit. Results: Statistically significant reductions of up to 82% in depressive scores were observed between baseline and 1, 7, and 21 days after AYA administration, as measured on the Hamilton Rating Scale for Depression (HAM-D), the Montgomery-Åsberg Depression Rating Scale (MADRS), and the Anxious-Depression subscale of the Brief Psychiatric Rating Scale (BPRS). AYA administration resulted in nonsignificant changes in Young Mania Rating Scale (YMRS) scores and in the thinking disorder subscale of the BPRS, suggesting that AYA does not induce episodes of mania and/or hypomania in patients with mood disorders and that modifications in thought content, which could indicate psychedelic effects, are not essential for mood improvement. Conclusions: These results suggest that AYA has fast-acting anxiolytic and antidepressant effects in patients with a depressive disorder. .

Study on the Quality Standard for Peganum Harmala Alkaloids Cream / 中国药师

Objective:To establish the quality standard for peganum harmala alkaloids cream ( CAPH) . Methods: The general quality of CAPH was inspected according to the general notices described in Chinese Pharmacopoeia volumeⅠ2010 edition. The qual-itative identification was carried out by TLC with harmine and harmaline as the index ingredients. The content determination was carried out by HPLC methods with harmine, harmaline and vasicine as the index ingredients. Results:The inspection items were all met the requirements. The experimental samples and the reference substances in TLC showed the identical spots with the same color and shape at the same position. The calibration curve of harmaline, harmine and vasicine was linear within the concentration range of 3. 440-110. 000 μg·ml-1 , 3. 340-107. 000 μg·ml-1 and 1. 380-22. 000 μg·ml-1 , respectively. The recovery was 98. 1%, 99. 8% and 99. 3% with RSD of 1. 75%, 1. 78% and 1. 95%, respectively (n=6). Conclusion: The established quality control methods meet the requirements of methodology, and the results lay foundation for the quality standard for CAPH.

Preparation of cRGD-modified harmine hydrochloride stealth liposomes / 中国药学杂志

OBJECTIVE: To prepare cRGD-modified harmine hydrochloride long circulating liposomes, investigate its characteristics in vitro, and finally establish its best preparation method. METHODS: Firstly the content determination method was established, and secondly harmine hydrochloride liposomes were prepared through reverse phase evaporation method. Both active and passive drug loading methods were investigated to entrap the drug. Thirdly, cRGD-DSPE-2000 was prepared by conjugation method, and then cRGD modified harmine hydrochloride long circulating liposomes were prepared. Finally the particle sizes, Zeta potential and the entrapment efficiency were determined. The release of the liposomes in vitro was measured. RESULTS: The standard curves all had good linearities except for the one using 100% methanol as solvent. The particle sizes of the liposomes prepared by passive loading method, active loading method, and long circulating cRGD modified liposomes were 227.2, 246.3 and 241.9 nm, respectively, and the Zeta potentials were about 20-30 mV. The entrapment efficiency were (36.78 ± 6.82)%, (81.77 ± 7.61)%, and (80.02 ± 1.27)%, respectively. The release of cRGD liposomes was slower than the liposomes without cRGD and HM solution. CONCLUSION: cRGD-modified long circulating harmine hydrochloride liposomes can be made through reverse phase evaporation method and active loading method. KEY

Harmine has no lung-protective effects in a canine model of lung ischemia-reperfusion injury / 中国组织工程研究

BACKGROUND:Inflammatory cellactivation and the generation of oxygen free radicals are important factors of lung ischemia-reperfusion injury. Adding the drugs with anti-inflammation and antioxidant effects into the lung preservation solution used, can improve the protection fluid, and play a crucial role in the study of lung ischemia-reperfusion injury and protection of the function of transplanted lung. OBJECTIVE:To discussion the effect of harmine in canine model of pulmonary ischemia-reperfusion injury. METHODS:Twelve healthy hybrid dogs were randomly divided into two groups, with six rats in each group. A canine model of lung ischemia-reperfusion injury was established, and the protecting liquid was perfused with the clockwise irrigation method. Control group:low potassium dextran protective fluid;experimental group:low potassium dextran+harmine protective fluid. After 2 hours of ischemia, the left lung circulation was recovered. The left lung tissue and blood samples were col ected from two groups of animal models after reperfusion, and their cytokines levels and the lung wet/dry weight ratio were detected and calculated. Bronchoalveolar lavage fluid was col ected to observe the pathological indicators. The main pulmonary artery pressure, left and right pulmonary artery pressure were recorded by continuous monitoring. RESULTS AND CONCLUSION:There were no statistical y significant differences in the interleukin-17, tumor necrosis factorαand endothelin 1 content in the left lung tissue and the blood between the two groups at 2 and 4 hours after reperfusion (P>0.05). After 4 hours of reperfusion in both groups, the neutrophil number, the number of lymphocytes, alveolar edema index, and vascular wal damage in bronchoalveolar lavage fluid showed no statistical y significant differences (P>0.05). Through the analysis of variance, the main pulmonary artery pressure, left pulmonary artery pressure and right pulmonary artery pressure also had no statistical significance between the two groups (P>0.05). By the analysis of cytokines, pathological indicators, lung wet/dry weight ratio, and pulmonary arterial pressure, harmine has no significant lung protection effect in canine model of lung ischemia-reperfusion injury.

Research on quality specification of the seeds of Peganum harmala L. of a uygur traditional medicine / 中国药学杂志

OBJECTIVE: To develop the quality specification of seeds of Peganum harmala. METHODS: According to the Chinese Pharmacopoeia (2010 Version, Volume 1) and its appendix method, the water, total ash, acid insoluble ash, 50% ethanol extractives, and heavy metal were analyzed for seeds of P. harmala. TLC method was used to separate harmaline (HAL), harmine (HAR) and vasicine (VAS) in seed samples using mixture of ethyl acetate-methanol-ammonia water (10:1.5:0.5) as a developing solvent on high performance silica G pre-coated plate with 254 nm fluorescent (GF254) and to identify them inspected under UV 366 nm, 254 nm, visualized by spraying with both Dragendorff reagent and by bioautographic assay. In the HPLC method, HAL and HAR were separated on a C18 column with acetonitrile-ammonium acetate water (19:81) as the mobile phase and detected at 330 nm. The HPLC fingerprints were performed on the same C18 column and eluted by using a linear gradient of acetonitrile (A) and 0.1 mmol · L-1 ammonium acetate buffer under the flow rate at 0.7 mL · min-1 and detected at 280 nm. RESULTS: In the TLC procedures, 254 and 366 nm fluorescent, Dragendorff reagent, and bioautographic assay for the detection of acetylcholinesterase inhibitor can be used for qualitative identification of the active ingredients. For the HPLC quantitative method, the calibration curve of HAR displayed ideal linearity over the range of 1.97-198.68 μg · mL-1 with average recovery of 99.69% (RSD of 1.89%). HAL displayed ideal linearity over the range of 1.70-345.30 μg · mL-1 with average recovery of 100.66% (RSD of 1.78%). The contents of HAL and HAR in 11 batches of seeds of P. harmala were 3.234% and 3.755%. In the characteristic fingerprints of seeds of P. harmala, four common peaks were identified. CONCLUSION: The results indicated that the water, total ash, acid insoluble ash, and 50% ethanol extractives were not more than 9.0%, 8.0%, 1.0%, and 22.0%, respectively. The heavy metal of plumbum, cadmium, arsenic, mercury, and copper were not more than 5 × 10-6, 3 × 10-6, 2 × 10-6, 2 × 10-6 and 20 × 10-6, respectively. The content limit of the sum of HAL and HAR was not lower than 5.5%. With the peak of HAL as reference peak, the variance of relative retention time of the four common peaks, in the characteristic fingerprints of seeds off. harmala, should be fluctuated in the range of 5% of the specified value. The qualitative and quantitative method established was suitable for the quality evaluation and assessment of seeds of P. harmala.

Preparation of harmine hydrochloride liposome by active loading method / 中草药

Objective To prepare a harmine hydrochloride liposome formulation with high encapsulation efficiency. Methods Preformulation investigation was carried out to obtain the drug physicochemical properties such as solubility and lgD in buffers of different pH value. Harmine hydrochloride was encapsulated into liposomes by active loading method. Encapsulation efficiency of liposomes was determined after the free drug was separated from liposome by ultrafiltration. The influence factors on the encapsulation efficiency including drug-lipid weight ratio, incubation temperature, pH value of external water phase were investigated. Results As the pH value increasing, the solubility of harmine hydrochloride was decreased, while the apparent oil-water distribution coefficient was increased. By active loading method, the encapsulation efficiency could be over 80% when the drug to lipid weight ratio was under 1∶5. The pH gradient between intervesicle and intravesicle obviously influenced the encapsulation efficiency, while incubation temperature had little effect on encapsulation efficiency. Conclusion Active loading is suitable for preparing harmine hydrochloride liposome with high encapsulation efficiency.

Optimization of prescription and anti-pruritic effect of harmine hydrochloride cream / 中成药

AIM: To select the prescription of harmine hydrochloride cream and to study the anti-itching effect of harmine hydrochloride cream. METHODS: A comprehensive score system,including appearance,homogeneity,coating performance,viscosity,particle diameter,temperature-resistant test,and centrifugal effect were established as evaluative criteria. Then uniform design was used for selecting the prescription of harmine hydrochloride cream. The histamine phosphate and 5-hydroxytryptamine (5-HT) were used to induce mouse cutaneous pruritus. Then the anti-pruritic effect of harmine hydrochloride cream in contrast to Piyanping Ointment as control group. RESULTS: The optimized prescription was as follows: 8% of monostearin,5% of Vaseline,6. 1% of beeswax, 1. 1% of Span-60,1. 9% of Tween-80. The local application of harmine hydrochloride cream could alleviate the itching induced by histamine phosphate and 5-HT,respectively. And the anti-pruritic effect of harmine hydrochloride cream was stronger than the effect of the positive control group. CONCLUSION: The prescription of the harmine hydrochloride cream is available,reasonable,and reproducible. The harmine hydrochloride cream has the anti-pruritic effect.

Prescription Selecting of Harmine. HCl Ointment / 中成药

Objective: To select the prescription of Harmine HCl ointment. Methods: The orthogonal design was used for selecting the prescription with transdermal absorption rate constant (K) and flow energy of activition (E ?) as selecting standard. Results: The optimum prescription is as follows: Azone (2.0%), Span-80 ( 0.2%), Tweens-80 (0.4%), Glycerylmonostearate (2.5%), Vaselin (4.0%), Liquid (11%). Conclusion: The prescription design is available, and the ointment has a good stability.

Study on the Content of Hydrochloric Harmine and Heat Stability of Harmine Gelatin Microspheres / 中国药房

0.05) in surface HM;however,the total content obtained from RP-HPLC was smaller.The pyrolytic activation energy was 93.37kJ/mol,frequency factor was 2.304?1013/min.CONCLUS_ION:UV-spectrophotometry is more simple and RP-HPLC is more precise in detection.HM-GMS is considerably heat-stable.