Janus kinase 3: a novel target for selective transplant immunosupression

Existing immunosuppressants inhibit lymphocyte activation and T cell cytokine signal transduction pathways, reducing the rate of acute rejection episodes to < 10%. However, the widespread tissue distribution of their molecular targets engenders pleiotropic toxicities. One strategy to address this problem seeks to identify compounds that selectively inhibit a target restricted in distribution to the lymphoid system. Janus kinase (Jak) 3 is such a molecule; it mediates signal transduction via the gamma common chain of lymphokine surface receptors. Disruption of this lymphoid-restricted enzyme would not be predicted to produce collateral damage in other organ systems. Development of selective Jak3 inhibitors has been difficult due to crossreactivity with its homologue, Jak2. In contrast to all other putative antagonists, which are discussed in detail herein, one Jak3 inhibitor, NC1153, shows at least 40-fold greater selective inhibition for Jak3 than for Jak2, is robustly synergistic with calcineurin antagonists, and, either alone or in combination with cyclosporin, produces no adverse effects in rodents preconditioned to be at heightened risk for nephrotoxicity, bone marrow suppression, or altered lipid metabolism.     

Physician adherence to recommendations for duration of empiric antibiotic treatment for uncomplicated urinary tract infection in women: a national drug utilization analysis

BACKGROUND: Current guidelines for the empiric treatment of uncomplicated urinary tract infection in women recommend that first-line trimethoprim-sulfamethoxazole (TMP-SMX) or ofloxacin be given for 3 days and nitrofurantoin for 5 days. Increasing the duration of treatment raises costs, and perhaps, the incidence of adverse effects, without contributing to effectiveness. The aim of this study was to investigate physician adherence to these recommendations. METHODS: The electronic patients record system of a nationwide health management organization in Israel was reviewed for all primary care visits by adult women treated empirically for cystitis or urinary tract infection from January 2001 to June 2002 (n = 7738 patient-physician encounters). The proportion of cases treated according to the guidelines, with regard to duration, was calculated for each drug used. RESULTS: Rate of adherence was 3.36% for cases of TMP-SMX treatment (95%CI: 2.56%, 4.15%), 22.23% for nitrofurantoin (95%CI: 19.81%, 24.65%) and 4.08% for ofloxacin (95%CI: 2.88%, 5.28%). The crude rate of adherence for all cases of treatment with these drugs was 8.67% (95%CI: 7.82%, 9.52%). CONCLUSIONS: The high rate of nonadherence observed (91.33%) indicate a need for a remedial education program for physicians to improve empiric treatment of urinary tract infection in women. Since this issue is of global importance, we believe our evaluation can serve as model for other settings and countries.     

Prevention of renal ischemic reperfusion injury using FTY 720 and ICAM-1 antisense oligonucleotides

BACKGROUND: Renal damage secondary to ischemia-reperfusion injuries (I-R) is frequent in organ transplantation and adversely affects the graft survival. An important component of this damage is caused by initial adhesion of neutrophils and lymphocytes to endothelial cells. FTY 720, which induces lymphopenia, has previously been shown to display protective effects in models of I-R. The purpose of the present study was to evaluate the combination of FTY 720 and intracellular adhesion molecule and ICAM-1 antisense oligonucleotides (AS-oligos), an agent designed to block the adhesion process. METHODS: Sprague-Dawley rats underwent syngenic kidney transplantation after donor kidneys had been preserved in cold solution for 2 hours. The treatment groups included: (1) FTY 720 (1 mg/kg) before reperfusion, (2) AS-oligos (2 mg/kg) during kidney perfusion, and (3) the combination of FTY 720 and AS-oligos. All animals were followed daily after transplantation; some were sacrificed on the second day for histologic analysis. RESULTS: All treated groups showed a maximal serum creatinine that was significantly less than the control (group 1: 2.76 +/- 1.4, group 2: 2.44 +/- 2.05, group 3: 1.51 +/- 0.42, and control: 4.04 +/- 0.5; P <.01) and returned to the basal value earlier. Also, treated animals showed less histologic stigmata of acute tubular damage. FTY 720 and AS-oligos used in combination showed a mild additive effect. CONCLUSIONS: The use of FTY 720 and/or AS-oligos significantly prevents functional renal damage secondary to I-R, displaying a mild additive effect in this model. Both agents offer the advantage of use during the donor and the graft operations.     

A microarray minisequencing system for pharmacogenetic profiling of antihypertensive drug response

We aimed to develop a microarray genotyping system for multiplex analysis of a panel of single nucleotide polymorphisms (SNPs) in genes encoding proteins involved in blood pressure regulation, and to apply this system in a pilot study demonstrating its feasibility in the pharmacogenetics of hypertension. A panel of 74 SNPs in 25 genes involved in blood pressure regulation was selected from the SNP databases, and genotyped in DNA samples of 97 hypertensive patients. The patients had been randomized to double-blind treatment with either the angiotensin II type 1 receptor blocker irbesartan or the beta 1-adrenergic receptor blocker atenolol. Genotyping was performed using a microarray based DNA polymerase assisted 'minisequencing' single nucleotide primer extension assay with fluorescence detection. The observed genotypes were related to the blood pressure reduction using stepwise multiple regression analysis. The allele frequencies of the selected SNPs were determined in the Swedish population. The established microarray-based genotyping system was validated and allowed unequivocal multiplex genotyping of the panel of 74 SNPs in every patient. Almost 7200 SNP genotypes were generated in the study. Profiles of four or five SNP-genotypes that may be useful as predictors of blood pressure reduction after antihypertensive treatment were identified. Our results highlight the potential of microarray-based technology for SNP genotyping in pharmacogenetics.     

Long-term effects of irbesartan and atenolol on the renin-angiotensin-aldosterone system in human primary hypertension: the Swedish Irbesartan Left Ventricular Hypertrophy Investigation versus Atenolol (SILVHIA)

We examined long-term influence of the angiotensin II type 1-receptor blocker irbesartan and the beta1-adrenergic receptor blocker atenolol on some neurohormonal systems implicated in the pathophysiology of cardiac hypertrophy. Thus, 115 hypertensive patients with left ventricular hypertrophy were randomized to receive double-blind irbesartan or atenolol, with additional therapy if needed. Neurohormone measurements and echocardiography were performed at weeks 0, 12, 24, and 48. Left ventricular mass was reduced more by irbesartan than by atenolol (-26 g/m2 versus -14 g/m2, P = 0.024), despite similar reductions in blood pressure. Plasma renin activity and angiotensin II increased (P < 0.001) by irbesartan (0.9 +/- 0.7 to 3.4 +/- 4.2 ng/mL x h, and 3.0 +/- 1.6 to 13.0 +/- 17.7 pmol/L), but decreased (P < 0.01) by atenolol (1.0 +/- 0.6 to 0.7 +/- 0.6 ng/mL x h, and 3.4 +/- 1.6 to 3.2 +/- 2.2 pmol/L). Serum aldosterone decreased (P < 0.05) by both irbesartan (346 +/- 140 to 325 +/- 87 pmol/L) and atenolol (315 +/- 115 to 283 +/- 77 pmol/L). Changes in left ventricular mass by irbesartan related inversely to changes in plasma renin activity, angiotensin II, and aldosterone (all P < 0.05). Plasma levels and 24-hour urinary excretions of catecholamines, plasma leptin, proinsulin, insulin and insulin sensitivity remained largely unchanged in both groups. Thus, the renin-angiotensin aldosterone system appears to be an important non-hemodynamic factor in the regulation of left ventricular mass.     

Comparison of blood pressure measurements on the bare arm, below a rolled-up sleeve, or over a sleeve

BACKGROUND: This study examined the effect of measuring blood pressure below subjects' rolled-up sleeves, over the sleeve, or on the bare arm. This is an important day-to-day issue for the busy GP. METHODS: The sample consisted of 201 subjects in family practice clinics and residents of a senior citizens' home. A digital device was used in all cases. Each participant underwent three blood pressure measurements in each of the following conditions in random order: cuff on bare arm; cuff over the sleeve; and cuff below the rolled-up sleeve. Differences between measurements were plotted against the mean blood pressure. Confounding factors controlled for were age, sex, clothing pressure and skin-fold thickness. RESULTS: Differences in mean blood pressure readings between the clothed and bare arm were 0.5 mmHg (SD 7.5) for systolic pressure and 1 mmHg (SD 5) for diastolic pressure; neither difference was significant. However, in hypertensive subjects (>140 mmHg systolic), although the mean difference remained small (systolic pressure, 2 mmHg, SD 10), the range of difference for individual subjects was -32 mmHg to +22 mmHg. CONCLUSION: The degree of clothing under the sphygmomanometer cuff does not have a clinically important effect on the blood pressure measurement. In patients known or found to be hypertensive, measurement on the bare arm is recommended.     

Thymoglobulin,sirolimus, and reduced-dose cyclosporine provides excellent rejection prophylaxis for pancreas transplantation

BACKGROUND: We investigated a novel immunosuppressive protocol including thymoglobulin induction in combination with sirolimus and corticosteroids, followed by introduction of markedly reduced exposures to cyclosporine to prevent pancreas-transplant rejection. METHODS: A 7-day course of thymoglobulin (1.5 mg/kg per day) was begun on postoperative day (POD) 0, together with 15 mg of sirolimus on POD 1, and followed by 5 mg per day, targeting these doses to achieve a trough of 10 to 20 ng/mL. When the serum creatinine was below 2.5 mg/dL, cyclosporine was introduced at 50 mg twice daily with dose adjustment to maintain a trough concentration of 100 to 125 ng/mL. RESULTS: The 18 patients included 14 simultaneous pancreas-kidney and 4 pancreas-after-kidney transplant recipients. Two patients were African-American, three patients had a pretransplant panel reactive antibody greater than 20%, and the human leukocyte antigen (HLA) mismatch was 4.5+/-1 (mean+/-standard deviation). With a mean follow-up of 13.6+/-4.7 months, patient, kidney, and pancreas graft survivals are 100%, 100%, and 94%, respectively. A single pancreas graft was lost to thrombosis. There were no acute rejection episodes and no opportunistic infections. Mean hospital stay was 9+/-3 days. At 3 months posttransplantation, the mean value of serum creatinine was 1.2+/-0.3 mg/dL, fasting glucose was 88+/-15 mg/dL, and sirolimus dose at month 3 was 6.3+/-3 mg per day and at month 12 2.7+/-1 mg per day. The average total daily cyclosporine A dose at month 3 was 208+/-62 mg per day and 133+/-13 mg per day at 1 year. CONCLUSIONS: This immunosuppressive regimen provided excellent prophylaxis against acute rejection with no opportunistic infections. We believe that careful monitoring of sirolimus and cyclosporine levels was critical to success of this protocol.     

Pharmacodynamics, pharmacokinetics, and safety of multiple doses of FTY720 in stable renal transplant patients: a multicenter, randomized, placebo-controlled, phase I study

BACKGROUND: FTY720, a novel immunomodulator, displays potent immunosuppressive activity in a variety of preclinical transplant models. This study examined the safety, pharmacodynamics, and pharmacokinetics of multiple doses of FTY720 in stable renal transplant patients. METHODS: This randomized, multicenter, double-blind, placebo-controlled, phase I study included adults who had been maintained on a regimen of cyclosporine A (CsA) microemulsion and prednisone (or its equivalent) for at least 1 year after renal transplantation. Patients received once-daily doses of 0.125, 0.25, 0.5, 1.0, 2.5, or 5.0 mg FTY720, or placebo for 28 days. After completion of study drug administration, the patients were monitored until day 56 by serial laboratory tests, clinical examinations, and recording of adverse events. The study includes 76 treatment courses (61 FTY720 and 15 placebo), with 65 patients enrolled once and 11 reenrolled. RESULTS: FTY720 doses greater than or equal to 1.0 mg/day produced a significant reduction in peripheral blood lymphocyte count by up to 85%, which reversed within 3 days after discontinuation of study medication. Compared with placebo-treated patients, FTY720 subjects did not show a major increase in adverse events or a change in renal function. Pharmacokinetic measurements revealed that FTY720 displayed linear relations of doses and concentrations over a wide range, but had no effect on CsA exposure. CONCLUSIONS: At doses up to 5.0 mg/day for 28 days, stable renal transplant patients treated with FTY720 in combination with CsA and prednisone displayed a dose-dependent, reversible decline in peripheral blood lymphocytes without an enhanced incidence of collateral toxicities, except possibly bradycardia.