An insulin derivative according to claim 4, wherein one or more amino groups are substituted by the Fmoc radical and one or more carboxyl groups are substituted by the Fm radical herein N- Fmoc , C- Fm -insulin. Human insulin is now widely available and in theory it is expected that it is less immunogenic since the last distinction as porcine or bovine insulins, both of human insulin by one and three amino acids. Lipogenesis was terminated by adding toluene-based scintillation fluid 1. Circulating glucose levels in Group B were significantly lower. To test the effect of Phe Lys BBN- Fmoc 2-insulin in lowering blood glucose levels in experimental diabetic rats, STZ-treated rats, 9 days after induction of diabetes, received either a single s. Phosphotyrosine content in PolyGlu4Tyr was quantitated by a radio-immunoassay procedure using specific monoclonal antibodies to phosphotyrosine final dilution 1: , and 12 BSA-phosphotyrosine conjugate.
Bi B29 Fig. Unlike regular insulin have Ultralente insulins to a very slow onset and a prolonged "flat" peak of action. Rapid-acting soluble insulin is effective in lowering blood glucose levels in this experimental system over a period of several hours only not shown. For traditional and therapeutic purposes, doses and concentrations of insulin are in units U words, which are based on the amount that is required to induce normoglycemia in fasting rabbits. In electrophilic substitution reactions, fluorene is first attacked at the 2 position and, generally, the nature of the substituent at position 9 CH20CO-OSu has no effect on the orientation of the substitution. The new concept of the invention for slow-release drugs includes their derivatization a new, more hydrophobic in general, drug derivatives.
Virtually all peptidic drugs and protein drugs do not follow due to their intrinsic hydrophilic non-lipophilic and polar properties and metabolic instability these criteria and must usually be administered by injection.
Further, as these molecules are rapidly degraded in the body by diverse mechanisms, notably proteolysis, they are usually short-living species. Unlike regular insulin have Ultralente insulins to a very slow onset and a prolonged "flat" peak of action. The invention further relates to pharmaceutical compositions comprising a prodrug according to the invention and a pharmaceutically acceptable carrier. Non-peptidic drugs, such as antibiotics, although relatively long-lived, have to be administered several times a day over a period of a week, or longer, to maintain the desired continuous circulating levels. The pharmaceutical composition may be presented in any suitable form, for example as an oral formulation or for subcutaneous injection. The resulting crude solid 95 mg was utilized further without any purification.
Thus, a single administration of N- Fmoc 3-insulin gave prolonged and satisfactory antidiabetic actions lasting over a period of four days following a delayed onset of approximately two days.
This long-lasting effect in vivo may be explained by the ability of the derivative to escape receptor-mediated endocytosis and also by its resistance to proteolysis. At the indicated time points aliquots were withdrawn for determining the biological potency relative to native insulin. If rapid-acting and long or intermediate- acting insulin preparations are coadministered together, a common disadvantage of such combinations is that, upon mixing, some of the rapid-acting insulins can be complexed with excess Zn 2 or protamine of the long or intermediate-acting preparation, being thus converted into an intermediate and even long-acting insulin. Preparation of N-Fmoc-insulin derivatives Synthesis Human insulin mg, So often a stabilization of the biologically active structure concomitantly with the desire to improve the metabolic stability, been tried.
To evaluate the long-acting capacity of Sulfmoc 2-insulin, downstream to subcutaneous absorption, normal rats were received a single intraperitoneal injection of native insulin, NPH-insulin, or Sulfmoc 2-insulin. Precipitation with dry ether afforded 62 mg of N- Fmoc 3, C- Fm n-insulin. The reaction mixture, which became clear after 1 h, was stirred overnight at room temperature, during which it turned turbid. An insulin derivative according to claim 4, wherein one or more amino and hydroxyl groups are substituted by the Fmoc radical and one or more carboxyl groups are substituted by the Fm radical herein N,O- Fmoc , C- Fm -insulin. They are suspensions of tiny Zinkinsulinteilchen and only differ in particle size, which determines the duration of their action.
The powder was dried in vacuum over P and then treated for 1 h at room temperature with 5 ml of trifluoroacetic acid to effect removal of N-terminal t-Boc protecting groups. Two preparations most frequently used are NPH Insulin NPH stands for Neutral Protamine Hagedom , a suspension of zinc-insulin crystals in phosphate buffer modified by the addition of protamine sulfate, and Lente Insulin, a suspension of insulin in acetate buffer modified by the addition of zinc chloride to minimize solubility of insulin. If necessary, the treatment with the Fmoc-insulin derivative is completed by daily administration of rapid-acting insulin. On the other hand it was shown that Fmoc-amino acids, including Fmoc-leucine, a low toxicity index in experimental animal models have Burch et al. The monomodified N-Fmoc-insulin derivatives can be separated from the di and tri modified derivatives by washing the crude solid with isopropanol. However, when highly purified, all three insulins have low but measurable capacity to stimulate the immune response.
At present, insulin is the predominant drug for diabetes mellitus, a group of syndromes characterized by hyperglycemia, altered metabolism of lipids, carbohydrates and proteins and an increased risk of complications from vascular diseases. Gennaro, ed.
Main efforts are currently focused on those goals. Da erwartet wird, dass kurz-wirkendes Insulin einen Zeitraum von 0,4—7 Stdn.