Syntheses and Pharmacological Activity of Some 17-[（2＇-Substituted）-4＇-pyrimidyl]androstene Derivatives As Inhibitors of Human 17α-Hydroxylase／C17,20-Layse＊

17-Heterocyclic substituted androstene derivatives have been found to be potent inhibitors for human testicular microsomal 17α-hydroxylase/C17.20-layse, which have potential usage in the treatment of benign prostatic hypertrophy(BPH) and prostatic cancer. In order to further investigate their structure-activity relationships, seven new 17-[(2‘-substituted)-4‘-pyrimidyl]androstene derivatives were designed and synthesized. The structures of the compounds were confirmed by IR, ^1H NMR, elemental analysis or MS measurements. The results of the pharmacological activity tests showed that‘compound 5 is a potent inhibitor for P45017α with IC50 225 nmol.L^-1.     

Exploratory screening study on anti-depresive activity of Helicid WN7-6 and its derivatives

AIM： To search some compounds that have anti-depressive activity from Helicid WN7-6 and its derivatives, and to explore their structure-function relationship. METHODS： 21 compounds including WN7-6, the lead compound were screened for antidepressant effect by the behavioral despair models （tail suspension test and the forced swimming test）. And to study their stimulative effect on central nerve system（CNS） by mice autonomic actest. RESULTS： Results shown that 11 of these compounds could decrease immobile time in suspend tail test and 6 compounds had the similar effect in force swimming test, compared with that of control group, the immobile time was obviously reduced （ P 〈 0.05, P 〈 0.01 ）. The results of autonomic activity test indicated that only 7 compounds could inhibit the activities of CNS （ P 〈 0.05, P 〈0.01）.[第一段]     

疟疾防治药物的研究——ⅩⅤ.双-(2,4-二氨基喹唑啉-6-取代氨甲基)芳香类化合物的合成及其抗疟作用

A series of bis(2,4-diamino-quinazol-6-yl-substituted aminomethyl) aromatic derivatives were synthesized. Compounds Ⅱ_1～5 were synthesized by two ways. In the first, 2,4,6-triaminoquinazoline was condensed with the corresponding aryl dialdehydes to form Schiff bases which were reduced with sodium borohydride; the second used a new method in which 2,4,6-triaminoquinazoline was directly condensed with the corresponding aralkyl dihalides to afford the same products. The derivatives Ⅱ_6～16 were prepared by formylation, nitrosation or methylation respectively.The inhibition activities of the title compounds on dihydrofolate reductase in rat liver were assayed. The activities of formylated or nitrosated products were 6-9 times, and of methylated products were twofold as active as that of the parent compounds. Ⅱ₆, Ⅱ₉, Ⅱ₁₀ and Ⅱ₁₄ were nearly as potent as pyrimethamine. Primary antimalarial screening in mice showed that no one possessed significant activity.     

Effects of resveratrol,curcumin and their derivatives on the activation of microglia induced by LPS

Objective To determine the inhibitory effects of 21 resveratrol derivatives and 3 natural curcuminoids on lipopolysaccharide(LPS)-induced Nitric oxide(NO)and tumor necrosis factor-alpha(TNF-α)production in microglia and their structure-activity relationships.Methods Cell viability was evaluated by the MTT reduction assay.Accumulation of nitrite(NO2-)in culture supernatant fluids was measured by the Griess reaction.Sodium nitroprusside(SNP)(2.5 mM)solution was used to determine the scavenging activities of these compounds.The levels of TNF-α in the culture medium were measured by using an ELISA kit.Semi-quantitative RT-PCR analysis was used to determine the mRNA levels of inducible NOS(iNOS)and TNF-α.Results It was found,for the first time,that certain resveratrol derivatives that have 3,5-dimethoxyl groups in the A-ring,such as(E)-4-(3,5-dimethoxystyryl)phenol(pterostilbene,compound 2),or have substituted the B-ring of resveratrol with quinolyl,such as(E)-5-[2-(quinolin-4-yl)vinyl]benzene-1,3-diol(compound 18)and(E)-4-(3,5-dimethoxystyryl)quinoline(compound 19),strongly inhibited NO production.Compounds 2,18,and 19 reduced LPS-induced protein and mRNA expression of inducible NO synthase(iNOS),but did not display direct NO-scavenging activity up to 30 μM in sodium nitroprusside(SNP)solution.Moreover,compounds 2,18,and 19 could also significantly inhibit the production of TNF-α by LPS-activated microglia.Furthermore,we found the demethoxy derivatives of curcumin have more potent inhibition activity on NO and TNF-α releasing in activated-microglia.Conclusions In the present study we compared the activated-microglia inhibition effect of resvertrol,curcumin and their derivatives and provided a glance of the structure-activity relationships of these compounds,the information is beneficial to design new potent compounds which can provide better therapeutic implications for various neurodegenerative diseases.     

Central pharmacological effect of Chinese Materia Medica Cynanchum Chinese R. Br

Aim： To study the Central pharmacological effect of the water - and chloroform - extract compounds from Cynanchum Chinese R. Br. Methods： The minimal neurotoxicity of the extract - compounds were measured by rotorod test. The Independent activity test and the hypnotic synergism test by under threshold hypnotic dosage of pentobarbital were employed to evaluate the central pharmacological action of the extract - compounds. All the extract - compounds were evaluated for anticonvulsant activity by maximal electroshock （MES） and subcutaneous metrazol （MET） induced seizure. Result： The extract - compounds significantly inhibited the spontaneous motor activity dose - dependently in mice after intraperitoneal administration. The two extract - compounds promoted the hypnotic effect by under threshold hypnotic doses of pentobarbital, and the ED50 values were 2.36 g/kg and 0. 75 g/kg, respectively. Meanwhile, the water - extract compounds exhibited significant protection after intraperitoneal （ip） administration in MET - induced seizures and the ED50 value was 2.34 g/kg; however, the chloroform - extract compounds did not produced protective effect in this seizure model. On the other hand, the chloroform - extract compounds exhibited significant protection in MES and the ED50 value was 1.34 g/kg; the water- extract compounds had no protective effect. Both extract -compounds showed no neurotoxicity as compared with phenytoin. Conclusion ： The extract compounds from Cynanehum Chinese R. Br show inhibition effect on CNS, and the water - and chloroform - extract compounds show different anticonvulsant activities in different seizure models in mice.     

Design, synthesis and pharmacological evaluation of 4-[2-alkylthio-5（4）-（4- substitutedphenyl）imidazole-4（5）yl]benzenesulfonamides as selective COX-2 inhibitors

Aim： To design and synthesize a series of benzenesulfonamide derivatives, 4-[2- alkylthio-5 （4）-（4-substitutedphenyl）imidazole-4（5）-yl]benzenesul fonamides （4a-4j）, which are intended to act as cyclooxygenase-2 （COX-2） inhibitors with good COX-2 inhibitor activity, and which will exert anti-inflammatory activities in vivo. Methods： Benzenesulfonamide derivatives were designed and synthesized through multi-step chemical reactions. All the synthesized compounds were evaluated in an in vitro assay. The active compound 4a-4f was selected for further evaluation in a carrageenan-induced rat paw edema model. Results： Docking studies showed that compound 4 bind into the primary binding site of COX-2 with the sulfonamide SO2NH2 moiety interacting with the secondary pocket amino acid residues. In the in vitro assay, compound 4 inhibited COX-2 with an inhibition concentration IC50 value of 1.23-8 nmol/L, compared to celecoxib with IC50 value of 1.5 nmol/L. Compound 4b and 4c had good potency and selectivity in comparison to the celecoxib. In the in vivo model, compound 4a-4f exhibited a moderate potency to inhibit 50% carrageenan-induced paw edema with value of 1.58-4.3 mg/kg. In the latter experiment, compound 4c was the most active compound. Conclusion： The antiinflammatory effects obtained for compound 4a-4j could be due to the presence of fluorine or hydrogen substituents in the para position of the phenyl ring of these compounds.     

Anticarcinogenic and antioxidant activity of diindolylmethane derivatives

To investigate the synthesis methods and the bioactivity of diindolylmethane (DIM) derivatives. METHODS:1) A 3D-Quantitative Structure-Active Relationships (QSAR) Comparative Molecular Field Analysis (CoMFA) study of 14 DIM derivatives was investigated to predict their anticarcinogenic activity. 2) Based on CoMFA model, a series of new derivatives of DIM were designed and synthesized. 3) Their free radical scavenging and antioxidant potentials were tested using in-vitro DPPH radical scavenging and ~-carotene antioxidant models. 4) The anticarcinogenic activities of some compounds were tested by using microculture tetrazolium assay (MTT) and sulforhodamine B (SRB) proteochromosomic assays. RESULTS: 1) The CoMFA model derived from DIM analogues proved a good predictive ability with q2 value of 0.827. 2) New designed compounds 3c and 4c exhibited 3-fold more potent radical scavenging activity than reference substance Vitamin E in DPPH model expressed by IC50 values. 3) The primary antitumor screening essay showed that some DIM derivatives designed exhibited the inhibitory activities to some tumor cell growth at relatively high concentration, and DIM was the most effective among them. CONCLUSION: DIM‘s 3D-QSAR model is reliable. According to it, eleven DIM derivatives weresynthesized, and two derivatives of them possess potent radical scavenging activities and some showed the inhibitory activities in primary anticancer assay in vitro.     

Design,synthesis and cytotoxic activity of novel 1-aroyl-4-(2-chloroethyl)semicarbazides

A series of aroyl derivatives of 4-(2-chloroethyl)semicarbazide were designed and synthesized to explore their antiproliferative activity against human brain carcinoma (U251) and human liver carcinoma (Hepg2) cell lines. The synthesized compounds were characterized by elemental analyses and spectroscopic data. It was established that compounds in which semicarbazide fragments are substituted with a (2-indolyl)carbonyl moiety showed a higher cytotoxic activity than the corresponding benzoyl derivatives. 1-[(5-Benzyloxy-1H-indol-2-yl)carbonyl]-4-(2-chloroethyl)semicarbazide (24) showed the highest cytotoxic activity against Hepg2 (IC₅₀= 21 μg/ml), while 4-(2-chloroethyl)-1-[(5-methoxy-1H-indol-2-yl)carbonyl]semicarbazide (23) was the most active compound against U251 (IC₅₀ = 8 μg/ml). __________ Translated from Khimiko-Farmatsevticheskii Zhurnal, Vol. 41, No. 4, pp. 12–15, May, 2007.     

N-取代多亚甲基二甲酰胺类癌细胞分化诱导剂的合成

N-Alkyl polymethylene dicarboxamides are known potent inducers of erythroid differentiation in murine erythroleukemia cells. The most active inducer N, N, N', N'-tetramethyl-1, 6-hexane-dicarboxamide has the same effectiveness as HMBA which has entered clinical trials as a differentiating agent. These compounds also have inducing activity in HL-60 human promyelocytic leukemia cells. In this paper, the synthesis of a series of N,N'-bis[2-(2-thiazolinyl)], N,N'-bis [5-(1-methyl-2-pyridonyl)], N, N'-bis [3-(1- phenyl- 5- pyrazolonyl )] polymethylene dicarboxamides and 3,3'-(polymethylene dicarbonyl)bis(1-methyl-2-imidazolidine-thiones) is reported. The inducing activities of the compounds were evaluated in vitro with HL-60 human promyelocytic leukemia cell line. Among them, N, N'-bis[-2- (2-thiazolinyl)]-1,8-octamethylene-dicarboxamide (Ⅰ₄) and N, N'-bis[5- (1-methyl- 2-pyridonyl)]-1,6-hexamethylenedicarboxamide (Ⅱ₃) were shown to be relatively effective inducers of differentiation.     

二氢叶酸还原酶抑制剂:5-取代苯硫基(硒基)-2,4-二氨基嘧啶类化合物的合成和抑酶活性

According to the principle of isosterism the-CH₂-group of 5-(substituted benzyl)-2, 4-diaminopyrimidine was modified by—S—and—Se—and some 5-(substituted phenyl)thio-2, 4-diaminopyrimidines and 5-(substituted phenyl)seleno-2,4-diaminopyrimidines were synthesized. Their inhibitory activities on L. casei and chicken liver dihydrofolate reductase were determined. Preliminary data showed that the inhibitory activities of these compounds were less than those of the corresponding 5-(substituted benzyl)-2, 4-diaminopyrimidines. Their selectivities are also decreased.     

抗肿瘤药物的研究:N′,N″-二螺三哌嗪类化合物的合成

In order to search for new antitumor drugs, sixteen N′, N″-dispirotripiperazine derivatives were synthesized from N′, N″-dispirotripiperazinium dichloride dihydrochloride by substitution, acylation and Mannich reaction. Six compounds were selected for preliminary pharmacological test. The result showed that five compounds possess inhibitory action against carcinoma S₃₇ in rats. The inhibitory activity of compounds Ⅵ and Ⅹ was 55.0% and. 41.9% respectively.     

Contrast study on optimizing antibacterial lead compounds from three kinds of derivates in vitro

AIM： To choose at least one kind of better derivate from 81 novel compounds for further structure-function relationship researches by contrast study on antibacterial activity of three kinds of derivates developed from decanoyl acetaldehyde, a certain kind of ethnodrug and pure synthetics ： Three quality control strains, staphylococcus aureus ATCC25923, bacillus coli ATCC25922 and bacillus aeruginosus ATCC27853 were used in the preliminary screening in vitro. Some active derivates were selected to carry out the minimal bactericidal concentration test, killing curve test, and the effect of cultivation parameters on minimal inhibitory concentration on clinical separated staphylococcus aureus and MRSA. Acute toxicity test of a few promising derivates were tested preliminarily by intragastric administration to mice RESULTS： There were significant differences of the antibacterial activity for the three different kinds of derivatives compared to the quality control groups. Results shown that 15 percent of the 53 compounds developed from decanoyl acetaldehyde had special activity of inhibiting staphylococcus aureus ATCC25923. Some optimized derivatives had better activity to clinical separated staphylococcus aureus and MRSA compared to the positive control drug decanoyl acetaldehyde.[第一段]     

Discovery and characterization of novel small-molecule agonists of G protein-coupled receptor 119

Aim： GPR119 is a G protein-coupled receptor （GPCR） that is highly expressed in pancreatic β-cells and intestinal L-cells and facilitates glucose-stimulated insulin secretion （GSIS）. GPR119 may represent a novel target for the treatment of metabolic disorders. Here, we sought to identify novel small-molecule GPR119 agonists.
Methods： A cell-based high-throughput screening assay was established using HEK293 cells stably expressing GPR119 and pCRE-luc reporter plasmid （HEK293/GPR119/pCRE-luc）. A compound library composed of 1440 compounds was screened. Mouse β-cell line MIN-6 and isolated mouse islets were used to evaluate the effects of candidate compounds on GSIS in vitro.

Results： Three compounds with novel structures （ZSY-04, -06, and -13） were found to activate GPR119-mediated signaling and to induce GPR119 desensitization. The EC50 values of ZSY-04, -06, and -13 in stimulating intracellular cAMP accumulation in HEK293/GPR119 cells were 2.758, 3.046, and 0.778 μmol/L, respectively. Furthermore, all three compounds displayed high selectivity for GPR119, and did not activate other 9 GPCRs tested. Moreover, all three compounds significantly increased GSIS in both MIN-6 mouse β-cells and isolated mouse islets at concentration of 10 μmol/L.

Conclusion： Three novel small-molecule GPR119 agonists （ZSY-04, -06, and -13） with high receptor selectivity and capacity to induce GSIS in vitro were discovered. These compounds are potential candidates to be structurally optimized into drugs for the treatment of type 2 diabetes.     

Molecular modeling,quantum polarized ligand docking and structure-based 3D-QSAR analysis of the imidazole series as dual AT~ and ETA receptor antagonists

Aim： Both endothelin ETA receptor antagonists and angiotensin AT1 receptor antagonists lower blood pressure in hypertensive patients. A dual AT1 and ETA receptor antagonist may be more efficacious antihypertensive drug. In this study we identified the mode and mechanism of binding of imidazole series of compounds as dual AT1 and ETA receptor antagonists. Methods： Molecular modeling approach combining quantum-polarized ligand docking （QPLD）, MM/GBSA free-energy calculation and 3D-QSAR analysis was used to evaluate 24 compounds as dual AT1 and ETA receptor antagonists and to reveal their binding modes and structural basis of the inhibitory activity. Pharmacophore-based virtual screening and docking studies were performed to identify more potent dual antagonists. Results： 3D-QSAR models of the imidazole compounds were developed from the conformer generated by QPLD, and the resulting models showed a good correlation between the predicted and experimental activity. The visualization of the 3D-QSAR model in the context of the compounds under study revealed the details of the structure-activity relationship： substitution of methoxymethyl and cyclooctanone might increase the activity against AT~ receptor, while substitution of cyclohexone and trimethylpyrrolidinone was important for the activity against ETA receptor; addition of a trimethylpyrrolidinone to compound 9 significantly reduced its activity against AT~ receptor but significantly increased its activity against ETA receptor, which was likely due to the larger size and higher intensities of the H-bond donor and acceptor regions in the active site of ETA receptor. Pharmacophore-based virtual screening followed by subsequent Glide SP, XP, QPLD and MM/GBSA calculation identified 5 potential lead compounds that might act as dual AT1 and ETA receptor antagonists. Conclusion： This study may provide some insights into the development of novel potent dual ETA and AT1 receptor antagonists. As a result, five compounds are found to be the best dual antagonists against AT1R and ETA receptors.     

双苄基异喹啉类生物碱的心血管药理作用

Tetrandrine, dauricine, daurisoline and neferine are bisbenzylisoquinoline alkaloid derivatives isolated from Chinese traditional medicine and herbs. The cardiovascular pharmacological effects and the mechanism of actions of these compounds were reviewed. Tetrandrine isolated from Stephania tetrandra S Moore possesses antihyper-tensive and antiarrhythmic effects. The antihypertensive effects of tetrandrine have been demonstrated in experi-mental hypertensive animals and in hypertensive patients. Recent studies showed that in addition to its calcium antagonistic effect, tetrandrine interacted with M receptors. Modulation by M receptor is one of the pharmacologi-cal mechanisms of cardiovascular effects of tetrandrine. Dauricine and daurisoloine were isolated from Menispermum dauricum DC. The antiarrhythmic effects of dauricine have been verified in different experimental arrhythmic models and in cardiac arrhythmic patients. Dauricine blocked the cardiac transmembrane Na ,K and Ca2 ion currents. Differin     

Study on pharmacodynamics of recombinant interferon α-2b suppository

Objective To investigate the antiviral activity of recombinant interferonα-2b suppository(IFNα-2b)in vivo and in vitro.Methods The cytopathic-effect inhibition assay was applied in this study to investigate the antiviral activity of this drug as well as yingtelong and axiluowei as positive control.The guinea pig model of vaginitis and skin infection caused by HSV-2 infection were established,treated with IFNα-2b suppository at dosages of 60000、180000、540000 IU,using IFNα-2b injection 180000 IU·kg-1 as controls.Score the pathological changes of appearance and skin,the virus activities of vaginal secretion and tissue sections of viginae were assayed after treatment.Results The TD50 of IFN α-2b and yingtelong for Vero cells was(>100)μg·mL-1 and(>100000)IU·mL-1,respectively.The IC50 of IFN α-2b and yingtelong and axiluowei for Herpes virus type 1 was(0.29±0.08)μg·mL-1 and(185.0±28.8)IU·mL-1 and(0.19±0.03)μg·mL-1,respectively.The mean scores for vaginal and skin lesion of the treated groups were lower than those of untreated group.Among these concentrations,the IFNα-2b suppository of 540000 IU·kg-1 group.Showed highest anti-viral activity.The virus activity in vaginal secretion of treated group was lower than that of untreated group too(P<0.01 or P<0.05).Tissue sections of viginae after treatment with IFNα-2b suppository showed significantly therapeutical effects on the degrees of vaginal lesion.At the same dosage,The anti-HSV activity of IFNα-2b suppository was also compared with IFNα-2b injection,the results showed that the activity of suppository of 540000 IU·kg-1 group was similar to that of the injection.Conclusions The IFNα-2b suppository has anti-viruses function both in vivo and in vitro.     

A cell-based,high-throughput homogeneous time- resolved fluorescence assay for the screening of potential K-opioid receptor agonists

Aim： The aim of this study was to identify K-opioid receptor （KOR） agonists from a library of 80 000 small-molecule compounds and provide the experimental basis for the development of new analgesic candidates. Methods： The cell-based, high-throughput screen for human KOR agonists was based on the LANCETM cAMP assay. Preliminary structure-activity relationship （SAR） analysis was applied according to the compounds＇ structures. An acetic acid twisting experiment was used to verify the pharmacodynamics. Results： In total, 31 compounds were identified as KOR agonists after preliminary and secondary screening. Of these compounds, five demonstrated significant KOR-stimulating activity that was comparable to U-50,488, a selective KOR agonist. The ECho values for I-7, I-8, 1-10, 11-5, and 11-8 were 13.34±1.65, 14.01±1.84, 9.57±0.19, 14.94±0.64, and 8.74±0.72 nmol/L, respectively. Based on SAR studies, the stimulating activity of compounds with 5-phenyl-7-（trifluoromethyl）-4,5,6,7-tetrahydropyrazolo [1, 5-a] pyrimidine （group I） and 3,4-dimethoxy-N-（2-oxoethyl）-N-p-tolylbenzenesulfonamide （group II） parent structures were higher than the compound with a 5-hydroxy-2-methylbenzofuran-3-carboxylic acJd （group III） parent structure. PharmacodynarnJc experiments JndJcated that 20-40 μg/kg ip of compounds 1-10 and 11-8 significantly decreased the number of writhes induced by acetic acid; this finding is consistent with the SAR studies. Furthermore, the analgesic effects of compounds I-10 and II-8 were significantly antagonized in the presence of the selective KOR antagonist nor-BNI. Conclusion： These findings collectively indicate that compounds I-10 and II-8 exhibit significant analgesic activities, providing evidence, at least in part, for their clinical application as new analgesic drugs.     

疟疾防治药物的研究——Ⅸ.2,4-二氨基-6-[N-(取代苄基)-N-(取代氨基甲基)氨基]-喹唑啉类衍生物的合成

In searching for new antimalarial agents ten new compounds of 2,4-diamino-6-[N-(substituted benzyl)-N-(substituted aminomethyl)-amino]-quinazoline have been synthesized. Preliminary screening results showed that only one (Ⅳ₆) of these compounds displayed a slight degree of antimalarial activity against plasmodium berghei in mice. The intermediate compound (Ⅰ₂) showed suppressive effect on plasmodium berghei in mice and prophylactic activity against plasmodium gallinaceum in chicken.     

Synthesis and bioassay of β-（1,4）-D-mannans as potential agents against Alzheimer＇s disease

Aim： Oligomannurarate 971 derived from a marine plant has shown neuroprotective effects. In this study we synthesized a series of truncated derivatives of the oligosaccharide, and investigated the effect of these derivatives against Aβ peptide toxicity in vitro. Methods： The sulfoxide method was applied to synthesize the derivatives. SH-SY5Y human neuroblastoma cells were treated with Aβ1-40 （2 pmol/L）, and the cell viability was detected using a CCK8 assay. Results： A series of β-（1,4）-D-mannosyl oligosaccharide, ranging from the disaccharide to the hexasaccharide, were synthesized. Addition of 10 pmol/L β-（1,4）-D-mannobiose 6, β-（1,4）-D-mannotriose 9 or β-（1,4）-D-mannotetraose 12 in SH-SY5Y cells significantly attenuated Aβ1-40-induced toxicity. The efficacies were similar to those caused by 10 pmol/L oligomannurarate 971 or alzhemed. Other oligosaccharides including oligomaltoses and oligocelluloses were less active. Conclusion： Synthetic homogeneous short chain β-（1,4）-D-mannans shows neuroprotective effect against Aβ peptide toxicity similar to that of heterogeneous oligomannurarate 971 and alzhemed.     

抗癌药物的研究:4-氯苯氧乙酰胺衍生物的合成

Since compounds Ⅲ and Ⅳ possess retinoid-like action and analogues of Ⅰ inhibit some cancer cells ,seventeen derivatives of chlorophenoxy acetamide were synthesized.Of the seven compounds screened ,two exhibit cytostatic activity(1 and　2).Mass spectra showeda　special expulsion of SO2 and CO from the sulfonamide compounds,Nuclear magnetic resonancesplitting patterns of these compounds also showed interesting features.