In vitro activity of cycloSal-nucleoside monophosphates and polyhydroxycarboxylates against orthopoxviruses.

Because variola virus might be used as a pathogen in biological attacks, there is an urgent need to provide effective antiviral drugs for the treatment of orthopoxvirus infections. Thus, the aim of the present study was to test the antiviral activity of 3 pro-nucleotides of the acyclic nucleoside analogues aciclovir (ACV), 3 of penciclovir (PCV) and 38 of the cyclic nucleoside analogue brivudin (BVDU), on the basis of cycloSaligenyl-nucleoside monophosphate approach against vaccinia virus and cowpox virus in vitro. In further experiments, 13 synthetic humic acid-like polymers, so-called polyhydroxycarboxylates, were examined. Antiviral screening was performed by means of the plaque reduction assay and for quantification of the cytotoxicity of the test compounds the XTT-based tetrazolium reduction assay EZ4U was used. As result, three cycloSal-monophosphate derivatives of ACV proved to be potent inhibitors of both vaccinia virus and cowpox virus replication in vitro. Among the tested monophosphate derivatives of cycloSal-PCV and cycloSal-BVDU, selected substances showed a promising antiviral activity against vaccinia virus and cowpox virus. For the polyanionic compounds, no relevant antiviral activity was detected. In conclusion, by the delivery of nucleoside monophosphates from neutral, membrane-permeable prodrugs on the basis of the cycloSaligenyl-nucleotide concept, different ACV, PCV and BVDU derivatives can act as potent and selective inhibitors of orthopoxvirus replication. However, most of the cycloSal-monophosphate derivatives of BVDU had a higher cytotoxicity than their parent nucleosides.

Selective inhibitory activity of polyhydroxycarboxylates derived from phenolic compounds against human immunodeficiency virus replication.

Polyhydroxycarboxylates (MW: 3,800-14,000) derived from phenolic (PDP) compounds have been found to inhibit the cytopathicity of HIV-1 and HIV-2 in MT-4 cells at concentrations that are not toxic to the host cells. The PDP compounds also inhibited syncytium formation in cocultures of MOLT-4 cells with HIV-1- or HIV-2-infected HUT-78 cells. They also interfered with the binding of OKT4A/leu3a monoclonal antibody (mAb) to the CD4 receptor, the binding of anti-gp120 mAb to HIV-1 gp120, and attachment of HIV-1 virions to MT-4 cells. The anti-HIV activity in this series of compounds can be ascribed to inhibition of the gp120-CD4 interaction and seems to depend on the presence of the anionic carboxylate groups. Their mechanism of action is similar to that of the heterogeneous polymer aurintricarboxylic acid (ATA).

Therapeutic effect of pyrophosphate analogues on cutaneous herpes simplex virus type 1 infection in guinea pigs.

We investigated the influence of disodium phosphonic formate (PFA-Na2) and trisodium thiophosphonic formate (TPFA-Na3), in comparison with acyclovir (Zovirax) and trisodium phosphonic formate (PFA-Na3) (Triapten) ointment, on the course of primary cutaneous herpes simplex virus infection in a guinea pig skin model. PFA-Na2 at 3.0%, TPFA-Na3 at 0.5% and PFA-Na3 at 0.5% as well as Triapten ointment (2.0% PFA-Na3) completely inhibited virus infection. Zovirax cream (5.0% acyclovir), applied five times (15 min., 4, 20, 24, and 28 h) after virus inoculation did not prevent virus infection. Similarly, application of Zovirax cream 5 times daily for 5 days did not prevent a vesicle formation following cutaneous herpes simplex virus infection of the guinea pig.

Inhibitory effect of cycloSaligenyl-nucleoside monophosphates (cycloSal-NMP) of acyclic nucleoside analogues on HSV-1 and EBV.

The in vitro antiviral activity of a new series of cycloSal-pro-nucleotides derived from the acyclic nucleoside analogues aciclovir and penciclovir against herpes simplex virus type 1 (HSV-1), thymidine kinase deficient (TK-) HSV-1, and Epstein-Barr virus (EBV) was evaluated. Using the XTT-based tetrazolium reduction assay EZ4U, the cycloSal derivatives were examined for their antiviral and cytotoxic effects in HSV-1 as well as HSV-1-TK--infected Vero cells. The anti-EBV activity was assessed by means of an EBV DNA hybridization assay using a digoxigenin-labeled probe specific for the Bam H1-W-fragment of the EBV genome and by measuring viral capsid antigen (VCA) expression in P3HR-1 cells by indirect immunofluorescence. Among the new cycloSal-phosphotriesters the three aciclovir monophosphates proved to be potent and selective inhibitors of HSV-1 replication, EBV DNA synthesis and EB-VCA expression. Of interest is the retention of activity of the aciclovir monophosphates in HSV-1-TK--infected cells. Particularly 3-methyl-cycloSal-aciclovir monophosphate retained the same effectiveness, as compared to the wild type virus strain. In contrast to the aciclovir pro-nucleotides the penciclovir cycloSal-phosphotriesters exhibited at best only a marginal antiviral effect on HSV and EBV replication.

Anti-HSV-1 activity of synthetic humic acid-like polymers derived from p-diphenolic starting compounds.

A panel of ten humic-acid-like polymers was synthesized by oxidation of p-diphenolic compounds and characterized by relative molecular weights, FT-IR spectra and functional group analysis. Using the XTT-based tetrazolium reduction assay EZ4U, both the low-molecular starting compounds and the synthesized polymers were examined for antiviral and cytotoxic activities in HSV-1-infected Vero cells. With the exception of hydroquinone, 2,5-dihydroxytoluene and 2,5-dihydroxybenzoquinone, the starting compounds failed to inhibit herpesvirus replication. The polymeric oxidation products, however, developed anti-HSV-1 activity with EC50 values in the range of 0.65 (2,5-DHPOP) and 322 microg/ml (2,5-DHBQOP). The CC50 values of the polymers varied among 32.0 (TMHYDROP) and >512 microg/ml (2,5-DHBQOP, HYDSULFOP). The most effective polymers were found to be 2,5-DHPOP 2,5-DHTOP and GENOP (EC50: 0.65, 1.6 and 2.2 microg/ml, respectively, and SI: > or = 400, > or = 80 and > or = 58, respectively). Functional group analysis revealed that increasing numbers of carboxyl groups together with a high content of hydroxyl groups tend to enhance the antiviral activity of polymers derived from p-diphenolic compounds.

In vitro activity of polyhydroxycarboxylates against herpesviruses and HIV.

The antiviral activity of 17 polyhydroxycarboxylates derived from phenolic compounds was evaluated against herpesviruses and HIV. When present during virus adsorption several of the polymers exhibited potent activity against herpes simplex virus type 1 (HSV-1), HSV-2, thymidine kinase deficient HSV-1, human cytomegalovirus (HCMV) and HIV-1 and HIV-2 at concentrations that were not toxic to the host cells. A close correlation was found between the 50% inhibitory concentrations of the polyhydroxycarboxylates against HCMV-induced cytopathicity, their inhibitory effect on the expression of HCMV-specific immediate early antigens and their inhibitory effects on HCMV adsorption to the cells. The antiviral activity of the phenolic polymers was dependent on the presence of a sufficient number of carboxylic groups. The mechanism of antiviral action of the polyhydroxycarboxylates can thus be ascribed to inhibition of virus adsorption. This type of compound may have potential in a vaginal gel to prevent sexual transmission of HSV and HIV.

Antiviral activity of enzymatically oxidized caffeic acid against herpesvirus hominis type 1 and type 2.

The cytotoxicity and antiviral activity of the caffeic acid oxidation product (KOP), a higher molecular polyphenolic compound of strong antiviral activity against herpesvirus hominis type 1 and type 2 (HVH 1, HVH 2), were tested in 6 cell cultures: rabbit kidney primary cells (RKP), rabbit testis primary cells (RTP), primary human embryo lung fibroblasts (LF), calf testis primary cells (CTP), FL- and HEp-2 cells. A marked inhibitory effect on the multiplication of HVH 1 and 2 has been observed in all cell systems at non-cytotoxic concentrations of 0.1-20 micrograms/ml KOP. The adsorption of the virus to cell surface was the most KOP-sensitive phase of herpesvirus multiplication cycle.

[The effect of phenolic polymers on retroviruses]. / Zur Wirkung von Phenolkörperpolymerisaten auf Retroviren.

Phenolic polymers synthesized by enzymatic oxidation of coffeic acid, chlorogenic acid, and gentisinic acid were found to strongly inhibit RNA-dependent DNA polymerase (revertase) of retroviruses. Except of two type C retroviruses inhibition became reversible by the addition of bovine serum albumin to the exogenous revertase test. The phenolic polymers tested did not influence the propagation of retroviruses in the cell culture. The replication of Rauscher leukemia virus in mice was diminished by a short-time preincubation of virus suspension with coffeic acid polymer (KOP). In contrast, the preincubation of a virus-containing serum with KOP increased the leukemogenic effect of the virus. KOP given to mice at a high dose subsequently to virus inoculation resulted in high revertase activities and in an elevation of spleen weights too.

[Penetration kinetics of potential virustatics into the human skin]. / Penetrationskinetik eines potentiellen Virustaticums in die menschliche Haut.

Caffeic acid oxidation product (KOP), a substance effective against various human viruses, penetrates quickly from a 1% W/O-emulsion into the skin and forms a reservoir in the horny layer. In the epidermis and dermis approximately 30 min after external application KOP concentrations of 1 to 3% of the applied total quantity are achieved, which remain nearly unchanged even after longer penetration time. In addition to references for therapy derivable from this the results permit to draw conclusions about drug resorption that can be proved in animal experiments under in-vivo conditions.

[Biotransformation of pyridazines. 1. Pyridazine and 3-methylpyridazine]. / Biotransformation von Pyridazinen. Teil 1: Pyridazin und 3-Methylpyridazin.

Pyridazin (1) and 3-methylpyridazine (6) undergo oxidative biotransformation in an unexpected high degree. Beside the unchanged compounds, after administration of 1 two isomeric monohydroxylated products (2, 3), 4,5-dihydrodihydroxypyridazine (4) and 4,5-dihydroxypyridazine (5) and after administration of 6 one ringhydroxylated 6-derivative (7), 3-hydroxymethylpyridazine (8), one ringhydroxylated 3-hydroxymethylpyridazine derivative (9) and 4,5-dihydroxy-3-methylpyridazine (10) were suggested as urinary metabolites in rats. 2 and 7 are the main metabolites of 1 and 6, respectively.

[Comparison of the in vitro activities of ammonium humate and of enzymically oxidized chlorogenic and caffeic acids against type 1 and type 2 human herpes virus (author's transl)]. / Vergleich der in-vitro-Wirksamkeit von Ammoniumhumat und enzymatisch oxidierter Chlorogen- und Kaffeesäure gegenüber Herpesvirus hominis Typ 1 und 2.

Ammonium humate (prepared from marsh water humic acids) as well as enzymically oxidized diphenolic compounds such as the oxidation products of chlorogenic and caffeic acids have strong antiviral activities in vitro. The effective concentrations lie considerably outside the cytotoxic range of the compounds mentioned.

[Intermediate term results of treatment with the Francobal endoprosthesis in osteoarthritis]. / Mittelfristige Behandlungsergebnisse mit der Francobal-Endoprothese bei Rhizarthrose.

Results one to eight years with a mean of five years after implantation of sixty Francobal-endoprostheses for total trapeziometacarpal joint replacement in fifty patients with osteoarthritis are presented. 91% of the patients achieved clinically good or very good results using a new standardized evaluation score. A worrying problem is the radiologic finding of asymptomatic lucencies in 43%, mainly around the stem component. Loosening, predominantly of the cup, has occurred in 18% and has been followed by two revision operations so far. Heavy manual work increases the risk of loosening, and such patients have to be excluded from this method.

[Nodular fasciitis of the upper arm in a sports marksman]. / Fasciitis nodularis am Oberarm eines Sportschützen.

Nodular fasciitis is a benign, fibroproliferative, tumorlike lesion. Histological differentiation from sarcomas can be difficult. Surgical excision is most effective, recurrences are rare. This case report deals with the onset and natural course of nodular fasciitis in the upper arm of a marksman. When instructing young people in shooting, he had to pace different weapons against the upper third of his arm instead against the shoulder. Here a tender swelling developed. After extirpation he reported pain relief. A recurrence has not been observed so far.

[Preliminary report of the results of syndactyly surgery]. / Vorläufige Ergebnismitteilung von Syndaktylieoperationen.

In the course of the past nine years, 68 patients (42 boys, 26 girls) were surgically treated for syndactylism in 84 hands. In doing so, 135 syndactylies occurring mostly in the form of severe malformations of the hand were separated. In most cases the web spaces were made according to the surgical techniques developed by Blauth. With the exception of eight cases where secondary corrections had to be made due to shallow web spaces or to scarring, all results were highly satisfactory. The authors tend to evaluate any evidence of such disadvantageous scarring as a change which should be considered as "recurring syndactylism".

[Long-term results of treatment of rare aseptic necrosis of the scaphoid bone (Preiser disease) with the Swanson implant]. / Längerfristiges Behandlungsergebnis der seltenen aseptischen Kahnbeinnekrose (M. Preiser) mit Swanson-Implantat.

In 1911 Preiser described the avascular necrosis of the scaphoid bone, which is regarded as a relatively rare clinical and radiological condition. The diagnosis is not established easily, and a small, avascular proximal fragment, e.g. in a scaphoid pseudarthrosis, should not be confused with Preiser's disease. Two histologically diagnosed cases are described: one of thema at operation 42-year old woman received a scaphoid Swanson implant in 1980. At follow-up eight years later, she presented with very satisfying clinical and radiological findings.

[Osteoid osteoma of the hand skeleton]. / Zum Osteoid-Osteom am Handskelett.

Two cases of osteoid osteoma in the hand are reported. One shows a rare localisation in the trapezium. Errors in the first diagnosis and the duration of the disease emphasize the difficulties one can encounter making the correct diagnosis of this tumor. The particular symptoms of osteoid osteoma when located in cancellous bone are discussed as well as the diagnostic aids of tomography and bone scan.

[Destruction of the saddle joint in scleroderma with multiple calcium deposits (Thibièrge-Weissenbach syndrome)]. / Sattelgelenkzerstörung bei Sklerodermie mit multiplen Kalkablagerungen (Thibièrge-Weissenbach-Syndrom).

A case of Thibièrge-Weissenbach-syndrome is reported, representing a special form of scleroderma combined with subcutaneous and periarticular calcifications, both belonging to the group of collagen diseases. Because of painful calcifying deposits in both hands a 53 year-old woman was examined in our out-patient department. Besides extirpation of massive calcifications, fusion of the first carpometacarpal joint was performed because of destruction of this joint.

[Sudeck syndrome of the hand. Historical review, treatment concept and results]. / Das Sudeck-Syndrom der Hand. Historischer Uberblick, Behandlungskonszept und Ergebnisse.

The literature on the etiology, pathogenesis, and therapy of Sudeck's atrophy is reviewed. The authors present their treatment regimen for reflex dystrophic hands that has been used successfully for more than twenty years. This program consists of a combination including drugs as well as physical and occupational therapy. Patients have to be guided psychologically. In-patient treatment is preferred. The ultimate aim of therapy is to restore the functional integrity of the affected hand. The choice of therapy depends on the stage of the disease. Removing pain and edema is the most important aim in stage I. This is achieved by immobilization of the affected extremity in an upward position, cooling the hand with ice, and careful physiotherapy supported by antiphlogistic drugs. In stage II the physiotherapy has to be intensified and should be supplemented by special balneologic (bathing) measures and functional splints. The ipsilateral shoulder can be affected and has to be treated adequately. In stage III additional surgical treatment might be helpful such as arthrolysis, arthroplasty, or arthrodesis of finger joints. The authors report on their results in seventy-seven dystrophic hands in a long-term follow-up between one and fourteen years. The results depend on the begin of the treatment in the different stages of the disease. Eighty-three percent of the patients were cured in stage I, only thirty-one percent in stage II, and no patient in stage III. The authors' experience using Calcitone shows that it has no influence on the functional results. Comparing their results to those obtained by others, the authors conclude that physical and occupational therapy are decisive in dealing with dystrophic hands.