Delivery of cytokines by liposomes. III. Liposome-encapsulated GM-CSF and TNF-alpha show improved pharmacokinetics and biological activity and reduced toxicity in mice.

In an attempt to potentiate the therapeutic index of cytokines, recombinant mouse granulocyte/macrophage colony-stimulating factor (GM-CSF) and recombinant human tumor necrosis factor alpha (TNF-alpha) were encapsulated in large (0.3-2.2 microns) multilamellar vesicles composed of various lipids, using several encapsulation methods. Liposomal cytokine activity was tested in vitro and in vivo and was compared with that of the soluble cytokines. The main observations were as follows: (a) The mean encapsulation efficiency, as determined by bioassays, was 49-79%, depending on the formulation, for GM-CSF, and 48% for TNF-alpha; (b) some of the entrapped cytokine preparations displayed high stability at 4 degrees C, with < 30% loss of biologic activity during a 4-month period; (c) release of TNF-alpha, but not of GM-CSF, from the liposomes was required for their biological activity in vitro; (d) plasma half-lives (t1/2 alpha, t1/2 beta) and the area under the curve (AUC) of the entrapped cytokines were 10-20 times greater than those of the soluble cytokines; (e) the toxicity of liposomal TNF-alpha was one-third and one-seventh that of soluble TNF-alpha in normal and tumor-bearing mice, respectively; (f) administration of liposomal GM-CSF (5 x 10(4)-2 x 10(5) U, one to five times) to normal and 5-fluorouracil-treated mice led to a two- to fourfold increase in the absolute number of peritoneal and spleen leukocytes and of GM colony-forming cells in the spleen, as compared with the levels obtained using soluble GM-CSF; and (g) under the experimental conditions used, neither free nor liposomal GM-CSF significantly increased the absolute number of blood leukocytes, although liposomal GM-CSF markedly (threefold) enhanced the level of blood granulocytes. Collectively, these findings suggest that liposome-entrapped GM-CSF and TNF-alpha may be more efficacious immunomodulators than the soluble cytokines.

[Treatment experience with male hypogonadism patients]. / Opyt lecheniia bol'nykh muzhskim gipogonadizmom.

A noticeable masculinizing effect with a high level of testosterone (T) in the blood up to its level in healthy males was obtained during treatment of 10 patients with primary (hypergonadotropic) hypogonadism using sustanon-250 at a dose of 1 ml i. m. once in 3-4 weeks. However no decrease in the initially elevated levels of the follicle-stimulating and luteinizing hormones was observed. The treatment of 21 patients with hypogonadotropic hypogonadism by small doses of chorionic gonadotropin (CG) (up to 1500 Units i. m. 3 times a weeks) turned out ineffective. The use of high doses of CG (3000-4000 units i. m. 3 times a week) or CG combined with menopause gonadotropin in most patients of this group produced a short-term clinical effect with an elevated T level in the blood. Hypogonadotropic hypogonadism combined with an acute decrease in the response of the testes to CG treatment even at large doses was found in 4 patients under 6 years with cryptorchidism who had undergone surgery for descent of the testes into the scrotum.