DE4336641A1 - Use of superoxide dismutase (SOD) for producing pharmaceuticals with an antiviral action - Google Patents

Abstract

The invention relates to the use of superoxide dismutase (SOD), of coupled superoxide dismutase and of superoxide dismutase packaged in liposomes for the production of pharmaceuticals with an antiviral action, in particular of pharmaceuticals for treatment of HIV infections.

Description

The invention relates to the use of superoxide dis mutase (SOD) of liposomally packaged superoxide dismutase and of coupled superoxide dismutase for the production of Medicines with an antiviral effect, in particular from Medicines used to treat HIV infection.

Superoxide dismutases (EC 1.15.1.1) are found in all aerobes occurring metal proteins in the organism for the Ent toxic oxygen species are responsible, especially the "super oxide" (O₂⁻). They catalyze that Disproportionation of the highly reactive and destructive Radical ions superoxide in hydrogen peroxide and Oxygen: 2O₂⁻ + 2H⁺ SOD H₂O₂ + O₂.

SOD from bovine erythrocytes (MG 31300) contains in its two identical subunits each 151 amino acid residues known sequence and a copper and a zinc atom and is very stable against denaturation. Various such Cu₂Zn₂-SOD can be found in almost all eukaryon tissues ten.

Manganese and iron SOD occurs in prokaryotes and plants (MG approx.40,000) and in liver mitochondria (MG 80,000) in front.  

SOD isolated from bovine blood or genetically engineered (e.g. Peroxinorm ®, Grünenthal) is used as an anti-rheumatic used cumulatively. Also anti-inflammatory properties of SOD are described (DE-OS 39 12 642 A1).

Surprisingly, it has now been found that SOD, liposomal packaged SOD and coupled SOD for therapy and pro prevention of infections caused by DNA viruses such as e.g. B. the herpes simplex virus, the cytomegalovirus, Pa pilloma viruses or the Epstein-Barr virus or in particular by retroviruses such as the oncoviruses HTLV-I and II and the Lenti virus Visna and human immunodeficiency virus HIV-I or -II are caused.

Isolates or genes isolated from bovine blood are particularly suitable technically manufactured SOD (MG approx. 32,000) for treatment the clinical manifestations of retroviral HIV infection tion in humans such as the advanced stage of AIDS-related complex (ARC) and the full clinical picture of AIDS.

Surprisingly, SOD can specifically inhibit the NF-κB-dependent virus expression the multiplication of Affect retroviruses. Of special therapeutic Interest is the inhibition of virus production in HIV infected cells, also in AZT-resistant cell lines.

Only 3′-azido-3′- deoxythymidine (DE-A 36 08 606) approved in AIDS patients sen. However, toxic side effects of AZT make up for that Bone marrow in about fifty percent of those treated Patients need blood transfusions.

SOD, liposomally packaged SOD and coupled SOD are effective tiviral without cyto in pharmacologically relevant doses  to be toxic. They also replace missing superoxide dismutase activity in HIV-infected cells. The invent Compounds according to the invention have an anti-AZT effect viral to both chronically and acutely infected Cells.

Good antiretroviral activity has been particularly noted in coupled SOD detected. Coupled SOD means in Sense of the invention that SOD is coupled to substances, which prolong the half-life in blood plasma. Come here in particular polyethylene glycol (PEG) coupled SOD, acylated SOD, glycosylated SOD (e.g. extracellular SOD EC type A, B, C) and protein-modified SOD in question, in particular is also low-density lipoprotein (LDL) or coupled to oxidized low-density lipoprotein (ox-LDL) SOD works well.

Liposomally packaged SOD is also extremely effective incorporated into the liposomes in the usual way becomes.

The drugs containing SOD, liposomally packaged SOD and coupled SOD are produced in the usual way by using the SOD with usual carrier and / or Formulated excipients and tablets, capsules, coated tablets, Produces syrups, solutions or suspensions.

Embodiments example 1 Manufacture of SOD coupled to PEG (PEG-SOD)

PEG-4 is with 10 molar excess of CDI (1,1'-carbonyl diimidazole) in dioxane for 2 hours, against H₂O dialyzed at 4 ° C and the CDI-PEG thus contained lyophilized. 1 µM SOD with 40 mM CDI-PEG in 10 mM Borate buffer, pH 8.5 implemented for 72 hours. Yield: 40% PEG-SOD.

Example 2 Production of SOD packaged liposomally with LDL

100 µl liposomes from 1: 1 (w / w) N- {1 (2, 3- dioleyloxy) propyl} -n, n, n-trimethylammonium chloride and dioleylphosphatidylethanolamine with 100 ul a 5 mM Solution of SOD in H₂O shaken for 24 hours at 25 ° C. The incorporation is 40%.

Example 3 Production of an oxidized low-density lipoprotein (ox- LDL) coupled SOD

1 mM LDL are 24 hours with 5 µM CuSO₄ in H₂O at 25 ° C. incubated. The Ox-LDL solution comes with equimolar amounts Incubated glutaraldehyde and SOD for 12 hours. Not vice versa set GA, GA coupled SOD and GA coupled Ox-LDL are separated by gel filtration.

Claims (8)

1. Use of superoxide dismutase (SOD) for Manufacture of antiviral drugs. 2. Use according to claim 1, characterized in that native or genetically engineered SOD used becomes. 3. Use according to one of claims 1 or 2, characterized characterized in that coupled SOD is used, the is coupled to substances that the half-life in Prolong blood plasma. 4. Use according to claim 3, characterized in that SOD coupled to polyethylene glycol (PEG) SOD, glycolysed SOD (e.g. excellular SOD EC type A, B, C) or protein-modified SOD is used. 5. Use according to claim 3, characterized in that low-density lipoprotein (LDL) or oxidized LDL coupled SOD is used. 6. Use according to one of claims 1 or 2, characterized characterized that liposomally packaged SOD is used  becomes. 7. Use according to one of claims 1 to 6 for Manufacture of drugs with antiretroviral Effect. 8. Use according to claim 7 for the production of Medicines used to treat AIDS and ARC.