betadex and beta-cyclodextrin-tetradecasulfate

Control of selectivity in the enantiomeric separation of three aromatic amino acids (phenylalanine, tyrosine and tryptophan) is demonstrated by electrokinetic capillary chromatography utilising temperature variations coupled with the use of sulphated-beta-cyclodextrin (s-beta-CD) as a pseudostationary phase. The concentration of s-beta-CD and temperature were used as experimental variables to control the observed selectivity. A double-coated capillary was used and proved very robust with reproducibility of migration times being <2.0% R.S.D. between runs and <2.6% on using a new capillary. The system was modelled successfully using an artificial neural network (ANN) comprising one input layer, two hidden layers and one output layer. The model accurately described the observed separations with a correlation coefficient of 0.999 being observed between predicted and observed migration times. Selectivity optimisation was achieved using the normalised resolution product and minimum resolution criteria, with both providing optima at different experimental conditions. The selectivity changes observed also allowed the estimation of electrolyte temperatures within the capillary at high operating currents (>100 microA). Using a 50 microm i.d. capillary and an electrolyte comprising 20 mM phosphate and 15 mM s-beta-CD, a temperature of 52 degrees C was calculated within the capillary at an applied voltage of +30 kV.

Topics: Amino Acids, Aromatic; beta-Cyclodextrins; Cyclodextrins; Electrophoresis, Capillary; Indicators and Reagents; Neural Networks, Computer; Phenylalanine; Spectrophotometry, Ultraviolet; Stereoisomerism; Temperature; Tryptophan; Tyrosine

Beta-Cyclodextrin tetradecasulfate binds fibroblast growth factors and possesses anticoagulant properties. This study was designed to assess the separate and combined effects of local intramural delivery and intravenous administration of beta-cyclodextrin tetrade-casulfate on neointimal formation and arterial damage following angioplasty.. Fifty-two pigs randomized into four groups underwent coronary artery angioplasty: 1) control, 2) continuous intravenous infusion of 100 mg/kg/d of beta-cyclodextrin tetradecasulfate, 3) intramural delivery of 1250 mg beta-cyclodextrin tetradecasulfate, 4) intramural delivery of 1250 mg beta-cyclodextrin tetradecasulfate followed by continuous intravenous infusion of 100 mg/kg/d. Fourteen days after injury, morphometric analysis revealed that arteries randomized to the intravenous beta-cyclodextrin tetradecasulfate groups had a decreased normalized neointima area: control, 3.03 +/- 0.75 mm(2); intravenous, 1.67 +/- 0.73 mm(2) (40% decrease; P < 10(-7)); intravenous plus local, 1.95 +/- 0.76 mm(2) (30% decrease; P < 10(-5)). There was no difference in neointimal response following local beta-cyclodextrin tetradecasulfate delivery only (2.82 +/- 1.14 mm(2)). Coronary arterial damage, defined as aneurysm, dissection, adventitial rupture, and retromedial hematoma was similar in all groups (12% in control and local groups, 10% in the intravenous group, 14% in the intravenous plus local; NS). Bleeding complications were more frequent in the intravenous and intravenous plus local groups compared to the local and control groups (23%vs 7.6% and 0%, respectively; P < 0.05).. Continuous intravenous administration of beta-cyclodextrin tetradecasulfate substantially reduced intimal hyperplasia, while intramural delivery had no effect, indicating that a single bolus of beta-cyclodextrin tetradecasulfate did not reduce intimal hyperplasia. There was no additive effect of local intramural delivery of beta-cyclodextrin tetradecasulfate. However, local delivery of beta-cyclodextrin tetradecasulfate induced less bleeding complications and did not lead to additional arterial injury, indicating that local delivery of an anticoagulant does not cause additional arterial injury.

Topics: Angioplasty, Balloon, Coronary; Animals; beta-Cyclodextrins; Coronary Vessels; Cyclodextrins; Electrocardiography; Hyperplasia; Infusions, Intravenous; Injections, Intravenous; Random Allocation; Swine; Tunica Intima

Neointimal formation is a major cause of restenosis after interventional vascular procedures. Beta-cyclodextrin tetradecasulfate (beta-CDT) has been shown to inhibit fibroblast growth factor activity and we hypothesized that beta-CDT would reduce intimal formation.. Three studies were performed: (1) pharmacokinetics of oral and intravenous beta-CDT and determination of optimal dose, (2) determination of efficacy of oral and intravenous beta-CDT in reducing neointimal formation after balloon-overstretch injury and (3) determination of the effect of beta-CDT on cellular proliferation, factor Xa activity, activated clotting time, activated partial thromboplastin time and thrombus formation.. Pharmacokinetics were determined in eight domestic swine following administration of oral beta-CDT and intravenous beta-CDT at three doses each. In the efficacy study, balloon-overstretch injury of 37 pigs (69 arteries) was performed and randomized into three groups (n = 23 arteries/group): control, oral administration of 300 mg beta-CDT/kg per day or intravenous infusion of 100 mg beta-CDT/kg per day. Animals were sacrificed 14 days later. Cellular proliferation and mural thrombus were determined in six arteries/group at 5 days and endothelial coverage was evaluated at 5 and 14 days.. Oral and intravenous beta-CDT reduced the intimal hyperplasia area normalized to injury index by 24 and 48%, respectively: control, 3.03 +/- 0.75 mm2, oral, 2.31 +/- 0.83 mm2 (P = 0.004) and intravenous, 1.67 +/- 0.73 mm2 (P = 0.0000002). beta-CDT reduced cellular proliferation (control, 55 +/- 18%, oral, 35 +/- 7%, P = 0.03 and intravenous, 30 +/- 12%, P = 0.01) and mural thrombus formation (control, 0.84 +/- 0.4 mm2, oral, 0.44 +/- 0.14 mm2, P = 0.04, intravenous, 0.42 +/- 0.09 mm2, P = 0.03). Endothelial coverage was increased in the experimental groups (P = 0.008, oral versus control, P < 0.0001, intravenous versus control). Factor Xa activity was inhibited 9-10 fold following intravenous administration while oral administration demonstrated no effect.. Both oral and intravenous formation of beta-CDT reduced intimal hyperplasia with the greatest reduction in the intravenous group. We postulate that beta-CDT was effective by the combination of increasing endothelial coverage, reducing mural thrombus formation, inhibiting factor Xa activity and reducing cellular proliferation.

Topics: Animals; beta-Cyclodextrins; Blood Glucose; Coronary Thrombosis; Coronary Vessels; Cyclodextrins; Disease Models, Animal; Dose-Response Relationship, Drug; Drug Evaluation; Endothelium, Vascular; Factor Xa; Magnesium; Models, Cardiovascular; Oligosaccharides; Randomized Controlled Trials as Topic; Statistics as Topic; Swine; Treatment Outcome; Tunica Intima; Whole Blood Coagulation Time

The purpose of this study was to determine whether the wall thickening observed in vein grafts after they were placed into the arterial circulation could be inhibited by periadventitial delivery of an insoluble sulfated polymer of beta-cyclodextrin (P-CDS) capable of tightly binding heparin binding growth factors.. Thirty-four New Zealand white rabbits underwent implantation of reversed autologous jugular vein interposition grafts in the common carotid artery and were randomized to receive either 20 mg P-CDS (n = 18) topically around the graft or no additional therapy (n = 16). Before being killed at 28 days, animals were given bromodeoxyuridine to assess smooth muscle cell proliferation. Histomorphometric analyses were performed after perfusion fixation.. Compared to controls, treatment with P-CDS was associated with reduced mean intimal thickness (24 +/- 3 vs 38 +/- 4 microns; mean SEM, p < 0.01) and intimal area (0.25 +/- 0.03 vs 0.54 +/- 0.09 mm2; p < 0.01). There was also significantly less medial thickness in the P-CDS group (45 +/- 3 vs 63 +/-3, p < 0.001). There was no significant difference in intimal or medial smooth muscle cell proliferation between P-CDS-treated and control vein grafts at 28 days. The polymer persisted in the adventitia with a mild foreign body reaction.. Periadventitial placement of P-CDS, a novel, insoluble, sulfated carbohydrate polymer, inhibits intimal and medial thickening of vein bypass grafts in this model of vein grafting. The persistence of P-CDS in vivo for prolonged periods, and the ease of topical application of P-CDS during vascular bypasses may have important implications for its future use in vascular surgery.

Topics: Administration, Topical; Animals; beta-Cyclodextrins; Carotid Artery, Common; Cell Division; Cyclodextrins; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Foreign-Body Reaction; Giant Cells; Jugular Veins; Male; Muscle, Smooth, Vascular; Rabbits; Tunica Intima; Tunica Media

Smooth muscle cell migration and proliferation are important regulatory processes in the development of intimal thickening after vascular injury. beta-cyclodextrin tetradecasulfate, an orally active synthetic heparin mimic, is effective in inhibiting rabbit aortic smooth muscle cell proliferation in vitro and in limiting restenosis in an experimental angioplasty restenosis model in rabbits (Hermann et al., 1993). However, its effects on migration are unknown, as are its effects on human vascular smooth muscle cell biology in general. Using a Transwell assay system, we demonstrated a dose dependent inhibition of human vascular smooth muscle cell random migration for beta-cyclodextrin tetradecasulfate with half maximal inhibition between 100-500 micrograms/ml and 85 +/- 1% inhibition at 10(3) micrograms/ml (P < .001, n = 4). At this latter concentration, inhibition of migration was also demonstrable using a linear under-agarose assay, where the mean velocity for beta-cyclodextrin tetradecasulfate-treated cells (8 +/- 2 microns/h, +/- S.E.) was significantly less than for control cells (21 +/- 4, P < .01), but equaled that of control cells 48 h after drug withdrawal (21 +/- 1, P = NS). Single cell analysis over 17 h using time lapse video microscopy revealed significant inhibition of total migration pathway distance for beta-cyclodextrin tetradecasulfate-treated smooth muscle cells compared with control smooth muscle cells (0.40 +/- 0.03 mm vs. 0.73 +/- 0.03, P < .01). We also demonstrated a dose-dependent inhibition of serum-induced proliferation by beta-cyclodextrin tetradecasulfate in both cultured human umbilical vein and coronary artery smooth muscle cells. To assess whether beta-cyclodextrin tetradecasulfate had any direct cellular toxicity, we measured the release of intracellular LDH at 6 or 24 h. At 10(3) micrograms/ml or less, there was no increase in LDH release compared with untreated cultures. Thus, beta-cyclodextrin tetradecasulfate may be an effective agent in inhibiting intimal thickening after vascular injury by limiting both smooth muscle cell migration and proliferation.

Topics: beta-Cyclodextrins; Cell Division; Cell Movement; Cells, Cultured; Cyclodextrins; Humans; L-Lactate Dehydrogenase; Muscle, Smooth, Vascular

Intimal thickening after vascular injury may be modulated in part by heparin binding growth factors. We hypothesized that placement of a therapeutic polymer in the periadventitial space capable of tightly binding growth factors might alter the vascular response to injury. We first demonstrated that incubation of rat aortic smooth muscle cells with an insoluble, sulfated polymer of beta-cyclodextrin (P-CDS) was associated with a dose-dependent inhibition of proliferation induced by fetal calf serum, fibroblast growth factor-2 (FGF-2), platelet-derived growth factor BB, or epidermal growth factor. Preincubation studies of P-CDS with FGF-2 revealed a very rapid removal of mitogenic activity. Using radiolabeled FGF-2 (0.25 microg/ml), we observed a very rapid association rate (0.34 +/- 0.07 min-1, n=4) and a very slow dissociation rate (3.3 +/- 0.2 X 10(-7) min-1) at 37 degrees C, suggesting a high affinity interaction. Using both Transwell and linear under-agarose assays, we demonstrated a significant inhibition of random migration (chemokinesis) by P-CDS. Unsulfated polymeric beta-cyclodextrin (P-CD) had little if any of these effects, suggesting that the high negative charge density of P-CDS was important for the effects. Finally, rats undergoing carotid artery balloon injury were randomized to treatment with periadventitial P-CDS or no treatment, and were killed at 4 (n=20), 14 (n=59), and 88 d (n=14). Morphometric analysis demonstrated significant and sustained inhibition of intimal thickening in P-CDS-treated rats at 14 (P < 0.01) and 88 d (P < 0.05) using absolute intimal area or intima/media area ratios. No inhibition was seen in a group of rats treated with P-CD. In P-CDS-treated rats, bromodeoxyuridine labeling studies revealed fewer labeled smooth muscle cells in the intima at 14 d (P=0.01), while staining with Evans blue revealed enhanced late endothelial cell regrowth. Thus, periadventitially applied sulfated beta-cyclodextrin polymer, which can tightly bind heparin binding growth factors, inhibits intimal thickening in vivo in a sustained fashion without using an additional delivery system. These studies suggest that cellular processes mediated by heparin binding growth factors may be modulated by P-CDS.

Topics: Angioplasty, Balloon; Animals; Aorta; Becaplermin; beta-Cyclodextrins; Carotid Arteries; Cell Division; Cyclodextrins; Dose-Response Relationship, Drug; Epidermal Growth Factor; Fibroblast Growth Factor 2; Growth Substances; Kinetics; Organ Culture Techniques; Platelet-Derived Growth Factor; Polymers; Proto-Oncogene Proteins c-sis; Rats; Recombinant Proteins; Thymidine; Tunica Intima

A novel method for quantitating angiogenesis and its inhibition has been developed for the chick embryo. The method is based on the vertical growth of new capillary blood vessels into a collagen gel through two parallel nylon meshes which align the capillaries for counting. Angiogenesis is induced by basic fibroblast growth factor contained within the gel and slowly released by aluminum sucrose octasulfate (sucralfate) or by tumor cells implanted on the gel. The potency of four different angiogenesis inhibitors was compared at concentrations of 0.6 to 600 nmole. This technique may facilitate the discovery and development of angiogenesis inhibitors for clinical application.

Topics: Allantois; Animals; beta-Cyclodextrins; Betamethasone; Biological Assay; Blood Vessels; Chick Embryo; Chorion; Cyclodextrins; Cyclohexanes; Fibroblast Growth Factor 2; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Protamines; Sesquiterpenes; Sucralfate

Tenon's fibroblast (TF) bleb encapsulation is a common cause of filtering surgery failure. Various pharmacologic agents have been used in an attempt to inhibit this response. The effects of three angiogenesis inhibitors were studied. AGM-1470, a fumagillin analog and hydrocortisone 21-phosphate (HC21P) complexed with heparin or with the heparin analog beta-cyclodextrin tetradecasulfate (BCD-TDS) in modulating human tenon's fibroblasts in cell culture. It has been clinically demonstrated that well vascularized blebs are associated with a poorer prognosis. In addition to reducing bleb neovascularization, angiogenesis inhibitors may have an additional therapeutic role in decreasing fibroblast activity. Drug effects were assessed by studying cell growth rates by cell Coulter counting and rate of wound closure. TF's were grown in DMEM with 10% FBS and 1% PCN-strep with fungizone. Angiogenesis inhibitors were added to growth medium in varying concentrations: AGM-1470 alone, HC21P complexed with heparin and HC21P complexed with BCD-TDS. Controls were grown in control medium alone, medium with added ethanol, or with added BCD-TDS, heparin or HC21P. We found a dose related inhibition of cell growth with all of the angiogenesis inhibitor combinations, which was not seen in the control groups. Tenon's fibroblast proliferation was significantly inhibited by AGM-1470 as seen in the four higher concentrations (P < 0.05) with insignificant inhibition at the lowest concentration of AGM-1470 (P = 0.38). The heparin:HC21P and heparin:BCD-TDS combinations demonstrated significant inhibition at all concentrations (P < 0.05). Wound closure was significantly inhibited by all the added agents, except AGM-1470 and the controls. The rate of wound closure was significantly reduced by the highest concentrations of the heparin:HC21P and heparin:BCD-TDS combinations (P < 0.05), although it was not significantly affected by lower concentrations (P > 0.05). Rank order of potencies of these agents for inhibition of TF proliferation was AGM-1470, BCD-TDS:HC21P, heparin:HC21P, HC21P, while the rank order of potencies for these agents for inhibition of wound closure was HC21P, heparin:HC21P, BCD-TDS:HC21P, AGM-1470. These selected angiogenesis inhibitors appear to have marked inhibitory effects on TF proliferation and migration, which may have a potential clinical role in modulating wound healing associated with glaucoma filtering surgery.

Topics: beta-Cyclodextrins; Cell Division; Cells, Cultured; Cyclodextrins; Cyclohexanes; Dose-Response Relationship, Drug; Eye; Fibroblasts; Glaucoma; Humans; Hydrocortisone; Neovascularization, Pathologic; O-(Chloroacetylcarbamoyl)fumagillol; Sesquiterpenes; Time Factors; Wound Healing

The fluorescence emission of a single tryptophan residue present in both FGF-1 and FGF-2 was used as a structural probe to directly assess the interaction of the growth factors with heparin or beta-cyclodextran tetradecasulfate. About 20-25% of the fluorescence of either FGF-1 or FGF-2 is quenchable, and is dependent on sulfation of the ligands. The quenchable fluorescence is associated with about 20% of total FGF, suggesting the presence of two fluorospectrometric forms of the protein. The equilibrium dissociation constants, determined by this method, for heparin or beta-cyclodextrin tetradecasulfate binding to FGF-1 are about 1 nM, whereas the values for FGF-2 are 1 and 23 nM, respectively. The method provides a direct tool to evaluate FGF-ligand interaction and assess the structural integrity of the proteins.

Topics: Animals; beta-Cyclodextrins; Binding Sites; Cyclodextrins; Fibroblast Growth Factor 1; Fibroblast Growth Factor 2; Heparin; In Vitro Techniques; Kinetics; Ligands; Protein Binding; Spectrometry, Fluorescence; Sulfates; Swine; Tryptophan

Folkman and coworkers have described angiostatic steroids that markedly inhibit neovascularization of the rabbit cornea when given topically with beta-cyclodextrin tetradecasulfate (beta-CD), yet have minimal or no glucocorticoid or mineralocorticoid activity. Our objective was to extend these observations to another species, the rat. We induced neovascularization by cauterizing rat corneas with silver nitrate/potassium nitrate; drugs were applied topically four times per day for 4 days in most experiments. Submicron sized emulsions of lipid-soluble dexamethasone and the angiostatic steroids 17 alpha-hydroxyprogesterone (1 or 10 mg ml-1) and cortexolone (1 or 10 mg ml-1) were prepared by lecithin encapsulation of drug microcrystals. The vehicle for water-soluble hydrocortisone 21-phosphate (HCP) +/- beta-CD (Molecusol; Pharmatec, Inc) was 10% Tween 20 in Tris-buffered 0.9% saline. Angiogenesis was significantly inhibited only by 1 mg ml-1 dexamethasone (-63.2% when compared with controls), 0.5 mg ml-1 HCP + 1 mg ml-1 beta-CD (-33.4%), and 1 mg ml-1 HCP (-40.2%). HCP (0.5 mg ml-1) or beta-CD (1 or 2 mg ml-1) alone had no significant effect on neovascularization; the inhibition by 1.0 mg ml-1 HCP was not potentiated by 2 mg ml-1 beta-CD. We also tested HCP and tetrahydro-S (TH-S) using 1.5% hydroxypropyl methylcellulose vehicle and beta-CD from Takeda Chemical Industries, Ltd., to simulate the procedure of Folkman and coworkers.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: 17-alpha-Hydroxyprogesterone; Animals; beta-Cyclodextrins; Cornea; Cortodoxone; Cyclodextrins; Dexamethasone; Drug Therapy, Combination; Female; Hydrocortisone; Hydroxyprogesterones; Male; Neovascularization, Pathologic; Nitrates; Potassium Compounds; Rats; Rats, Sprague-Dawley; Silver Nitrate; Steroids

Topics: Acetates; Aged; AIDS Vaccines; American Medical Association; Amines; Animals; Anticonvulsants; beta-Cyclodextrins; Cells, Cultured; Clinical Trials as Topic; Cochlear Implants; Complementary Therapies; Constriction, Pathologic; Coronary Artery Bypass; Cyclodextrins; Cyclohexanecarboxylic Acids; Depressive Disorder; Drug Therapy, Combination; Epilepsy; Extracorporeal Membrane Oxygenation; Felbamate; Fetus; Gabapentin; gamma-Aminobutyric Acid; Genetic Therapy; Health Care Costs; HIV Infections; Humans; Hyperlipoproteinemia Type II; Lamotrigine; Muscle, Smooth; National Institutes of Health (U.S.); Oxygen; Phenylcarbamates; Propylene Glycols; Respiration, Artificial; Respiratory Distress Syndrome; Triazines; United States; Vascular Surgical Procedures

Tetrahydrocortisol, beta-cyclodextrin tetradecasulfate, and minocycline used alone or in combination are not very cytotoxic toward EMT-6 mouse mammary tumor cells growing in monolayer. Tetrahydrocortisol (100 microM, 24 h) and beta-cyclodextrin tetradecasulfate (100 microM, 24 h) protected EMT-6 cells from the cytotoxicity of CDDP, melphalan, 4-hydroperoxycyclophosphamide, BCNU, and X-rays under various conditions of oxygenation and pH. Minocycline (100 microM, 24 h) either had no effect upon or was additive with the antitumor alkylating agents or X-rays in cytotoxic activity toward the EMT-6 cells in culture. The combination of the three modulators either had no effect upon or was to a small degree protective against the cytotoxicity of the antitumor alkylating agents or X-rays. The Lewis lung carcinoma was chosen for primary tumor growth-delay studies and tumor lung-metastases studied. Tetrahydrocortisol and beta-cyclodextrin tetradecasulfate were given in a 1:1 molar ratio by continuous infusion over 14 days, and minocycline was given i.p. over 14 days, from day 4 to day 18 post tumor implantation. The combination of tetrahydrocortisol/beta-cyclodextrin tetradecasulfate diminished the tumor growth delay induced by CDDP and melphalan and produced modest increases in the tumor growth delay produced by cyclophosphamide and radiation. Minocycline co-treatment increased the tumor growth delay produced by CDDP, melphalan, radiation, bleomycin, and, especially cyclophosphamide, where 4 of 12 animals receiving minocycline (14 x 5 mg/kg, days 4-18) and cyclophosphamide (3 x 150 mg/kg, days 7, 9, 11) were long-term survivors. The 3 modulators given in combination produced further increases in tumor growth delay with all of the cytotoxic therapies, and 5 of 12 of the animals treated with the 3-modulator combination and cyclophosphamide were long-term survivors. Although neither tetrahydrocortisol/beta-cyclodextrin tetradecasulfate, minocycline, nor the three modulator combination impacted the number of lung metastases, there was a decrease in the number of large lung metastases. Treatment with the cytotoxic therapies alone reduced the number of lung metastases. Addition of the modulators to treatment with the cytotoxic therapies resulted in a further reduction in the number of lung metastases. These results indicate that agents that inhibit the breakdown of the extracellular matrix can be useful additions to the treatment of solid tumors.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; beta-Cyclodextrins; Cell Division; Cell Hypoxia; Cyclodextrins; Lung Neoplasms; Male; Mammary Neoplasms, Experimental; Mice; Mice, Inbred C57BL; Minocycline; Tetrahydrocortisol; Tumor Cells, Cultured

Heparin inhibits smooth muscle cell proliferation in vitro, a property that makes it potentially useful in preventing restenosis after angioplasty. Its utility in this setting is limited by the inability to use high doses (secondary to anticoagulant effects) and the need for subcutaneous administration. We tested the ability of beta-cyclodextrin tetradecasulfate (CDT), a nonanticoagulant synthetic heparin mimic, to inhibit smooth muscle cell proliferation in vitro and tested its efficacy when orally administered for the prevention of angioplasty restenosis in a rabbit atherosclerosis model. Vascular smooth muscle cells were cultured from rabbit aortas by the explant technique. Passaged cells were plated at low density in microtiter plates in the presence or absence of varying concentrations of heparin or CDT in culture medium containing 10% fetal calf serum. Using both 3H-thymidine incorporation and total protein assays, both heparin and CDT caused a similar dose-dependent inhibition of proliferation. We next tested the effect of orally administered CDT in the prevention of restenosis in focal femoral artery arteriosclerotic lesions created in hypercholesterolemic New Zealand White rabbits by air-dessication endothelial injury and subsequent peripheral angioplasty. Animals were followed up for 1 month and were fed normal chow supplemented by tap water with or without CDT. In animals receiving the highest concentration of CDT (2 mg/mL drinking water), the percentage of arterial cross-sectional area with intimal hyperplasia decreased from 50.5 +/- 1.7% (control) to 26.9 +/- 2.2% (p < 0.001), with the intimal/medial ratio being decreased from 1.4 +/- 0.4 to 0.5 +/- 0.2 (p = 0.056).(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Angioplasty; Animals; Arteriosclerosis; beta-Cyclodextrins; Cell Division; Constriction, Pathologic; Cyclodextrins; Disease Models, Animal; Muscle, Smooth, Vascular; Rabbits; Vascular Diseases

The growth of new blood vessels plays an important role in the pathogenesis of several diseases including cancer, diabetes, and arthritis. Beta-cyclodextrin tetradecasulfate, when administered with an appropriate steroid inhibits angiogenesis, and can stimulate angiogenesis when given alone. The regulation of angiogenesis is not well understood, and the mechanism of action of beta-cyclodextrin tetradecasulfate is similarly not well defined. Ecto-protein kinase activity that utilizes extracellular ATP has recently been reported on several types of cells. Human neutrophils appear to possess two distinct ecto-protein kinase activities; one that phosphorylates exogenous substrates including vitronectin and basic fibroblast growth factor, and one that phosphorylates endogenous cell-surface proteins. This report shows that beta-cyclodextrin tetradecasulfate inhibits the phosphorylation of the exogenous substrates casein, vitronectin (the major ecto-protein kinase substrate in serum), and basic fibroblast growth factor by human neutrophil ecto-protein kinase activity. In contrast, beta-cyclodextrin tetradecasulfate had no effect on the phosphorylation of endogenous cell-surface proteins by the neutrophil ecto-protein kinase activity. Ecto-protein kinase activity that was inhibited by beta-cyclodextrin tetradecasulfate was also detected on porcine aortic and human umbilical vein endothelial cells. The effects of beta-cyclodextrin tetradecasulfate on ecto-protein kinase activities may play a role in its effects on angiogenesis.

Topics: beta-Cyclodextrins; Caseins; Cyclodextrins; Fibroblast Growth Factor 2; Glycoproteins; Humans; Hydrocortisone; Neovascularization, Pathologic; Neutrophils; Phosphorylation; Protein Kinases; Vitronectin

Our goal has been to provide a quantitative analysis of the random motility and chemotaxis of microvessel endothelial cells (MEC) in order to understand the role of these functions in the development of new capillaries. A major difference of our work from previous investigations has been our use of mathematical analysis to interpret experimental results, and to relate in vitro migration measurements to in vivo angiogenesis observations. In this paper we present some of our methods and their rationale, with recent results from both experiment and mathematical models.

Topics: Adipose Tissue; beta-Cyclodextrins; Cell Movement; Cells, Cultured; Chemotaxis; Cyclodextrins; Endothelium, Vascular; Fibroblast Growth Factor 1; Humans; Hydrocortisone; Mathematics; Microcirculation; Models, Cardiovascular; Neovascularization, Pathologic

The formation of a blood supply (angiogenesis) is critical to the growth of solid tumors. The naturally occurring steroid tetrahydrocortisol, the synthetic cyclodextrin derivative beta-cyclodextrin tetradecasulfate, and the tetracycline derivative minocycline have antiangiogenic activity. Tetrahydrocortisol and beta-cyclodextrin tetradecasulfate in a 1:1 molar ratio by continuous infusion over 14 days and minocycline administered i.p. over 14 days from day 4 to day 18 postimplantation of the Lewis lung carcinoma significantly increased the growth delay of the primary tumor after treatment with cis-diamminedichloroplatinum(II), melphalan, cyclophosphamide, Adriamycin, bleomycin, and radiation therapy administered in standard regimens. Addition of the antiangiogenic agents to treatment with the cytotoxic therapies not only reduced the number of lung metastases formed from the primary tumor but also reduced the number of large metastases. Five of 12 animals treated with the antiangiogenic modulators and cyclophosphamide were long-term survivors (> 120 days). Thus, antiangiogenic therapies can potentiate the efficacy of standard anticancer therapies.

Topics: Animals; Antineoplastic Combined Chemotherapy Protocols; beta-Cyclodextrins; Bleomycin; Cisplatin; Cyclodextrins; Doxorubicin; Drug Screening Assays, Antitumor; Drug Synergism; Lung Neoplasms; Mice; Mice, Inbred C57BL; Minocycline; Neoplasm Transplantation; Tetrahydrocortisol; Tumor Cells, Cultured

It is known that hydrocortisone can be converted to a potent angiogenesis inhibitor by coadministration with heparin or with a sulfated cyclodextrin. The activity of tetrahydrocortisol-S, a purely angiostatic corticosteroid, can be potentiated by beta-cyclodextrin tetradecasulfate as shown in this study. This drug "pair" and other pairs of corticosteroids and beta-cyclodextrin tetradecasulfate can be applied topically to inhibit corneal neovascularization. Endotoxin-induced corneal neovascularization in rabbits was treated with beta-cyclodextrin tetradecasulfate coadministered with either: hydrocortisone, tetrahydrocortisol-S, or 6-alpha-fluoro-17,21-dihydroxy-16-beta-methyl-pregna-4,9(11),diene,3, 20-dione. When optimal ratios of steroid and cyclodextrin were used, neovascularization was reduced to 13%, 26%, and 28% of untreated controls for the three steroids, respectively. Hydrocortisone-cyclodextrin drug pairs suppressed virtually all inflammatory cell infiltration (induced by endotoxin), whereas tetrahydrocortisol-cyclodextrin pairs only partially reduced inflammation. These results demonstrate that corneal neovascularization and corneal inflammation are separable processes and that the neovascularization may be treated specifically using angiostatic steroids without inflammatory activity.

Topics: Administration, Topical; Angiogenesis Inducing Agents; Animals; beta-Cyclodextrins; Betamethasone; Corneal Neovascularization; Cortodoxone; Cyclodextrins; Disease Models, Animal; Drug Synergism; Hydrocortisone; Male; Rabbits

Topics: beta-Cyclodextrins; Cyclodextrins; Dose-Response Relationship, Drug; Erythrocytes; Hemoglobins; Hemolysis; Heparin; Humans; Spectrophotometry

Topics: beta-Cyclodextrins; Cell Movement; Cells, Cultured; Cyclodextrins; Endothelium, Vascular; Fibroblast Growth Factor 1; Humans; Hydrocortisone; Microcirculation; Neovascularization, Pathologic

beta-Cyclodextrin tetradecasulfate was found to have a very strong affinity for fibroblast growth factor (FGF) and could substitute for heparin in FGF purification. Basic FGF was purified about 200,000-fold from a rat chondrosarcoma using a method of biaffinity chromatography in which the beta-cyclodextrin tetradecasulfate polymer was mixed with copper-Sepharose. This method takes advantage of the strong affinity of FGF for both beta-cyclodextrin tetradecasulfate and copper.

Topics: Animals; beta-Cyclodextrins; Cells, Cultured; Chromatography, Affinity; Cyclodextrins; Dextrins; Electrophoresis, Polyacrylamide Gel; Fibroblast Growth Factors; Humans; Mice; Mice, Inbred BALB C; Protein Binding; Rats; Recombinant Proteins; Starch