Resistance-modifying agents. 8. Inhibition of O(6)-alkylguanine-DNA alkyltransferase by O(6)-alkenyl-, O(6)-cycloalkenyl-, and O(6)-(2-oxoalkyl)guanines and potentiation of temozolomide cytotoxicity in vitro by O(6)-(1-cyclopentenylmethyl)guanine.

A series of O(6)-allyl- and O(6)-(2-oxoalkyl)guanines were synthesized and evaluated, in comparison with the corresponding O(6)-alkylguanines, as potential inhibitors of the DNA-repair protein O(6)-alkylguanine-DNA alkyltransferase (AGT). Simple O(6)-alkyl- and O(6)-cycloalkylguanines were weak AGT inactivators compared with O(6)-allylguanine (IC(50) = 8.5 +/- 0.6 microM) with IC(50) values ranging from 100 to 1000 microM. The introduction of substituents at C-2 of the allyl group of O(6)-allylguanine reduced activity compared with the parent compound, while analogous compounds in the O(6)-(2-oxoalkyl)guanine series exhibited very poor activity (150-1000 microM). O(6)-Cycloalkenylguanines proved to be excellent AGT inactivators, with 1-cyclobutenylmethylguanine (IC(50) = 0.55 +/- 0.02 microM) and 1-cyclopentenylmethylguanine (IC(50) = 0.39 +/- 0.04 microM) exhibiting potency approaching that of the benchmark AGT inhibitor O(6)-benzylguanine (IC(50) = 0.18 +/- 0.02 microM). 1-Cyclopentenylmethylguanine also inactivated AGT in intact HT29 human colorectal carcinoma cells (IC(50) = 0.20 +/- 0.07 microM) and potentiated the cytotoxicity of the monomethylating antitumor agent Temozolomide by approximately 3- and 10-fold, respectively, in the HT29 and Colo205 tumor cell lines. The observation that four mutant AGT enzymes resistant to O(6)-benzylguanine also proved strongly cross-resistant to 1-cyclopentenylmethylguanine indicates that the O(6)-substituent of each compound makes similar binding interactions within the active site of AGT.

Dimethyldioxirane converts benzene oxide/oxepin into (Z,Z)-muconaldehyde and sym-oxepin oxide: modeling the metabolism of benzene and its photooxidative degradation.

Oxidation of 7-oxabicyclo[4.1.0]hepta-2,4-diene (benzene oxide)/oxepin with dimethyldioxirane (DMDO) gave mainly (Z,Z)-muconaldehyde, with complete diastereoselectivity. Similarly, 2-methyl-7-oxabicyclo[4.1.0]hepta-2,4-diene (toluene 1,2-epoxide)/2-methyloxepin gave (Z,Z)-1,6-dioxohepta-2,4-diene, while 2,6-dimethyl-7-oxabicyclo[4.1.0]hepta-2,4-diene (1,2-dimethylbenzene 1,2-epoxide)/2,7-dimethyloxepin gave (Z,Z)-2,7-dioxo-3,5-octadiene. By monitoring the DMDO oxidation of benzene oxide/oxepin by 1H NMR spectroscopy, a significant byproduct was identified as 4,8-dioxabicyclo[5.1.0]octa-2,5-diene (sym-oxepin oxide). This observation supports the hypothesis that the route to (Z,Z)-muconaldehyde proceeds from oxepin via 6,8-dioxabicyclo[5.1.0]octa-2,4-diene (oxepin 2,3-oxide), with a minor pathway leading to sym-oxepin oxide. The DMDO oxidations described provide model systems for the cytochrome P450-dependent metabolism of benzene and for the atmospheric photooxidation of benzenoid hydrocarbons.

Chemistry of muconaldehydes of possible relevance to the toxicology of benzene.

(Z,Z)-Muconaldehyde reacted with primary amines to give N-substituted-2(2'-oxoethyl)-pyrroles, which were reduced to N-substituted-2-(2'-hydroxyethyl)-pyrroles by sodium borohydride. The pyrrole-forming reaction is exhibited by valine and its methyl ester, and is being developed with terminal valine in hemoglobin as a means of dose monitoring (Z,Z)-muconaldehyde, a putative metabolite of benzene. Reactions in aqueous solution between (Z,Z)-muconaldehyde and adenosine, deoxyadenosine, guanosine, or deoxyguanosine leading to pyrrole-containing adducts are described. The elucidation of the structures of the adducts was assisted by the study of reactions between (Z,Z)-muconaldehyde and both nucleoside derivatives and a model compound for guanosine. Reactions of (Z,Z)-muconaldehyde are complicated by its isomerization to (E,Z)- and (E-E)-muconaldehyde. The kinetics of this process have been studied in benzene, acetonitrile, and dimethylsulfoxide.

Studies on the molecular toxicology of buta-1,3-diene and isoprene epoxides.

Reactions of ethenyloxirane with amino (RNH2) and thiol (R'SH) nucleophiles occur by an SN2 mechanism involving competing ring cleavage at C-2 and C-3. In contrast, 2-ethenyl-2-methyloxirane reacts with amino (RNH2) and thiolate (R'S-) nucleophiles in methanol by regioselective SN2 attack at C-3 ("neo-pentyl" position). However, in pure water or methanol SN1 reaction occurs mainly at C-2.

Mechanisms of formation of adducts from reactions of glycidaldehyde with 2'-deoxyguanosine and/or guanosine.

Convenient synthesis of rac-glycidaldehyde from rac-but-3-ene-1,2-diol and (R)-glycidaldehyde from D-mannitol are described. (R)-Glycidaldehyde (1) reacts with guanosine in water (pH 4-11, faster reaction at higher pH) to give initially 6(S)-hydroxy-7(S)-(hydroxymethyl)-3-(beta-D-ribofuranosyl)-5,6,7- trihydroimidazo[1,2-alpha]purin-9(3H)-one (7a) and 6(S),7(R)-dihydroxy-3-(beta-D-ribofuranosyl)-5,6,7,8- tetrahydropyrimido[1,2- alpha]purin-10(3H)-one (8a). The former decomposes to 7-(hydroxymethyl)-5,9-dihydro-9-oxo-3-(beta-D-ribofuranosyl)imidazo[1,2- alpha]purine (3a), 5,9-dihydro-9-oxo-3-(beta-D-ribofuranosyl)imidazo[1,2-alpha]purine (5a, 1,N2-ethenoguanosine), and formaldehyde, while the latter adduct is relatively stable. The position of the hydroxymethyl group on the imidazo ring of 7-(hydroxymethyl)-5,9-dihydro-9-oxo-3-(beta-D-ribofuranosyl)imidazo-[1,2 - alpha]purine was proved by 13C NMR analysis of adducts derived from [1-15N]guanosine and [amino-15N]guanosine. At longer reaction times, the adduct 7,7'-methylenebis[5,9-dihydro-9-oxo-3-(beta-D-ribofuranosyl)imidazo[1,2- alpha]purine (4a) is formed from guanosine and glycidaldehyde. The structure analysis of this adduct was also aided by 13C NMR analysis of the 15N-labeled adduct derived from [1-15N]guanosine. Analogous adducts were obtained from the reaction between glycidaldehyde and deoxyguanosine. Mechanisms of formation of the adducts from glycidaldehyde and guanosine/deoxyguanosine are proposed and supported by model studies with simple amines. The formaldehyde produced in the reactions described reacts with guanosine to give the known adduct N2-(hydroxymethyl)guanosine (9).

The achievement of isoeffective bronchial mucosal dose during endobronchial brachytherapy.

PURPOSE: The use of endobronchial brachytherapy in the treatment of lung cancer is increasing due to the more widespread availability of high dose rate afterloading equipment. The complications include small airway (segmental and small lobar bronchi) fibrosis, stenosis, and obstructive complications in addition to hemorrhage. A progressive reduction in the diameter of the bronchial lumen occurs at each division of the bronchial tree. If uniform dwell times along a bronchial catheter treatment length are used, this will result in higher doses being given to the bronchial mucosa in the distal part of the treatment volume where the brachytherapy source mucosa distances are smaller, and underdosage proximally, where the source mucosa distances are larger. METHODS AND MATERIALS: The known mathematical relationships of the sequential reductions in the diameter of the bronchial lumen have been incorporated into two methods of optimization, which have been compared to uniform dwell times along a treatment length from trachea to segmental bronchus. RESULTS: The resulting isodose plots are presented, and demonstrate the extent of the overdosage distally, and the underdosage proximally when using uniform dwell times, and the achievement of isoeffective mucosal doses when using differential dwell times. CONCLUSION: This refinement in brachytherapy technique offers the potential for reduced normal tissue complications and possibly improved tumor control by reducing overdosage and underdosage, respectively.

Radiotherapy and chemotherapy for inoperable non-small cell lung cancer.

Non-small cell lung cancer is a major cause of mortality and significant morbidity in the UK. The majority of patients are inoperable and the optimum management of these patients requires a multidisciplinary approach involving the cooperation of respiratory physicians, thoracic surgeons and clinical oncologists (radiotherapists). Treatment techniques are constantly being refined and new approaches developed.

Non-uniform dwell times in line source high dose rate brachytherapy: physical and radiobiological considerations.

The ability to vary source dwell times in high dose rate (HDR) brachytherapy allows for the use of non-uniform dwell times along a line source. This may have advantages in the radical treatment of tumours depending on individual tumour geometry. This study investigates the potential improvements in local tumour control relative to adjacent normal tissue isoeffects when intratumour source dwell times are increased along the central portion of a line source (technique A) in radiotherapy schedules which include a relatively small component of HDR brachytherapy. Such a technique is predicted to increase the local control for tumours of diameters ranging between 2 cm and 4 cm by up to 11% compared with a technique in which there are uniform dwell times along the line source (technique B). There is no difference in the local control rates for the two techniques when used to treat smaller tumours. Normal tissue doses are also modified by the technique used. Technique A produces higher normal tissue doses at points perpendicular to the centre of the line source and lower doses at points nearer the ends of the line source if the prescription point is not in the central plane of the line source. Alternatively, if the dose is prescribed at a point in the central plane of the line source, the dose at all the normal tissue points are lower when technique A is used.

A mathematical model of intraluminal and intracavitary brachytherapy.

The adaptation of the linear-quadratic model to allow for the effect of tumour regression and clonogen repopulation between initial teletherapy and subsequent brachytherapy has been extended to include the geometrical conditions encountered in intraluminal and intracavitary brachytherapy. For a radiation line source placed at the centre of a lumen or cavity, regression of any endoluminal tumour towards its mural origin will not result in any change in the minimum brachytherapy-tumour dose with time. In contrast, regression of transmural tumour will cause a potentially advantageous increase in the minimum brachytherapy-tumour dose with time. The latter effect will be opposed by tumour clonogen repopulation. The log(e) cell kill due to brachytherapy has been calculated for tumours of diameters 2, 4 and 6 cm at completion of teletherapy. The centres of the tumours were assumed to be at distances of 0, 1 and 2 cm from the radiation source. Tumour linear regression rates (lambda) ranging from 0.025 to 0.25 per week and tumour clonogen doubling times (Tp) of 2.5, 5 and 15 days were used in the calculations. The results demonstrate the critical importance of the distance of the tumour centre from the line source as well as the influence of tumour diameter, lambda and Tp. In some instances, both maximum and minimum values of log(e) cell kill occur. Calculations of tumour cure probabilities reveal that these variations in log(e) cell kill predicted by the model can produce highly significant differences in tumour control rates. Where the relevant parameters can be assessed directly or estimated from previous experience, the model provides a basis for the design of future intraluminal or intracavitary brachytherapy protocols.

Effect of overall time when radiotherapy includes teletherapy and brachytherapy: a mathematical model.

By expanding the linear quadratic equations to allow for the interval of time between teletherapy and brachytherapy in a model which assumes continual exponential tumour regression following teletherapy, the loge cell kill resulting from brachytherapy is found to depend on radiosensitivity, tumour regression rate (lambda) and tumour clonogen doubling time (Tp). When lambda is relatively small the precise timing of brachytherapy following teletherapy is shown to influence the tumour cure probabilities. The model predicts a reduction in tumour control if brachytherapy is delayed when lambda values are small and Tp values are short, but when Tp values are long, deferral of brachytherapy may be advantageous. By differential calculus the occurrence of a minimum value of log cell kill can be found. Application of these concepts may improve the results of clinical radiotherapy.

Differential free-radical activity after successful and unsuccessful thrombolytic reperfusion in acute myocardial infarction.

BACKGROUND: Free-radical generation after successful thrombolysis in acute myocardial infarction may jeopardize ischaemic but viable myocardium, thus limiting the optimal benefits of reperfusion. METHODS: Circulating free-radical activity was assessed in 25 consecutive patients with acute myocardial infarction. Those who successfully reperfused (Group A) were compared with those who did not (Group B). We also compared patients who had or had not developed Q waves and patients with and without previous angina or myocardial infarction. All patients presented within 6 h of the onset of chest pain and received standard intravenous streptokinase therapy. Free-radical activity in serial serum samples collected over 72 h was measured using the percentage molar ratio (PMR) of the concentrations of 9,11-linoleic acid to 9,12-linoleic acid, and malonaldehyde concentration. RESULTS: Throughout the study period Group A (n = 11) showed significantly greater change in serum PMR and malonaldehyde levels compared with Group B (n = 14) (P < 0.01). PMR differences between the two groups were most pronounced at 3 and 12 h (P < 0.001). Patients with non-Q-wave myocardial infarction (n = 5) showed significantly greater changes in serum PMR and malonaldehyde levels (P < 0.01) compared with those with Q-wave infarction (n = 20). A history of previous infarction or angina had no apparent effects on the changes in serum free-radical activity. CONCLUSIONS: Successful early reperfusion and non-Q-wave myocardial infarction are both associated with a significantly greater increase in the levels of markers of serum free-radical activity immediately after infarction. The results support present concepts of free-radical-mediated reperfusion injury. Use of these assays may identify those patients who may be at risk from free-radical-mediated reperfusion injury.

Reactions of muconaldehyde isomers with nucleophiles including tri-O-acetylguanosine: formation of 1,2-disubstituted pyrroles from reactions of the (Z,Z)-isomer with primary amines.

(Z,Z)-Muconaldehyde reacts with primary amines, including valine and lysine (epsilon-amino), to afford N-substituted-2-(oxoethyl)pyrroles, which were reduced with sodium borohydride to the more stable N-substituted-2-(hydroxyethyl)pyrroles. The formation of the pyrrole aldehydes was performed in a variety of solvents including aqueous methanol. With tri-O-acetylguanosine, the putative pyrrole aldehyde derived from reaction at NH2 condenses with N-1 (guanine component) to afford a bicyclic adduct: 4,5-dihydro-5-hydroxy-10-beta-D-tri-O- acetylribosylpyrrolo[1',2':3,4]pyrimido[1,2-a]purin-7(10H)-one (5a). This was hydrolyzed to 4,5-dihydro-5-hydroxy-10-beta-D- ribosylpyrrolo[1',2':3,4]pyrimido[1,2-a]purin-7(10H)-one (5b). The structures of 5a and 5b were primarily based on NMR evidence and comparison with 4,5-dihydro-5-hydroxy-9-methylpyrrolo[1',2':3,4]pyrimido[2,1-b] pyrimidin-7-one (7), obtained from reacting (Z,Z)-muconaldehyde with 2-amino-4-hydroxy-6-methylpyrimidine. (E,Z)-Muconaldehyde also reacted with primary amines to give N-substituted-2-(oxoethyl)pyrroles, whereas (E,E)-muconaldehyde gave bis-imines. The results described are discussed in the context of the carcinogenicity of benzene.

Accuracy of diagnostic coding of hospital admissions for cryptogenic fibrosing alveolitis.

To determine the accuracy of diagnostic coding of cryptogenic fibrosing alveolitis, the case notes of 166 admissions to four hospitals were reviewed. These consisted of all admissions that had been coded as "idiopathic fibrosing alveolitis" (ICD code 516.3: 97 admissions) or as "postinflammatory pulmonary fibrosis" (ICD code 515.9: 69 admissions). Of 88 available records of admissions coded as idiopathic fibrosing alveolitis, 70 (80%) patients had definite cryptogenic fibrosing alveolitis, and six (7%) possible cryptogenic fibrosing alveolitis according to predetermined conventional clinical criteria. Only seven (8%) admissions were clearly coded wrongly. Sixty four records were available for patients coded as having postinflammatory pulmonary fibrosis; 16 (25%) of these patients had definite cryptogenic fibrosing alveolitis, a further 12 (19%) had possible cryptogenic fibrosing alveolitis or fibrosing alveolitis with a connective tissue disorder, and the remainder had a very wide range of diagnoses. In this study the idiopathic fibrosing alveolitis (ICD 516.3) code was relatively reliable, but a substantial proportion of admissions coded under postinflammatory pulmonary fibrosis (ICD 515.9) also had cryptogenic fibrosing alveolitis and code 515.9 was of little diagnostic value. The data are inadequate for case finding, though in respect of cryptogenic fibrosing alveolitis may be adequate for planning purposes. There continues to be a need for more medical input into the process of diagnostic coding.