[Cysteine proteinases and their inhibitors in the development of mouse HA-1 hepatoma and antineoplastic therapy]. / Tsisteinovye proteinazy i ikh ingibitory pri razvitii myshinoi HA-1 gepatomy i protivoopukholevoi terapii.

Development of murine HA-1 hepatoma was accompanied by increased activity of cathepsin B (in ascitic cells), cathepsin D (in ascitic fluid) and increased activity of procathepsin B. There were some changes of cysteine proteinases in liver and spleen, not involved directly into tumor growth. The most prominent changes included the decreased level of cysteine proteinase inhibitors cystatin C and stefin A in ascitic cells (and to a lesser degree in liver tissue). During tumor development serum cystatin C concentration decreased by 3-times compared to intact mice. Treatment by antitumor drug Ukraine increased life span of mice with HA-1 hepatoma (transplanted intravenously), decreased the increment of tumor weight. In ascite such treatment caused a decrease of number of tumor cells and an increase of number of macrophages. Ukraie (administered once or 5-times in a dose of 0.5 mg per mice) increased cystatin C level, revealing protective mechanism of action.

Clinical aspects of cancer treatment and new biochemical mechanisms of the drug Ukrain.

A random group of 50 patients in tumor stages T1-3N0-2M0 was selected from breast cancer patients and given Ukrain therapy by intravenous injection. Twenty-five patients received a total dose of 50 mg Ukrain (5 mg every second day, 10 injections altogether). Twenty-five patients received a total dose of 100 mg Ukrain (10 mg every second day, 10 injections altogether).

Preliminary results of individual therapy of chronic hepatitis C by Ukrain and interferon-alpha.

The effects of Ukrain and recombinant human interferon-alpha 2b (IFN) on the state of the thiol-disulfide ratio (SH/SS) of the blood (Russian Federation patent no. 2150700) were studied in vitro using the amperometric titration method. The blood of 73 chronic hepatitis C (CHC) hepatitis C virus (HCV)-RNA-positive patients was examined. Ukrain was tested in doses of 0.05-2.0 micrograms/ml and IFN in 20-1000 U/ml of blood. After in vitro examination, 59 patients were treated: 28 with Ukrain and 31 with IFN. The first group of 16 patients (including eight with HCV genotype 1b) was treated with individually selected optimal doses of Ukrain (0.5-2.5 mg every second day). The second group of 12 patients was treated with doses of 2.5 mg Ukrain independent of in vitro test results. The third group of 31 patients was treated with individually selected optimal doses of IFN (0.5-2 MU 3 times a week). It was found that 79.4% of CHC patients were sensitive to Ukrain in vitro and 65.1% were sensitive to IFN. CHC patients with genotype 1b were sensitive to IFN only in 16.7% of cases while the figure for Ukrain was 92.3%. CHC patients with other HCV genotypes (3, 1a, 2) were sensitive to Ukrain in 86.7% of cases and to IFN in 70.6%. After 1 month of individual therapy with Ukrain, 87.5% of CHC patients, including six of eight cases with HCV genotype 1b, became PCR-HCV negative. In the group receiving the standard dose of Ukrain, virological response was only 33.3%. After 1 month, 74.2% of CHC patients treated with individual doses of IFN became PCR-HCV negative and after 3 months 90.3% were PCR-HCV negative. The prognostic significance of the method for screening preparations for the treatment of CHC patients was 89.8%. Treatment with Ukrain was without serious negative effects and the number of side effects of IFN in individual therapy was significantly reduced. Ukrain can be used in the treatment of CHC patients, alone or in combination with IFN preparations; in the cases with HCV genotype 1b Ukrain seems more promising than IFN. Individual therapy with Ukrain and IFN increased the efficacy of treatment 2.5-fold in comparison with standard monotherapy with the same preparations, significantly decreased the number of side effects and dramatically improved cost-effectiveness.

Cysteine proteinase inhibitor level in tumor and normal tissues in control and cured mice.

Cystatin C is the best known extracellular endogenous cysteine proteinase inhibitor and has been studied as a possible index of tumor growth and as a marker of the effectiveness of antitumor therapy. The aim of this study was to evaluate cystatin C concentrations in murine tumor tissues (compared with other organs not directly involved with tumor development, such as the liver and spleen) during treatment with several antitumor drugs (Ukrain and/or cyclophosphane). Cystatin C concentrations in murine tissues and biological fluids was determined by enzyme-linked immunosorbent (ELISA) assay. The cystatin C ELISA test is a sandwich immunoassay, which uses immobilized rabbit antihuman cystatin C Pab and mouse antihuman cystatin C Mab-HRP (monoclonal antibodies, conjugated with horseradish peroxidase). We observed decreased serum cystatin C concentrations compared with controls in all nontreated tumor models: HA-1 hepatoma (solid and ascitic forms), lung adenocarcinoma (solid and ascitic forms) and LS lymphosarcoma. In the ascitic fluid of mice with HA-1 hepatoma the cystatin C concentration was much lower than in the serum of the same mice (about 20-fold lower). In the HA-1 model of hepatoma cells cystatin C concentration decreased about 2-3-fold compared with the control (intact liver) and Ukrain significantly increased the cystatin C concentration. Cyclophosphane treatment of LS lymphosarcoma significantly increased the cystatin C concentration in serum. Cyclophosphane treatment (50 mg/kg, single injection) increased cystatin C by up to 8-fold more in tumor issue. Ukrain treatment of LS lymphosarcoma was also followed by increased levels of cystatin C in tumor tissue (4-fold); cyclophosphane plus Ukrain had a similar positive effect. In the group with LS lymphosarcoma Ukrain or cyclophosphane plus Ukrain treatment induced a significant increase in cystatin C concentration in liver. Liver cystatin C concentration decreased in the HA-1 hepatoma group and treatment with Ukrain or carboxymethylated beta-1, 3-glucan (CMG) increased this index in both groups. Spleen cystatin C concentrations decreased about 5-fold in LS lymphosarcoma compared with controls and combined treatment with cyclophosphane plus Ukrain restored the index to the normal value. We can conclude that both murine tumors studied were characterized by low cystatin C concentrations in tumor tissues and decreased cystatin C concentrations (to a lesser degree) were also observed in liver and spleen as a result of the "toxic" effect of tumor bearing. Effective treatment in all cases (especially with Ukrain or a combination of cyclophosphane plus Ukrain) induced a significant increase in cystatin C. Obviously, the decrease in cystatin C concentration predominantly in tumor tissue was connected with tumor development and restoration of cystatin C level may be used as a marker of efficacy of antitumor therapy.

Influence of Ukrain on patients with surgically treated breast cancer (introductory remarks).

Studies were undertaken to evaluate the influence of Ukrain on the postoperative period in patients with breast cancer. The group under Ukrain medication was comprised of 10 patients in stage T1-3NO-1MO. The control group was comprised of 22 patients in the same clinical and laboratory staging as those treated with Ukrain in the first year of postoperative follow-up metastatic lymph nodes were observed in one patient from the control group. It is planned to enlarge the groups and to continue the observation up to the third year to obtain reliable and more statistically significant data.

Influence of Ukrain on breast cancer.

The influence of Ukrain on breast cancer was studied in ten women and monitored in each patient by clinical examination, mammography, ultrasonography, histopathology and immunofluorescence in the tissue material obtained after radical mastectomy performed ten days after discontinuation of the treatment. Observations accumulated during the study and analytical data support the view that Ukrain, by changing the antigenic expression of malignant cells, make them recognizable for the immune system which rejects them in a host-versus graft manner.

Influence of Ukrain on patients with surgically treated breast cancer. Part I. Clinical and laboratory parameters.

Studies were undertaken to evaluate the influence of Ukrain on clinical and laboratory parameters in ten patients with breast cancer treated with the drug in the preoperative phase. The control group was composed of eight patients of similar age and advancement of the disease who did not receive Ukrain before mastectomy. Data from the present studies indicate that the drug is distinctly helpful from the surgical viewpoint by increasing tumour hardness and enlargement of metastatic lymph nodes. Furthermore, treatment with it is fully safe, with no side effects or allergic reactions.

Influence of Ukrain on patients with surgically treated breast cancer. Part II. Hormonal profile.

The present study was undertaken to evaluate the influence of Ukrain on the hormonal status of ten patients with breast cancer, treated in the preoperative phase by Ukrain. A control group consisted of eight patients of similar age and advancement of the disease, who did not receive the drug before mastectomy. The data were compared to those obtained in ten healthy women of similar age. The results indicate that Ukrain only slightly influences the hormonal dynamics and its destructive mode of action on neoplastic tissue does not significantly affect hormonal activities.

Influence of Ukrain on patients with surgically treated breast cancer. Part III. The immune system.

The present study was undertaken to evaluate the influence of Ukrain on immune parameters in ten patients with breast cancer, treated with the drug in the preoperative phase. The control group consisted of eight patients of similar age and advancement of the disease, who did not receive Ukrain before mastectomy. The data were compared with the parameters obtained in 15 healthy blood donors. It was found that treatment with Ukrain distinctly regulates cellular and, slightly, humoral immunity, non-specific immunity and also complement activity.

Influence of Ukrain on patients with surgically treated breast cancer. Part IV. Electromicroscopic and cytochemical evaluation.

The present studies were undertaken to evaluate by light and by electron microscopy the influence of Ukrain on the morphology of breast cancer. The studies were carried out on material obtained from ten patients with breast cancer, treated preoperatively with Ukrain. Control material for the studies was obtained from patients of similar age and advancement of the disease, who did not receive Ukrain. The data obtained in the present studies indicate that Ukrain is responsible for severe changes in morphology of tumour cells. Histological examination by both light and electron microscopy revealed cells characteristic of those undergoing apoptosis. The stromal changes of the tumour were characterized by intensive proliferation of the connective tissue accompanied by an immune reaction expressed by mononuclear infiltrates.

Amino acids and their derivatives in blood plasma of patients with breast cancer treated with Ukrain. Part V.

The present study was undertaken to evaluate the influence of Ukrain on the free amino acids pool in blood plasma in ten patients with breast cancer, treated in the preoperative phase with the drug. The control group consisted of five patients of similar age and advancement of the disease, who did not receive Ukrain before mastectomy. The data obtained from these studies indicate that Ukrain positively influences the metabolism of amino acids and their derivatives. The most characteristic changes were increase of proline, taurine and glutamic acid.

Amino acids and their derivatives in tumour tissue from patients with breast cancer treated with Ukrain. Part VI.

The present study was undertaken to evaluate the influence of Ukrain on the amino acids pool and their derivatives in tumour tissue of ten patients with breast cancer, treated in the preoperative phase with Ukrain. The control group consisted of five patients of similar age and advancement of the disease, who did not receive Ukrain. The data obtained indicate that Ukrain influences cancerous tissue, as demonstrated at the level of amino acids. The most characteristic changes observed in cancerous tissue after treatment with Ukrain were increases in the levels of proline, taurine and glutamic acid.

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part I: Preliminary studies of bone parameters.

Ukrain, thiophosphoric acid alkaloid derivatives from Chelidonium majus L. was administered intraperitoneally in a dose of 28 mg/kg (equivalent to 0.1 LD50) every other day for six months to female rats with ovariectomy-induced early osteoporosis. Administration of Ukrain was started on the second day after the surgical operation. At the end of the long-term treatment with Ukrain each rat was tested for the strength of both humeri and some parameters of rat femur were measured. The body weight of ovariectomized rats was also examined. The present results show that the decrease in the mechanical strength of the humeral bones and some changes in the femur caused by ovariectomy were prevented by the six-month treatment with Ukrain. However, in both ovariectomized groups and in ovariectomized rats pretreated with Ukrain an increase of body weight was observed.

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part II: Preliminary studies of peripheral blood parameters.

Studies on Wistar rats have demonstrated that six months after ovariectomy no changes were observed in peripheral blood morphology other than a decreased leukocyte count. A similar effect was noted in the sham-operated group. The present results show that Ukrain treatment started on the second postoperative day and continued every other day for six months significantly increased leukocyte count in comparison with the untreated ovariectomized and sham-operated groups. The activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were also estimated in this study. In none of the groups studied was any change of ALT activity noted. The activity of AST was significantly increased in the sham-operated and untreated ovariectomized rats. After six-month treatment with Ukrain, the activity of this enzyme was significantly decreased in the serum of untreated ovariectomized rats. In rats pretreated with Ukrain some of the studied parameters of peripheral blood were similar to those of the control group.

Effect of six-month treatment with Ukrain on early osteoporosis induced by ovariectomy in rats. Part III. Preliminary studies of some hormone levels.

The effect of six-month treatment with Ukrain on the level of some hormones was investigated in ovariectomized rats. The serum levels of prolactin, adrenocorticotropic hormone (ACTH), corticosterone, aldosterone and progesterone were measured by radioimmunoassay (RIA). Six months after ovariectomy, a decrease in prolactin and progesterone levels was observed. No changes were noted in the levels of ACTH, corticosterone and aldosterone in the serum of the ovariectomized animals. On the other hand, six-month treatment with Ukrain increased the levels of prolactin and progesterone and decreased the levels of corticosterone and aldosterone in the ovariectomized rats. The progesterone level in the treated group was four times higher than in the ovariectomized group. The six-month treatment with Ukrain had no effect on the level of ACTH of the serum of the ovariectomized rats. The present results show that prolonged treatment with Ukrain significantly affects hormone secretion after ovariectomy.

Antinociceptive effect of ten day administration of Ukrain in mice and interaction of the treatment with morphine.

The present study shows that Ukrain, administered i.c. in doses of 9.5 and 19 mg/kg once daily for 10 days in mice, produced evident antinociceptive action in the writhing syndrome and hot-plate tests. Moreover, we found that as a result of 10-day treatment with Ukrain the antinociceptive action of morphine (0.1 mg/kg s.c. administered after the last dose of Ukrain) was significantly decreased. These results and the observed interaction between Ukrain and morphine seem to be contra-indications to their combined usage.

Does the Ukrain preparation protect mice against lethal doses of bacteria?

Ukrain, a semi-synthetic preparation obtained from Chelidonium majus L, is used in the treatment of cancer diseases. It has been observed to exert a protective influence in mice infected by influenza viruses. Recently, the influence of the preparation on the survival of mice infected by lethal doses of E. coli and S. aureus has been estimated. This preparation was administered to Balb/c mice subcutaneously in doses of 0.04, 0.4 and 4.0 mg/kg of body weight. Ukrain was given every second day during 20 days, or a short-term before-and-after method at 48, 24 and 2 h before the infection and or 2, 24 and 48 h after the infection of mice. The mice were infected intraperitoneally with E. coli or S. aureus in doses equivalent to 2LD50. Increased survival of mice, depending on the dose of the preparation and the kind of infecting bacterium was observed. The highest survival (50%) occurred in mice infected with E. coli and receiving the amount of the preparation corresponding to 0.4 mg/kg. The lowest survival was observed in mice infected by S. aureus and receiving the preparation in the amount of 4.0 mg/kg. Higher protective effectiveness of the Ukrain preparation was observed in mice when the preparation had been administered during 20 days as compared to the short-term before-and-after regime.

Interaction between Ukrain and morphine in their ten-day treatment in mice in the writhing syndrome test.

The antinociceptive properties of Ukrain and of morphine in their 10 day treatment in mice were studied by the writhing syndrome test. It was found that the antinociceptive action of Ukrain after 10 days of treatment was similar to that of morphine (0.1 mg/kg s.c.) in the same period. Results indicate that the combined treatment with Ukrain and morphine for 10 days completely reduced their antinociceptive action.

Biophysiological effects of Ukrain therapy in a patient with breast cancer (case report).

The effects on different biophysiological parameters and subjective impressions were studied in a patient with breast cancer who was not previously given any therapy before receiving Ukrain. Daily measurements of pulse, blood pressure, temperature and various laboratory examinations were carried out. Development and course of subjective and objective phenomena seem to be typical for patients in whom Ukrain could induce long-term complete remission. The patient described here has had to data 12 years without any oncopathological symptoms.

Potential therapeutic efficacy of Ukrain (NSC 631570) in AIDS patients with Kaposi's sarcoma.

In this study two case reports are presented of therapy with Ukrain, a semi-synthetic thiophosphoric acid compound of alkaloids isolated from Chelidonium majus L, for the treatment of AIDS patients with Kaposi's sarcoma. Ukrain was injected i.v. in the dose of 5 mg every other day for a total of 10 injections. During treatment the Kaposi's sarcoma lesions diminished in size, showed decolouration and no lesion appeared in the 30-day interval after the beginning of treatment. Both patients tolerated Ukrain well and showed an improved immunohaematological status: an increase in total leukocytes, T-lymphocytes and T-suppressor numbers. In one case T-helper lymphocytes were also increased. Future investigations are necessary for the evaluation of Ukrain efficacy in the treatment of this type of patients.