Anisodine hydrobromide alleviates oxidative stress caused by hypoxia/reoxygenation in human cerebral microvascular endothelial cells predominantly via inhibition of muscarinic acetylcholine receptor 4

Background:Anisodine hydrobromide(AT3),an anti-cholinergic agent,could be delivered to the brain across the blood-brain barrier and has been used clinically for the treatment of cerebral ischemia/reperfusion injury.Endothelial dysfunction can be caused by hypoxia/reoxygenation(H/R)via oxidative stress and metabolic alterations.The present study investigated whether AT3 regulates the production of nitric oxide(NO)and reactive oxygen species(ROS),and the HIF-1αpathway via regulation of muscarinic acetylcholine receptors(mAChRs)in brain microvascular endothelial cells after H/R exposure.Methods:Under H/R conditions,hCMEC/D3 cerebral microvascular endothelial cells were treated with AT3.Specific inhibitors of M2-and M4-mAChRs were used to explore the mechanism by which AT3 influences oxidative stress in endothelial cells.Then,mAChRs expression was detected by western blotting and NO production was detected by Greiss reaction.The intracellular ROS level was measured using DCFH-DA probes.The expression of hypoxia-inducible transcription factor 1α(HIF-1α)was also detected.Results:While H/R induced the expression of M2-and M4-mAChRs,AT3 suppressed the H/R-upregulated M2-and M4-mAChRs.H/R also induced the production of NO,ROS,and apoptosis.AT3 and M4-mAChR inhibitors inhibited the H/R-induced production of NO and ROS and apoptosis.HIF-1αwas induced by H/R,but was suppressed by AT3.Conclusion:Thus,the in vitro evidence shows that AT3 protects against H/R injury in cerebral microvascular endothelial cells via inhibition of HIF-1α,NO and ROS,predominantly through the downregulation of M4-mAChR.The findings offer novel understandings regarding AT3-mediated attenuation of endothelial cell apoptosis and cerebral ischemia/reperfusion injury.

Total Synthesis of Anisodine

Aim To design a practical synthetic route of anisodine. Methods Starting from3α-hydroxy-6β-acetyltropine, anisodine was synthesized in 11 steps. Result Anisodine wasobtainded with an overall yield of 2.6 % . Conclusion Total synthetic route of anisodine wasachieved which may afford a possible route for commercial preparation of anisodine.

Asymmetric Synthesis of Anisodine

Anisodine was synthesized from 3α-hydroxy-6β-acetyltropine in 11 steps. Laevorotary isomer of anisodine was prepared from the asymmetric dihydroxylation of compound 10 via the osmium catalyzed process employing p-chlorobenzoyl dihydroquinidine as the chiral ligands.

Clinical study of Qiming granule combined with compound anisodine injection in the treatment of diabetic retinopathy

Objective:To investigate the clinical study of Qiming granule combined with compound anisodine injection in the treatment of diabetic retinopathy.Methods:104 patients with nonproliferative diabetic retinopathy admitted to department of ophthalmology in Xishui Hospital Affiliated to Hubei University of science and technology from September 2016 to May 2018 were selected as the study subjects.According to the principle of treatment,they were divided into 52 cases in the control group and 52 cases in the treatment group,the clinical efficacy,blood sugar and lipid levels,hemorheological changes and adverse reactions were compared between the two groups.Results:The total effective rate difference between the treatment group(92.31%)and the control group(71.15%)was statistically significant(χ^(2)=7.792,P=0.005).After treatment,the two groups of fasting plasma glucose(FPG),hemoglobin A1c(HbA1c),total cholesterol(TC),low density lipoprotein-cholesterol(LDL-C),whole blood high-viscosity,whole blood cut viscosity,whole blood low-cut viscosity,plasma viscosity,number of microangioma and area of the hemorrhage were significantly lower than before treatment(P<0.05),and the levels of FPG,HbA1c,TC,LDL-C,whole blood highviscosity,whole blood cut viscosity,whole blood low-cut viscosity,plasma viscosity,number of microangioma and area of the hemorrhage in the treatment group were significantly lower than those in the control group(χ^(2)=0.122,P=0.727).Conclusions:Qiming granule combined with compound anisodine injection can effectively improve the clinical efficacy of patients with non-proliferative DR,and its mechanism may be related to improve blood sugar and blood lipid levels and correct blood rheology disorder,reduce microangioma and hemorrhage.It has clinical promotion significance.

Neuroprotective Effect of Compound Anisodine in a Mouse Model with Chronic Ocular Hypertension

Background：Compound anisodine （CA） is a compound preparation made from hydrobromide anisodine and procaine hydrochloride.The former is an M-choline receptor blocker with the function of regulating the vegetative nervous system,improving microcirculation,and so on.The latter is an antioxidant with the activities ofneuroprotection.This study aimed to investigate the potential neuroprotection of CA,which affects the degeneration of the retinal ganglion cells （RGCs） in an animal model with chronic ocular hypertension.Methods：Female C57BL/6J mice （n =24） were divided randomly into four groups：Normal control group without any treatment （Group A,n =6）;CA control group with feeding the CA solution （Group B,n =6）;microbeads （MBs） control group with injecting MB into the anterior chamber （Group C,n =6）;CA study group with MB injection and with feeding the CA solution （Group D,n =6）.Intraocular pressure （IOP） was measured every 3 days after MB injection.At the 21st day,neurons were retrograde-labeled by Fluoro-Gold （FG）.Animals were sacrificed on the 27th day.Retinal flat mounts were stained immunohistologically by β-Ⅲ-tubulin.FG-retrograde-labeled RGCs,β-Ⅲ-tubulin-positive RGCs,and β-Ⅲ-tubulin-positive nerve fibers were quantified.Results：Mice of Groups C and D expressed the incidence of consistent IOP elevation,which is above the IOP level of Group A with the normal one.There is no significant difference in IOP between Groups A and B （P ＞ 0.05）.On the 27th day,there were distinct loss in stained RGCs and nerve fibers from Groups C and D compared with Group A （all P ＜ 0.00l）.The quantity was significantly higher in Group D as compared to Group C （all P ＜ 0.00l） but lower than Group A （all P ＜ 0.001）.There was no significant difference in the quantity of RGCs and nerve fibers between Groups A and B （all P ＞ 0.05）.Conclusions：These findings suggest that CA plays an importantly neuroprotective role on RGCs in a mouse model with chronic ocular hypertension.

STRUCTURE AND ABSOLUTE CONFIGURATION OF ANISODINE HYDROBROMIDE

Anisodine is a noval anticholinergic drug extracted from the Datura plant Scoplia tangutic. Its chemical characteristics and pharmacological effects have been investigated. in detail.

An in vitro study on interaction of anisodine and monocrotaline with organic cation transporters of the SLC22 and SLC47 families

Current study systematically investigated the interaction of two alkaloids, anisodine and monocrotaline, with organic cation transporter OCT1, 2, 3, MATE1 and MATE2-K by using in vitro stably transfected HEK293 cells. Both anisodine and monocrotaline inhibited the OCTs and MATE transporters. The lowest IC50 was 12.9 μmol·L-1 of anisodine on OCT1 and the highest was 1.8 mmol·L-1 of monocrotaline on OCT2. Anisodine was a substrate of OCT2(Km = 13.3 ± 2.6 μmol·L-1 and Vmax = 286.8 ± 53.6 pmol/mg protein/min). Monocrotaline was determined to be a substrate of both OCT1(Km = 109.1 ± 17.8 μmol·L^-1, Vmax = 576.5 ± 87.5 pmol/mg protein/min) and OCT2(Km = 64.7 ± 14.8 μmol·L^-1, Vmax = 180.7 ± 22.0 pmol/mg protein/min), other than OCT3 and MATE transporters. The results indicated that OCT2 may be important for renal elimination of anisodine and OCT1 was responsible for monocrotaline uptake into liver. However neither MATE1 nor MATE2-K could facilitate transcellular transport of anisodine and monocrotaline. Accumulation of these drugs in the organs with high OCT1 expression(liver) and OCT2 expression(kidney) may be expected.