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D Hoffmann

Publications and source records attributed to D Hoffmann.

466 records · Page 26Linked to original sources
PubMedDate not supplied

Attachment of rhodosaminylanthracyclinone-type anthracyclines to the hinge region of monoclonal antibodies.

We have found that a maleimidobenzoyl spacer attached to OH-4' of the rhodosamine moiety of rhodosaminylanthracyclinone-type anthracyclines is most suitable for the attachment of these drugs to carriers, providing important advantages: The spacer is selectively and most readily introduced into the rhodosamine moiety of the drugs, is stable enough for proper handling of the derivatives, and can easily be attached to thiol groups of carrier systems such as reduced monoclonal antibodies. The anthracyclines can be liberated from the conjugates by mere hydrolysis, requiring neither hydrolytic enzymes nor acidic pH. Liberation of the drugs can, moreover, be affected by the presence of the appropriate substituents Z on the phenylene ring of the spacer, thus allowing slowed or enhanced liberation of the cytostatically active drug. The corresponding p-maleimidobenzoyl derivatives of beta-rhodomycin I, N,N-dimethyldaunorubicin, and rodorubicin have been attached to thiol groups of the hinge region of reduced monoclonal antibody BW 494/32, directed against a pancreatic cancer associated glycoprotein antigen, resulting in MoAb BW 494/32 conjugates, carrying 4.8-6.8 mol of cytotoxic residues/mol of MoAb. Rodorubicin was similarly attached to MoAb BW 575/931/2, directed against a small cell lung cancer associated antigen and to MoAb BW 431/26, recognizing an epitope detectable on carcinoembryonic antigen. The results provide evidence that the newly developed method of coupling of anthracyclines to the hinge region of monoclonal antibodies may be of broader use.

Anthracyclines↗
PubMedDate not supplied

Supercritical fluid extraction in the determination of tobacco-specific N-nitrosamines in smokeless tobacco.

A new approach to the analysis of the carcinogenic, tobacco-specific N-nitrosamines (TSNA) in moist snuff tobacco is based on the extraction of tobacco with methanol-modified supercritical carbon dioxide. Extracted TSNA are trapped across a glass cartridge filled with Tenax GR, from which they are subsequently released by thermal desorption and analyzed by capillary gas chromatography with a thermal energy analyzer. The analytical recoveries for the major TSNA range from 83 to 98%; the detection limits are below 2 ng/g. The methodology is fast, reproducible, highly selective, and sensitive. The supercritical fluid extraction (SFE) releases up to 7 times more of the highly carcinogenic 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) from tobacco than has been determined after conventional solvent extraction. Studies have confirmed that this is not an artifact. In contrast, the cyclic N-nitrosamines, N'-nitrosonornicotine, N'-nitrosoanabasine, and N'-nitrosoanatabine, showed no significant quantitative differences whether determined by the SFE method or the conventional solvent extraction method.

Carbon Dioxide↗
PubMedDate not supplied

Rat liver metabolism of benzo[b]naphtho[2,1-d]thiophene.

Thioarenes, sulfur-containing polycyclic aromatic hydrocarbons, have been detected in a number of environmental sources. The metabolism of one thioarene, benzo[b]naphtho[2,1,d]thiophene ([2,1]BNT), by F344 rat liver 9000g supernatant (S-9) was studied. [2,1]BNT which is structurally analogous and has similar carcinogenic potency to chrysene, was metabolized to six ethyl acetate-extractable metabolites when incubated with S-9 from Aroclor 1254-treated F344 rats. Each metabolite was collected from reverse-phase HPLC, and their identities were determined by analysis of MS and NMR data. In order of elution from HPLC they are as follows: (1) trans-1,2-dihydroxy-1,2-dihydrobenzo[b]naphtho[2,1-d]thiophene, (2) benzo[b]naphtho[2,1-d]-thiophene sulfone, (3) benzo[b]naphtho[2,1-d]thiophene sulfoxide, (4) trans-3,4-dihydroxy-3,4-dihydrobenzo[b]naphtho[2,1-d]thiophene, (5) 8- or 9-hydroxybenzo[b]naphtho[2,1-d]thiophene, and (6) 7-hydroxybenzo[b]naphtho[2,1-d]thiophene. In addition, the identities of metabolites 1, 2, 3, and 4 were confirmed by comparison to standards. The syntheses of the sulfone and sulfoxide of [2,1]BNT are reported here. The syntheses of the dihydrodiols were reported previously. Metabolite 5, a hydroxy[2,1]BNT, was the major metabolite formed by liver S-9 from untreated F344 rats. Microsomal preparations from these rats also produced significant amounts of the dihydrodiols, 1 and 4, and the sulfoxide, 3. Microsomes prepared from Wistar rats produced dihydrodiols and the sulfone and sulfoxide of [2,1]BNT. Therefore, [2,1]BNT is metabolized by both ring oxidation and sulfur oxidation in these two strains of rats.

Animals↗
PubMedDate not supplied

Chemical studies on tobacco-specific N-nitrosamines in the smoke of selected cigarettes from the U.S.A., West Germany, and France.

Studies on the variations of biologically important smoke constituents of popular cigarettes in several European countries and in the United States include investigations on the types and levels of volatile (VNA) and tobacco-specific nitrosamines (TSNA). GLC-TEA and HPLC-TEA methods are used to determine the VNA and TSNA in mainstream and sidestream smoke of cigarettes. This report presents data on the smoke of some West German cigarettes which demonstrate that the VNA levels are about the same as those in the smoke of U.S. cigarettes; but that they are lower than those in the smoke of French cigarettes, made of dark tobacco types. The tobacco-specific nonvolatile N-nitrosamines determined in the mainstream smoke of commercial U.S. blended cigarettes (N-nitrosoanatabine, N-nitrosonornicotine and 4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone together) ranged from 680 to 830 ng per cigarette, while those for commercial West German products ranged from 340 to 780 ng per cigarette in nonfilter brands and from 180 to 450 ng in filter-tipped cigarettes. The analyzed French products contained 1010 to 1380 ng of these compounds in the smoke of each cigarette.

Chromatography, Gas↗
PubMedDate not supplied

Quinoline and methylquinolines in cigarette smoke: comparative data and the effect of filtration.

A method has been developed to determine quantitatively quinoline, isoquinoline, and each of the methylquinoline isomers in cigarette smoke. This method was applied to the smoke of several different commercial cigarettes with and without filter tips. Quantitative analyses were accomplished by capillary gas chromatography, using a 50-m OV-101 fused silica capillary column and a 50-m Dexsil 300 fused silica column. Quinoline, isoquinoline and all seven of the methylquinolines were found in cigarette smoke. When cigarettes were analyzed with and without filter tips, these compounds were reduced by filtration, but not as much as tar was reduced. The significance of these data is discussed in view of the biological activity of specific isomers of these environmental pollutants.

Chromatography, Gas↗
PubMedDate not supplied

Comparison of in vitro and in vivo properties of rhirudin (HBW 023) and a synthetic analogous peptide.

Recombinant hirudin and a shortened synthetic analogue, with the amino acid sequence of D-Phe-Pro-Arg-Pro-(Gly)4-Asn-Gly-Asp-Phe-Glu- Glu-Ile-Pro-Glu-Glu-Tyr-Leu, are specific thrombin inhibitors which in a concentration-dependent manner inhibit thrombus formation as well as clot propagation both in vitro and in vivo. In comparison to the analogue, lower molar concentrations of rhirudin affected doubling of aPTT and TT as well as inhibition of thrombin amidolytic activity or thrombin-induced platelet aggregation in vitro. In the rat wire coil-induced thrombosis model, a 50% thromboprotective effect may be brought about with doses of 0.043 mumol/kg of rhirudin and 1.43 mumol/kg of the synthetic peptide. However, doubling of bleeding times is caused, on average, by dosages of between 0.143 and 0.43 mumol/kg rhirudin or approximately 0.143 mumol/kg of the analogue. Treatment groups included animals revealing significant prolongation of bleeding times as well as nonresponders. Despite the 10-fold longer impact on aPTT after application of rhirudin, the extent of mean bleeding time prolongation is identical to that of the analogue.

Amino Acid Sequence↗
PubMedDate not supplied

Comparative response of normal and of human papillomavirus-16 immortalized human epithelial cervical cells to benzo[a]pyrene.

Laboratory evidence suggests synergism of human papillomavirus (HPV) infection with cigarette smoking behaviors in enhancing the risk of cervical cancer. In this preliminary investigation, we tested the hypothesis that HPV infection may alter the metabolic activation of tobacco smoke carcinogens, such as benzo[a]pyrene (B[a]P), thereby playing a role in the etiology of cervical cancer. We examined in vitro the metabolism and DNA adduct formation of [3H]B[a]P in normal and HPV-16 immortalized human epithelial cervical cells in culture, and investigated the effect of [3H]B[a]P on growth of these cells. Cultures of normal human cervical cells and of HPV-16 immortalized cervical epithelial cells were exposed to 0.2 microM [3H]B[a]P for 24 and 48 h. [3H]B[a]P inhibited growth of both normal and HPV-16 immortalized cervical cells. However, the growth inhibition of normal cells was more profound than that of HPV-16 immortalized cells. Comparison of the metabolism of [3H]B[a]P in these cells indicated that they both metabolize [3H]B[a]P predominantly to [3H]trans-9,10-dihydroxy-9,10-dihydrobenzo[a]pyrene ([3H]B[a]P-9, 10-diol), [3H]r-7,t-8, 9,c-10-tetrahydroxy-7,8,9, 10-tetrahydrobenzo[a]pyrene ([3H]trans-anti-B[a]P-tetraol), and unknown polar products. Enzymatic hydrolysis of water-soluble metabolites indicated that the levels of glucuronide and sulfate conjugates in these cells are negligible. Similarly, both cell lines form similar [3H]B[a]P-DNA adducts. However, the level of the (+)[3H] anti-B[a]P diol epoxide (BPDE)-deoxyguanosine adduct in HPV-16 immortalized cells after 24 and 48 h exposures was 3.8 and 3. 1 pmol/mg DNA, respectively, which is 2.2-fold and 2.6-fold greater than the level of this adduct in normal cells. Under the conditions and within the time frame employed in these assays, both the cell growth and DNA damage induced by [3H]B[a]P appear to be higher in HPV-16 immortalized cells than those detected in normal cells. The results, although preliminary, suggest that HPV-16 immortalized cervical cells are more susceptible to DNA damage by BaP which, in part, may enhance their transformation to malignant cells.

Benzopyrenes↗
PubMedDate not supplied

Metabolites of phenanthridine formed by rat liver homogenate.

The ethyl acetate-extractable metabolites of phenanthridine as formed in vitro with Aroclor-induced rat liver homogenate were isolated and structurally identified. The relative amounts of these metabolites were determined using [6-14C]phenanthridine. The major metabolites of phenanthridine formed under these incubation conditions were identified as phenanthridine-N-oxide, 1,2-dihydroxy-1,2-dihydrophenanthridine, and 9,10-dihydroxy-9,10-dihydrophenanthridine. Phenanthridone and 2-hydroxyphenanthridine were identified as minor metabolites. These data were obtained using an identical incubation mixture as employed in mutagenicity assays. It is suggested that metabolites in addition to phenanthridone are likely to contribute to mutagenicity of phenanthridine observed in assays performed with metabolic activation.

Animals↗
PubMedDate not supplied

Indication of a certain molecular correlation between rat IgE and human IgE.

The PCA-activity of rat sera sensitized with Ascaris suum antigen can be absorbed by antihuman IgE and by rabbit or sheep antihuman IgE convalently bound to sephadex particles or paper discs respectively. In vitro, rat immune sera show positive results using the human RIST-system abolished by preheating for 2 hours at 56 degrees C and therefore excluding an interaction of IgG 4. In contrast, no rat sera activity was seen using the human PRIST-system. These results agree with the concept of a single common antigenic determinant in human and rat IgE since in the human RIST and PRIST-system the binding of rat IgE to antihuman IgE could only be demonstrated on the principle of competitive inhibition (RIST) but not on that of a sequential binding (PRIST). The possibility of a certain heterogeneity in the IgE population can not be excluded.

Animals↗
PubMedDate not supplied

Preparation and evaluation of PEG-bound thrombin inhibitors based on 4-amidinophenylalanine.

The dipeptide Mtr-Asp-D-Adf-Pip 10 represents a potent thrombin inhibitor. In comparison to NAPAP, 10 exhibited improved tolerability and a longer half-life in vivo, i.e., 20 +/- 5 min. We have coupled aminopolyethyleneglycolmonomethylether of various molecular weights to the carboxyl moiety of 10 and evaluated their biological properties. First, Mtr-Asp-OBut was coupled to the amino group of the PEG employing TOTU as an activating agent. This was followed by the removal of the OBut protecting group and coupling of D-Adf-Pip using TOTU as well. The PEG-bound thrombin inhibitors showed inhibition constants vs. thrombin in the subnanomolar range, i.e., they were more active than the parent molecule 10. Moreover, the pegylated inhibitors exhibited a longer lasting effect in vivo. In rats the half-life of Mtr-Asn (PEG10000-OMe)-D-Adf-Pip 14 was determined to be 63 min. Mtr-Asn(PEG10000-OMe)-D-Adf-Pip 14 showed a half-life of 120 min in pigs. It could be concluded that these PEG-bound thrombin inhibitors may be employed as versatile drugs for parenteral administration in treating thrombotic disorders.

Animals↗
PubMedDate not supplied

Uptake and metabolism of carcinogenic levels of tobacco-specific nitrosamines by Sudanese snuff dippers.

It was recently reported that toombak, a type of snuff used in the Sudan, contained unusually high levels of tobacco-specific nitrosamines. To estimate the internal dose of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) received by individuals who use this type of tobacco, urine from a group of users was analyzed for 2 metabolites of NNK, 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) and its O-glucuronide, NNAL-Gluc. NNK is a strong lung carcinogen believed to contribute to human lung cancer. NNAL is also a lung carcinogen. NNAL and NNAL-Gluc were analyzed by gas chromatography with a nitrosamine selective detector. The average levels detected were 0.39 +/- 0.14 (SD) nmol/ml urine (n = 7) and 0.88 +/- 0.50 nmol/ml urine (n = 7), respectively. In a 24-h period, these individuals would excrete from 0.12 to 0.44 mg of these two metabolites (expressed per mg NNAL). Therefore, assuming chronic toombak use, the minimum daily dose of NNK to which these users were exposed was 0.12-0.44 mg. This is the highest documented uptake of a nonoccupational carcinogen. Two diastereomers of NNAL-Gluc were present in all urine samples analyzed. Previously, these two diastereomers were identified in the urine of an NNK-treated patas monkey but only one was detected in the urine of NNK-treated rats. The level of the 4-hydroxy-1-(3-pyridyl)-1-butanone releasing hemoglobin adduct was also quantified in these individuals. This adduct is believed to be a measure of NNK activation. The levels ranged from 68 to 323 fmol/g hemoglobin [mean, 148 +/- 104 (SD)]. The wide range of adduct levels which were observed suggests that despite similar levels of NNK exposure, there are significant differences in the ability of individuals in this population to activate NNK, as well as potential differences in their cancer risk.

2-Hydroxypropyl-beta-cyclodextrin↗
PubMedDate not supplied

Urinary trans,trans-muconic acid as an indicator of exposure to benzene in cigarette smokers.

Epidemiological studies have shown an association between cigarette smoking and increased risk of myeloid leukemia in smokers. In evaluating this link it is important to note that cigarette smoke contains benzene, among other carcinogens. Since chronic benzene, among other carcinogens. Since chronic benzene exposure causes acute myeloid leukemia in humans, we aimed to determine the uptake and metabolic activation of benzene from cigarette smoke in smokers by measuring the levels of the urinary benzene metabolite, trans,trans-muconic acid (t,t-MA). The method used involved a clean-up procedure, followed by high-performance liquid chromatography with UV detection. The levels of urinary t,t-MA ranged from 0.02 to 1.3 mg/g creatinine, resulting in a mean of 0.29 +/- 0.04 mg/g creatinine in 42 male smokers, and corresponding values in nonsmokers ranged from "nondetectable" to 0.52 mg/g creatinine with an average of 0.09 +/- 0.02 mg/g creatinine. In the current study, the levels of t,t-MA in smokers were about 3 times higher than those in nonsmokers (P = 0.0001), and a significant correlation between concentration of t,t-MA and levels of cotinine in smokers was observed (r = 0.55; P = 0.0001; 95% confidence interval, 0.30-0.93), suggesting that urinary t,t-MA can be utilized as a biochemical marker to quantitate benzene exposure due to cigarette smoking.

Animals↗
PubMedDate not supplied

Effects of 1,4-phenylenebis(methylene)selenocyanate, phenethyl isothiocyanate, indole-3-carbinol, and d-limonene individually and in combination on the tumorigenicity of the tobacco-specific nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone in A/J mouse lung.

In this study we examined whether chemopreventive agents that had each been shown to be effective against lung tuorigenesis induced in A/J mice by 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were more effective when applied together as a "cocktail" than as individual compounds. Groups of A/J mice were fed a diet containing 1,4-phenylenebis(methylene)selenocyanate (p-XSC; 5 ppm as selenium, 0.0005%), phenethyl isothiocyanate (PEITC; 0.008%), indole-3-carbinol (I3C; 0.18%), d-limonene (d-L, 0.63%), or a mixture of all four at the above levels. Mice were fed experimental diets (AIN-76A plus a chemopreventive agent, or a mixture of the four chemopreventive agents) for 17 weeks. One week after beginning the experimental diets, the animals received a single i.p. injection of 10 mumol NNK (2.07 mg) in 0.1 ml saline. Sixteen weeks after the NNK application the bioassay was terminated. Dietary p-XSC, PEITC, I3C, d-L, and their admixture reduced significantly the number of lung tumors per mouse from 8.1 in the positive control to 3.2, 3.7, 4.9, 2.4, and 2.5, respectively (p < 0.05). The inhibition of lung tumor multiplicity in mice fed either the mixture or d-L alone was also significantly stronger than in those fed the diet containing only I3C. However, neither individual agents nor their mixture had a measurable effect on lung tumor incidence. Although the effect of the mixture on lung tumor incidence in this assay remained imperfect, this preliminary investigation provides some basis for the future design of chemoprevention studies.

Animals↗