Hepatic Uptake Clearance Research Articles

Synthesis and biological evaluation of gallium polyaminothiols (PAT)

Two polyaminothiol ligands, one hexadentate, N,N′,N″-tris[2-methyl(2-propanethiol)]-1,4,7-triazacyclononane (TACN), and another potentially heptadentate, tris[2-methyl(2-propanethiol)]aminoethylamine (TMAE), were synthesized and characterized using spectroscopic and analytical methods. Both ligands were labeled with gallium-67 at pH 3.0–3.5 in high yields. The resulting gallium chelates were lipophilic. The biodistribution studies for both the chelates showed hepatic uptake and biliary clearance. The total % ID activity for [ 67Ga]TACN in liver and intestine at 5 min was 33.86 increasing gradually to 61.4% at 30 min. The chelates were in vivo stable to plasma-transferrin transchelation as confirmed by scintigraphic images obtained by [ 67Ga]TACN in rats and by plasma incubation of the chelates. These results indicate that Ga-TACN is a potentially useful tracer for hepatobiliary imaging using PET. No brain uptake was found.

Distinction between the depletion of opsonins and the saturation of uptake in the dose-dependent hepatic uptake of liposomes.

Opsonins play a role in the hepatic uptake of particles such as bacteria, lipid emulsion, and liposomes. The objective of this study was to distinguish between opsonin depletion and uptake saturation in the dose-dependent hepatic uptake of liposomes. The uptake of opsonized and unopsonized liposomes was determined in the isolated perfused liver. Serum (2.9 mL) was required to opsonize 1 mumol liposomes fully, indicating that a rat (250 g with 10 mL of serum) can opsonize 3.5 mumol liposomes. Next the dose effect on hepatic uptake of opsonized and unopsonized liposomes was examined. Saturation of uptake was found only for the opsonized liposomes. On the other hand, the hepatic uptake clearance decreased dose dependently from 4.31 to 0.79 (mL/min), with increasing doses from 0.075 to 17 mumol/250 g, respectively, after i.v. administration. Thus, the decrease in the hepatic uptake clearance at the medium dose was due to the saturation of uptake alone, and at the high dose it was due to opsonin depletion as well. These results show that the saturation of liposomal uptake in the liver and the depletion of opsonins occurred at different liposome dosage levels.

KINETIC ANALYSIS OF LIPOSOME UPTAKE BY RES IN COMPARISON TO THEIR DEGRADATION IN BLOOD AND RES

Kinetic analysis of the hepatic uptake of liposomes was performed in comparison to the degradation of liposomes in blood and liver. The degradation clearance of liposomes in blood was shown to depend on the size of liposomes. In vitro kinetic study on the degradation of liposomes suggested the mechanism of this size dependency due to the Increased affinity of liposomes for complement. There was two kinds of hepatic uptake components of liposomes, one is a size dependent opsonized pathway, and the other is a size Independent unopsonized pathway. The size dependent opsonization of liposomes was shown to result from the size dependent complement activation and this component explained the size dependended hepatic uptake clearance of liposomes in vivo. The kinetic analysis of the hepatic degradation of liposomes was also performed by labeling liposomes with both 125I-albumin and 3H-cholesterylhexadecylether. Although the mechanism of hepatic uptake are considered to be different depending on the size of liposomes, the kinetics of hepatic degradation of liposomes was same for each size of liposomes and lay in the order of hour.

Nonlinear pharmacokinetics of hepatobiliary transport of rose bengal in rats after i.v. bolus administration with varying doses.

To investigate the nonlinear kinetics in the hepatobiliary transport of rose bengal (RB), the time profiles of plasma concentration and biliary excretion rate after its i.v. administration at various doses were measured in rats. The total body clearance decreased remarkably with increased dose. The hepatic uptake clearance also showed a similar dose dependency, and saturation of hepatic uptake at least partly accounts for the dose-dependent change in total body clearance. The peak biliary excretion rate approached the transport maximum (approximately 150 nmol min-1 kg-1) with increased dose. To further clarify which process in RB hepatobiliary transport has nonlinearity, we analysed thus obtained data based on a three-compartment model. The hepatic uptake and sequestration rate constants decreased remarkably with increased dose. The initial hepatic uptake rates assessed from the plasma disappearance rate during the early phase fit well to the Michaelis-Menten equation with a saturable and a nonsaturable component. The maximum uptake velocity and Michaelis constant were 4.7 mumol min-1 kg-1 and 360 microM, respectively. That hepatic uptake has a much higher capacity (about 30 fold) than biliary excretion suggests that biliary excretion can be a rate-determining process in the overall hepatobiliary transport of RB. We conclude that the saturation of both hepatic uptake and biliary excretion could be the main causes for the nonlinear pharmacokinetics of hepatobiliary transport of RB.

Disposition characteristics of macromolecules in tumor-bearing mice.

As part of the strategy for the design of macromolecular carriers for drug targeting, the disposition characteristics of macromolecules were studied in mice bearing tumors that served as target tissues. Eight kinds of macromolecules including four polysaccharides and four proteins with different molecular weights and electric charges were used; tissue distribution and tumor localization after intravenous injection were studied. Pharmacokinetic analysis revealed that the tissue radioactivity uptake rate index calculated in terms of clearance was different among the tested compounds; especially, the urinary radioactivity excretion clearances and the total hepatic radioactivity uptake clearances varied widely. Compounds with low molecular weights (approximately 10 kD) or positive charges showed lower tumor radioactivity accumulation; radioactivity was rapidly eliminated from the plasma via rapid urinary excretion or extensive hepatic uptake, respectively. On the other hand, large and negatively charged compounds, carboxymethyl dextran, bovine serum albumin, and mouse immunoglobulin G, showed higher radioactivity accumulation in the tumor (calculated total amounts were 15.6, 10.8, and 20.8% of the dose, respectively) and prolonged retention in the circulation. These results demonstrated that the total systemic exposure rather than the uptake rate index was correlated with total tumor uptake. Molecular weight and electric charge of the macromolecules significantly affected their disposition characteristics and, consequently, determined radioactivity accumulation in the tumor. It was concluded that a drug-carrier complex designed for systemic tumor targeting should be polyanionic in nature and larger than 70,000 in molecular weight.

Macromolecule-macromolecule interaction in drug distribution: Effect of .ALPHA.-globulin concentration on the hepatic uptake of fractionated 3H-heparin by perfused rat liver.

The effect of alpha-globulin, the dominant binding protein for fractionated 3H-heparin, on the hepatic uptake of 3H-heparin was studied by liver perfusion experiments in rats. Fractionated 3H-heparin concentration in the recirculated perfusate decline one-exponentially with time for each of six initial concentration levels of alpha-globulin. The hepatic uptake clearance of fractionated 3H-heparin was 0.154 ml/min/g liver in the absence of alpha-globulin, and it decreased with increasing alpha-globulin concentrations. This result indicates that the hepatic uptake rate of alpha-globulin-bound fractionated 3H-heparin is lower than that of unbound fractionated 3H-heparin. On the other hand, it was indicated that almost all fractionated 3H-heparin binds to alpha-globulin at 8 mg/ml of alpha-globulin in in vitro study. However, the hepatic uptake clearance of the heparin at the concentration was of a certain value that could not be ignored. It was suggested that alpha-globulin-bound fractionated 3H-heparin also contributed to the hepatic uptake of fractionated 3H-heparin. Therefore, a protein-mediated transport system, which has been reported for some low molecular weight drugs, may also exist in the hepatic uptake of such a high molecular weight compound as fractionated 3H-heparin.

Importance of glycosylation for hepatic clearance of renal renin.

Three differently glycosylated forms of renin (renin A, B-1, and B-2) were highly purified from rat kidneys by pepstatin-aminohexyl-Sepharose affinity chromatography and by serial lectin affinity chromatography on concanavalin A (con A) and lentil lectin-Sepharose, and the role of glycosylation of renin was investigated. Renin A and renin B-1 were loosely and tightly bound to con A, respectively, but did not bind to lentil lectin. Renin B-2 bound to both con A and lentil lectin. These three forms of renin were all similar in their physicochemical characteristics, including molecular weight, isoelectric point, specific activity, Km, optimum pH, and antigenicity. Each form of renin, labeled with 125I and given intravenously to anesthetized rats, disappeared from the circulation at different rates (metabolic clearance rates of 5.05 +/- 1.02, 17.1 +/- 2.5, and 36.0 +/- 4.1 ml.min-1.kg-1 for renins A, B-1, and B-2, respectively). Labeled renin A distributed to a similar extent in the liver and kidney (21.2 +/- 0.2 and 15.2 +/- 0.8% of the injected dose, respectively), whereas renins B-1 and B-2 were distributed predominantly in the liver (56.3 +/- 1.2 and 72.3 +/- 3.7% of the injected dose, respectively) and to a lesser extent in the kidney (4.3 +/- 0.3 and 2.1 +/- 0.2%, respectively). Deglycosylation of renin B-1 with endoglycosidase F resulted in no loss of its enzymatic activity or antigenicity but greatly reduced the metabolic clearance rate to 18% (from 17.1 +/- 2.5 to 3.09 +/- 0.17 ml.min-1.kg-1). Deglycosylation of renin B-1 greatly decreased its uptake by the liver (from 56.3 +/- 1.2 to 3.3 +/- 0.2%) and increased its uptake by the kidney (from 4.3 +/- 0.3 to 23.9 +/- 0.9%). These studies indicate the importance of glycosylation of renin for its hepatic uptake and metabolic clearance rate.

Cyclotron isotopes and radiopharmaceuticals—XXXIII Synthesis and structural effects of selective biliary excretion of halogenated indotricarbocyanines

Fine indotricarbocyanines of structure I were synthesized and labeled with 131I in the 5 position. The substituent, R, was varied ▪ R  H, F, Cl, Br and I. The dynamics of hepatic uptake and blood clearance of the labeled compounds were determined in mice. Kidney uptake in all cases was negligible. The hepatic excretion displayed 2 components. The initial hepatic disappearance rates of R  H and R  F were 7.3 and 4.5%/min, respectively. The maximum liver activity in mice occured at 2–5 min and the % remaining in the liver at 0.5 h was 3.6, 8.5, 19, 29 and 47% for R  H, F, Cl, Br and I substitution. It is notable that a small change in the substituent at the 5′ position in the molecule has such a pronounced effect. Whether electronic and/or steric effects are controlling the mechanism of hepatobiliary clearance is not obvious. A correlation of the liver activity with the covalent radius of R was noted. The comparative studies were at an injected dose of 0.4–0.6 μmol/kg, and typically at a specific activity of 200 mCi/mol. Loading dose effects were not appreciable under these conditions. Scintigraphic results are reported for dogs and a rabbit. The results suggest that indotricarbocyanines of structure (I) labeled with 123I or with 18F are potential radiopharmaceuticals for dynamic hepatobiliary function studies.

HEPATIC UPTAKE AND BLOOD CLEARANCE OF FREE CO57B12 AND OF CO57B12 BOUND TO GASTRIC JUICE AFTER INTRAVENOUS INJECTION IN MAN.

In 9 normal subjects, hepatic uptake and blood clearance of radioactivity were measured for 7–10 days after the intravenous administration of a tracer dose of Co57B12 alone. In 5 of these subjects, the measurements were repeated after intravenous administration of Co57B12 bound to normal human gastric juice (NHGJ). Administration of B12 bound to NHGJ resulted in an augmented hepatic uptake of the vitamin at 1 hr. significantly greater than that for B12 alone. This difference became less marked with time and was not significant at the end of 1 week. Blood clearance during the first 24 hr. following administration was significantly slower for NHGJ-bound B12 than for B12 injected alone.