Showing papers by "Eli Lilly and Company published in 1985"

Hybrid antibodies can target sites for attack by T cells

Abstract: It would be advantageous in the case of certain diseases to be able to focus a strong T-cell response at a chosen target, for example, in treating cancer or infections that have escaped the normal host response. At present, it seems inconceivable that we could use antigen-specific lines or clones of effector T cells for this purpose because of complications due to the major histocompatibility restriction of T-cell specificity and the problem of rejection of transplanted effector cells. Here we describe a novel technology which combines the power of T lymphocytes in eliminating unwanted cells and causing beneficial inflammatory reactions with the great advantages of monoclonal antibodies (their specificity and availability). We show that heteroconjugates of monoclonal antibodies (referred to hereafter as hybrid antibodies), in which one of the component binding sites is anti-T-cell receptor and the other component binding site is directed against any chosen target antigen, can focus T cells to act at the targeted site. Monoclonal antibodies directed against the T-cell receptor, such as the anti-allotype used here, are mitogenic for resting T cells and can be used to induce effector T cells carrying the T-cell receptor determinant which can then be directed against the target by a hybrid antibody.

Isolation, sequence determination and expression in Escherichia coli of the isopenicillin N synthetase gene from Cephalosporium acremonium

Abstract: The enzyme isopenicillin N synthetase (IPS) catalyses the oxidative condensation of δ-(L-α-aminoadipyl)-L-cysteinyl-D-valine (LLD-ACV) to isopenicillin N, which is a central reaction in the pathway to clinically important penicillins and cephalosporins. Here we report the cloning, characterization and expression in Escherichia coli of the gene encoding the IPS protein in Cephalosporium acremonium. The IPS gene was identified by purifying IPS protein, determining the first 23 amino-terminal amino acids, preparing a set of synthetic oligonucleotides encoding a portion of the determined amino-acid sequence, and probing a cosmid genome library with the mixed oligonucleotides. A cosmid hybridizing with the probe was isolated and the IPS gene was localized and sequenced. The IPS gene encodes a polypeptide of relative molecular mass (Mr) 38,416. When this open reading frame was cloned into an E. coli expression vector and inserted into E. coli, the recombinant E. coli produced a new protein co-migrating with authentic IPS as the major protein of the cell (∼20% of cell protein). Crude cell extracts condensed LLD-ACV to a penicillinase-sensitive molecule whose antibacterial activity indicated that it was isopenicillin N.

Chronic effects of fluoxetine, a selective inhibitor of serotonin uptake, on neurotransmitter receptors

Abstract: Fluoxetine administration to rats at a dose of 10 mg/kg i.p. daily up to 12 or 24 days failed to change the concentration-dependent binding of [3H]WB4101, [3H]clonidine and [3H]dihydroalprenolol to alpha 1-, alpha 2- and beta-adrenergic receptors, respectively; [3H]quinuclidinyl benzilate to muscarinic receptors; [3H]pyrilamine to histamine H1 receptors and [3H]naloxone to opiate receptors. Persistent and significant decreases in receptor number (Bmax value) without changes in the dissociation constant (KD value) of [3H]5-HT binding in cortical membranes were observed upon chronic treatment with fluoxetine administered either by intraperitoneal injection or incorporation in the diet. A detectable reduction of 5-HT1 receptor number occurred after once-daily injections of fluoxetine at 10 mg/kg i.p. within 49 hours. After pretreatment for 3 days with p-chlorophenylalanine, an inhibitor of 5-HT synthesis, followed by repeated administration of fluoxetine, 5-HT1 receptor numbers were higher than those of normal rats, suggesting a dependence on synaptic concentration of 5-HT for fluoxetine to affect a receptor down-regulation. These studies provide further evidence for the selectivity of fluoxetine as an inhibitor of 5-HT reuptake, resulting in a selective down-regulation of 5-HT1 receptors in the cerebral cortex of rat brain.

Antitumor properties of vindesine-monoclonal antibody conjugates.

Abstract: The anticancer alkaloid vindesine (VDS) was conjugated to four mouse monoclonal antibodies recognizing human tumor-associated antigens. The antibodies were 96.5 (antimelanoma, IgG2a); 791T/36 (antiosteogenic sarcoma, IgG2b); 11.285.14, and 14.95.55 (anticarcinoembryonic antigen, IgG1 and IgG2a respectively). Conjugates VDS-96.5 and VDS-791T/36 were tested in vitro and shown to be specifically cytotoxic for target cells expressing the appropriate antigen. The in vivo effects of the antibodies and conjugates were tested against human tumor xenografts in athymic or immunodeprived mice using multiple treatments. Conjugate VDS-96.5 retarded the initial growth of a melanoma xenograft, whereas free antibody was without effect. Similarly, VDS-791T/36 but not free antibody retarded the growth of osteogenic sarcoma 791T. The most marked antitumor effects observed were those obtained with VDS conjugates of the anti-CEA antibodies against a colorectal tumor xenograft. Antibody 14.95.55 suppressed tumor growth both alone and as a VDS conjugate, whereas 11.285.14 produced only a slight effect alone but an almost complete and lasting suppression of tumor growth as a VDS conjugate. Free VDS had little effect at nontoxic levels. Acute studies showed that VDS-11.285.14 conjugate was considerably less toxic than free VDS in Balb/c mice.

Stabilized insulin formulation

Abstract: This invention provides an insulin formulation stabilized against aggregation containing a hydroxybenzene and a polyethylene glycol-polypropylene glycol polymer of the formula ##STR1## having an average molecular weight of about 8350 and in which the average number of ethyleneoxy units per molecule, designated by the sum of a and c, is about 150, and the average number of propyleneoxy units per molecule, designated as b, is about 30.

Inhibition of serotonin uptake by optical isomers of fluoxetine

Abstract: The optical isomers of fluoxetine, a selective inhibitor of serotonin (5-hydroxytryptamine, 5HT) uptake, have been compared pharmacologically. As inhibitors of 3H-5HT uptake in cortical synaptosomes and of 3H-fluoxetine binding in cortical membranes, the (+)-isomer of fluoxetine was slightly more potent than the (−)-isomer. The potencies to inhibit 3H-5HT uptake ex vivo in synaptosomal preparations of cerebral cortex and brain stem were about the same, except that the (−)-isomer had a much shorter duration of action than the (+)-isomer in rats.

Selectable marker for development of vectors and transformation systems in plants

Abstract: The present invention discloses functional and selectable micro-Ti plasmids. The hygromycin phosphotransferase (aphIV) gene from Escherichia coli was inserted between the 5' promoter and associated amino terminal region-encoding sequence of an octopine synthetase gene and the 3' terminator signal sequence of a nopaline synthetase gene. These constructs were assembled between T-DNA border fragments in a broad-host-range vector and used to create antibiotic-resistant plant cells.

Single-dose and steady-state pharmacokinetics of tomoxetine in normal subjects.

Abstract: A pharmacokinetic profile of tomoxetine, a selective norepinephrine uptake inhibitor, was developed in human volunteers following single and multiple oral administrations. Following the administration of a single 90-mg oral dose of tomoxetine to four normal volunteers, the plasma half-life was 4.3 +/- 0.5 hours. Mean plasma clearance was 0.60 +/- 0.14 L/Kg/hr, and the mean volume of distribution was 3.7 +/- 0.9 L/kg. Multiple doses of tomoxetine (20 mg bid and 40 mg bid) for seven days were administered to an additional seven subjects. The data appeared to have a bimodal distribution. The mean plasma half-life determined following the last dose was 4.6 +/- 0.5 hours in five subjects. The other two subjects, one at each dose level, demonstrated accumulation of tomoxetine occurring from the first to last dose where tomoxetine disappeared from plasma with a mean half-life of 19 hours.

Sustained release tablets

Abstract: The present invention provides sustained release tablets in unit dosage form comprising an active agent which has low aqueous solubility.

Computed Tomography in the Prediction of Outcome in Head Injury

Abstract: To determine the prognostic significance of computed tomographic (CT) findings in head injury, retrospective analysis was performed in 128 randomly selected severe head-injury patients managed with a standardized protocol. The minimal criterion for entry into this study was that the patients were unable to obey simple commands or utter formed words. Serial CT was performed on admission and 3-5 days, 2 weeks, 3 months, and 1 year after injury. A scale of severity of abnormalities was devised taking into account the size of the traumatic lesions on CT. The CT findings using the proposed scale were correlated with the clinical outcome and analyzed using linear logistic regression. Other characteristics such as midline shift, multiplicity, and corpus callosum and brainstem lesions were not included in the analysis either because they did not affect the prognosis or because too few of these lesions were present for statistical analysis. The correct prediction rate of outcome using the proposed scale for CT findings alone was found to be 69.7%. When CT findings were combined with the Glasgow Coma Scale score this rate was increased to 75.8%.

Isolation of a58365a and a58365b, angiotensin converting enzyme inhibitors produced by streptomyces chromofuscus

Abstract: A58365A and A58365B, angiotensin converting enzyme inhibitors, were isolated from the culture filtrate of Streptomyces chromofuscus NRRL 15098. A58365A and A58365B are homologous nitrogen-containing bicyclic structures of molecular formulae C12H13NO6 and C13H15NO6.

6-substituted-4-dialkylaminotetrahydrobenz(c,d)indoles

Abstract: (±)-4-substitutedamino-6-alkoxy, benzyloxy, acyloxy, or hydroxy-1,3,4,5-tetrahydrobenz[c,d]indoles, pharmaceutically acceptable salts thereof are, anti-depressants.

Water-in-oil microemulsions for cosmetic uses

Abstract: The present invention provides water-in-oil microemulsions suitable for cosmetic uses containing moisturizing agents or sunscreens.

The structure of A201A, a novel nucleoside antibiotic.

Abstract: The structure of the novel nucleoside antibiotic A201A has been determined by a combination of chemical and spectroscopic methods. It is composed of 6-dimethylaminopurine, 3-amino-3-deoxyribose, p-hydroxy-α-methylcinnamic acid, a novel unsaturated hexofuranose and 3, 4-di-O-methylrhamnose. Structures have also been assigned to several related minor factors simultaneously isolated from the fermentation broth. These unique nucleosides have very interesting similarities and differences in structure with the known antibiotics puromycin and hygromycin A.

Anti-diabetic compounds

Abstract: A class of compounds having insulin-like activity is described. These compounds have the formula ##STR1## in which X is --OH, -Arg-Arg-OH, or -Arg-Arg-Glu-Ala-Glu-Asp-Leu-Gln-Val-Gly-Gln-Val-Glu-Leu-Gly-Gly-Gly-Pro-Gly-Ala-Gly-Ser-Leu-Gln-Pro-Leu-OH.

Intravenous nizatidine kinetics and acid suppression.

Abstract: The effectiveness of intravenous nizatidine in suppressing gastric acid secretion was evaluated by different methods of inducing secretion in two single-blind studies. In study 1, seven subjects were given single, 20-min intravenous infusions of nizatidine (6.25, 25, 75, 150, or 250 mg) before modified sham feeding (MSF). Gastric acid suppression after nizatidine was contrasted with that after placebo and MSF. All doses of nizatidine reduced secretion; the 150- and 250-mg doses of nizatidine suppressed secretion for at least 2.5 hr. In study 2, eight subjects received one 5-min intravenous infusion of placebo, cimetidine (300 mg), or nizatidine (25, 50, 100, or 250 mg) weekly for 6 wk. Secretion was induced by infusing pentagastrin (2 µg/kg/hr) 45 min before the study drug was dosed and for 3.5 hr thereafter. Again, all doses of nizatidine reduced gastric acid, chiefly by decreasing volume. Nizatidine induced a clear dose-response effect; nizatidine (100 mg) and cimetidine (300 mg) had approximately equal suppressive effects. Nizatidine (100 mg) and cimetidine (300 mg) reduced acid output by 62% and 63% and reduced volume of secretion by 48% and 51% over the 3.5-hr period. Gastric acid suppression and plasma nizatidine concentrations were directly related. Nizatidine kinetics were linear and proportional to dose. The t½ was 1.3 hr (range 0.7 to 2.1 hr), the volume of distribution was 1.2 ± 0.5 l/kg, and clearance was 0.6 ± 0.2 l/kg/hr. Laboratory abnormalities and side effects were minor in both studies. Clinical Pharmacology and Therapeutics (1985) 37, 162–165; doi:10.1038/clpt.1985.29

Long-Term Permeation Kinetics of Estradiol: (III) Kinetic Analyses of the Simultaneous Skin Permeation and Bioconversion of Estradiol Esters

Abstract: The skin permeation system developed earlier in this laboratory was utilized to study the kinetics of the simultaneous skin permeation and bioconversion of 5 estradiol esters. The equilibrium solubility of estradiol esters in the lipophilic silicone fluid and in hydrophilic PEG 400/saline solution was found to be dependent upon the alkyl chain length of the esters. Estradiol-3,17-diacetate had a greater solubility in silicone fluid and a lower solubility in PEG 400/saline solution than estradiol-17-acetate. The (skin/silicone fluid) partition coefficients were observed to decrease as the alkyl chain increased in length. During the course of skin permeation, the estradiol esters were metabolized by esterase to regenerate estradiol. The rate of appearance of estradiol from the estradiol esters was observed to be dependent upon the ester concentration on stratum corneum surface and to follow the order of: diacetate > valerate > heptanoate > cypionate > acetate. From the dermal uptake and metabolism s...