Synthesis and antibacterial activity of novel 3-substituted carbacephems

A series of novel 3-heterocyclothio substituted carbacephems having phenylglycyl side chain have been prepared starting from 3-H carbacephem. The compounds exhibit better chemical stability than the corresponding cephalosporin and strong activity against Gram-negative and Gram-positive organisms including Enterococcus faecalis.     

罗氟司特中间体3-环丙基甲氧基-4-二氟甲氧基苯甲醛的合成

目的研究罗氟司特中间体的合成。方法 3,4-二羟基苯甲醛依次与Boc酸酐和氯甲基环丙烷反应后得3-环丙基甲氧基-4-O-Boc-苯甲醛,脱掉Boc后再与二氟一氯甲烷醚化制得罗氟司特中间体3-环丙基甲氧基-4-二氟甲氧基苯甲醛。结果总收率约62%。结论本合成路线操作简单,成本低,有利于大规模制备。     

Antibacterial activities of new cephems against clinically isolated organisms (author's transl)

The antibacterial activities of cefotaxime (CTX), cefoperazone (CPZ), ceftizoxime (CZX), cefmenoxime (CMX), latamoxef (LMOX), cefotiam (CTM), cefazolin (CEZ), gentamicin (GM) and cefsulodin (CFS) were investigated. All causative organisms were isolated from patients with urinary tract infections treated in Tokai University Hospital. The results were as follows. 1) The MICs of CMX, CTX, and CZX against most of clinically isolated strains of E. coli, K. pneumoniae, Indole (-) Proteus sp. were 0.1 microgram/ml and lower. And then CTM, LMOX and CPZ showed similar antibacterial activities. 2) LMOX and GM showed potent antibacterial activities against C. freundii which was considered to be causative organisms of infections in rare cases. 3) Against S. marcescens, CMX, CTX, CZX, and LMOX showed very potent antibacterial activities. 4) Against P. aeruginosa, CFS, GM and CPZ showed moderate antibacterial activities. 5) Against Enterobacter sp., GM and CMX showed potent antibacterial activities.     

Aminothiazolylglycyl derivatives of carbacephems. I. Synthesis and antibacterial activity of novel carbacephems with substituted aminothiazolyl groups

A series of new carbacephem compounds which have substituted aminothiazolylglycyl side chain have been prepared starting from corresponding carbacephems with aminothiazolylmethoxyimino group. Among them, the compound having 3,4-dihydroxybenzoyl group showed very sharp activity against Pseudomonas aeruginosa. Moreover, the optical resolution of alpha carbon of aminothiazolylglycyl moiety was carried out through preparation of optically active side chain and the (S)-isomer (KT-4380) was found to be the most active against Pseudomonas sp. as well as other Gram-negative strains.     

人工佐剂三棕榈酰-S-甘油酰-半胱氨酸(P3C)及其中间体Fmoc-Lys P3C的合成

三棕榈酰 S 甘油酰 半胱氨酸 (P3C)是一种B细胞分裂原 ,并能在体内激活细胞毒T淋巴细胞 .它是一种无毒佐剂 .对该佐剂进行了合成 ,重点进行了化合物Fmoc LysP3C的合成方法研究 .所合成化合物的纯度和产率均较高 ,具有制备意义 .     

Effects of 1,25-dihydroxyvitamin D3 analogue 1,24(OH)2-22-ene-24-cyclopropyl D3 on proliferation and differentiation of a human megakaryoblastic leukemia cell line.

The novel analogue of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3), 1,24(OH)2-22-ene-24-cyclopropyl D3 (calcipotriol, MC903), exhibits similar effects on cell proliferation and cell differentiation in a newly established human megakaryoblastic leukemia cell line (HIMeg). MC903 was found to inhibit cell proliferation and induce cell differentiation in a liquid culture system at concentrations comparable to those of 1,25(OH)2 D3. Colony formation assay showed that MC903 or 1,25(OH)2 D3 markedly diminished the colony-forming ability of HIMeg cells at concentrations of 10(-6) M to 10(-10) M. Cell cycle analysis demonstrated that, as seen with 1,25(OH)2 D3, MC903 also altered the cell cycle distribution; the fraction of cells in G0 + G1 increased while those in S and G2 + M decreased. It can be concluded from these findings that 1,25(OH)2 D3 and its analogue MC903 have approximately equipotent effects on cells of megakaryoblastic lineage and are potentially useful in studying the cellular processes that are responsible for megakaryocytopoiesis.     

(3-Aminocyclopentyl)methylphosphinic acids: novel GABA(C) receptor antagonists

Our understanding of the role GABA(C) receptors play in the central nervous system is limited due to a lack of specific ligands. Here we describe the pharmacological effects of (+/-)-cis-3- and (+/-)-trans-3-(aminocyclopentyl)methylphosphinic acids ((+/-)-cis- and (+/-)-trans-3-ACPMPA) as novel ligands for the GABA(C) receptor showing little activity at GABA(A) or GABA(B) receptors. (+/-)-cis-3-ACPMPA has similar potency to (1,2,5,6-tetrahydropyridin-4-yl)methylphosphinic acid (TPMPA) at human recombinant rho1 (K(B)=1.0+/-0.2microM) and rat rho3 (K(B)=5.4+/-0.8microM) but is 15 times more potent than TPMPA on human recombinant rho2 (K(B)=1.0+/-0.3microM) GABA(C) receptors expressed in Xenopus oocytes. (+/-)-cis- and (+/-)-trans-3-ACPMPA are novel lead compounds for developing into more potent and selective GABA(C) receptor antagonists with increased lipophilicity for in vivo studies.     

Determination of 6-chloro-3-(3-cyclopropyl 1,2,4-oxadiazol-5-yl)-5-methyl-imidazo less than 1,5-a greater than-quinoxalin-4(5h)-one in rat serum, urine and brain by solid-phase extraction and liquid chromatography.

A simple, rapid, and accurate liquid chromatographic method with ultraviolet detection and solid-phase extraction is described for the quantitation of 6-chloro-3-(3-cyclopropyl 1,2,4-oxadiazol-5-yl)-5-methyl-imidazo less than 1,5-a greater than-quinoxalin-4(5h)-one (I, U-80447) in rat serum, urine and brain. Linear calibration curves were obtained in the concentration ranges of 5 ng ml-1-20 micrograms ml-1 (serum), 20 ng ml-1-20 micrograms ml-1 (urine), and 50 ng g-1-200 micrograms g-1 (brain). Intra- and inter-assay precision and accuracy were all found to be less than 10% at the three concentrations evaluated. The absolute extraction recovery each from serum, urine and brain was greater than or equal to 90%. Application of this method to the quantitation of the title compound in rat serum and brain for a pharmacokinetic study is reported.     

Comparison of in vitro antibacterial activities of new cephems against clinical organisms (isolated between June 1983 and January 1984)

We compared the in vitro antibacterial activities of ceftizoxime (CZX), cefotaxime (CTX), cefmenoxime (CMX), cefoperazone (CPZ), ceftazidime (CAZ), latamoxef (LMOX) and cefotetan (CTT) against 2,729 strains of 11 organisms freshly isolated from 10 medical institutions in Japan between June 1983 and January 1984 and obtained the following results: Against S. pyogenes, LMOX and CTT, which have the methoxy group at the 7 position, were less active than the other drugs. LMOX inhibited 80% of S. pyogenes at 0.78 micrograms/ml; CTT, at 1.56 micrograms/ml; but CZX and CTX inhibited 100% at 0.025 micrograms/ml or lower; CMX, at 0.05 micrograms/ml; and CPZ and CAZ, at 0.20 micrograms/ml. Against H. influenzae, E. coli, K. pneumoniae, P. mirabilis and indole-positive Proteus, these test antibiotics, especially CZX, CTX and CMX, which have the aminothiazolyl methoxyimino group, were potently active. Against S. marcescens CZX and CAZ were more active than the other drugs and against P. aeruginosa CAZ was more active than the other drugs. The test organisms did not tend to acquire resistance to these cephems when our results were compared with the results obtained at the development period.     

Antimalarial drugs inhibit human 5-HT(3) and GABA(A) but not GABA(C) receptors

BACKGROUND AND PURPOSE: Antimalarial compounds have been previously shown to inhibit rodent nicotinic acetylcholine (nACh) and 5-HT(3) receptors. Here, we extend these studies to include human 5-HT(3A), 5-HT(3AB), GABA(A) alpha1beta2, GABA(A) alpha1beta2gamma2 and GABA(C) rho1 receptors.EXPERIMENTAL APPROACH: We examined the effects of quinine, chloroquine and mefloquine on the electrophysiological properties of receptors expressed in Xenopus oocytes.KEY RESULTS: 5-HT(3A) receptor responses were inhibited by mefloquine, quinine and chloroquine with IC(50) values of 0.66, 1.06 and 24.3 microM. At 5-HT(3AB) receptors, the potencies of mefloquine (IC(50)=2.7 microM) and quinine (15.8 microM), but not chloroquine (23.6 microM), were reduced. Mefloquine, quinine and chloroquine had higher IC(50) values at GABA(A) alpha1beta2 (98.7, 0.40 and 0.46 mM, respectively) and GABA(A) alpha1beta2gamma2 receptors (0.38, 1.69 and 0.67 mM, respectively). No effect was observed at GABA(C) rho1 receptors. At all 5-HT(3) and GABA(A) receptors, chloroquine displayed competitive behaviour and mefloquine was non-competitive. Quinine was competitive at 5-HT(3A) and GABA(A) receptors, but non-competitive at 5-HT(3AB) receptors. Homology modelling in combination with automated docking suggested orientations of quinine and chloroquine at the GABA(A) receptor binding site.CONCLUSIONS AND IMPLICATIONS: The effects of mefloquine, quinine and chloroquine are distinct at GABA(A) and GABA(C) receptors, whereas their effects on 5-HT(3AB) receptors are broadly similar to those at 5-HT(3A) receptors. IC(50) values for chloroquine and mefloquine at 5-HT(3) receptors are close to therapeutic blood concentrations required for malarial treatment, suggesting that their therapeutic use could be extended to include the treatment of 5-HT(3) receptor-related disorders.     

Characterization of Apolipoprotein C3 (Apo C3) LNA/DNA Impurities and Degradation Products by LC-MS/MS

Apolipoprotein C3 (Apo C3) LNA/DNA gapmer was evaluated under various stress and formulation conditions for the purpose of its development as a potential biotherapeutic for low density lipoprotein (LDL) lowering. Using ion-pairing (IP) reversed-phase (RP) liquid chromatography ultra-high resolution (UHR) tandem mass spectrometry (IP-RPLC-MS/MS), a combination of accurate mass measurements and collision-induced dissociation enabled in-depth characterization of Apo C3 LNA/DNA oligonucleotide, in particular the inherent impurities following synthesis and degradation products after exposure to stress conditions. In this study, oligonucleotide samples were stressed under different pH and UV exposure conditions. The primary impurities in Apo C3 LNA/DNA were losses of nucleotide moieties from both the 5′- and 3′-terminus leading to n-1, n-2, etc. species. Desulfurization and depurination were observed in Apo C3 LNA/DNA after a week under UV light stress conditions at low pH. Guanine oxidation and dimerization were the primary degradation products detected under UV light exposure for 1 week at high pH. The effect of antioxidants on the levels of these degradation products was evaluated under neutral pH conditions. In the presence of all antioxidants, levels of guanine oxidation and desulfurization under tested conditions were the same as those in the unstressed sample, except for sodium ascorbate. The thorough understanding of the Apo C3 LNA/DNA oligonucleotide structure, its impurities, and degradation products laid the foundation for the successful formulation development of this novel biotherapeutic modality.     

华支睾吸虫病患者血清中免疫球蛋白和C_3含量的变化

本文分别采用 ELISA 双抗体夹心法和琼脂单向免疫扩散法检测了华支睾吸虫病患者血清中各类免疫球蛋白和 C_3的含量。结果显示:患者血清中 I gG、I gM 和 I gE 的水平明显高于正常对照组,I gA 的水平与正常人相比无明显改变;而 C_3的含量显著低于正常人。提示:人体感染华支睾吸虫病后,I gG、I gM 和 I gE 与相应的特异性抗体呈同步增高的趋势;补体和抗体协同参与华支睾吸虫病的免疫学发病机理,在此过程中消耗补体,使 C_3含量降低。     

囊虫病患者血清中免疫球蛋白和C_3含量的检测

应用ELISA双抗体夹心法和单向琼脂扩散法检测了30例囊虫病患者和正常献血者血清中免疫球蛋白及C_3的含量。结果:囊虫病患者血清中IgG和IgE的水平明显高于正常人,IgA和C_3的含量显著低于正常人;IgM略有增高,但与正常对照组间无显著差异。显示人体患囊虫病后,血清中以IgG和IgE增高为主,IgA和C_3呈降低趋势。     

Chemotactic effects of the complement-derived peptides C3a,C3ai and C5a (classical anaphylatoxin) on rabbit and guinea-pig polymorphonuclear leukocytes

Summary The complement-derived peptides C3a, C3a_i and C5a (= classical anaphylatoxin) were purified from hog serum and examined for chemotactic activity on rabbit and guinea-pig polymorphonuclear leukocytes (PMN) with the Boyden chamber technique (with filters of 3,0 m pore size). When media containing albumin or serum were used all peptides induced chemotaxis of both cell species. Only C3a showed a pronounced species dependence in that it was much more active on rabbit than on guinea-pig PMN. No gross differences were found between the influence of 0.5% BSA and 10% heated (56°, 30 min) homologous serum added to the medium. In the absence of protein chemotaxis did not occur.Abbreviations C3a, C5a cleavage peptides derived from the third and fifth component of complement respectively - C3a_i spasmogenically inactive form of C3a - PMN polymorphonuclear leukocytes - BSA bovine serum albumin     

Comparative induction of hepatic zinc-thionein and increase in tissue calcium by bacterial endotoxin in endotoxin-sensitive (C3H/HeN) and endotoxin-resistant (C3H/HeJ) mice

Induction of zinc-thionein (Zn-Th) by endotoxin was studied in mice using an endotoxin-sensitive C3H/HeN strain and an endotoxin-resistant C3H/HeJ strain to find a relation between the sensitivity of these strains to endotoxin and the inducibility of hepatic Zn-Th by the endotoxin. Both strains of female mice were injected with endotoxin at two doses and the increase in hepatic Zn-Th levels was examined after 24 h. At the lower dose (0.25 mg/kg body weight), C3H/HeJ mice induced Zn-Th at a markedly lower level than C3H/HeN mice. However, both strains exhibited a comparable amount of Zn-Th when a higher dose (10 mg/kg body weight) of endotoxin was used. A parallel increase in hepatic calcium concentration was observed with the induction of hepatic Zn-Th in both strains. The injection of a spleen supernatant fraction from C3H/HeJ mice into C3H/HeN mice did not reduce the Zn-Th induction by endotoxin in C3H/HeN mice.     

Response of isolated hepatocytes from carcinogen sensitive (C3H) and insensitive (C57BL) mice to signals inducing replication or apoptosis

The mouse strain C3H shows high incidence of liver tumors in carcinogenicity testing, while the strain C57BL exhibits low incidence. The F1 generation hybrids, B6C3F1, which are widely used in long-term carcinogenesis bioassays, are of intermediate sensitivity. We asked whether this strain difference could be due to different susceptibility of the parenchymal cells to signals inducing replication or apoptosis. Hepatocytes were isolated and cultured according to standard protocols. We tested (1) for the induction of DNA synthesis by epidermal growth factor (EGF), (2) for its inhibition by TGF-beta1, and (3) for the induction of apoptosis by TGF-beta1. Basal rates of DNA synthesis in untreated hepatocytes cultured from C3H and B6C3F1 mice were 6.5 and 3.5 times higher, respectively, than in hepatocytes from C57BL on day 3. Moreover, addition of EGF (10 ng/ml) increased DNA synthesis on day 3 in hepatocytes from C3H (4.2-fold) and B6C3F1 (2.7-fold) more strongly than in hepatocytes from C57BL. Treatment with TGF-beta1 inhibited basal and EGF-stimulated DNA synthesis dose-dependently. Inhibition was maximal at 1 ng TGF-beta1/ml in cultures from C57BL mice, and at 0.3 ng/ml in hepatocytes from C3H mice. In untreated hepatocytes from both strains virtually no apoptotic figures (condensed or fragmented nuclei, Hoechst 33285 staining) were found. After treatment with TGF-beta1 the incidence of apoptotic nuclei in hepatocytes from C57BL was higher than in cells from C3H mice (1.7% vs 3% on day 3). Thus it appears that hepatocytes from C57BL mice possess a lower growth potential, as indicated by a low basal rate of DNA synthesis and low inducibility by EGF, but a higher sensitivity to induction of apoptosis by TGF-beta1 than hepatocytes of the C3H strain. These findings may be helpful to explain the different susceptibility to induction of hepatocarcinogenesis in C3H and C57BL mice.