A method for determination of Ukrain in blood plasma for monitoring and pharmacokinetic study.

We developed a method using high-performance liquid chromatography for the determination of the main fluorescent component of Ukrain, a novel antitumor and immune-stimulating drug. Our method was based on ion-pair separation of Ukrain from perchloric acid extracts using reversed-phase column, buffer with high molarity (0.5 M potassium phosphate, pH 2.65), high concentration of ion-pair reagent in the mobile phase (10 mM octylsulfonic acid), controlled temperature of the separation (45 degrees C) and detection by fluorescence (360/455 nm). Under the above conditions a peak of the main Ukrain compound was resolved from fluorescent peaks of the sum of alkaloids of Chelidonium majus L. although several peaks of alkaloids were retained in Ukrain as traces. The height of this main peak was nearly constant, while the alkaloid peaks varied depending on the series of the preparation; chelidonine and thio-triethylenephosphoramide gave no peaks. Analytical recovery for Ukrain from human plasma was 98.0 +/- 4.5%. Therefore, Ukrain possesses neither significant stable binding to plasma proteins nor adsorption in blood cells.

The dynamics of concentration of the main fluorescent component of Ukrain in the tissues and blood plasma of rats with W-256 tumor after a single intravenous injection.

We determined the blood plasma and tissue concentrations of the main fluorescent component of free Ukrain after a single intravenous injection of this substance (0.25 mg/kg) to intact and carcinosarcoma W-256 bearing rats using ion-pair high-performance liquid chromatography with fluorescent detection. The plasma and tissue concentrations of Ukrain were lower than expected, assuming its uniform tissue distribution and first-order kinetics. W-256 tumor tissue was the only tissue that showed accumulation of Ukrain (its concentration at 45 min after injection was up to 2.84-fold higher than in the blood plasma). Based on the elevation of Ukrain concentration in tissues at approximately 15 min after injection and taking into account its high molecular weight as well as the high dimensions of the Ukrain molecule or ion at physiological pH values, we conclude that the preparation penetrates into tissues by active transport or by favored diffusion. Due to the existence of such transport systems together with excretion of Ukrain and probably also its biotransformation, the disappearance of the preparation from the blood cannot be described using a one-compartment model. A relatively higher affinity for Ukrain was observed in tumor tissue and liver, while affinity was lowest in the brain and muscles. The presence of tumors decreased Ukrain concentrations in plasma and normal tissues in comparison with those in control animals.

New biochemical mechanisms of the anticancer effect of Ukrain in the treatment of cancer of the urinary bladder.

The aim of this study was to elucidate the mechanisms of the anticancer effect of Ukrain by comparing the processes of formation of the pool of free amino acids and their derivatives in the blood plasma and tumor biopsy specimens and unchanged bladder tissue in 28 patients with T1N0M0 bladder cancer. The examination was carried out before and after Ukrain treatment (10 mg i.v./day, for 20 days), which was combined with systemic chemotherapy for bladder cancer. Twenty-eight patients served as controls and received systemic chemotherapy only. Compared with healthy donors, the blood plasma of patients with urinary bladder cancer showed decreased concentrations of thiol-containing free amino acids and glutamine (Gln) and increased levels of nonessential (glutamic acid, proline, alanine) and aromatic (phenylalanine) free amino acids. In contrast to conventional chemotherapy, treatment with Ukrain eliminated the blood plasma amino acid imbalance in patients with bladder cancer, concomitantly enriching the pool of free amino acids and their derivatives in unchanged urinary bladder tissue and decreasing concentrations of Gln and leucine (Leu), regulators of malignant cell proliferation and differentiation, by 30-50%. In this situation, the concentrations of Gln and Leu in tumor tissue and the surrounding healthy urinary bladder tissue correlated highly significantly and negatively (r = -0.95). In conclusion, Ukrain prevents active free amino acid transport into urinary bladder tumor tissue, inhibiting the activities of protein biosynthesis, gluconeogenesis and energy production. The combined decrease in Gln and Leu levels in urinary bladder tumor tissue is a specific sign of the antitumor effect of Ukrain and a mechanism of its cancerostatic action by controlling the processes of amino acid pool formation in the tumor.

Results of Ukrain monotherapy of prostate cancer.

This study included 15 patients with newly diagnosed prostate cancer with an average age of 71 years (62-85 years). The patients received Ukrain at a total dose of 100 mg (10 mg intravenously every second day, 10 injections altogether). After two to three injections of Ukrain, all the patients noted considerable subjective improvements in their state. Ukrain increased the amount of total T-lymphocytes, including "active" T-lymphocytes, decreased the content of T-suppressors and increased that of T helpers, correspondingly raising the T helper/T-suppressor ratio. Our results undoubtedly indicate the efficacy of Ukrain in the treatment of prostate cancer.

Comparison of the efficacy of different doses of Ukrain in the combined treatment of breast cancer.

The aim of this study was to compare the efficacy of different Ukrain doses in the combined treatment of 75 patients with breast cancer. The patients were divided into three groups: groups I and II (25 patients each) were treated with 50 mg and 100 mg of Ukrain, respectively, before surgery; group III (25 patients) served as control (without Ukrain treatment). Clinical observations, biochemical, hormonal and immunologic indices indicated that both doses of Ukrain had a similar beneficial effect on patient outcome and may be indicated in the presurgical treatment of patients with breast cancer.

Comparative evaluation of the efficiency of various Ukrain doses in the combined treatment of breast cancer. Report 1. Clinical aspects of Ukrain application.

This study was carried out in 75 patients with histologically confirmed breast cancer. Patients were divided into three groups of 25. The control group received symptomatic corrective therapy prior to mastectomy. The two other groups were given neoadjuvant therapy with Ukrain injections. The first group received a total course dose of 50 mg Ukrain at single doses of 5 mg injected every second day (a total of 10 injections) and the second group received a total Ukrain dose of 100 mg but with single doses of 10 mg. Five to seven days after the last injection patients from all groups were subjected to mastectomy according to Halsted, Patey or Madden. No allergic reactions or adverse effects were observed after the first course of injections or the whole course of Ukrain therapy, regardless of the dose. After five to six injections some patients noticed slight burning sensations and insignificant morbidity in the tumor area, which, according to a number of authors, testifies to the therapeutic activity of the preparation. Practically all patients who were administered Ukrain noticed remarkable positive changes in the second half of treatment: improvement in appetite, normalization of sleep, disappearance of general weakness and the appearance of confidence in recovery. After the course of treatment with Ukrain, the contours of the tumorous node became more clearly defined, which facilitated mastectomy. Changes in the tumor tissue were one-sided in their qualitative differences in comparison to the control group and were not dose-dependent. Qualitative and quantitative reactions to Ukrain by both intact lymphatic nodes and those affected by the metastatic process contribute to more quantitative and radical performance of the most important stage of mastectomy--removal of the regional cellulose together with the lymphatic nodes. The results of this study showed the efficiency of both doses (50 and 100 mg) of Ukrain with neither performing significantly better than the other.

Comparative evaluation of blood plasma and tumor tissue amino acid pool in radiation or neoadjuvant preoperative therapies of breast cancer with the antitumor drug Ukrain.

This study comparatively evaluated free amino acid pool formation in patients with T1-3N0-2M0 breast cancer treated with the drug Ukrain (25 patients, i.v. 100 mg/course) in combination with preoperative radiation or neoadjuvant therapies (25 subjects, total dose 20 Gy). All the patients underwent radical mastectomy. Preoperative radiation did not essentially change the range of the blood plasma parameters studied. However, we observed decreased concentrations of blood plasma ornithine and citrulline and a reduced content of aminobutyric acid, as compared with levels on admission, which may indicate an acceleration of detoxication processes in the liver. In comparison with healthy mammary gland tissue, the tumor tissue of the patients subjected to radiation therapy showed 1.5- to twofold increased concentrations of cysteate, taurine, aspartate, glutamate, proline, glycine, alanine, valine, tyrosine and histidine, which substantiates the idea of tumor tissue being a trap for numerous energy and plastic substrates and indicates active transport of the above compounds into the tumor. The application of Ukrain had virtually no influence on concentrations of the majority of blood plasma amino acids and derivatives: the total concentration of the compounds studied as well as the essential and nonessential amino acid pools remained unchanged. As compared with healthy breast tissue, the considerably increased levels of thiol-containing amino acids, such as methionine, cystine, cysteate and taurine, in the tumor tissue of patients receiving neoadjuvant therapy with Ukrain, indicates high activity of trans-sulfuration processes in this tissue. Simultaneously, in contrast to radiation therapy, Ukrain induced a marked dose-dependent increase in the concentration of proline in breast tumor tissue. The above changes were consistent with the results of the morphological study which confirmed the emergence of numerous foci of necrosis in the tumor and indicated activation of Ukrain-induced proteolytic and degradation processes in the tumor. The results obtained have led us to conclude that a mechanism of Ukrain's cancerostatic effect is to control the transport and reactions of intermediate amino acid metabolism as well as to activate proline biosynthesis in the tumor, causing enhanced development of connective tissue. It is suggested that an important practical conclusion from the present study is the lack of damaging effect of preoperative radiation therapy in the above regimen and the favorable (normalizing) action of Ukrain, at a course dose of 100 mg, on the amino acid pool formation in the organism of patients with breast cancer.

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).

Ukrain: a novel antitumor drug.

A review of the recent literature on the new anticancer drug Ukrain is provided herein. We review Ukrain, a thiophosphate derivative of alkaloids from Chelidonium majus L., its capacity to exert selective cytotoxic and cytostatic effects on tumor cells, simultaneously acting as an immune response modifier, its good tolerance and lack of side effects even after long-term application, perspectives of the application of this drug in oncology.