Showing papers by "Eli Lilly and Company published in 1984"

Derivative of the tryptophan operon for expression of fused gene products

Abstract: The present invention comprises improved derivatives of the tryptophan operon useful for expressing fused gene products in E. coli and relate organism. Two of the improved derivatives disclosed are encoded on 0.43 and 0.55 kb EcoRI restriction fragments from plasmids pCZ20 and pLEBGH2 (in strains NRRL B-15881 and NRRL B-15882), respectively. The modified derivatives have been placed on recombinant DNA cloning and expression vectors. A variety of expression vectors have been constructed that drive expression of fused gene products. Two novel gene sequences, encoding insulin-like growth factors I and II, have been expressed with the modified tryptophan operon system.

The effects of biosynthetic human proinsulin on carbohydrate metabolism.

Abstract: Large quantities of biosynthetic human proinsulin have recently become available through recombinant DNA technology. Since the in vivo effects of human proinsulin have not been studied in man, we compared the dose-response relationship for stimulation of glucose disposal and suppression of hepatic glucose output by proinsulin and insulin. Ten normal subjects were studied using the euglycemic glucose clamp technique. The human proinsulin and insulin infusion rates were chosen to achieve steady-state proinsulin levels 10-fold higher than insulin levels on a molar basis, based on previous observations that porcine proinsulin has approximately 10% the potency of insulin. Proinsulin infusion rates of 2.75, 7.5, 22.5, and 45 micrograms/m2/min were compared with insulin infusion rates of 0.63, 1.67, 5, and 10 micrograms/m2/min. Primed, continuous infusions of insulin yielded steady-state levels within 25 min, whereas proinsulin levels did not reach a steady state for 120-180 min. The metabolic clearance rate of insulin was 11-12 ml/kg/min at the lower infusion rates but fell to 8.4 ml/kg/min at the highest infusion rate. The metabolic clearance rate of proinsulin was 3.0-3.5 ml/kg/min at all infusion rates. Dose-response analysis demonstrated that proinsulin-mediated glucose disposal was approximately 8% that of insulin. In contrast, proinsulin-mediated suppression of hepatic glucose output was approximately 12% that seen with insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Association of parvoviruses with rheumatoid arthritis of humans

Abstract: A small virus resembling parvoviruses in its morphological and physicochemical properties was derived from synovial tissue of a patient with severe rheumatoid arthritis. This virus, designated RA-1, elicits a syndrome in neonatal mice that includes neurological disturbances, permanent crippling of limbs, dwarfism, alopecia, blepharitis, "masking," and a rigid curvature of the thoracic spine. Polyclonal antibodies against RA-1 display high virus neutralizing activity and in immunoassays detect reactive antigen in synovial cells from different rheumatoid arthritis patients but not persons with osteoarthritis. Putative parvoviruses isolated from several other rheumatoid arthritis patients are only weakly pathogenic for newborn mice but can generate RA-1 virus-specific antigens in tissues of these animals. It has not been established that RA-1 and existing parvoviruses of mammalian species are related.

In vitro and in vivo anti-Candida activity and toxicology of LY121019.

Abstract: LY121019 (N-p-octyloxybenzoylechinocandin B nucleus) is a semisynthetic antifungal antibiotic that possesses potent anti-Candida activity. The MIC50 and the MIC90 for both LY121019 and amphotericin B were 0.625 and 1.25μg/ml, respectively. Only an 8-fold increase in the MIC against C. albicans occurred during 34-day exposure to subinhibitory concentrations indicating that LY121019 has a low potential for causing resistance development. Scanning electron microscopic studies revealed that LY121019 caused severe damage to the C. albicans cell. The ED50's for LY121019 and amphotericin B administered parenterally to mice were 7.4 and 2.5mg/kg, respectively. Parenterally administered LY121019 at doses of 6.25mg/kg significantly reduced the recovery of C. albicans from infected mouse kidneys. Orally administered 50 and 100mg/kg doses of LY121019 were effective in eliminating C. albicans from the gastrointestinal tract of infected mice. Topical application of 5% LY121019 was as effective as 3% nystatin in the treatment of superficial C. albicans infections. Local administration of LY121019, nystatin, or miconazole was effective against rat vaginal candidiasis. LY121019 was administered intravenously to dogs at doses up to 100mg/kg/day, 5 days a week for 3 months; all dogs survived. Compound related effects included a histamine-like reaction, increased serum alkaline phosphatase and SGPT, fatty vacuolization of the liver, and some tissue damage at the injection site. The no effect dose in dog was 10mg/kg. LY121019 had no more than 1/20 the toxicity of amphotericin B in the dog.

Process for purifying proteins and compounds useful in such process

Abstract: This invention describes a process for separating a biologically active polypeptide or protein in the form of its precursor from a mixture containing said precursor and impurities, which comprises contacting said precursor with a resin containing immobilized metal ions, said precursor comprising the biologically active polypeptide or protein covalently linked directly or indirectly to an immobilized metal ion chelating peptide, binding said precursor to said resin, and selectively eluting said precursor from said resin. Such precursor compounds are also described.

Vacuum vial infusion system

Abstract: An apparatus for connecting a vial into the delivery tube of an intravenous administration set, the set including a container of fluid connected to one end of a delivery tube with the other end of the delivery tube adapted for insertion into the patient. The apparatus includes a penetrating spike penetrating an end of the vial which provides two pathways into the interior of the vial with two sections of the delivery tube connected to the two pathways of the penetrating spike. The apparatus also includes a frame for maintaining the two sections of the delivery tube adjacent to each other and clamp for releasably closing the adjacent sections of the delivery tube to temporarily prevent flow of fluid through the penetrating spike.

Disposition and metabolism of a new benzothiophene antiestrogen in rats, dogs and monkeys

Abstract: A new benzothiophene-derived antiestrogen (LY156758) when orally administered was well absorbed in rats and monkeys while approx. 20% was absorbed in dogs. In the rat the compound was subject to first-pass hepatic metabolism which led to low levels of parent drug in the systematic circulation together with a small amount as the glucuronide conjugate. In monkeys the compound occurred primarily as the glucuronide conjugate of parent drug with very little circulating free drug. The systemic bioavailability of free parent drug in plasma was 39% in rats, 17% in dogs and 5% in monkeys. Excretion of the drug in rats and dogs was primarily via the bile. Approx. 1% of the dose was excreted in the urine of rats and dogs after oral dosing. In rats, at least 50% of an oral dose was excreted in bile as the glucuronide conjugate of parent drug.

Growth hormone release factor analogs

Abstract: A class of compounds having growth hormone releasing activity is described. These compounds have the formula A compound having the formula H-Tyr-Ala-Asp-Ala-Ile-Phe-Thr-Asn-Ser-Tyr-Arg-Lys-Val-Leu-Gly-Gln-Leu-Ser-Ala-Arg-Lys-Leu-Leu-Gln-Asp-Ile-X-Ser-Arg-Gln-Gln-Gly-Glu-Ser-Asn-Gln-Glu-Arg-Gly-Ala-Arg-Ala-Arg-Leu-Y in which Y is OH or NH2 and X is Met(O), Met(S-Me), Hse, Gln, Asn, Ser, Thr, Leu, lie, Ala, Gly, Pro, Val, Phe, Trp, Tyr, Cys, Lys, Arg, His, Glu, or Asp.

Actaplanin, new glycopeptide antibiotics produced by actinoplanes missouriensis the isolation and preliminary chemical characterization of actaplanin

Abstract: Actaplanin (A4696), a new complex of broad spectrum Gram-positive antibiotics is produced by Actinoplanes missouriensis. High performance liquid chromatography was used to show that this complex is composed of several actaplanins. Hydrolytic experiments with acetaplanins A, B1, B2, B3, C1 and G showed that these actaplanins were composed of the same peptide core, an amino sugar and varying amounts of glucose, mannose and rhamnose. The neutral sugar content was determined for each actaplanin. A bioautographic study of aglycone formation during hydrolysis of the actaplanin complex showed that within a short time a simple mixture of two antimicrobially active hydrolysis products was obtained. These substances retained the antimicrobial spectrum and a high percentage of the antibiotic activity of the parent actaplanin complex. Methanolysis of the acetaplanin complex as well as the individual actaplanins resulted in the selective loss of the neutral sugar moieties and the isolation of actaplanin psi (pseudo)-aglycone--the core peptide which still retained an amino sugar group. The 1H NMR spectrum of this substance indicated a similarity to many features of ristocetin psi-aglycone. Hydrolytic studies showed that the amino sugar present in actaplanin was identical with L-ristosamine. It is concluded that the aglycone of actaplanin is a complex peptide composed of aromatic amino acids, and that the actaplanins each possess this aglycone and L-ristosamine but are differentiated by their neutral sugar composition.

Interactions of three inotropic agents, ASL-7022, dobutamine and dopamine, with α- and β-adrenoceptors in vitro

Abstract: Three inotropic agents, ASL-7022, dobutamine and dopamine, were evaluated for their α-and β-adrenoceptor mediated effects in vitro in a variety of isolated organs and in radioligand binding studies. All compounds were α1-adrenoreceptor agonists in rat and guinea pig aortae, but the rank orders of potency were exactly opposite in these two tissues. Only the rank potency order of dobutamine>ASL-7022>dopamine obtained in rat aorta was consistent with the results obtained in radioligand binding studies to α1-adrenoreceptors in rat cerebral cortex and to previous results obtained in vivo in the pithed rat. The results obtained in guinea pig aorta did not parallel the radioligand binding studies in rat brain or our previous results in pithed rat, and suggests that species differences exist between postsynaptic vascular α1-adrenoreceptors in rat and guinea pig aorta, consistent with previous conclusions. ASL-7022 was found to be a potent α2-adrenoreceptor agonist in field-stimulated guinea pig ileum, and was approximately 10-fold more potent than dobutamine in this respects, which was also confirmed by radioligand binding studies to α2-adrenoreceptors in rat cerebral cortex. The β1-adrenoreceptor mediated effects of these compounds were evaluated in guinea pig atria, where the rank order of potency was dobutamine>ASL-7022>dopamine. An identical rank order of affinity was established for these compounds by displacement of 3H-dihydroalprenolol from β1-adrenoreceptors in rat cerebral cortex. The β1-adrenoreceptor mediated effects of dobutamine and ASL-7022 in guinea pig atria were completely direct in nature and not secondary to the release of endogenous catecholamines. In contrast, a major component of the β1-adrenoreceptor mediated tachycardia produced by dopamine in guinea pig atria was indirect in nature as evidenced by the marked attenuation in potency that occurred following catecholamine depletion with reserpine. All three compounds elicited β2-adrenoreceptor mediated inhibition of tone in rat uterus, with the rank order of potency being ASL-7022>dobutamine>dopamine. Again, this rank order of β2-adrenoreceptor potency was also reflected in β2-adrenoreceptor affinity as assessed by displacement of 3H-dihydroalprenolol from β2-adrenoreceptors in rat cerebellum. Based on these results, it may be concluded that for α-adrenoceptors, dobutamine is a selective β2-adrenoreceptor agonist, ASL-7022 is a selective α2-adrenoreceptor agonist, and dopamine is a nonselective α-adrenoceptor agonist. For β-adrenoceptor mediated effect, ASL-7022 is a selective α2 agonist, while dobutamine and dopamine are nonselective β-adrenoceptor agonists. It is likely that the complex inotropic and hemodynamic activities of ASL-7022, dobutamine and dopamine result from the sum of their individual effects at the α-and β-adrenoceptor subtypes.

A-21978C cyclic peptides

Abstract: A-21978C cyclic peptides of the formula ##STR1## wherein R is selected from the group consisting of hydrogen, an amino-protecting group, 8-methyldecanoyl, 10-methylundecanoyl, 10-methyldodecanoyl, the specific C10 -alkanoyl group of A-21978C factor C0 and the specific C12 -alkanoyl groups of A-21978C factors C4 and C5 ; R1 and R2 are, independently, hydrogen or an amino-protecting group, and salts thereof, are prepared by enzymatic deacylation of an antibiotic selected from A-21978C complex, A-21978C factors C0, C1, C2, C3, C4 amd C5 and blocked A-21978C complex and factors C0, C1, C2, C3, C4 and C5, using an enzyme produced by the Actinoplanaceae, preferably by Actinoplanes utahensis. The A-21978C cyclic peptides and salts thereof are useful intermediates to prepare new semi-synthetic antibacterial agents.

Pharmacologic and toxic effects in animals of dextropropoxyphene and its major metabolite norpropoxyphene: a review.

Abstract: 1953, Pohland and Sullivan published on a series of newly synthesized drugs that had structural similarity to methadone (Pohland & Sullivan, 1953). One of these compounds (a-d,l-4-dimethylamino-1,2-diphenyl-3-methyl-2propionyloxybutane hydrochloride) was found to be a potent analgesic in animals (Robbins, 1955) and man (Gruber et al., 1955) and was later named propoxyphene. The dextrorotatory and laevorotatory stereoisomers of propoxyphene

Selective method for blocking 5ht2 receptors

Abstract: Method of blocking 5HT without effect on alpha receptors with 1-loweralkyl-6-straight chain alkyl-8β-hydroxycycloalkyloxycarbonylergolines.

The discovery, fermentation, isolation, and structure of antibiotic A33853 and its tetraacetyl derivative

Abstract: The structure of antibiotic A33853, isolated from the culture broth of Streptomyces sp., NRRL 12068, is reported. The structure was deduced from an X-ray crystallographic study of its tetraacetyl derivative. Tetraacetyl A33853 is unique because it contains an anhydride moiety, an unexpected product from the reaction of A33853 with acetic anhydride and pyridine.

Method of inhibiting aromatase

Abstract: This invention provides a method of inhibiting aromatase and treating or preventing estrogen-dependent diseases in mammals by administering certain azole derivatives.

N-demethylvancomycin, a novel antibiotic produced by a strain of Nocardia orientalis. Taxonomy and fermentation.

Abstract: A novel vancomycin analog, N-demethylvancomycin, is produced by a soil isolate collected in Yucatan, Mexico. Taxonomic studies indicated this microorganism, designated NRRL 15232, is a strain of Nocardia orientalis. Unlike some glycopeptide antibiotics, virtually none of the N-demethylvancomycin synthesized remained bound to the cells of the producing culture. Antibiotic production was markedly depressed by the addition of orthophosphate to the fermentation medium. Enrichment of the medium with tyrosine, p-hydroxyphenylglycine, p-hydroxyphenylglyoxylic acid, or leucine, all putative precursors of the aglycone, stimulated the biosynthesis of N-demethylvancomycin.

Incorporation of amino acid-derived carbon into tylactone by Streptomyces fradiae GS14.

Abstract: Washed cells from 72-h cultures of Streptomyces fradiae GS14 were used to examine the distribution of radiolabel from 14C-amino acids and related compounds into tylactone, CO2, and cells. Test compounds were categorized according to products of their oxidative degradation. Those compounds known to produce propionyl-coenzyme A by direct catabolic oxidation were designated as group I. Group II included those compounds oxidized to methylmalonyl-coenzyme A via succinyl-coenzyme A and the tricarboxylic acid cycle. Group III contained compounds known to be oxidized to acetoacetyl-coenzyme A. The total amount of label recovered after 60 min ranged from 3 to 65%. Although label from all test compounds except proline (group II) and lysine (group III) was incorporated into tylactone after 60 min, label from group I and group III compounds was incorporated at levels five times greater than label from group II compounds. From 55 to 75% of the recovered label from propionate (I), asparagine (II), glutamine (II), glutamate (II), alpha-ketoglutarate (II), and succinate (II) was recovered as 14CO2. From 75 to 95% of the recovered label from the remaining compounds tested was located in the cells. Based on the data, a pathway for the role of amino acids in the biosynthesis of tylactone is proposed.

Cloning of a DNA fragment from Cephalosporium acremonium which functions as an autonomous replication sequence in yeast.

Abstract: A fragment of DNA which functions as an autonomous replication sequence in yeast was cloned from Cephalosporium acremonium. Mitochondrial DNA (mtDNA) was isolated from an industrial strain of C. acremonium (08G-250-21) highly developed for the production of the antibiotic, cephalosporin C. Size, 27 kb, and restriction pattern indicated this DNA was identical to mtDNA previously isolated (Minuth et al. 1982) from an ancestral strain (ATTC 14553) which produces very low amounts of cephalosporin C. A 1.9 kb Pst1 fragment of the Cephalosporium mtDNA was inserted into a Pst1 site of the yeast integrative plasmid, Ylp5, to produce a 7.5 kb plasmid, designated pPS1. The structure of pPS1 was verified by restriction analysis and hybridization.

Cytotoxic compositions of transferrin coupled to vinca alkaloids

Abstract: There is provided by this invention cell-targeting cytotoxic compositions comprising transferrin covalently coupled to vinca alkaloid.

Autoreactivity to collagen in a murine lupus model.

Abstract: MRL/1 mice exhibit many characteristics of human systemic lupus erythematosus including antinuclear antibodies, circulating immune complexes, glomerulonephritis, and death secondary to renal failure. In addition, these mice have elevated levels of rheumatoid factor and spontaneously develop arthritis that has many similarities to human rheumatoid arthritis. Our present studies indicate that, with age, they also develop reactivity to types I and II collagen. The levels of antibodies against native or denatured types I and II collagen in the sera of 4–5-month-old MRL/1 mice are significantly higher than those in the sera of age-matched Balb/c or MRL/n mice. The specificity of these antibodies for collagen was demonstrated by a competitive binding assay. The T cells from 1- or 2-month-old MRL/1 mice exhibited a significant proliferative response in the presence of type I collagen and a mild or no response to type II collagen. Both antigenic and mitogenic responses decreased with age. The results suggest that the development of autoimmunity to collagen may play an important role in the perpetuation of arthritis, vasculitis, and glomerulonephritis in MRL/1 mice.