Search

Intratracheal (i.t.) or intravenous (i.v.) administration of FR901451, a potent inhibitor of human leukocyte elastase (HLE) prevented HLE-induced lung hemorrhage in hamsters with ED50 values of 10.5 micrograms/site and 8.1 mg/kg, respectively. alpha 1-Antitrypsin (alpha 1-AT) also showed inhibitory effect in this model. However, the ED50 value by i.t. injection of FR901451 was 20-fold lower than that of alpha 1-AT. Moreover, FR901451 i.t. significantly modulated porcine pancreas elastase (PPE)-induced changes of the respiratory mechanics in hamsters. The ED50 values were 529 micrograms/site and 244 micrograms/site, which were expressed by static lung compliance (Cst) and vital capacity (VC) of the lungs, respectively. These results suggest that FR901451 could be clinically useful agent for the treatment of the destructive lung disease such as pulmonary emphysema.

WF11899A, B and C, novel water-soluble lipopeptides related to the echinocandins, possess potent anti-Candida activities. The IC50s of the compounds against four clinical isolates of Candida albicans ranged from 0.004 to 0.03 microgram/ml by microbroth dilution assay. These compounds mildly suppressed the growth of Aspergillus fumigatus and A. niger. WF11899A, B and C showed a potent in vivo anti-Candida activity. Particularly, WF11899A was superior to cilofungin, and equal to fluconazole. 1,3-beta-glucan synthase was inhibited by these compounds at the IC50s of 0.7, 0.7 and 1.8 micrograms/ml for WF11899A, B and C, respectively. However, they hemolysed mouse red blood cells in vitro at the concentration of 62 micrograms/ml.

WS79089A, B and C, which are novel endothelin converting enzyme (ECE) inhibitors have been isolated from the fermentation broth of Streptosporangium roseum No. 79089. These inhibitors were purified from an acetone extract of whole culture broth followed by Silicar CC-4 column chromatography and HPLC. WS79089A, B and C showed highly selective ECE inhibition activity with IC50 values of 0.73 microM 0.14 microM and 3.42 microM, respectively. On the basis of spectroscopic and chemical evidence, the tentative structures of WS79089A, B and C have been proposed, they have benzo[a]naphtacen chromophores.

Publisher Summary This chapter describes the screening for immunosuppressants. There are several types of cells responsible for specialized functions in the immune system that act through hormone-like substances, much as in the nervous system. It was found that T cells could be cultured indefinitely in the presence of immunological hormone interleukin 2. It has been demonstrated time and again that this interleukin plays a crucial role in the immune response. On the one hand, the immune system protects the body against diseases from invasion by foreign organisms, but on the other hand, it can attack a lifesaving organ transplant or mistakenly respond to the body's own molecules that, in turn, results in autoimmune diseases such as multiple sclerosis, rheumatoid arthritis, and juvenile diabetes. A large accumulation of data shows that activated T cells are associated directly with damage to the tissues during graft rejection and autoimmune diseases. This finding rationalizes the idea that activated T cells can be treated as a pathogen, similar to pathogenic microbes in an infectious disease, and this is leading to new therapeutic approaches for abnormal immune reactions. Depressed function of activated T cells can lead to the suppression of an abnormal immune response. One of the targets for the depression of cell function is IL-2. Selective inhibition of IL-2 production could inhibit T cell growth, which in turn, would result in suppression of the immune response.

In the course of our search for non-steroidal androgen-receptor antagonists of microbial origin, Pseudomonas sp. No. 2838 was found to produce an inhibitor of androgen binding to its receptor. This compound, named WB2838, was isolated and identified as 3-chloro-4-(2-amino-3-chlorophenyl)-pyrrole. The IC50 value of WB2838 for partially purified rat prostate cytosol receptor was 8.0 x 10(-7) M. However, the IC50 value of WB2838 against estrogen-receptor binding was about 90-fold greater than that against androgen-receptor binding. WB2838 inhibited the growth of androgen-responsive mouse mammary carcinoma SC-3 cells in the presence of 10(-8) M testosterone at IC50 value of 4.1 x 10(-7) M. This inhibition was reversed by adding 10(-5) M testosterone to the culture medium. WB2838 also showed the inhibitory activity against the growth of the ventral prostate induced by testosterone propionate in castrated immature rats. Therefore, it was concluded that WB2838 was a non-steroidal androgen-receptor antagonist.

R1128 B (1,3,6-trihydroxy-8-n-butylanthraquinone), a new antibiotic produced by Streptomyces sp. No. 1128, inhibited estrogen binding to its receptor. The IC50 value of R1128 B for partially purified rat uterine cytosol receptor was 1.2 x 10(-7) M. However, the IC50 value of R1128 B against androgen-receptor binding was about 50-fold greater than that against estrogen-receptor binding. R1128 B was a competitive inhibitor against estrogen-receptor binding. R1128 B inhibited the growth of estrogen-responsive human mammary adenocarcinoma MCF-7 cells in soft agar. This inhibition, however, was reversed when estradiol was added to the culture medium. R1128 B showed antitumor activities against MCF-7 both xenografted to nude mice and implanted in subrenal capsule of mice (SRC assay). The potency of R1128 B was about 8-fold lower than that of tamoxifen both in vitro and in vivo.

A novel human leukocyte elastase inhibitor, FR901277 was discovered in the fermentation broth of Streptomyces resistomycificus No. 7622. FR901277 has a molecular weight of 961 and a molecular formula of C47H63N9O13. The mode of inhibition is competitive, with a Ki of 1.2 x 10(-8) M. Oral administration of FR901277 at doses from 32 to 320 mg/kg significantly prevented human leukocyte elastase-induced foot edema in mice.

FK-506, a novel immunosuppressant from Streptomyces tukubaensis, was a selective anti-T lymphocyte agent like ciclosporin (CS). This agent suppressed both cell-mediated and humoral immunity, without depressing the bone marrow. Futhermore, it was effective in a rat skin allograft model. These results provided a strong motivation to develop FK-506 for clinical application to organ transplantation.

Data from several studies suggest that tachykinins may play an important role in the pathophysiology of airway diseases, especially asthma. Our aim is to discover tachykinin antagonists which exhibit therapeutically useful anti-asthmatic activity. In our search for activities inhibiting the binding of [3H]substance P to guinea-pig lung membrane preparations, we have found that the fermentation product, WS9326A, isolated from Streptomyces violaceusniger, is a potent tachykinin receptor antagonist.

WS-7528, produced by Streptomyces sp. No. 7528, was extracted from cultured broth, purified by solvent extraction followed by chromatography on silica gel and then isolated as pale yellow powder (C16H14O5, mp 95-98 degrees C). WS-7528 inhibited estrogen binding to its receptor protein in rat uterine cytosol. The IC50 value of WS-7528 for partially purified rat uterine cytosol receptor was 5.7 x 10(-8) M. This compound was found to induce the growth of the estrogen dependent cell line MCF-7. WS-7528 was tested orally and subcutaneously in immature rats to confirm its effect on the growth of the uterus. WS-7528 has also weak anti-inflammatory activity on the carrageenin paw edema of the rat model.

Fermentation, isolement structure et proprietes biologiques de FR 900403, un antifongique, produit par kernia Sp. F-19849. Les proprietes physicochimiques sont exposees. On etudie l'activite antifongique contre Candida albicans in vitro et chez des souris infectees

Streptoverticillium fervens produced a new antibiotic, FR-900848, which has a high specific activity against filamentous fungi. Purified by solvent extraction and column chromatography, the compound appears as colorless crystals. Its structure is C32H43N3O6, which consists of 5'-amino-5'-deoxy-5, 6-dihydrouridine with an unsaturated fatty acid having unprecedented four serial and one isolated cyclopropane rings.

WS1358A1, a novel inhibitor of renal dehydropeptidase (DHP), augmented the urinary recovery of a carbapenem antibiotic imipenem and improved its protective effect in experimental infections when simultaneously administered to mice with the antibiotic. WS1358A1 was a competitive DHP inhibitor with a Ki value of 1.6 x 10(-7) M.

WSl358Al (FR104007) and Bl (FR104008), new potent inhibitors of renal dehydropeptidase, were isolated from the culture broth of strain No. 1358 which was identified as Streptomyces parvulus subsp. In vitro inhibitory activities (IC50 value) of WSl358Al and Bl against porcine renal DHP were 3 and 600 nM, respectively.

FR109615, a new antibiotic active against Candida, was isolated from Streptomyces setonii No. 7562. Based on the spectroscopic data, the structure of FR109615 was elucidated as cis-2-aminocyclopentane-1-carboxylic acid (1). The compound showed the excellent in vivo efficacy in a generalized infection test of mice.

FK-565 restored the number of granulocytes in the bone marrow in mice treated with mitomycin C (MMC). Granulocyte counts recovered to the normal level 3 days after MMC treatment at a dose of 0.1 mg/kg of FK-565. The restorative activity of FK-565 was more effective than that of muramyldipeptide (MDP) or lipopolysaccharide (LPS).

WS-9659 A, a novel phenazine, produced by a Streptomyces sp., had testosterone 5 alpha-reductase inhibition activity on rat, dog and human prostates. However, WS-9659 A did not show any inhibitory activities for aldose reductase on rabbit lenses and lactate dehydrogenase on pig hearts. WS-9659 A was a competitive inhibitor against testosterone 5 alpha-reductase on rat prostates by use of testosterone as a substrate. Radio receptor binding assay of androgen receptor of rat prostates revealed that WS-9659 A had no affinity for this receptor. WS-9659 A was tested subcutaneously in immature castrated rats to confirm its effect on the growth of the ventral prostates induced by testosterone propionate.

An interesting immunoactive peptide, FK-156, has been isolated from the fermentation broth of Streptomyces olivaceogriseus sp. nov. and Streptomyces violaceus. The compound was purified by column chromatography with activated carbon, ion exchange Sephadex and cellulose powder. FK-156, obtained as white powder, exhibits a wide variety of immunostimulatory activity in vivo and in vitro with experimental animals. Pretreatment of mice or rats with this peptide protected the animals against lethal infection with Escherichia coli and resulted in prolongation of life span of tumor bearing animals.

A complex of the new antitumor antibiotics (WF-1360, WF-1360A, B, C, D, E and F) was produced by Rhizopus sp. No. F-1360. Structural studies of these compounds suggested that they were novel 16-membered-ring lactones having an oxazole ring in their structures. WF-1360 was found to be identical with rhizoxin (1) and WF-1360B, C, E and F were de-termined to be homologues of 1 with structures 2, 3, 4 and 5, respectively. These compounds were cytotoxic when tested on P388 leukemia cells in vitro. WF-1360 was highly active against leukemia L1210 and melanoma B16. They also exhibited potent antifungal activities, but weak antimicrobial activities against some Gram-positive or negative bacteria.

FR-900483 is a new immunoactive substance produced by a fungus, Nectria lucida F-4490. Concanavalin A-stimulated lymphocyte proliferation, which had been suppressed by addition of immunosuppressive factor, was restored to a normal level by the addition of FR-900483. Furthermore, FR-900483 restored the capacity of immunosuppressed mice to produce antibody against sheep red blood cells.

Vinigrol, produced by a fungal strain identified as Virgaria nigra, was extracted from the cultured mycelium, purified by solvent extraction followed by chromatography on silica gel and then isolated as crystals (C20H34O3, mp 108 degrees C). Vinigrol decreased arterial blood pressure of anesthetized normotensive rats dose-dependently when administered intravenously. Vinigrol inhibited platelet activating factor and epinephrine induced platelet aggregation.

FK409, a novel vasodilator with anti-platelet aggregation activity, has been isolated from the acid-treated fermentation broth of Streptomyces griseosporeus No. 16917, which was cultured on a medium containing NaNO3 for 4 days. FK409 was purified from the culture-filtrate by extraction with ethyl acetate after adjusting the pH to 3.0 with HC1, followed by silica gel chromatography. The molecular formula of this compound was determined to be C8H13N3O4. In vitro, FK409 showed a potent relaxation activity on noradrenaline induced contraction of rat aorta. In addition to the vasodilating activity, this compound also showed potent anti-aggregation activities towards rabbit platelets. In vivo, intravenously administered FK409 resulted in marked blood pressure lowering in rats.

A new antitumor antibiotic, FR900840, was isolated from a culture broth of Streptomyces strain No. 8727 as a pale yellowish prism and the molecular formula was determined to be C7H11N3O5. The antibiotic exhibited prominent antitumor effects on human tumor cell lines both in vitro and in vivo.

The structure of novel phosphonic acid antibiotics, FR-33289, FR-31564, and FR-32863, produced by strains of Streptomyces, have been established as I, II, and III, respectively, on the basis of spectroscopic and chemical evidences.

FR-900520 and FR-900523, novel neutral macrolide immunosuppressants, were isolated from the cultured broth of Streptomyces hygroscopicus subsp. yakushimaensis No. 7238. Their molecular formulae were determined as C43H69NO12 and C42H67NO12, respectively. The compounds suppressed immune response in vitro. IC50 values of FR-900520 and FR-900523 for mouse mixed lymphocyte reaction were 0.55 nM and 1.6 nM, respectively. FR-900520, the major component, clearly prolonged skin allograft survival in rats.

FR-900130 is a new antibiotic having an acetylene moiety and is very unstable in aqueous solution. The antibiotic was isolated as the acetyl derivative by ion-exchange resin and adsorption chromatography. The structure has been determined to be L-2-amino-3-butynoic acid by spectral data of its derivatives.

Using Cellophane-sheet techniques, changes in nucleotide pool size during conidia formation of Penicillium chrysogenum on agar plates were studied. On conidia-forming medium (nutritionally poor medium), the pool size of nucleoside triphosphates, especially GTP, decreased dramatically at mid-growth phase, concomitantly with the exhaustion of extracellular phosphate. Onset of conidia formation was observed just after the GTP pool decrease.

Many strains of yeast which can utilize n-alkanes as the sole source of carbon were isolated from flowers and fruits. Among them, a strain, OH23, identified as Candida tropicalis, formed acidic substances from n-alkanes. The principal products from n-alkanes with odd and even numbers of carbons were identified as glutaric and adipic acids, respectively. The culture conditions for their formation were investigated. n-Pentadecane and n-hexadecane were the best substrates for the formation of glutaric and adipic acids, respectively. Yields of 170 mg of glutaric and 64 mg of adipic acid were obtained from 100 ml of media containing 4% (v/v) n-pentadecane and n-hexadecane, respectively, and 0.5% casamino acids.

Candida tropicalis OH23 capable of forming glutaric acid or adipic acid from n-alkanes, was found to form two acidic compounds from glucose which were different from the above compounds. The compounds were isolated and their structures were examined by elementary analysis, infrared spectroscopy, NMR spectroscopy and study of decomposed products. The compounds were proved to be identical with N- acetyl- L -alanine and N-acetylglycine.

FR-900490 is a new type of immunoactive substance produced by a fungus Discosia sp. F-11809. The colony forming units in culture (cfu-c) in bone marrow cells, which were suppressed by immunosuppressive factor obtained from the serum of sarcoma 180 tumor bearing mouse, was restored to normal level by the addition of FR-900490 in vitro. Furthermore, in mitomycin C (MMC)-treated mice the subsequent administration of FR-900490 caused a significant increase of cfu-c in bone marrow cells depressed by MMC.

Ceftizoxime (CZX) was evaluated for absorption and excretion as well as for therapeutic effectiveness in neonates and premature infants. The following results were obtained. 1. Serum CZX concentrations were determined in 8 neonates or premature infants who were not more than 6 days old. Serum concentrations of the drug were examined in 6 neonates and/or premature infants after intravenous administration of about 20 mg/kg body weight. Average concentration at 1/2, 2, 4 and 6 hours after administration were 52.3, 36.4, 26.7 and 16.7 micrograms/ml, respectively. Serum concentrations in the other 2 infants who were given 29.7 and 25.1 mg/kg, were as high as 71 and 94 micrograms/ml at 1/2 hour and 22.1 and 39 micrograms/ml at 6 hours, respectively. Serum half-lives in 5 of the 6 mature neonates ranged from 2.36 to 3.34 hours, with averaged 2.75 hours, but was exceptionally long, 7.92 hours, in the other one. Half-lives in the 2 premature infants were 4.14 and 4.90 hours. 2. The therapeutic effectiveness on bacterial infection was evaluated for 10 newborn infants. Intravenous doses of 16.9 to 24.6 mg/kg were given in b.i.d. or t.i.d. regimen to 4 cases with pneumonia and 2 with septicemia, urinary tract infection and fetal infection each. To 1 infant with septicemia complicated with cephalohematoma, higher doses ranged from 21.8 to 49.8 mg/kg were given t.i.d. or q.i.d. Therapeutic efficacies were assessed as "Excellent" in 3, "Good" in 6, and "Poor" in 1, with an efficacy rate of 90.0%. Eradication of bacteria was complete in 2 infants each with Escherichia coli-induced septicemia or urinary tract infection. 3. For prophylactic use, the drug was given to 8 newborn infants in intravenous doses of 17.5 to 29.1 mg/kg b.i.d. or t.i.d. and no infection occurred in 7 cases. 4. No adverse reactions were obtained. Slight and transient increases in platelet count, GOT and GPT in 1 case and eosinophilia in another were observed. 5. These results suggested that CZX in an intravenous dose of 20 mg/kg b.i.d. or t.i.d. regimen in newborn infants up to 7 days of age would be effective and safe for the treatment of neonatal bacterial infections.

WF-3681 was isolated from a cultured filtrate of Chaetomella raphigera as a novel inhibitor of aldose reductase. It was extracted with ethyl acetate and then purified with silica gel chromatography. Its molecular formula was determined to be C13H12O5 by elemental analysis and high resolution electron impact mass spectrometry. IC50 of WF-3681 was 2.5 X 10(-7) M for partially purified aldose reductase of rabbit lens.

A new species of Streptomyces which produces a new cell wall-inhibitory antibiotic, FR-9000981) containing phosphonic acid in its molecule, is named and described. The species name proposed, Streptomyces rubello, nurinus, refers to the aerial mass color. Streptomyces rubellonturinus subsp. indigoferus also produces FR-900098 and the related compound FR-332892). FR-900098 related compounds, FR-32863) and FR-315642) are produced by Streptomvices lavendulae.

Three phosphonic acid antibiotics were found to be produced in the fermentation broths of Streptomyces. FR-31564 and FR-32863 were produced by Streptomyces lavendulae. FR-33289 was produced by a strain of Streptomyces, identified as Streptomyces rubellomurinus subsp. indigoferus. They are distinct from, but resemble FR-900098 which was reported in our preceding paper, in their chemical and biological characteristics.

WF-10129 is an angiotensin converting enzyme (ACE) inhibitor produced by Doratomyces putredinis. IC50 of the compound is 1.4 X 10(-8) M for the ACE activity. WF-10129 was purified from cultured filtrate by successive ion exchange chromatography and HPLC. The chemical structure 1 was elucidated on the basis of spectroscopic and chemical evidence. The compound is a dipeptide composed of L-tyrosine and a novel amino acid. WF-10129 inhibits the pressor response of angiotensin I when administered intravenously at 0.3 mg/kg in rats.

A strain of Streptomyces, isolated from a soil sample and identified as Streptomyces xanthocidicus, has been found to produce FR-900148, a new antibiotic containing chlorine in its molecule. The antibiotic inhibits both Gram-positive and negative bacteria. However, it is not effective against wild type of Pseudomonas aerguinosa. Its antibacterial action is considered to result from cell wall synthesis inhibition since it causes spheroplast formation from susceptible cells.