RU2482844C2 - Silibilin component for treating hepatitis - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: claimed are: application of silibinin component of general formula

(I) for obtaining medication for parenteral introduction for trweatment of viral hepatitis, with medication optionally containing cyclodextrin and/or phospholipid, and set of similar purpose, which includes said silibinin component and other medication, representing one or several pharmaceutical agents from: arginine, glutamate, silymarin, citiolone, epodemiol, ornithine oxoglurate, tidiacic arginine, myoinosite, methionine and N-acetyl methionine, choline, ornithine aspartate, cyanidanol, thiopronin, betaine, cyanocobalamin, leucine, levolose, acyclovir, idoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valaciclovir, cidofovir, penciclovir, valganciclovir, brivudin, interferon. Medication preferably does not contain silidianin, and/or silicristin, and/or isosilibinin.

EFFECT: reduction of viral strain and reactivation of patients after parenteral introduction of claimed silibinin component.

21 cl, 12 dwg, 5 ex

Description

The present invention relates to the use of a silibinin component for the manufacture of a medicament for the treatment of viral hepatitis, preferably hepatitis B or C, in particular for reducing viral load. Preferably, the drug is adapted for parenteral administration. Preferably, the silibinin component is an ester of silibinin.

Silibinin {3,5,7-trihydroxy-2- (3- (3-hydroxy-4-methoxyphenyl) -2- (hydroxymethyl) -2,3-dihydrobenzo [b] [1,4] dioxin-6-yl ) chroman-4-one, or according to the nomenclature of the European Pharmacopoeia (2R, 3R) -3,5,7-trihydroxy-2 - [(2R, 3R) -3- (4-hydroxy-3-methoxyphenyl) -2- ( hydroxymethyl) -2,3-dihydro-1,4-benzodioxin-6-yl] -2,3-dihydro-4H-1-benzopyran-4-one}, is the main component of silymarin and the main flavonoid extracted from milk thistle ( Silybum marianum Gaertneri).

Silibinin has the structure:

The diastereoisomers silibinin A and silibinin B are separately presented in the literature:

Silibinin is the main constituent of silymarin (in a 50:50 mixture of silybin A and silybin B). Other constituent parts include isosilibinin (isosilibin A and isosilibin B), silidianin, silikristin, isosilikristin, taxifolin and others. Methods for isolating silibinin are known in the prior art (e.g., US 4,871,763).

Silibinin and silymarin have been studied and described in detail. In this regard, we can refer to the work of N.-C. Kim et al., Org. Biomol. Chem. 1, 2003, ss. 1684-1689; D.Y.W. Lee et al., J. Nat. Prod. 66, 2003, pp. 1171-1174; D.J. Kroll et al., Integrative Cancer Therapies, 6, 2007, pp. 110-119; Z. Wen et al., DMD Fast Forward, doi: 10.1124 / dmd.107.017566; and US 4871763.

The history of the medicinal plant Silybum marianum dates back to about 2000 years. Silymarin - an extract of milk thistle seeds, is an ancient medicine for the treatment of various disorders of the liver and gall bladder, including hepatitis, cirrhosis, as well as a hepatoprotector that protects the liver from poisoning by wild mushrooms, alcohol, chemicals and toxins from the environment. The mechanism of action of silymarin is diverse. The largest randomized controlled trial conducted in the 1970s showed that prolonged use of silymarin can reduce mortality in patients with chronic cirrhosis (P. Ferenci et al., J Hepatol 9, 1989, pp. 105-113). However, the role of this drug in treating liver diseases remains controversial (S. Verma et al., Clinical Gastroenterology and Hepatology 5, 2007, pp. 408-416; F. Rainone, Am Fam Phys 72 (7), 2005, cc. 1285-1288). In part, this uncertainty is caused by limited information on pharmacokinetics and on optimal dosage regimens. Silymarin is poorly soluble in water, and oral administration results in limited bioavailability.

Pharmaceutical uses of silibinin are also known. Silibinin has strong antioxidant properties (see A. Pietrangelo et al., Gastroenterology 109, 1995, pp. 1941-1499; M.I. Lucena et al., Int J Clin Pharmacol Ther 40, 2002, cc. 2-8; L. Mira et al., Biochem Pharmacol 48, 1994, cc.753-759) and antifibrotic properties (see G. Boigk et al., Hepatology 26, 1987, pp. 643-649; C. Dehmlow et al., Hepatology 23, 1996, pp. 749-754), therefore, it is potentially applicable as a medicine for the treatment of chronic liver diseases. The pure substance silibinin is administered intravenously, for example, when poisoning the liver with a pale toadstool (toxins amenitin, phalloidin) to prevent liver destruction (see K. Hruby et al., Hum Toxicol 2, 1983, pp. 138-195). The effect of fungal poisoning is partially explained by the stimulation of the nucleolar polymerase A, which increases the synthesis of ribosomal protein and inhibits lipid peroxidation (J. Connenbichler et al., Prog Clin Biol Res. 213, 1986, cc. 319-331). Clinical studies have also been successful in the prevention and treatment of certain types of cancer (L. Varghese et al., Clin Cancer Res 11 (23), 2005, pp. 8441-8447; K. Letschert et al., Toxicological Sciences 91, 2006, pp. .140-149).

Silibinin ester is prepared as an infusion solution, for example Legalon® SIL in the Federal Republic of Germany.

Viral hepatitis refers to infections that infect the liver and are caused by viruses. Viral hepatitis is being actively investigated worldwide. Viral hepatitis not only causes high mortality, but also depletes medical resources and can have dire economic consequences. Most of all viral hepatitis can be prevented.

The concept of "viral hepatitis" refers to five different diseases that are caused by at least five different viruses. Hepatitis A and hepatitis B (infectious hepatitis and serum hepatitis, respectively) represent different diseases and both are diagnosed by a specific serological test. Hepatitis C and E represent a third category that is different in type, with hepatitis C being transmitted parenterally and hepatitis E being transmitted enterally. Hepatitis D, or delta hepatitis, is a disease caused by a different type of virus depending on hepatitis B infection. This form of hepatitis can be in the form of superinfection in a hepatitis B carrier or in the form of co-infection in an individual with acute hepatitis B.

Hepatitis C is a human infectious disease caused by the hepatitis C virus (HCV). Hepatitis C virus infection can lead to severe liver damage, such as inflammation of the liver parenchyma, liver fibrosis, cirrhosis and liver carcinoma. In more than 80% of infected patients, HCV infection becomes chronic. HCV transmission usually occurs through the parenteral route through the blood.

It has been estimated that approximately 170 million people worldwide are infected with hepatitis C virus (HCV). For decades, infected patients have had no symptoms until they develop cirrhosis and / or hepatocellular carcinoma. In about 40–50% of cases in the United States, liver transplantation is caused by HCV infections. Six HCV genotypes (HCV1-HCV6) were identified, which differ in geographical distribution and in their response to treatment.

HCV proteins have been shown to induce STAT-3 activation through oxidative stress and Ca ²⁺ signal transduction (K. Koike et al., Hepatol Res 34, 2006, pp. 65-73; G. Waris et al., J Virol 79, 2005, pp. 1569-1580), as well as lipid peroxidation products and antioxidant gene expression (M. Okuda et al., Gastroenterology 122, 2002, pp. 366-375). It is believed that the balance of oxidative and reduction potentials in the cell (cellular redox potential) has pronounced effects on the metabolic pathways of signal transduction (YMJanssen et al., Am J Physiol 273, 1997, pp. 799-796), including attenuated IFN- alpha signal transduction (D.Di Bona et al., J Hepatol. 45, 2006, pp. 271-279).

HCV infection is divided according to ICD10 (WHO, version 2007) into acute (B17.1) and chronic hepatitis C (B18.2).

HCV is one of the most important causes of acute or chronic hepatitis. However, the course of the disease can vary greatly and is highly susceptible to variability. Therefore, one cannot talk about the typical course of the disease, since HCV infection manifests itself in a wide range of clinical manifestations, i.e. variable symptoms, different clinical pictures and various secondary liver diseases and lesions outside the liver.

In approximately 20% of patients with acute hepatitis, liver inflammation is a consequence of HCV infection. However, in the acute phase, hepatitis C is usually asymptomatic and therefore is not diagnosed in approximately 85% of cases. In some cases, there are only non-specific flu-like symptoms. Typically, infection does not occur during the acute phase.

Hepatitis C becomes chronic in approximately 85% of patients with acute HCV infection. Such a high degree of transition of the disease to the chronic phase is the result of high variability of HCV, i.e. the gene that encodes the envelope of HCV mutates with high frequency. Due to the high variability of the virus and, in particular, the high variability of the antigenic epitope of HCV, the mutant HCV avoids recognition by the human immune system. In approximately 25% of patients, chronic liver inflammation results in the formation of cirrhosis and an increased risk of liver carcinoma (see, e.g., J.N. Hoofnagle, Hepatology 1997, 26, appendix 1, pp. 15S-20S; M.I. Memon et al., Journal of Viral Hepatitis 9, 2002, pp. 84-100; SLTan et al., Nature Reviews, Drug Discovery 1, 2002, cc.867-881).

HCV infected patients usually receive standard combination treatment with pegylated interferon-α2a or interferon-α2b and ribavirin. For HCV infections with genotypes 2 or 3 (HCV2 or HCV3 infections), this combination treatment is carried out for 24 weeks. In HCV infections with genotype 1 (HCV1), combined treatment is performed for HCV1-positive patients for 48 weeks. Many HCV-infected patients, however, discontinue treatment due to existing side effects and / or because of rejection of parenteral administration and duration of use. In addition, only about 50% of patients with HCV1 have a positive outcome for such a long treatment, i.e. the remaining do not respond to such treatment, i.e. are non-responders (see, e.g., R.E. T. Smith, Nature Reviews, Drug Discovery, 5, 2006, p. 715). Treatment for pegylated interferon plus ribavirin caused by hepatitis C virus disease is ineffective in about half of patients with genotype 1. Treatment failure is either due to the lack of response of the body to treatment (minimal reduction in virus titer), or due to relapse (strong initial response to treatment replaced by a new increase in virus titer during or after treatment). Such different treatment consequences may depend on various factors, including the host’s genetic status, immune response, and the genetic differences of the viruses (see MWFried et al., New England Journal of Medicine 347, 2002, pp. 975-982; HSConjeevaram et al. Gastroenterology 131, 2006, pp. 470-477; MPManns et al., Lancet 358, 2001, pp. 958-965; DBStrader et al., Hepatology 39, 2004, p. 114771; SJHadziyannis et al., Ann Intern Med, 140, 2004, pp. 346-355). Genetic differences in viruses can include either differences that existed prior to treatment or differences that occur during treatment due to the evolution of the virus in response to the pressure of the treatment being applied.

New treatment options are being developed, including optimizing existing standard treatment with pegylated interferon plus ribavirin, especially targeted antiviral therapy for HCV, new immunomodulating agents, and treatment aimed at reducing fibrosis (see REStauber et al., Drugs 68 (10), 2008, p.1347).

To date, there is no vaccine against HCV. Standard methods of treatment are very expensive, have little effect in terms of eradicating HCV infection, and sometimes cause significant side effects (SLTan et al., Nature Reviews, Drug Discovery 1, 2002, p. 867; R. Bartenschlager, ibid., P. 911 )

There is a need for drugs to treat viral hepatitis, especially hepatitis B and C.

The problem solved in the present invention is to obtain an effective drug for the treatment of viral hepatitis, in particular hepatitis B or C, which has advantages compared with drugs of the prior art. If possible, the drug should not have side effects or have mild side effects, and be effective, for example, for patients with hepatitis C who do not significantly respond to traditional combination therapy with peginterferon / ribavirin. In addition, the drug must have pronounced antiviral properties and, thus, reliably reduce the load of the virus.

Achieving this objective is set forth in the claims of the present invention.

It has been unexpectedly found that silibinin, its pharmaceutically acceptable salts and / or derivatives are applicable for the treatment of inflammation, viral liver diseases, especially hepatitis C. Thus, in patients with hepatitis C who do not respond to treatment with immunomodulating / antiviral combination therapy (non- responders), for example peginterferon / ribavirin, which is currently the standard treatment for hepatitis C, a significant reduction in the load of the virus can be achieved by administration, preferably parent oral administration, silibinin component. It has also been shown that pre-treatment with the silibinin component increases the response of patients to the later administration of interferon and ribavirin.

Studies relating to the treatment of HCV infections, especially regarding the suppression of HCV infections by administering silymarin, are described in the prior art (see, for example, R. Sailer et al., Drugs 61 (14), 2001, pp. 2035-2063; K .E. Mayer et al., Journal of Viral Hepatitis, 12, 2005, cc. 559-67; US 2005/0123628; SJPolyak et al., Gastroenterology 132, 2007, cc. 1925-1936).

R.Sailer et al. Report that although the effect of silymarin on viral replication has not been established, pharmacological estimates suggest that it inhibits inflammation and the cytotoxic cascade of events triggered by a viral infection. Oral administration of the silibinin-phosphatidylcholine complex (IdB1016, 240 mg of silibinin twice a day) in a short-term placebo-controlled experimental study of 20 patients with chronic hepatitis showed that the evolution of AST levels was significantly reduced in the silibinin group in the absence of significant differences between the other studied liver function indices ( see A. Vailati et al., Fitoterapia, 64 (3), 1993; G. Buzzelli et al., Int. J. Clin. Pharmacol. Ther. Toxicol. 31, 1993, cc. 456-460).

K.E. Mayer et al. Describe that oral treatment with silymarin leads to a decrease in serum transaminases compared with baseline in four studies and with placebo in only one study. However, there is no evidence that silymarin affects the load of the virus or improves liver histology in hepatitis B or C (see MLChavez, J. Herb. Pharmacother. 1 (3), 2001, pp. 79-90; LBSeeff et al., Hepatology, 34 (3), 2001, pp. 595-603). It was concluded that silymarin compounds probably lower serum transaminases in patients with chronic viral hepatitis, but do not presumably affect the load of the virus or histology of the liver.

US 2005/0123628 relates, inter alia, to the preparation and oral administration of compositions comprising glycyrrhizin, magnolia vine, ascorbic acid, L-glutathione, silymarin, lipoic acid and D-alpha tocopherol. It is reported that these compositions are useful for reducing oxidative stress and lipid peroxidation, for treating chronic liver disease, chronic viral hepatitis C infection and non-alcoholic fatty hepatitis. Numerous studies report the hepatoprotective properties that silymarin exhibits against a large number of different toxins, including acetaminophen, ethanol, carbon tetrachloride and D-galactosamine, as well as against ischemic damage, radiation and iron toxicity. During the first twenty weeks of an open, non-randomized, single-center clinical trial, subjects were given orally administered twice daily for a total of 1000 mg of glycyrrhizin; three times a day in a total amount of 1500 mg of lemongrass plant extract; three times a day in a total amount of 6000 mg of ascorbic acid; twice a day in a total amount of 300 mg of L-glutathione; three times a day in a total amount of 750 mg of milk thistle extract; twice a day in a total amount of 300 mg of lipoic acid; and once a day in the amount of 800 ME D-alpha-tocopherol. During the first ten weeks of the study, four different parenteral compositions were administered to the subjects twice weekly by intravenous injection, none of which contained silymarin. After 10 weeks, 12.0% of subjects, after 20 weeks, 24.0% of subjects showed a 1 log reduction in virus load. In US 2005/0123628 there is not even a hint that silymarin, or that only silybinin, may be responsible for a relatively small decrease in viral load.

S.J. Polyak et al. Compare in vitro standardized silymarin extract (MK-001) with commercial silymarin preparations. Both drugs are reported to exhibit antiviral effects when using cell culture-based models, although the effects of commercial drugs are not as strong as MK-001. MK-001 inhibits the expression of tumor necrosis factor alpha in anti-CD3 stimulated peripheral blood mononuclear cells and nuclear factor-dependent kappa B transcription in Huh7 human hepatoma cells. In addition, the dose of MK-001 suppresses infection of Huh7 and Huh7.5.1 cells with JFH-1 virus in a dose-dependent manner. MK-001 has an effect against HCV infection in isolated cells and, when combined with interferon-α, inhibits HCV replication to a greater extent than inhibition with interferon-α alone. To compare the anti-HCV effects of MK-001 with commercial silymarin preparations, the Ultrathistle® product (Natural Wellness, Montgomery New York) and the Silybinin® product (Indena SpA, Milan) are also tested. However, it is reported that MK-001 has a stronger antiviral effect compared to Ultrathistle® and Silybinin®. Based on in vitro studies, it was concluded that since there is an anti-HCV effect, the standardized silymarin MK-001 extract is superior to two commercial products. S.J. Polyk does not report anything about the parenteral administration of purified silibinin, not to mention the treatment of non-responders. In addition, the conclusions made by Polyak et al. Contradict clinical studies that showed no exposure to silymarin on HCV in patients with chronic hepatitis C (MDTanamly et al., Dig Liver Dis. 36, 2004, pp. 752-759; E Gabbay et al., World J Gastroenterol. 13, 2007, pp. 5317-5323).

It has now surprisingly been found that administration, especially parenteral administration, preferably of the pure silibinin component, reduces the in vivo load of the virus in patients with viral hepatitis. Thus, the silibinin component is able to lower the viral load. This fact allows you to optimize the dose of silibinin in the absence of other components of silymarin, which can cause unwanted side effects.

The decrease in viral load by parenteral administration of the silibinin component is especially unexpected, since clinical studies show that silymarin does not affect HCV in patients with chronic hepatitis C (M. Torres et al., PRHealth Sci J 23 (2), 2004, pp. 69 -74; MDTanamly et al., Dig Liver Dis., 36, 2004, pp. 752-759; A Gordon et al., J Gastroenterol Hepatol. 21, 2006, cc. 275-280; E. Gabbay et al. , World J Gastroenterol. 13, 2007, cc. 5317-23; LBSeeff et al., Hepatology, 80 (11), 2008, cc. 1900-1906).

M.Torres et al. Report a clinical study in which patients aged 21-65 years with chronic hepatitis C who were not exposed to antiviral therapy were invited to participate. 34 patients were randomized to treat S.marianum 160 mg orally three times a week for four weeks or no treatment was performed (control). Research has shown that S. marianum does not act as an antiviral agent.

M.D. Tanamly et al. Report a clinical study in which 177 patients with chronic hepatitis caused by hepatitis C virus were randomly divided to receive either orally silymarin or multivitamin supplements. The study showed that the recommended dose of silymarin does not affect hepatitis C virusemia.

A. Gordon et al. Report a clinical trial involving 24 subjects with chronic hepatitis C, namely a randomized, double-blind, placebo-controlled trial. Subjects received 12 weeks of S.marianum (or 600 mg, or 1200 mg / day) and placebo. Conducted biochemical, virological, psychological research, as well as an analysis of the quality of life. Seventeen patients completed the study. The study showed that the average changes in HCV RNA titers did not differ significantly in subjects treated with S. marianum compared with those who received placebo.

E. Gabbay et al. Report a clinical trial involving 100 patients with chronic HCV infection for whom treatment with interferon was unsuccessful and who were randomized to receive seven different antioxidants along with 250 mg silymarin capsules three times a day. The main endpoints of the study were liver enzymes, HCV RNA levels, and histology. Research has shown that antioxidant therapy does not affect viral load.

LBSeeff et al report a long-term antiviral treatment for cirrhosis in hepatitis C with cirrhosis (HALT-C), which includes subjects with chronic hepatitis C who are non-responders to primary antiviral therapy but are willing to participate in long-term treatment with pegylated interferon . No beneficial effect of silymarin on hepatitis C virus RNA (HCV) levels was found. Thus, those receiving silymarin had similar HCV levels when compared with those who did not receive such treatment.

In addition, it was unexpectedly discovered that the introduction, especially parenteral administration, of the silibinin component promotes conventional treatment with peginterferon / ribavirin. It has been established that the component silibinin activates (reactivates) the sensitivity of patients to traditional treatment with peginterferon / ribavirin and / or enhancers of the antiviral effect of traditional therapy with peginterferon / ribavirin.

Brief Description of the Figures

Figure 1. Example 1, study 1: oxidative stress parameters before and after infusion of 10 mg / kg silibinin component for 4 hours (p-RCM = derivatives of reactive oxygen metabolites, BAP = biological antioxidant potential).

Figure 2. Example 1, Study 1: HCV RNA (log IU / ml; mean ± SD) before (1 day) and after (7 days) intravenous administration of 10 mg / kg of the silibinin component / day.

Figure 3. Example 1, study 1: changes in HCV-RNA after intravenous administration of 10 mg / kg / day of the silibinin component for 7 days, followed by combination therapy with peginterferon alfa-2a / ribavirin and 140 mg of silymarin three times a day.

Figure 4. Example 1, study 2: changes in HCV-RNA during the administration of the silibinin component in different doses for 14 days, followed by combination therapy with peginterferon alfa-2a / ribavirin, started on day 8.

Figure 5. Example 1, study 2: mean ± SD of the decrease in HCV-RNA during 7 days of monotherapy in the form of intravenous administration of the silibinin component and 7 days of intravenous administration of the silibinin component with peginterferon alpha 2a / ribavirin in different doses.

6. Example 1, study 2: changes in HCV-RNA after completion of intravenous administration of silibinin (2 weeks) in 14 patients receiving silibinin in the amount of 15 or 20 mg / kg / day. Combination therapy with peginterferon alfa-2a / ribavirin is started on day 8 and 280 mg of silymarin three times a day on day 15.

7. Example 2, specific patient, changes in HCV-RNA after intravenous administration of 20 mg / kg / day of the silibinin component during two administration intervals, including 14 consecutive days, the first interval in intravenous administration starts on day 24, and the second interval in administration, starting with 35 weeks, during continuous combination therapy with 180 mcg peginterferon alfa-2a / ribavirin for 60 weeks.

Fig. 8. Example 2, specific patient, changes in HCV-RNA after intravenous administration of 20 mg / kg / day of the silibinin component during the administration interval of 14 consecutive days, and starting from the 32nd week during continuous combination therapy using 180 μg peginterferon alpha 2a / ribavirin for 60 weeks.

Fig.9. Example 2, specific patient, changes in HCV-RNA after intravenous administration of a 20 mg / kg / day silibinin component during an administration interval of 14 consecutive days and starting from the 72nd week during continuous combination therapy using 180 μg peginterferon alfa-2a / ribavirin for 80 weeks.

Figure 10. Various methods for the combined use of ribavirin and / or pegylated interferon alpha and a drug containing a silibinin component are shown schematically.

11. Data from an in vitro NS5B inhibition study for six purified silymarin components.

Fig. 12. Data from the in vitro NS5B inhibition study for silibinin bis (hydrogen succinate).

The present invention relates to the use of a silibinin component for the manufacture of a medicament, preferably a virusostatic or antiviral, more preferably reducing a viral load, for the treatment of viral hepatitis, in particular hepatitis B or C, preferably infections of chronic or acute viral hepatitis C, preferably parenteral administration.

In the context of the present invention, the term “drug” is preferably synonymous with “drug treatment”.

In a preferred embodiment, the present invention relates to the use of a silibinin component for the manufacture of a medicament which is substantially free of silidianin and / or silicristin and / or isosilibinin for the treatment of viral hepatitis, preferably hepatitis B or C.

In a preferred embodiment of the present invention, the treatment of viral hepatitis, in particular hepatitis B or C, is carried out by reducing the viral load. It has been established that silibinin components are able to reduce the viral load in patients with hepatitis B or C. This is especially surprising since there is no evidence in the prior art that silymarin, a mixture of which contains a certain amount of silibinin, affects the viral load or improves liver histology in hepatitis B or C (see K.E. Mayer et al., Journal of Viral Hepatitis, 2005, 12, 559-567).

In another preferred embodiment of the present invention, the treatment of viral hepatitis, in particular hepatitis B or C, is carried out in patients who have or who have undergone liver transplantation. Patients who have undergone liver transplantation due to viral hepatitis are at risk of resuming viral hepatitis in the transplanted liver. Typically, the virus is not completely removed from the body when the infected liver is removed surgically and the remaining viruses in the body re-infect the transplanted liver. In chronic hepatitis C, infected patients are newly infected after liver transplantation in 100% of cases. It has been unexpectedly found that silibinin can reduce the viral load, the risk of re-infection after liver transplantation can be significantly reduced by the introduction, preferably parenteral administration, of the silibinin component.

Forms of viral hepatitis are known to those skilled in the art.

At present, at least six different forms have been established: hepatitis A, B, C, D, E, and G. The causative organisms of these infections are hepatotropic viruses. They belong to different families of viruses in each case and have a genome from DNA or RNA. Transmission occurs either through food or through tissue fluids such as sperm and blood. There are also differences in different forms regarding the course of the disease and its severity. Although hepatitis A and E are usually acute, hepatitis B, C and D can lead to a chronic course of the disease, and in some cases, to serious complications.

In the present description, the term "viral hepatitis" preferably includes hepatitis B and C.

In a preferred embodiment, the present invention is treated by reducing the viral load of one or more viruses selected from the group consisting of the genotypes of HCV1, HCV2, HCV3, HCV4, HCV5 and HCV6, preferably HCV1, but not only one of them.

If the genotype is the HCV1 genotype, subtypes 1a, 1b, 1c, 1d, 1e, 1f, 1g, 1h, 1i, 1j, 1k and 1l are preferred. If the genotype is the HCV2 genotype, subtypes 2a, 2b, 2c, 2d, 2e, 2f, 2g, 2h, 2i, 2j, 2k, 2l, 2m, 2n, 2o, 2p and 2q are preferred. If the genotype is the HCV3 genotype, subtypes 3a, 3b, 3c, 3d, 3e, 3f, 3g, 3h, 3i, 3j and 3k are preferred. If the genotype is the HCV4 genotype, subtypes 4a, 4b, 4c, 4d, 4e, 4f, 4g, 4h, 4i, 4j, 4k, 4l, 4m, 4n, 4o, 4p, 4q, 4r and 4t are preferred. If the genotype is the HCV5 genotype, subtype 5a is preferred. If the genotype is HCV6 genotype, subtypes 6a, 6b, 6c, 6d, 6e, 6f, 6g, 6h, 6i, 6j, 6k, 6l, 6m, 6n, 6o, 6p and 6q are preferred. For the nomenclature of hepatitis C virus genotypes and subtypes, see, for example, P. Simmonds et al., Hepatology, 42, 2005, 962-973.

In a preferred embodiment, the present invention relates to the use of a silibinin component for the manufacture of a medicament which is preferably adapted for parenteral administration for the treatment of viral hepatitis, preferably hepatitis C, to patients who do not respond to conventional immunomodulating / antiviral combination therapy, for example ribavirin / interferon therapy ( Non-responders), and / or to patients who partially respond to traditional immunomodulating / antiviral combination therapy, for example, ribavirin / interferon therapy (“partial responders”), and / or patients who have a clear initial response, followed by the resumption of viral titers during or after therapy (“relapses”).

The present invention also relates to the treatment of viral hepatitis C with a silibinin component, and this treatment follows the traditional combination therapy with ribavirin / interferon. Preferably, treatment with the administration of the silibinin component begins after failure of treatment with ribavirin / interferon (either at first or after a certain period of treatment).

In connection with the traditional treatment of hepatitis C by introducing ribavirin / interferon, the concepts of non-responders, partial responders, and patients with relapse are known to specialists in this field. Currently, treatment of viral hepatitis C with pegylated interferon together with ribavirin is unsuccessful for about half of patients with genotype 1. Failure in treatment manifests itself either in the absence of a response to treatment (minimal reduction in virus titers) or in the development of relapse (initial clear answer to treatment is replaced by restoration of virus titers during or after treatment).

For the purpose of specification, non-responders are preferably called patients who do not have a decrease in viral load of <2 log ₁₀ IU / ml (i.e. 100 times) with the administration of ribavirin / interferon (usually peginterferon α), preferably for 12 weeks. In a preferred embodiment of the present invention, for non-responders, virus titers are reduced by ≤2.1 log ₁₀ IU / ml and absolute titers ≥4.62 log ₁₀ IU / ml at a minimum value.

For the purpose of specification, partial responders are preferably called patients who do not have a decrease in viral load of ≥2 log ₁₀ IU / ml by week 12 with detectable HCV RNA for 24 weeks.

For the purpose of specification, patients with relapse are preferably called patients in whom the decrease in virus titers is ≥2.8 log ₁₀ and the absolute titers temporarily fall below the detection limit (2.78 log ₁₀ IU / ml).

In the present description, the term "dosage form" is preferably synonymous with the concept of "injectable form" or "dosage form". For example, if the dosage form is intended for oral administration, for example, in the form of a tablet, such a tablet preferably represents a dosage form for administration, which contains a dose of the silibinin component, intended for administration at the appropriate time within the framework of the introduction. If the dosage unit includes one tablet, the dosage unit corresponds to the form administered. However, the dosage form can also be divided into a number of injectable forms, for example, a series of tablets, which in each case contain only part of the dose, but together they contain a whole dose of the silibinin component, intended for administration at the appropriate time as part of the administration schedule (such tablets are dosed units are intended for almost simultaneous administration).

In the present invention, the term "silibinin component" preferably refers to silibinin, including all stereoisomers thereof, for example silibinin A and silibinin B, its pharmaceutically acceptable salts and / or derivatives, in particular esters. Preferred esters are derivatives of inorganic acids, for example phosphoric acid or sulfuric acid; or organic acids, for example formic acid, acetic acid, propionic acid, citric acid, malic acid, mandelic acid, etc.

Among dicarboxylic acid hemiesters, hemiesters, for example malic acid, glutaric acid, succinic acid, adipic acid, suberic acid, azelaic acid, sebacic acid, fumaric acid, maleic acid, itaconic acid, phthalic acid, terephthalic acid, and isophthalic acid, are particularly preferred. Preferred hemiesters are digemisuccinates, which may be present in the form of free acids or their salts, for example sodium, potassium or ammonium salts. One or more hydroxyl groups of silibinin can be esterified. Preferably, 1, 2, 3, 4 or all of the silibinin hydroxyl groups are esterified.

In a preferred embodiment of the present invention, the silibinin component is silibinin-C-2 ', 3-bis (hydrogen succinate) or a pharmaceutically acceptable salt thereof, for example sodium salts, potassium salts, ammonium salts, etc., as well as mixtures thereof. Disodium salt is particularly preferred.

Gluconic acid esters are also suitable esters.

Preferably, the silibinin component is a compound of general formula (I)

wherein

R ₁ , R ₂ , R ₃ , R ₄ and R _{5 are} independently selected from the group consisting of —H, —SO ₃ H, —PO ₃ H ₂ , —CO — C ₁ —C ₈ -alkylene-OH , -CO-C ₁ -C ₈ -alkylene-CO ₂ H, -CO-C ₁ -C ₈ -alkylene-SO ₃ H, -CO-C ₁ -C ₈ -alkylene-OPO ₃ H ₂ , -CO- C ₁ -C ₈ alkylene-PO ₃ H ₂ , - (C ₂ -C ₃ alkylene-O) _n -H, where n = 1 to 20, -CO-C ₁ -C ₈ alkylene-N ( C ₁ -C ₃ -alkyl) ₃ ⁺ X ^- , where X ^- represents a pharmaceutically acceptable anion,

or their pharmaceutically acceptable salts. Preferably, R ₁ , R ₂ and R ₅ are —H.

More preferably, the silibinin component of the general formula (I) has the stereochemical general formula (I-A) or (I-B)

In a preferred embodiment of the present invention, the compound of general formula (I-A) is mixed with the compound of general formula (I-B) in any relative mass ratio, for example 50 ± 5: 50 ± 5. In a preferred embodiment of the present invention, however, the diastereomeric excess (di) of the compound of general formula (IA) is at least 50% di, more preferably at least 75% di, even more preferably at least 90% di, even more preferably at least 95% di, most preferably at least 98% di, and particularly preferably at least 99% di. In another preferred embodiment of the present invention, the diastereomeric excess of the (di) compound of general formula (1-B) is at least 50% di, more preferably at least 75% di, even more preferably at least 90% di, even more preferably at least 95% di, most preferably at least 98% di, and particularly preferably at least 99% di.

Other preferred silibinin components are described in patent WO 03/090741, the essence of which is referred to in the present description.

Preferably, the silibinin component in pure water at room temperature has better solubility compared to silibinin per se.

In a preferred embodiment, the present invention relates to the use of a silibinin ester for the manufacture of a medicament which is preferably processed for parenteral or oral administration for the treatment of viral hepatitis, in particular hepatitis B or C. Preferably, the medicament is substantially free of silidianin and / or silikristin , and / or isosilibinin.

In a preferred embodiment of the present invention, the drug is processed for parenteral administration. Parenteral administration can be carried out, for example, subcutaneously, intravenously, intramuscularly, intraarterially, intraperitoneally, intradermally, intraarticularly, intrathecally, intracardially, into the vitreous body of the eye, retrobulbar, intrapulmonary and intraosseous.

Particularly preferably, the drug is processed for injection or infusion, in particular for intravenous or intra-arterial administration.

Suitable drugs suitable for injection or infusion are known to those skilled in the art. For more details see, for example, book: K.N.Bauer et al. “Lehrbuch der Pharmazeutischen Technologie [Textbook of Pharmaceutical Technology]”, 1999, 6th ed., WVG Stuttgart.

Medicines suitable for injection are conventional sterile solutions, emulsions or suspensions which are prepared by dissolving, emulsifying or suspending the active substance and, optionally, also excipients in water, in an appropriate liquid, in addition to water, which should not be sterilized, if there is a justification for this, or mixtures of these solvents.

Medicines for infusion are conventional sterile aqueous solutions or emulsions with water as a constant phase.

Medicines for injection or infusion may optionally contain additional excipients. Excipients of this type are preferably solvents, for example lecithin and poloxamer 188, isotonic agents, for example sodium chloride, glucose and mannitol, buffers, for example acetate, phosphate and citrate, antioxidants, for example ascorbic acid, sodium metahydrogen sulfite, sodium sulfite and sodium hydrogen sulfite chelating agents, for example disodium edithate, preservatives, for example, hydroxybenzoic acid esters, benzyl alcohol and cholesterol, as well as emulsifiers, for example lecithin, fatty alcohols, sterol s, sorbitol fatty acid esters, polyoxyethylene sorbitol fatty acid esters, polyoxyethylene fatty acid glycerides, polyoxyethylene fatty acid esters, polyoxyethylene fatty alcohol esters, glycerol fatty acid esters and poloxamers.

A particularly preferred drug is a powder for the preparation of an infusion solution comprising silibinin-C-2 ', 3-bis (hydrogen succinate), preferably in the form of a disodium salt, and optionally inulin as an excipient. Containers containing 598.5 mg of silibinin-C-2 ', 3-bis (hydrogen succinate) disodium salt and inulin powder are adapted to prepare an infusion solution and are manufactured in Germany under the brand name Legalon® SIL. In a preferred embodiment of the present invention, the medicament of the present invention is bioequivalent to this composition.

In another preferred embodiment of the present invention, the medicament is processed for oral administration. Preferably, the medicament is an oral administration form selected from the group consisting of tablets, capsules, sugar coated tablets, pills and sachets.

When the silibinin component is administered orally, it must be ensured that the bioavailability of the silibinin component in the oral dosage form is sufficiently high. Therefore, the limiting factor is the pronounced lipophilicity of silibinin.

In a particularly preferred embodiment, the present invention relates to the use of a silibinin component for the manufacture of a medicament that is processed for oral administration and substantially free of silidianin and / or silicristin and / or isosilibinin for the treatment of viral hepatitis, preferably hepatitis B or C.

It seems that these additional components of silymarin also have a physiological effect (for example, can cause side effects), but for the treatment of viral hepatitis, silibinin (or its analogues) is most effective, especially to reduce the viral load. Thus, with the introduction of silymarin, i.e. mixtures of silibinin, silidianin, silicristin, isosilibinin and other components, the total dose of silymarin should be relatively high to provide a certain amount of silibinin. For example, if silymarin contains, for example, 42 wt.% Silybinin, the administration of 125 mg of silymarin provides only about 52 mg of silybinin and about 73 mg of other compounds that also have a physiological effect (but not the desired effect). The risk of unwanted side effects increases with a dose of a physiologically active substance. Thus, since the complication is the profile of undesirable side effects, the administration of 52 mg of highly pure silibinin is superior to the administration of 125 mg of silymarin with 42 wt% silibinin (see T. Ding et al., J. Pharm. Biomed. Anal. 26 (1), 2001, pp. 155-161).

The structures of silibinin (silibin), silidianin, silicristin, and isosilibinin (isosilibine), described below, are given by D.Y.-W. Lee et al., J. Nat. Prod. 66, 2003, pp. 1171-1174; N.-C. Kim et al., Org. Biomol. Chem., 1, 2003, ss. 1684-1689):

Suitable administration forms that are applicable for oral administration (oral medicaments) are known to those skilled in the art. For more details see, for example, book: K.N.Bauer et al. “Lehrbuch der Pharmazeutischen Technologie [Textbook of Pharmaceutical Technology]”, 1999, 6th ed., WVG Stuttgart.

The oral administration form is preferably selected from the group consisting of tablets, powders, pills, granules, tablets coated with sugar, syrups, juices, solutions, effervescent powders, effervescent granules, effervescent tablets, lyophilisates and capsules. A particularly preferred form of oral administration is a tablet, a sugar-coated tablet, granules, pills or powders, particularly preferably tablets.

Suitable excipients for compositions in oral dosage forms are known to those skilled in the art. See, e.g., H.P. Fiedler, Lexikon der Hilfstoffe für Pharmazie, Kosmetik und angrenzende Gebiete [Encyclopedia of excipients for pharmacy, cosmetics and related areas], Editio Cantor Aulendorf, 2001.

Tablets can be prepared, for example, by mixing the silibinin component with known excipients, for example inert diluents, for example calcium carbonate, calcium phosphate and lactose, disintegrants, for example corn starch and alginic acid, binders, for example starch and gelatin, lubricants, for example magnesium stearate or talc and / or agents to achieve a depot effect, for example, carboxymethyl cellulose, cellulose acetate phthalate or polyvinyl acetate. Tablets may also consist of a number of layers. In addition to these solvents, tablets may also contain additives, for example sodium citrate, calcium carbonate and dicalcium phosphate, together with various additional substances, for example starch, preferably potato starch, gelatin, etc. In addition, slip agents, for example magnesium stearate, can be additionally used for tabletting. , sodium lauryl sulfate and talc.

Sugar-coated tablets can be obtained, for example, by coating the cores, which are prepared in a manner similar to the method for producing tablets with agents commonly used to prepare sugar coatings for tablets, for example, collidone or shellac, gum arabic, talc, titanium dioxide or sugar. To avoid the effect of depot or incompatibility, the core may also consist of a number of layers. Sugar-coated tablets may also consist of a number of layers to obtain a depot effect, and can be obtained using the above tablet excipients.

Juices, syrups, emulsions, suspensions and oral solutions may additionally contain a sweetener, for example saccharin, cyclamate, glycerin or sugar, as well as a flavoring agent, for example a flavoring agent, for example vanillin or orange extract. In addition, they may contain additional suspension or thickening agents, for example, carboxymethyl cellulose sodium, moisturizing agents, for example, condensation products of fatty alcohols with ethylene oxide, or preservatives, for example p-hydroxybenzoic acid esters.

Capsules can be prepared, for example, by mixing the silibinin component with inert carriers, for example lactose or sorbitol, and placing them in gelatin capsules. Excipients that can be used are, for example, water, pharmaceutically acceptable organic solvents, such as paraffins (e.g., oil fractions), vegetable oils (e.g., peanut or sesame), mono- or polyfunctional alcohols (e.g., ethanol or glycerin) solvents, for example natural minerals (e.g. kaolins, clays, talc, chalk), ground synthetic materials (e.g. finely divided silicic acid and silicates), sugars (e.g. sucrose, lactose and dextrose), emu ligators (e.g. lignin, sulphite-alcohol stillage, methyl cellulose, starch and polyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc, stearic acid and sodium laurite sulfate).

The drug may release the silibinin component immediately or from a controlled form. If release from a controlled form occurs, preferably such a form is formarethard. Slow release in the present invention preferably implies a release profile over which the silibinin component is released over a relatively long period at a reduced rate for a prolonged therapeutic effect. This can be, in particular, achieved by oral administration. The term “at least partially delayed release” of the present invention includes any medicament that guarantees a modified release of the silibinin component in the dosage form. The drugs are preferably in coated or uncoated administration forms, which are prepared using special excipients, by certain processes, or by combining both forms, for a selectively modified release rate or release site. Due to the duration of the release course in the case of the drugs of the present invention, the following types are included: delayed release (extended release), repeated release release, sustained release, and sustained release. For more details see, for example, book: K.N.Bauer et al. “Lehrbuch der Pharmazeutischen Technologie [Textbook of Pharmaceutical Technology]”, 1999, 6th ed., WVG Stuttgart.

Appropriate measurements of the controlled release of the active compound are known to those skilled in the art. If the drug is in the form for oral administration, for example in the form of a tablet, delayed release can be achieved, for example, by immersing the silibinin component in a polymer matrix and / or film coating the oral administration form with a membrane.

Solid, semi-solid or liquid controlled release formulations may be used in the present invention. Solid drugs are preferred, for example, oral osmotic systems, coated tablets, matrix tablets, multilayer tablets, jacket tablets, sugar coated jacket tablets, diffusion control pills, adsorbates and soft gelatin capsules depot. Oral drugs with controlled release of the active compound are particularly preferred in the form of coated tablets (film tablets), jacket tablets or matrix tablets, matrix tablets are particularly preferred.

Drugs with controlled release of the active substance may contain a silibinin component in a soluble, suspended and / or solid, amorphous or crystalline form.

To obtain controlled release dosage forms of the active compound of the present invention with controlled release of the active compound, the silibinin component can be used in the form of particles of different sizes, for example, unmilled, ground particles or microforms.

In controlled-release drugs, the active compound, the silibinin component, is preferably contained in the form of particles containing the active substance, for example, pills, granules, microcapsules, tablets, extrudates or crystals, which are coated with a diffusion-control membrane.

Such controlled diffusion drugs are preferably composed of many particles, i.e. they preferably consist of a plurality of coated cores, for example, neutral pills, onto which a mixture of a silibinin component with a conventional binder and thickener is applied, optionally together with conventional softeners and solvents, and then coated with a diffusion glaze, softener and other excipients. Drugs with controlled diffusion of the present invention, in addition, may consist of homogeneous cores, including silibinin component, which are obtained, for example, by granulation, rotary granulation, agglomeration in a fluidized bed, tabletting, wet extrusion or extrusion of melting, optionally giving a spherical forms and glaze for diffusion, which may contain emollients and other excipients.

Particles containing a silibinin component may contain excipients, for example acids or buffering agents, which modify the pH and thereby affect the decrease in the dependence of the release of the silibinin component on the pH of the release medium.

The diffusion control membrane may also include additional excipients, which, due to pH-dependent solubility, affect the permeability of the membrane at different pH values and thus minimize the pH dependence of the release of the silibinin component.

The binders and thickeners used to form the coated neutral pills are preferably hydroxypropyl methylcellulose (HPMC) and polyvinylpyrrolidone (PVP). Other natural, synthetic or partially synthetic polymers, for example methyl cellulose (MC), hydroxypropyl cellulose (HPC), other hydroxyalkyl celluloses and hydroxyalkyl methyl celluloses, carboxymethyl celluloses and their salts, polyacrylic acids, polymethacrylates, gelatin, starch and derivatives, can be similarly used.

For the preparation of pills, particles and (mini) tablets containing a silibinin component, cellulose, microcrystalline cellulose, cellulose derivatives such as HPMC, HPC and low substitution hydroxypropyl cellulose (HPC-NHS), dicalcium phosphate, lactose, PVP and sucrose are preferably used. as binders and fillers, by granulation methods, agglomeration in a fluidized bed, wet extrusion and tabletting.

To obtain pills by extrusion melting, the silibinin component is immersed in thermoplastic excipients. Preferably, the appropriate thermoplastic excipients are preferably HPC, HPMC, ethyl cellulose, hydroxypropyl methyl cellulose acetosuccinate, PVP, vinyl pyrrolidone / vinyl acetate copolymer, polyethylene glycol, polyethylene oxide, polymethacrylates, polyvinyl galactinates, for example, polyisinyl alginate, polyisinyl alginate, polyisinyl alginate, fatty acids.

Particles that contain a silibinin component can also include pH modifying agents, such as acids, bases, and buffering agents. The addition of such substances can significantly reduce the pH dependence of the release of the silibinin component and its salts, hydrates, and solvates.

Excipients that are used to modify the pH in cores containing a silibinin component are, for example, adipic acid, malic acid, L-arginine, ascorbic acid, aspartic acid, benzenesulfonic acid, benzoic acid, succinic acid, citric acid, ethanesulfonic acid, 2 -hydroxyethanesulfonic acid, fumaric acid, gluconic acid, glucoronic acid, glutamic acid, potassium hydrogentartran, maleic acid, malonic acid, methanesulfonic acid, toluenesul acid, trometamol, tartaric acid. Preferably, citric acid, succinic acid, tartaric acid and potassium hydrogen hydrogen tartrate are used.

Ethyl celluloses (e.g., Aquacoat® or Surelease®) and polymethacrylates (e.g., Eudragit® NE, Eudragit® RS and RL) are especially preferred for the production of a diffusion glaze. However, other materials, such as cellulose acetate and cellulose acetate butyrate, can also be used as film-forming diffusion-controlling polymers.

In addition to the diffusion-controlling polymer, the sprayed glaze may also contain additional excipients with pH-dependent solubility, for example, enteric dissolution polymers, for example cellulose phthalate, in particular cellulose acetate phthalate and hydroxypropyl methyl cellulose phthalate, in particular cellulose succinates, in particular cellulose acetosuccinate or hydroxyacetate, for example Eudragit® product L). By adding such substances, the pH dependence of the release of the silibinin component can be reduced.

The softeners used are, for example, citric acid derivatives, phthalic acid derivatives, benzoic acid and benzoic acid esters, other aromatic carboxylic acid esters, aliphatic dicarboxylic acid esters, monoacetate, diacetate and triacetate glycerol, polyols, fatty acids and their derivatives, glycerides acetylated fatty acids, castor oil and other natural oils, migliol and fatty acid alcohols.

To prevent adhesion of the coated particles during the process and in the final product, viscosity reducing agents such as talc, magnesium stearate, glycerol monostearate and aerosil can be added to the glaze.

The release rate is controlled by the composition of the glaze and the thickness of the glaze layer. The permeability enhancing agents are pore forming agents that can be added to the glaze or to the particles that are coated and which contain the silibinin component. Spore forming agents are soluble polymers, for example polyethylene glycols, PVP, PVA, HPMC, GPC, hydroxyethyl cellulose (HEC), MC, carboxymethyl cellulose and their salts, dextrins, maltdextrins, cyclodextrins, dextrans or other soluble substances, for example, urea, potassium chloride, sodium , ammonium chloride, sucrose, lactose, glucose, fructose, maltose, mannitol, sorbitol, xylitol and lactitol.

Excipients with pH-dependent solubility, which may be components of a diffusible coating, are, for example, enteric polymers, for example cellulose phthalates, in particular cellulose acetate phthalate and hydroxypropyl methyl cellulose phthalate, in particular cellulose succinates, in particular cellulose acetosuccinate and acetylsuccyl acetate, for example Eudragit® product L).

In addition, the drug with controlled release of the silibinin component may be a coated administration form that contains one or more swellable excipients that increase greatly when liquid penetrates through the membrane and causes rupture of the membrane as a result of swelling and volume expansion. As a result of rupture of the shell, the release of the pharmaceutical agent from the drug becomes possible (pulsating release). Such drugs as swellable excipients preferably comprise polyvinylpyrrolidones, crospividones, cross-linked sodium carboxymethyl cellulose sodium, cross-linked sodium carboxymethyl starch sodium salt, polyethylene oxides, polymethacrylates, low substitution hydroxypropyl cellulose (HPC-nc). Suitable coating materials are preferably cellulose acetate, ethyl cellulose and polymethacrylates.

Drugs with a coating controlled by diffusion and pulsating release can be used directly as a finished pharmaceutical form. However, they can also be further processed, optionally with the addition of excipients, to obtain the final form of administration (eg, capsules, tablets, sachets). To achieve the desired release profile, various coated particles can also be combined with other particles in a pharmaceutical form, and an initial dose may be administered, for example, by combining quick release particles, for example uncoated pills, granules or powders.

Controlled release drugs that can be used are also formulations that contain a silibinin component in the matrix. Such matrix formulations release the silibinin component by diffusion and / or erosion. Preferably, these drugs are in the form of a tablet or a series of tablets, which, for example, can be encapsulated. Tablets may be coated (film tablets) or varnished. Such drugs are obtained, for example, by mixing the components and direct tableting, or by dry or wet granulation, followed by tableting.

The matrix forming agents may be water soluble, water swellable, or water insoluble. Preferably, the medicaments contain one or more water-swellable polymers.

The water-soluble and water-swellable matrix forming polymers used are preferably hydroxypropyl methyl cellulose (HPMC), hydroxyethyl methyl cellulose, hydroxypropyl cellulose (HPC), hydroxyethyl cellulose, methyl cellulose (MC), ethyl cellulose, hydroxy cellulose, hydroxymethyl cellulose and hydroxyalkyl gaur gum and carob bean flour, xanthan gum, polyethylene oxides, polyacrylic acids, polymethacrylate acid, polymethacrylate acid derivatives, polyvinyl alcohols (PVA), partially hydrolyzed polyvinyl acetate, polyvinylpyrrolidone, agar, pectin, acacia, tragacanth, gelatin, starch or starch derivatives and mixtures thereof. The use of HPMC is particularly preferred.

In addition, water-insoluble substances can be used as structure-forming agents, for example, unsaturated or saturated (hydrogenated) fatty acids and their salts, esters or amides, mono-, di- and triglycerides of fatty acids, waxes, ceramides, cholesterol derivatives and mixtures of these substances.

Medicines may also contain conventional tabletting excipients, preferably finely divided silicon (Aerosil® product), magnesium stearate, talc, PVP, lactose or microcrystalline cellulose.

In addition, substances can be incorporated into the matrix, which control the pH in the matrix. By the addition of such pH modifying excipients and / or by the addition of substances that increase the pH or are washed out by dissolution from the matrix and thus increase the porosity or permeability of the matrix and / or induce matrix erosion, it is possible for these preferred embodiments of the present invention to achieve the greatest degree of pH independent release.

The matrix containing the silibinin component can also be in special geometric shapes, the release from the matrix depending on the special geometry and surface of the matrix. The surface of the matrix and the release surface can be controlled, for example, by compression to obtain special formats (for example, ring-shaped tablets), and / or by applying a subzone, or by applying barrier layers using a multilayer press.

Compositions with different release properties can preferably be combined to formulate the pharmaceutical form in the form of multilayer tablets or tablets with a jacket core. For example, by using multilayer tablets comprising a quick release layer or jacquette core tablets having a quick release coating, a controlled release of the present invention is achieved with a high initial release of the silibinin component, although it can be achieved by using a jacquette core tablet with a quick release releasing core and increased release at the end.

Another drug with controlled release of the silibinin component is a drug in which the silibinin component is added to a matrix consisting of one or more physiologically acceptable excipients by a melting process. The release of the silibinin component from such “fused extrudates” occurs due to diffusion and / or erosion. Preferably, such controlled release formulations of the silibinin component are in the form of granules, pills or tablets. Forms obtained by melting extrusion include, in particular, pills and granules, which can be processed to obtain other pharmaceutical forms, for example, by encapsulation or tabletting, optionally with the addition of other traditional pharmaceutical excipients. In addition, the fused extrudates of the present invention can be milled and subsequently used in such milled form to produce other drugs, for example matrix tablets. Additional processing also includes a combination of formulations having different pharmaceutical release, for example, retard particles and quick release particles, to obtain a drug.

Fused extrudates and / or pharmaceutical forms that are prepared from fused extrudates may be coated or varnished. Fused extrudates are preferably obtained by mixing the silibinin component with at least one fusible physiologically acceptable (carrier) and optionally conventional additional pharmaceutical substances, melting at a temperature of from 50 to 250 ° C, preferably 60 to 200 ° C, by injection of the melt or extrudate and shaping. With this method, the components can be mixed before melting or during melting, or some of the components are fused and other components are added to this melt. The mixture of solvents, silibinin component and optionally additional substances is thermoplastic deformable and therefore can be extruded. Numerous methods, based on their names, give an idea of the method of shaping the mixture, for example, hot granulation, cold granulation, calendering, extrusion and deformation of the still plastic mass or rounding.

The thermoplastic carriers used, which preferably swell and dissolve in physiological environments, are preferably: polyvinylpyrrolidone (PVP), copolymers of N-vinylpyrrolidone (NVP) and vinyl esters, in particular vinyl acetate, copolymers of vinyl acetate and crotonic acid, partially hydrolyzed polyvinyl acetate, polyvinyl acetate, polyvinyl acetate, polyvinyl acetate, cellulose ethers, in particular methyl cellulose and ethyl cellulose, hydroxyalkyl cellulose, in particular hydroxypropyl cellulose, hydroxyalkyl methyl cellulose, in particular droksipropilmetiltsellyulozy and hydroxyethylmethyl cellulose, carboxymethyl cellulose, cellulose phthalates, in particular cellulose acetate phthalate and hydroxypropylmethylcellulose phthalate, cellulose succinate, in particular cellulose acetate succinate and hydroxypropylmethylcellulose acetate succinate, polyhydroxyalkyl acrylates, poligidroksialkilmetakrilaty, polyacrylates and polymethacrylates (Eudragit® type products), copolymers of methyl methacrylate and acrylic acid, polylactides , polyethylene glycols, polyethylene oxides and polysaccharides, for example galactomannans and ginovaya acid and its alkali metal salts and ammonium salts.

Preferred thermoplastic excipients for the preparation of drugs with controlled release of the silibinin component are hydroxypropyl cellulose, polyvinylpyrrolidone, vinylpyrrolidone / vinyl acetate copolymers, polymethacrylates, in particular Eudragit® L product, hydroxypropylmethyl cellulose acetate and succinate, and polyethylene polyethylene. Emollients that can be used to lower the glass transition temperature of the mixture are, for example, propylene glycol, glycerin, triethylene glycol, butanediols, pentanols, for example, pentaerythritol, hexanols, long chain alcohols, polyethylene glycols, polypropylene glycols, polyethylene / polypropylene glycols, for example, silicones , dimethyl phthalate, diethyl phthalate, dibutyl phthalate), benzoic acid and benzoic acid esters, other aromatic carboxylic acid esters (e.g. trimellitic acid esters slots), derivatives of citric acid (e.g. triethyl citrate, tributyl citrate, acetyl triethyl citrate), esters of aliphatic dicarboxylic acids (e.g. dialkyl adipates, esters of sebacic acid, in particular diethyl sebacinates, esters of tartaric acid), glycerol monoacetate, glycerol fatty acid, dihydric acid, derivatives (e.g. glycerol monostearates, acetylated fatty acid glycerides, castor oil and other natural oils, migliol), fatty acid alcohols (e.g. cetyl alcohol, cetearyl alcohol), sugar pa, polyhydric alcohols and sugar derivatives (e.g., erythritol, isomalt, lactitol, mannitol, maltitol, maltdekstrin, xylitol).

In addition to the silibinin component, carrier (s) and optionally softener (s), the extrudable mixture may also contain other pharmaceutical traditional additional substances, for example, slip agents and lubricant release agents, slip agents and flow agents, fillers and adsorbents, stabilizers, free radical traps, complexing agents, antioxidants, photostabilizers, propellants, surfactants, preservatives, colorants, sweeteners and flavorings.

Slip and mold release agents may include, for example, stearic acid and stearates, in particular aluminum, calcium and magnesium stearates, calcium behenate, sodium stearyl fumarate, talc, silicones, waxes, and mono-, di- and triglycerides, for example glycerol monostearate, glycerol distearate, glycerol dibehenate, glycerol monooleate, glycerol palmitine stearate.

The flow agents used are preferably animal and vegetable oils, preferably in hydrogenated form with a melting point of at least 50 ° C, waxes (e.g. carnauba wax), mono-, di- and triglycerides (e.g. glycerol monostearate, glycerol distearates, glycerol dibehenate , glycerol monooleate, glyceroyl palmitostearate), phosphatides, in particular lecithin.

Useful filler materials, for example, are titanium dioxide, alumina, magnesium oxide, silicic acid and silicates, stearic acid and stearates, cellulose derivatives (e.g. methyl cellulose), starch and starch derivatives, sugars, polyhydric alcohols and sugar derivatives.

Drugs with controlled release of the silibinin component can also be fused extrudates, which contain an excipient with pH-modifying properties and / or pH-dependent solubility. Using these excipients (for example, acids, bases, buffers of enteric polymers already described above), the pH dependence of the release of the silibinin component can be minimized.

Upon receipt of fused extrudates, there may be the formation of “solid solutions” in which the silibinin component is contained in the matrix in molecular dispersed form.

Drugs with controlled release of the silibinin component may also be osmotic pharmaceutical release systems. In principle, osmotic systems of this type are known in the prior art. In the present invention, a pharmaceutical release from a pharmaceutical form is usually based on osmotic pressure as a driving force.

The osmotic system preferably consists of a core that contains a silibinin component, optionally a hydrophilic swelling agent and optionally a water-soluble substance for inducing osmosis, and optionally also pharmaceutically acceptable excipients, and a shell that consists of a permeable material that is impermeable to core components and has at least one an outlet through which core components can be released.

The material from which the drug release coating of the present invention is formed with controlled release of the silibinin component is formed from a semi-permeable, i.e. permeable to water, aqueous media and biological fluids, and impermeable or very limited permeable to core components, and suitable for forming a film coating. The selectively semipermeable shell material is insoluble in tissue fluids, does not corrode, does not collapse in the gastrointestinal tract and is excreted unchanged, or it undergoes bioerosion only towards the end of the release period.

Typical coating materials for the osmotic system are preferably acylated cellulose derivatives (cellulose esters), which are from mono- to trisubstituted with acetyl groups or from mono- to disubstituted with acetyl groups and additionally an acyl radical instead of acetyl, for example cellulose acetate, cellulose triacetate, cellulose acetate / ethyl carbamate, cellulose acetate phthalate, cellulose acetate methyl carbonate, cellulose acetate succinate, cellulose acetate dimethylamino acetate, acetodiethylaminoac cellulose acetate, cellulose acetoethyl carbonate, cellulose acetochloroacetate, cellulose acetoethyl oxalate acetate, cellulose acetate methyl sulfonate, cellulose acetate propionate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, cellulose acetate, acetate

Ethyl cellulose, copolymers of alkylene oxide and alkyl glycidyl ether, polymer epoxides, polyglycols and polylactic acid derivatives are also suitable semi-permeable osmotic membrane membranes. In addition, mixtures of water-insoluble acrylates per se, for example, a copolymer of ethyl acrylate and methyl methacrylate can be used.

If necessary, the shell of the osmotic system may also contain emollients, for example emollients mentioned above, and other additional substances, such as pore forming agents. If necessary, photoprotective glaze can be used for a semi-permeable coating, which may consist, for example, of HPMC and HPC, and an appropriate emollient (e.g. polyethylene glycol) and pigments (e.g. titanium oxide, iron oxides).

In order to control the initial dose of the silibinin component, the osmotic system can also be provided with a coating that contains the silibinin component, from which the silibinin component is preferably quickly released upon contact with the release medium before the osmotically controlled release of the silibinin component from the core begins.

Suitable water-swellable polymers that may be contained in the core of the osmotic system are preferably polyethylene oxides (e.g., Polyox® product), xanthan gum, copolymers of vinyl pyrrolidone and vinyl acetate, polyvinyl pyrrolidones, crospovidones, cross-linked sodium carboxymethylmethyl cellulose cellulose hydroxymethylmethyl cellulose cellulose hydroxymethylmethyl cellulose with a low degree of substitution (GPC-NSZ), poly (hydroxyalkyl methacrylate), alginates and galactomannan mi as well as other hydrophilic polymeric swelling agents and their mixtures.

Suitable osmotically active substances that can be added to the core for the induction of osmosis are water-soluble salts of organic and inorganic acids or non-ionic organic substances with high solubility in water, for example carbohydrates, in particular sugars, or amino acids. As an example, we can cite several substances that can be included in the core of the osmotic system separately or in a mixture for the induction of osmosis: inorganic salts, for example chlorides, sulfates, sulfites, carbonates, bicarbonates, phosphates, disubstituted phosphates, monosubstituted phosphates of alkali metals and alkaline earth metals for example sodium, lithium, potassium, calcium or magnesium, organic acids, for example adipic acid, ascorbic acid, succinic acid, citric acid, fumaric acid, maleic acid, in foreign acid, benzoic acid and their alkali or alkaline earth metal salts, acetates, pentoses, for example arabinose, ribose or xylose, hexose, for example glucose, fructose, galactose or mannose, and saccharides, for example sucrose, maltose or lactose, trisaccharides, for example raffinose, polyols, for example mannitol, sorbitol, maltitol, xylitol or inositol, and urea. Sodium chloride and sodium carbonate are preferably used.

In addition, the osmotic system may include other conventional pharmaceutical adjuvants, for example, lubricating and separating agents, glidants, binders, colorants, thickeners, protective colloids, stabilizers and surfactants.

The osmotic release system is preferably obtained by standard methods, for example wet granulation or dry pressing, tabletting and subsequent organic coating.

The coating of the osmotic system has at least one open outlet through which the silibinin component is released, optionally together with other core components. The output can be made in the shells in various ways, for example, by perforation, machining, or laser drilling. The term “open” also includes biodegradable materials that protrude from the shell when administering the medicament of the present invention and result in the formation of open in situ exits.

In another embodiment of the present invention, for the controlled release of the silibinin component, the silibinin component may also be contained as an ion exchange complex (adsorbate).

Preferably, the drug is processed for administration once a day, twice a day, three times a day, or four times a day.

In a preferred embodiment of the present invention, from 0.5 to 75% by weight of the initially contained silibinin component is released from the drug after 1 hour in vitro. Appropriate conditions for determining the release of active substances in vitro are known to those skilled in the art. In this regard, information can be obtained, for example, in the European Pharmacopoeia. Preferably, the release is determined using a paddle stirrer in an artificial gastric juice (buffer with a pH value of 1.2) or in an artificial intestinal juice (buffer with a pH value of 7.6). The amount of released silibinin component can be investigated, for example, using HPLC and UV scopy.

Preferred release profiles A ₁ to A _{8 are} summarized in the table below.

By
after
time
[h] A ₁ A ₂ A ₃ A ₄ A ₅ A ₆ A ₇ A ₈ wt.% wt.% wt.% wt.% wt.% wt.% wt.% wt.% 0.5 5.0-34 6.0-33 7.0-32 9.0-31 11-30 13-30 15-29 17-28 one 12-53 15-52 18-50 20-48 22-46 24-44 27-42 30-40 2 25-74 27-71 29-68 31-65 33-62 36-60 39-58 42-56 3 33-85 36-82 39-79 42-76 45-73 48-71 50-69 52-67 four 41-92 44-89 47-86 50-83 53-81 55-79 58-77 60-75 6 52-98 55-97 58-96 60-94 63-92 66-90 69-88 72-86 8 > 62 > 65 > 68 71-99 74-98 76-98 78-97 80-97 12 > 70 > 73 > 76 > 79 > 82 > 84 > 86 > 88

In a preferred embodiment of the present invention, the medicament comprises cyclodextrin and / or phospholipid.

Pharmaceutical formulations containing silibinin and cyclodextrins are known in the art (see, for example, EP 422497). Preferably, silibinin forms an incorporated complex with cyclodextrin. Preferred cyclodextrins are α-, β- and γ-cyclodextrins, their O-C ₁ -C4-alkyl and hydroxy-C ₁ -C ₄ -alkyl derivatives.

Pharmaceutical compositions that contain silibinin and phospholipids are also known in the art (see US 4,764,508). Preferably, silibinin forms a complex with a phospholipid. Preferred phospholipids are phosphatidylcholine, phosphatidylethanolamine and phosphatidylserine. Preferred silibinin phospholipid complexes are tertiary complexes additionally containing vitamin E (tocopherol). Complexes of this type are known from the prior art and are referred to as “SPV complexes” (see A. Federico, Gut. 55 (6), 2006, pp. 901-902).

In addition to the silibinin component, the drug may contain one or more terpenes. Due to the action of terpene, the necessary conditions for adsorption, and the adsorption processes in general, can be improved. The terpenes can be natural or synthetic essential oils and / or their terpenoid components in the form of pure substances or mixtures, or derivatives of such pure substances. Among the essential oils, in particular, thyme oil, eucalyptus oil, pine needles oil, tea tree oil, cayeput oil, cardamom oil, peppermint oil, sage oil and rosemary oil, preferably thyme oil, can be noted. Of terpenes - substances also intended to be included, in particular, hemiterpenes can be used, which include, for example, isoprene, tiglinic acid, isovalerianic acid; monoterpenes, including acyclic monoterpenes, for example, 2,6-dimethyloctane, α-mirken, (E) -p -cimene, perylene, linalul, geraniol, (S) - (+) citronellol and monocyclic monoterpenes, for example cyclopropane monoterpenes and cyclobutane monoterpenes for example chrysanthemum acid or union, cyclopentane monoterpenes, for example iridoids or nonpetalactones or (-) - secologanin and (-) - oleuropein, monoterpenes cyclohexane, for example, o-menthane, cis- or trans-p-menthane, (R) - ( +) - limonene, terpinols, (-) - menthol, (+) - perillaldehyde, (-) - menton or (+) - carvone, bicyclic monoterpenes, for example, with oxygen-bridged terpenes 1,4-cineol, 1,8-cineole or ascaridole; cyclopropane bicycles of karana and tuyana, cyclobutane bicycles of pinan, and bicycloheptanes camphan and fenhan; sesquiterpenes, for example, famosan, bisabolan, germacran, elemen and gumilan. Particularly preferred terpenes are thymol, menthol, cineole, borneol, carvone, limenone and pinene, usually preferably thymol.

The drug contains a silibinin component. Silibinin is a component of silymarin. Preferably, in addition to silibinin or silibinin components, the drug does not contain any of the other components of silymarin. If the silibinin component is silibinin itself, the drug preferably does not contain any of the other silibinin components of silymarin. If the silibinin component is not silibinin itself, but, for example, silibinin ether, the drug preferably does not contain any of the other components of silymarin, i.e. also does not contain silibinin.

Preferably, one or more substances selected from the group consisting of isosilibinin, silidianin, silikristin, taxifolin, isosilikristin, silimonin, silandrin, siligermin and neosiligermin are absent in the drug, i.e. the drug preferably substantially does not contain at least one of the above substances. In this regard, the term "practically not contained" means that the residual content of the substance is preferably less than 2.0 wt.%, More preferably less than 1.0 wt.%, Even more preferably less than 0.5 wt.%, Most preferably less than 0 , 1 wt.% And especially less than 0.05 wt.% Of the total weight of the drug. Analytical methods for determining the residual amounts of these substances are known to those skilled in the art, for example, HPLC.

It has been established that the individual components of silymarin differ in chemical and physical properties and affect the pharmacological effect of silymarin to varying degrees, therefore, it is useful to prescribe silibinin, or its derivatives, and / or salts as the only component of silymarin, i.e. exclusively. Obviously, in this way, both patient efficiency and compliance can be improved.

In addition, it was unexpectedly found that the tolerance of the various constituent parts of silymarin is different, and that silybin is better tolerated, especially less toxic, compared to silymarin (i.e., compared to a mixture containing components other than silibinin).

In a preferred embodiment, the present invention relates to the use of a silibinin component for the manufacture of a medicament which is preferably processed for parenteral or oral administration and which, in addition to the silibinin component, does not contain any of the constituents of silymarin for the treatment of viral hepatitis, in particular hepatitis B and or C .

Particularly preferred drugs that are adapted for oral administration of the silibinin component are described below. All of these oral dosage forms in total comprise preferably a silibinin component in a substantially pure form, i.e. preferably in the absence of other components of silymarin, especially in the absence of isosilibinin and / or silicristin and / or silidianin.

Preferably, the oral dosage forms are oral immediate release dosage forms, i.e. the silibinin component is rapidly released from the dosage form, which leads to the rapid entry of the drug into the gastrointestinal tract. In a preferred embodiment of the present invention, 30 minutes after the administration of the oral dosage form, at least 75 wt.%, More preferably at least 80 wt.%, Even more preferably at least 85 wt.%, Most preferably at least 90 wt.% and especially at least 95 wt.% initially contained silibinin component is released from the oral dosage form.

In a preferred embodiment of the present invention, a medicament is provided as a solid solution. A solid solution is preferably obtained by immersing the silibinin component in a molecularly dispersed form of a highly soluble, preferably amorphous polymer matrix with a large specific surface area. The silibinin component may be contained in a molecular dispersion form, i.e. not in microcrystalline, and not in finely divided crystalline. A highly soluble amorphous state can be achieved by the use of highly soluble solid polymer solvents when extracting the silibinin or silibinin component from the silymarin extract. Such a technical pharmaceutical composition increases the solubility of the silibinin component and its dissolution rate.

An example of such a solid solution includes a silibinin component, an appropriate polymer (e.g., polyvinylpyrrolidone (PVP) or a polyvinylpyrrolidone copolymer, e.g., Kollidon® 25 product), and optionally dextrin (e.g., maltodextrin). The composition may contain additional excipients, for example, aerosol and / or talc.

Preferred solid solution options from B ₁ to B _{6 are} presented in the table below.

Wt% B ₁ B ₂ In ₃ B ₄ B ₅ B ₆ Silibinin component 1,0-50 2,5-20 8.0 ± 5.0 8.0 ± 4.0 8.0 ± 3.0 8.0 ± 2.0 PVP 1,0-97 10-80 64 ± 15 64 ± 12 64 ± 10 64 ± 7.0 Dextrin 1,0-70 5.0-50 22.8 ± 20 22.8 ± 15 22.8 ± 10 22.8 ± 7.0 Aerosil 0-10 0-7.5 4.0 ± 3.0 4.0 ± 2.5 4.0 ± 2.0 4.0 ± 1.5 Talc 0-5.0 0-2.5 1.2 ± 1.0 1.2 ± 0.7 1.2 ± 0.5 1.2 ± 0.3

The composition may be provided, for example, in a hard gelatin capsule.

In another preferred embodiment of the present invention, there is provided a drug in the form of a self-emulsifying microemulsion. Self-emulsifying lipid systems can be used as carriers and can lead to high bioavailability of the drug agent contained in them. The lipid system is colloidal in nature and allows the absorption of microparticles, especially of colloidal size, also through the lymphatic system in the gastrointestinal tract. Typically, the dissolved drug is saturated, but recrystallization does not occur. When administered orally, lipophilic drugs, such as the silibinin component, the microemulsion generally serves as an optimized carrier that increases the solubility of dissolved or highly dispersed drugs at the point of absorption. In other words, the lipid system acts as an adsorption enhancer.

An example of such a lipid system is a silibinin component, a corresponding first emulsifier (for example, lauroyl macrogolglyceride, for example Gelucire® 44/14), and an optional second emulsifier (for example, caprilocaprilm microgolglyceride, for example Labrasol®). The composition may further include excipients, for example polysorbate.

Preferred C ₁ to C ₆ solid solution options are presented in the table below.

Wt% C ₁ C ₂ C ₃ C ₄ C ₅ C ₆ Silibinin component 0.1-50 0.5-20 4.0 ± 3.5 4.0 ± 3.0 4.0 ± 2.5 4.0 ± 2.0 First emulsifier 1,0-99 5-97 54 ± 15 54 ± 12 54 ± 10 54 ± 7.0 Second emulsifier 0-70 0-70 41 ± 20 41 ± 15 41 ± 10 41 ± 7.0 Polysorbate 0-10 0-7.5 1.5 ± 1.0 1.5 ± 0.7 1.5 ± 0.5 1.5 ± 0.3

A composition, which may be solid or preferably semi-liquid, may be provided, for example, in a hard gelatin capsule or in a soft gelatin capsule.

In another preferred embodiment of the present invention, a medicament is provided in the form of a nanotechnological composition. The average particle size is preferably less than 1 micron. Nanoparticles are able to penetrate biological biological structures. The silibinin component is preferably adsorbed on the surface of said nanoparticles. Nanoparticles are preferably selected from the group consisting of inorganic nanoparticles and organic nanoparticles.

Inorganic nanoparticles include crystalline silicates, for example, of mineral origin or artificial silicates, for example metallosilicates, for example aluminosilicates (for example zeolites). Such inorganic nanoparticles are preferably chemically modified so that they carry electrostatic charges. Silicates are an ultrathin base for nanoparticles and the silibinin component is bonded (adsorbed) to the microporous surface of the nanoparticles.

Organic nanoparticles include clusters or agglomerates of low molecular weight proteins, or oligopeptides, or lipids. A suitable protein carrier is, for example, protamine.

Methods for producing nanoparticles are known to those skilled in the art. For example, colloidal nanoparticles as carriers for the oral release of a drug can be prepared by spraying the drug, i.e. silibinin component, together with the corresponding carrier materials under pressure at, for example, 60 ° C, through nozzles equipped with perforated sieves (matrices) in highly cooled towers. Spontaneous cooling forms an amorphous phase consisting of nanoparticles.

Solid lipid nanoparticles can be obtained, for example, by homogenization of high pressure and subsequent cooling by spraying. Preferably a drug, i.e. the silibinin component is used as a solution in an appropriate solvent or nominally in the form of sub microparticles. The silibinin component can be sprayed and homogenized by pressure, respectively, when mixed with a lipid solvent and a surfactant at a temperature of, for example, 60 ° C. After optionally adding finely divided fillers as an external phase, as well as glidants and other surfactants, the resulting gelatin capsules can be filled with the composition.

Examples of such dense lipid nanoparticles include the core of the silibinin component, the corresponding first emulsifier (for example, stearoyl macrogolglyceride, for example, Gelucire® 50/13 product), and optionally the corresponding macromolecular nonionic surfactant (for example, poloxamer). The composition preferably also contains an external phase (coating) comprising a first surfactant (e.g. Tween 20), aerosil and a second surfactant (e.g. glyceryl palmitostearate, e.g., Percirol® product).

Preferred embodiments of D ₁ to D ₆ solid solution are presented in the table below.

wt.% D ₁ D ₂ D ₃ D ₄ D ₅ D ₆ Silibinin component 0.1-30 0.5-20 4.5 ± 3.0 4,5 ± 2,5 4,5 ± 2,0 4,5 ± 1,5 First emulsifier 10-99 20-95 75 ± 20 75 ± 15 75 ± 10 75 ± 7.5 Macromolecular nonionic surfactant 0-50 0-40 15 ± 10 15 ± 7.5 15 ± 5 15 ± 2.5 First surfactant 0-10 0.1-7.5 1.5 ± 0.7 1.5 ± 0.5 1.5 ± 0.3 1.5 ± 0.2 Spray can 0-10 0.1-7.5 3.0 ± 2.0 3.0 ± 1.5 3.0 ± 1.0 3.0 ± 0.7 Second surfactant 0-10 0.1-7.5 1.5 ± 0.7 1.5 ± 0.5 1.5 ± 0.3 1.5 ± 0.2

Loaded nanoparticles achieve a much faster onset of drug delivery.

The medicament contains a silibinin component, preferably in a dose of at least 10 mg, at least 15 mg, at least 20 mg, at least 25 mg, at least 50 mg, at least 75 mg, at least 100 mg, at least 125 mg, at least 150 mg, at least 175 mg, or at least 200 mg; more preferably at least 225 mg, at least 250 mg, at least 275 mg, at least 300 mg, at least 325 mg, at least 350 mg, at least 375 mg, or at least 400 mg; even more preferably at least 425 mg, at least 450 mg, at least 475 mg, at least 500 mg, at least 525 mg, at least 550 mg, at least 575 mg or at least 600 mg ; most preferably at least 625 mg, at least 650 mg, at least 675 mg, at least 700 mg, at least 725 mg, at least 750 mg, at least 775 mg, or at least 800 mg; and especially at least 825 mg, at least 850 mg, at least 875 mg, at least 900 mg, at least 925 mg, at least 950 mg, at least 975 mg, or at least 1000 mg ; in each case, as an equivalent dose based on silibinin.

The medicament contains a silibinin component, preferably at a dose of at least 1.0 mg / kg, more preferably at least 2.5 mg / kg, even more preferably at least 5.0 mg / kg, most preferably at least 7, 5 mg / kg and especially at least 10 mg / kg, at least 12.5 mg / kg, at least 15 mg / kg, at least 17.5 mg / kg, at least 20 mg / kg at least 22.5 mg / kg, at least 25 mg / kg, at least 27.5 mg / kg or at least 30 mg / kg, based on the patient’s body weight and in each case in the form of an equivalent coagulant doses of silibinin. Preferably, said dose is a daily dose. Thus, if the drug is adapted, for example, for administration twice a day, the corresponding daily dose is divided into two parts of an equal amount. Similarly, for example, if the drug is adapted for administration three times a day, the corresponding daily dose is divided into three parts of an equal amount.

In a preferred embodiment of the present invention, the daily dose of the silibinin component is at least 5, more preferably at least 10, even more preferably at least 15, and most preferably at least 20 mg per kg body weight, based on an equal weight of silibinin.

In a preferred embodiment of the present invention, the daily dose of the silibinin component is 20 mg per kg body weight, based on an equal mass of silibinin. Thus, if the drug is adapted for once-daily administration, it preferably contains the entire amount of the silibinin component, for example 1400 mg of silibinin for a patient weighing 70 kg. If the drug is adapted for administration twice a day, it preferably contains half the amount of the silibinin component, for example 700 mg of silibinin for a patient weighing 70 kg. If the drug is adapted for administration three times a day, it preferably contains a third of the amount of silibinin component, for example 467 mg of silibinin for a patient weighing 70 kg. If the drug is adapted for administration four times a day, it preferably contains a quarter of the amount of the silibinin component, for example 350 mg of silibinin for a patient weighing 70 kg.

», где каждая цифра означает число часов и подчеркнутые цифры означают длительность инфузии, а неподчеркнутые цифры означают lag-фазу между двумя периодами инфузии. Предпочтительно режим лечения разнородный, т.е. за 24 ч все инфузии равно дозированы на протяжении равных периодов времени, и lag-фазы между следующими друг за другом инфузиями также идентичны.If the drug is adapted for parenteral administration, preferably for infusion, the preferred treatment regimen is 4 equal infusions, lasting 2 hours each. Preferably, after 4 hours, the same infusion is repeated such that a total of 4 infusions are carried out over 24 hours. This mode can be schematically abbreviated as "

”, Where each digit indicates the number of hours and the underlined numbers indicate the duration of the infusion, and the underlined numbers indicate the lag phase between the two infusion periods. Preferably, the treatment regimen is heterogeneous, i.e. after 24 hours, all infusions are equally dosed over equal periods of time, and the lag phases between consecutive infusions are also identical.

Based on the foregoing, the preferred modes of parenteral administration are summarized in the table below.

Once a day

Twice a day

Thrice a day

Four times a day

In a preferred embodiment of the present invention, the medicament is adapted for administration once, twice, three or four times a day so that the total daily dose administered by the established method contains at least 300 mg, at least 325 mg, at least 350 mg, at least 375 mg, or at least 400 mg; more preferably at least 425 mg, at least 450 mg, at least 475 mg, at least 500 mg, at least 525 mg, at least 550 mg, at least 575 mg, or at least 600 mg; even more preferably at least 625 mg, at least 650 mg, at least 675 mg, at least 700 mg, at least 725 mg, at least 750 mg, at least 775 mg, or at least 800 mg ; more preferably at least 825 mg, at least 850 mg, at least 875 mg, at least 900 mg, at least 925 mg, at least 950 mg, at least 975 mg, or at least 1000 mg ; most preferably at least 1050 mg, at least 1100 mg, at least 1150 mg, at least 1200 mg, or at least 1250 mg; and especially at least 1300 mg, at least 1350 mg, at least 1400 mg, at least 1450 mg, or at least 1500 mg; in each case, the equivalent dose is based on silibinin.

Preferred pharmacokinetic parameters AUC _0-t , AUC _t-∞ , AUC _0-∞ and AUC _0-∞ (corrected) (preferably after several infusions, for example after 11 infusions; single dose: 12.5 mg / kg; daily dose: 4 infusion; total dose: 11 infusions) are summarized as embodiments from E ₁ to E ₈ in the table below.

E ₁ E ₂ E ₃ E ₄ E ₅ E ₆ E ₇ E ₈ mcg h / ml mcg h / ml mcg h / ml mcg h / ml mcg h / ml mcg h / ml mcg h / ml mcg h / ml AUC _0-t 333 ± 200 333 ± 150 333 ± 125 333 ± 100 333 ± 80 333 ± 60 333 ± 40 333 ± 20 AUC _t-∞ 322 ± 200 322 ± 150 322 ± 125 322 ± 100 322 ± 80 322 ± 60 322 ± 40 322 ± 20 AUC _0-∞ 655 ± 200 655 ± 150 655 ± 125 655 ± 100 655 ± 80 655 ± 60 655 ± 40 655 ± 20 AUC _0-∞
(corrected) 414 ± 200 414 ± 150 414 ± 125 414 ± 100 414 ± 80 414 ± 60 414 ± 40 414 ± 20

In a preferred embodiment of the present invention, the medicament containing the silibinin component is adapted for adjuvant therapy, preferably for immunomodulating / antiviral combination therapy, for example interferon / ribovarin.

In a preferred embodiment of the present invention, in addition to the silibinin component, the drug contains an additional pharmaceutical agent, which is preferably applicable for the treatment of inflammatory liver diseases, especially preferably viral liver diseases, especially for the treatment of hepatitis B or C.

Preferably, the additional pharmaceutical agent is selected from the group comprising liver therapies, lipotropic agents [A05B]; nucleosides, nucleotides, selective reverse transcriptase inhibitors [J05AB]; interferons [L03AB] and monoclonal antibodies against HBV (hepatitis B virus). The designations in square brackets refer to the PBX index, preferably in the 2007 German version.

Particularly preferably, the other pharmaceutical agent is selected from the group consisting of arginine glutamate, cytiolone, epodemiol, ornithine oxoglurate, thidiacycaricinin, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiopronin, betaine, cyanocobal, cyanocobal , udoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valaciclovir, cidofovir, penciclovir, valganciclovir, brivudin, interferon alpha, interferon beta, interferon gamma, interferon alpha-2a, interferon alpha-2b, interferon alpha 1, interferon beta-1a, interferon beta-1b, interferon alfacon-1, PEGinterferon alpha-2b, PEGinterferon alpha-2a and interferon gamma 1b.

In a preferred embodiment of the present invention, the treatment of a patient with a silibinin component is intended to support and / or treat viral hepatitis, especially hepatitis B or C, followed by treatment with another pharmaceutical agent, which is preferably selected from the group consisting of arginine glutamate, silymarin, cytiolone, epodemiol, ornithine oxoglurate, thidiatsicarginine, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiopronin, betaine, cyanocobalamin, leucine, lavuloses, acyclovir, udoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valacyclovir, cidofovir, penciclovir, valganciclovir, brivudin, interferon alpha, interferon beta, interferon gamma, interferon alpha-2a, interferon alpha-2, interferon alpha-2, interferon alpha-2 -1a, interferon beta-1b, interferon alfacon-1, peginterferon alpha-2b, peginterferon alpha-2a and interferon gamma 1b.

Thus, it is preferable, after treatment of viral hepatitis, especially hepatitis B or C, with a drug agent that contains a silibinin component, to treat viral hepatitis, especially hepatitis B or C, with another drug agent.

In a preferred embodiment of the present invention, the drug is processed as part of a subsequent treatment, wherein the drug is first administered during the first period, preferably parenterally, and then the other drug is administered during the second period. Preferably, the first period includes at least 2 days, more preferably at least 3 days, even more preferably at least 4 days, most preferably at least 5 days and particularly preferably at least 6 days. Preferably, the second period includes more than a day than the first period. Preferably, the second period includes at least two days, more preferably at least 3 days, even more preferably at least 4 days, most preferably at least 5 days and especially at least 6 days. In a particularly preferred embodiment of the present invention, the second drug comprises a combination of ribavirin and pegylated interferon alfa, the second period lasting 24-48 weeks.

Preferably, the other drug contains one or more pharmaceutical agents selected from the group consisting of arginine glutamate, silymarin, cythiolone, epodemiol, ornithine oxoglurate, thidiacycarginine, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiop cyanocobalamin, leucine, levulose, acyclovir, udoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valacyclovir, cidofovir, penciclovir, valganciclovir, brivudine, interferon alpha, interferon beta, interferon gamma, and terferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon beta-1a, interferon beta-1b, interferon alfacon-1, PEGinterferon alpha-2b, PEGinterferon alpha-2a and interferon gamma 1b and especially HBV monoclonal antibodies, preferably interferon and / or ribavirin and / or silymarin. If the other drug contains interferon, then it is preferably pegylated interferon alpha (PEGinterferon alpha-2a or PEGinterferon alpha-2b).

In a particularly preferred embodiment of the present invention, the other drug comprises one or more pharmaceutical agents selected from the group consisting of isosilibinin, silidianin, silikristin, taxifolin, isosilicristin, silimonin, silandrin, siligermin and neosiligermin, more preferably only one pharmaceutical agent selected from the specified above the list. Preferably, the other drug includes a silibinin component, characterized in connection with the above-described drug agent, which is administered during the first period, and preferably practically does not contain at least one, preferably all of the above substances. In this regard, the term "practically does not contain" means that the residual content of the substance is preferably less than 2.0 wt.%, More preferably less than 1.0 wt.%, Even more preferably less than 0.5 wt.%, Most preferably less 0.1 wt.% And especially less than 0.05 wt.% Of the total weight of the drug.

Another drug may, in principle, be processed for parenteral or oral administration. According to the present invention, it is preferably processed for another route of administration as opposed to a drug that is administered during the first period. Particularly preferably, the other drug is processed for oral administration. In a particularly preferred embodiment of the present invention, a drug that is administered during the first period is adapted for parenteral, preferably intravenous, administration, and another drug that is administered during the second period following the first period is adapted for oral administration.

In a preferred embodiment of the present invention, the treatment regimen of the present invention includes two phases, one after the other, which are called the first period and the second period. Preferably, during the first period, a medicament containing a silibinin component is administered, preferably parenterally, but no other liver treatment is administered at the same time. During the second period, another drug is administered that preferably contains ribavirin and / or pegylated interferon alfa. In a preferred embodiment of the present invention, a medicament containing a silibinin component is also administered during the second period, preferably parenterally. In another preferred embodiment of the present invention, the medicament containing the silibinin component is not administered during the second period, i.e. only the indicated other drug is administered.

Preferred Embodiments F ₁ through F _{15 of} the biphasic administration regimen are summarized in the table below.

Day F ₁ F ₂ F ₃ F ₄ F ₅ F ₆ F ₇ F ₈ F ₉ F ₁₀ F ₁₁ F ₁₂ F ₁₃ F ₁₄ F ₁₅ First period ≥1 ≥1 ≥2 ≥2 ≥2 ≥3 ≥3 ≥4 ≥3 ≥4 ≥4 ≥5 ≥5 ≥7 ≥7 Second period ≥1 ≥2 ≥1 ≥2 ≥3 ≥2 ≥3 ≥3 ≥4 ≥4 ≥5 ≥4 ≥5 ≥7 ≥14

In another preferred embodiment of the present invention, the treatment regimen of the present invention includes three phases, one after the other, which are called the first period, the second period and the third period. Preferably, during the first period, a medicament containing a silibinin component is administered, preferably parenterally, but no other liver treatment is administered at the same time. During the second period, another drug is also administered that preferably contains ribavirin and / or pegylated interferon alfa and a drug containing a silibinin component, preferably parenterally. Preferably, said other drug is administered during the third period, which preferably contains ribavirin and / or pegylated interferon alfa, but the drug containing the silibinin component is not administered during the third period, i.e. only the indicated other drug is administered.

Preferred embodiments from G ₁ to G _{15 of a} three-phase treatment regimen are summarized in the table below.

Day G ₁ G ₂ G ₃ G ₄ G ₅ G ₆ G ₇ G ₈ G ₉ G ₁₀ G ₁₁ G ₁₂ G ₁₃ G ₁₄ G ₁₅ The first
period ≥1 ≥1 ≥2 ≥1 ≥1 ≥2 ≥2 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥14 ≥14 Second
period ≥1 ≥2 ≥1 ≥1 ≥2 ≥2 ≥1 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥7 ≥14 Third
period ≥1 ≥1 ≥1 ≥2 ≥2 ≥1 ≥2 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥7 ≥7

In yet another preferred embodiment of the present invention, the treatment regimen of the present invention includes three phases that follow one after another, namely a first period, a second period and a third period. Preferably, another drug is administered during the first period, which preferably contains ribavirin and / or pegylated interferon alfa, and no drug containing the silibinin component is administered during the first period. Said second drug is still administered during the second period, which preferably contains ribavirin and / or pegylated interferon alfa, and a drug containing the silibinin component is also administered (co-administered) during the second period, preferably parenterally. Preferably, said other drug is administered during the third period, which preferably contains ribavirin and / or pegylated interferon alfa, but no drug containing the silibinin component is administered during the third period, i.e. only the indicated other drug is administered. In other words, according to a preferred embodiment of the present invention, said other drug is continuously administered, which preferably contains ribavirin and / or pegylated interferon alpha, and a drug containing a silibinin component is preferably co-administered during the interim period (= second period), preferably parenterally.

Preferred embodiments of H ₁ to H _{15 of the} three-phase treatment period are summarized in the table below.

Day H ₁ H ₂ H ₃ H ₄ H ₅ H ₆ H ₇ H ₈ H ₉ H ₁₀ H ₁₁ H ₁₂ H ₁₃ H ₁₄ H ₁₅ The first
period ≥1 ≥1 ≥2 ≥1 ≥1 ≥2 ≥2 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥14 ≥14 Second
period ≥1 ≥2 ≥1 ≥1 ≥2 ≥2 ≥1 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥7 ≥14 Third
period ≥1 ≥1 ≥1 ≥2 ≥2 ≥1 ≥2 ≥2 ≥3 ≥4 ≥5 ≥6 ≥7 ≥7 ≥7

Figure 10 visualizes various methods for co-administration of ribavirin and / or pegylated interferon alpha and a drug containing a silibinin component (options a ₁ ) to m ₂ )). Each lane refers to the time period of administration. For example, in embodiment f ₁ , administration begins with ribavirin / pegylated interferon alpha and continues. During the intermediate period, the silibinin component is simultaneously administered.

Another aspect of the present invention relates to a medicament as described above, preferably adapted for parenteral administration, for the treatment of viral hepatitis as described above.

Another object of the present invention relates to a kit comprising at least one drug of the present invention, which includes a silibinin component, and at least one other drug. Both drugs of the present invention that contain a silibinin component and another drug are described above in such a way that the kit of the present invention is also used in all similar preferred embodiments of the present invention.

In a preferred embodiment of the present invention, the kit contains as many drugs (individual dosage units) as are necessary for continuous therapy, wherein the drug that contains the silibinin component is first administered during the first period and then the second drug is administered during the second period . Preferably, the first period includes at least 2 days, more preferably at least 3 days, even more preferably at least 4 days, most preferably at least 5 days and particularly preferably at least 6 days. Preferably, the second period includes more than a day compared to the first period. Preferably, the second period includes at least 2 days, more preferably at least 3 days, even more preferably at least 4 days, most preferably at least 5 days and particularly preferably at least 6 days.

In a particularly preferred embodiment, the present invention relates to the use of a silibinin component, preferably a silibinin ester, for the manufacture of a medicament which is processed for parenteral administration for the treatment of viral hepatitis C in non-responders in the treatment of ribavirin / interferon, i.e. in patients who do not respond to immunomodulating / antiviral combination therapy, such as ribavirin / interferon therapy.

Another object of the present invention relates to a silibinin component, preferably a silibinin ester, preferably for parenteral administration, for the treatment of viral hepatitis, preferably hepatitis C. Preferred embodiments of the present invention become apparent from the above description of preferred embodiments of other objects of the present invention and are not repeated accordingly.

Another object of the present invention relates to the treatment of viral hepatitis, preferably hepatitis C, comprising the administration, preferably parenteral administration, of a pharmaceutically effective amount of a silibinin component, preferably a silibinin ester, to a subject in need thereof. Preferred embodiments of the objects of the present invention become apparent from the above description of preferred embodiments of other objects of the present invention and, therefore, are not repeated.

The following examples further illustrate the present invention, but they do not limit its scope.

Example 1

The silibinin component is administered parenterally in the form of silibinin-C-2 ', 3-bis (hydrogen succinate) (Legalon Sil® product, Madaus, Cologne) (hereinafter referred to as silibinin).

Patients and methods

The patients Protocol 1 Protocol 2 N (men / women) 16 (14/2) 20 (17/3) Mean age (years ± standard deviation) 49.9 ± 9.7 52.7 ± 12.8 Genotype (1/2/4) 15 / - / 1 17/1/2 Stage fibrosis: 0-2 3 10 3-4 13 7 Absent - 3 Prior treatment * PEG-interferon alfa-2a / ribavirin fourteen eighteen PEG-interferon alfa-2b / ribavirin 2 four Log fall by week 12 of prior therapy: > 2 ** 3 four 1-2 four one <1 5 12 Absent 2 3 fourteen eighteen 2 four * some patients undergo more than one treatment cycle
** all were positive after 24 weeks

For these studies, patients who proved to be non-responders were selected for combination therapy with full doses of peginterferon / ribavirin. The lack of response to treatment manifests itself in the absence of a decrease in> 2 log viral load after 12 weeks of treatment, and / or in the absence of response to the end of treatment. Patients underwent a liver biopsy 2 years before inclusion in the present study. Standard inclusion / exclusion criteria are used for treatment with peginterferon / ribavirin.

Study protocol

During the screening phase, 35 days prior to the first dosing, the patient is evaluated for inclusion / exclusion criteria in a drug study. All patients should have at least one quantitative HCV-RNA assay for 6 months prior to the screening phase.

Protocol 1

First, patients receive daily 10 mg / kg of silibinin (Legalon Sil® product, Madaus, Cologne), which is administered by infusion over 4 hours for 7 consecutive days. On the first day, blood is taken for analysis to determine the initial indicators of oxidative stress, as well as every 30 minutes during infusion and 2 hours after completion of the infusion. On day 8, treatment is replaced with 140 mg of silymarin (Legalon product, Madaus, Cologne) three times a day orally in combination with 180 mcg / week peginterferon alfa-2a (PEGASYS® product, Roche, Basel) and 1-1.2 g / day ribavirin (COPEGUS® product, Roche, Basel).

Protocol 2

After obtaining the results according to the first protocol, silibinin is treated for 2 weeks and various doses of silibinin are administered. First, patients receive daily 5, 10, 15 or 20 mg / kg of silibinin by infusion for 4 hours for 14 consecutive days. On the 8th day of treatment, 180 μg / week of pegylated interferon alpha-2a and 1-1.2 g / day of ribavirin are started. After 14 days, patients receive 280 mg of silymarin (Legalon® product, Madaus, Cologne) orally three times a day. During the 14 days of the infusion period, blood is taken daily to determine the viral load.

In both protocols, in case of intolerance to pegylated interferon alfa-2a or ribavirin, standard dosage recommendations are used. Antiviral combination therapy is received for a total of 24 weeks (when stopping treatment of patients with a> 2 log decrease by 12 weeks); respondents for antiviral therapy for 24 weeks are invited to continue treatment for an additional 48 weeks. After the end of the infusion period, patients are evaluated after 2, 4 weeks and then monthly until the end of therapy after 24 weeks.

The protocol was approved by the Ethics Committee of the Medical University of Vienna. The details of the study are explained to patients, and they all give a subscription of consent to participate in the study.

Methods

Serum HCV RNA was determined by TaqMan PCR (Cobas Ampliprep / Cobas TaqMan VHC Test; detection limit 15 IU / ml, Roche Diagnostics).

Reactive oxidative metabolites in the blood are measured by derivatives of reactive oxygen metabolites (p-RCM; Diacron, Grosseto, Italy) and the amount of antioxidants by biological antioxidant potential (BAP; Diacron, Grosseto, Italy) using a portable system for determining free radicals ( FRAS 4, SEAC, Calenzano, Italy), before the silibinin infusion, every 30 min during the infusion (1 day) and 2 hours after the infusion. The p-RCM method measures reactive oxidative metabolites (mainly hydroperoxides) released from plasma proteins under the influence of an acid buffer, which in the presence of iron produce alkoxyl and peroxyl radicals, in accordance with the Fenton reaction. Such radicals, in turn, are capable of oxidizing an alkyl-substituted aromatic amine (N, N-diethylparaphenylenediamine), thereby obtaining a pink colored derivative that is quantitated at 505 nm. Results for reactive oxidative metabolites are expressed in Caratelli units (Ekar; norm: 250-300, 1 Ekar = 0.08 mg hydrogen peroxide / dl). Using the BAP method, the intensity of decolorization of an iron chloride solution mixed with a thiocyanate derivative is measured by adding a plasma sample photometrically at 505 nm, which is proportional to the ability to reduce iron ions by the amount of antioxidants present in the plasma (norm> 2200 pM). The manufacturer’s methods do not specify which substances are determined.

Statistics

First, the main resulting variable is the viral response, expressed as the percentage of patients who are PCR negative at the end of treatment (24 weeks). Secondary effective variables are the degrees of viral response after 12 weeks, the safety and tolerability of treatment with peginterferon / ribavirin / silymarin, quality of life relative to baseline, after 24 weeks, 48 weeks, 72 weeks (SF-36, digital pain rating scale), and oxidative status after infusion of silibinin. Due to the unexpectedly strong viral response after 7 days of silibinin infusion, the infusion is stopped and the study is redesigned based on the parameters of the viral response and using longer infusion periods and higher doses of silibinin. For the initial study, the sample size is determined based on Gehan’s two-stage plan. Based on previous studies, a response rate of> 10% seems to be justification for further study of the treatment regimen. 29 patients recovered in the first stage, respectively (probability of error β = 5%).

results

Protocol 1

Sixteen characterized non-responders are included (see table above for more details). All patients received a full dose of pegylated interferon (12 peginterferon alfa-2a, 2 peginterferon alfa-2b) and ribavirin (1000-1200 mg / day) for at least 12 weeks. The parameters of oxidative stress do not change during the infusion of silibinin (figure 1).

The content of serum HCV RNA decreases in all patients with intravenous monotherapy with silibinin (Fig. 2) (baseline: 6.59 ± 0.53, day 8: 5.26 ± 0.81 log IU / ml, [mean value ± standard deviation ], p <0.001) with an average log reduction of 1.32 ± 0.55 over one week. In parallel, ALT decreases from 162 ± 133 to 118 ± 107 U / L (p = 0.004). In all patients, HCV RNA remains detectable upon initiation of peginterferon / ribavirin treatment. Three patients refused combination therapy with peginterferon / ribavirin. In 11 of the remaining 13 patients, HCV-RNA again increases after completion of silibinin infusion despite the initiation of peginterferon / ribavirin treatment. By week 12, all patients remain HCV-RNA positive, but at 5, a drop> 2 log and they continue treatment (Fig. 3). None of them became HCV-RNA negative by week 24, one patient had a 5.5 log drop, and he continued treatment on his own.

Protocol 2

Twenty non-responders with certain attributes are included (see table above for more details). All patients receive a full dose of pegylated interferon treatment (18 PEGinterferon alfa-2a, 4 PEGinterferon alfa-2b; 2 patients received 2 courses of treatment) and ribavirin (1000-1200 mg / day) for at least 12 weeks.

Figure 4 shows the kinetics of the virus content in these patients. The load of the virus is constantly decreasing. After 7 days of monotherapy with silibinin at a dose of 5 mg / kg, minimal effectiveness is manifested (n = 3, log drop 0.55 ± 0.5), and doses of 10 mg / kg (n = 19 [including patients in protocol 1], log drop 1.41 ± 0.59), 15 mg / kg (n = 5, log drops 2.11 ± 1.15) and 20 mg / day (n = 9, 3.02 ± 1.01) lead to a high degree a significant reduction in the load of the virus (p <0.001).

After 1 week of combination therapy with silibinin and peginterferon / ribavirin, the virus load decreases further (drop log: 5 mg / kg: 1.63 ± 0.78; 10 mg / kg: 4.16 ± 1.28; 15 mg / kg 3, 69 ± 1.29; 20 mg / kg 4.8 ± 0.89; all p <0.0001 relative to the initial level) (Fig. 5). Two out of five patients in the dose administration group of 15 mg / kg and four out of nine patients in the administration group of the dose of 20 mg / kg have HCV-RNA <15 ME on day 15. HCV-RNA is <15 IU / ml in 8 and 7 patients for 4 weeks (protocol of a 5-week study) and 12 weeks (protocol of a 13-week study) after starting peginterferon / ribavirin, respectively. Antiviral combination therapy is continued for all patients (Fig.6).

Security

Silibinin is generally well tolerated. Five patients reported mild symptoms of a gastrointestinal upset (abdominal pain: 5, diarrhea: 2, nausea 1), two complained of a headache and one for joint pain. Patients are all rated moderate and fall after completion of the infusion; no changes in dosage are required. All patients in the 15 and 20 mg / kg groups report a sensation of warmth at the beginning of the infusion, which decreases within 30 minutes without treatment. No severe side effects. With monotherapy, no changes in hemoglobin, leukocytes, platelets and creatinine are detected. Typical side effects of antiviral combination therapy are noted (including shortness of breath in one patient due to Hemophilus influenzae-induced pneumonia, requiring discontinuation of peginterferon / ribavirin treatment for 8 weeks).

This example shows that parenteral administration of silibinin- (C-2 ', 3-bis (hydrogen succinate)) has a pronounced antiviral effect against hepatitis C. This observation shows that this drug can be used to treat chronic hepatitis C, especially in non correspondents.

Surprisingly, it was found that intravenously, silibinin- (C-2 ', 3-bis (hydrogen succinate)) is a potent antiviral agent in patients with chronic hepatitis C who do not respond to standard antiviral combination therapy. Silibinin is well tolerated intravenously, but there are side effects. The most common side effect is reported to be a temporary sensation of warmth. The antiviral effect is dose-dependent, but does not persist after the completion of the infusion period by oral administration of silymarin.

When comparing, it was found that similar amounts of silymarin administered orally do not affect the HCV load (A. Gordon et al., J Gastroenterol Hepatol. 21, 2006, pp. 275-80), reflecting differences in the bioavailability and metabolism of silibinin and leading to significantly lower plasma levels. After oral dosing, silymarin flavonolignans are rapidly gluconidated and eliminated rapidly with short half-lives (Z. Wen et al., Drug Metab Dispos. 36 (1), 2008, pp. 65-72).

Example 2

Patients are constantly treated with 180 mcg peginterferon alfa-2a and a weight-dependent amount of ribavirin. Despite this treatment, five HCV-RNA patients are positive after 24 weeks of treatment: three men and two women; four patients with HCV genotype 1 and one patient with HCV genotype 3a; three patients with cirrhosis.

Four patients can be rated as unexposed, but one patient can be rated as being in the relapse stage relative to the previous two therapies (24 and 48 weeks).

During the course of treatment, 180 μg of peginterferon alfa-2a and a weight-dependent amount of ribavirin are administered intravenously to all patients at least once for 14 consecutive days 20 mg / kg / day. During this period, combination therapy with peginterferon / ribavirin is continued.

All 5 patients become negative for HCV RNA.

7 shows the results for one specific patient (male, 55 years old). You can see that treatment with peginterferon / ribavirin causes a decrease in viral load of only about log 7 IU / ml to about log 4.5 IU / ml after 24 weeks. Co-treatment of 20 mg / kg / day with intravenous silibinin bis (hydrogen succinate) for 14 days, however, leads to a sharp decrease in the viral load from about log 4.5 IU / ml to a value below the sensitivity limit. After the first interval of parenteral silibinin bis (hydrogen succinate) administration, the viral load rises again to approximately 2 IU / ml, which nevertheless is constantly suppressed below the detection limit by the second joint intravenous administration of 20 mg / kg / day silibinin bms (hydrogen succinate) in for 14 days.

Fig. 8 shows the results for another specific patient (female, 44 years old). From the figure it follows that peginterferon / ribavirin rarely causes a decrease in viral load from about 7 IU / ml to about 5 IU / ml after 30 weeks. Joint intravenous treatment of 20 mg / kg / day with silibinin bis (hydrogen succinate) for 14 days after 30 weeks, however, leads to a sharp and constant decrease in viral load from about 4 IU / ml to a value below the detection limit.

Figure 9 shows the results for one specific patient (male, 52 years old). From the figure it follows that peginterferon / ribavirin causes an effective decrease in viral load from about 5 IU / ml to a value close to the detection limit of <15 IU / ml. Joint intravenous administration of 20 mg / kg / day silibinin bis (hydrogen succinate) for 14 days after 72 weeks causes a further decrease in viral load below the detection level.

These clinical studies show that parenteral treatment with the silibinin component over a relatively short period of time supports and significantly improves traditional treatment with peginterferon / ribavirin. It was shown that parenteral administration of the silibinin component activates (reactivates) the sensitivity of patients to the traditional treatment with peginterferon / ribavirin (Figs. 7 and 8) and / or increases the antiviral effect of the traditional treatment with peginterferon / ribavirin (Fig. 9).

Example 3

An in vivo study was performed to describe the plasma concentration / temporal profiles of silibinin in 8 patients with chronic hepatitis C who were given intravenous infusion for 20 days with 20 mg silibinin / kg body weight (Legalon® SIL). For multiple doses of 20 mg / kg body weight, plasma concentration / time and PK profiles are analyzed for parameters of free and total silibinin for 1 day (= conditions for one dose) and compared with indicators for 7 days (= expected steady state conditions).

Analytical method

Examine the samples using the well-known method HPLC-UV. Throughout the study, the analytical method is confirmed by two calibration curves for each analytical run. Analysis of chromatograms with the data presented on the calibration curves and quality control samples shows that the result of determining the concentrations of total and free silibinin A and silibinin B in such a study is reliable.

Pharmacokinetic parameters are summarized in the table below.

Total silibinin 1st day 7th day Silibinin A Silibinin B Silibinin A Silibinin B AUC (0-∞) [h ng / ml] 61733 ± 27489 13745 ± 76040 - -

Total silibinin 1st day 7th day Silibinin A Silibinin B Silibinin A Silibinin B AUC (0-t _z ) [h ng / ml] 50019 ± 20048 109038 ± 51342 - AUCss [h ng / ml] - - 84,299 ± 25,111 150,780 ± 47,780 C _min [h ng / ml] - - 1967 ± 831 3311 ± 1426 C _max [ng / ml] 4550 ± 928 9539 ± 2843 5791 ± 977 11083 ± 2269 C _av [ng / ml] - - 3512 ± 1046 6282 ± 1991 t _1/2 [ng / ml] 8.30 ± 2.26 8.29 ± 2.98 13.32 ± 3.66 12.02 ± 2.91 HVD [h] 9.28 ± 3.36 9.47 ± 3.29 15.20 ± 4.09 13.17 ± 3.62 MRT [h] 13.17 ± 3.74 13.44 ± 4.39 19.22 ± 5.28 17.34 ± 4.20 CL [ml / (h kg)] 0.435 ± 0.336 0.233 ± 0.237 0.269 ± 0.128 0.156 ± 0.091 Vz [ml / kg] 4.7 ± 1.3 2.4 ± 1.1 4.67 ± 0.73 2.43 ± 0.55 t _max [h] 4.14 ± 0.18 4.17 ± 0.18 - - % PTF [%] - - 121.89 ± 54.43 137.59 ± 52.51

Free silibinin 1st day 7th day Silibinin A Silibinin B Silibinin A Silibinin B AUC (0-∞) [h ng / ml] 3614 ± 1648 753 ± 397 - - AUC (0-t _z ) [h ng / ml] 3302 ± 1551 559 ± 339 - - AUCss [h ng / ml] - - 4095 ± 1942 104U627 C _min [h ng / ml] - - 59 ± 40 3.3 ± 8.7 C _max [ng / ml] 316 ± 108 90 ± 44 315 ± 119 120 ± 54 C _av [ng / ml] - - 171 ± 81 43 ± 26 t _1/2 [ng / ml] 4.58 ± 1.35 5.16 ± 4.96 6.85 ± 1.29 4.35 ± 1.66 HVD [h] 10.12 ± 4.29 6.09 ± 2.14 11.87 ± 2.64 7.17 ± 1.63 MRT [h] 8.49 ± 2.64 8.81 ± 6.12 9.88 ± 1.86 6.27 ± 2.39 CL [ml / (h kg)] 7.0 ± 4.2 44.8 ± 42.5 5.9 ± 2.9 26.8 ± 16.7 Vz [ml / kg] 51.4 ± 12.7 285.8 ± 163.9 55.3 ± 18.5 140.4 ± 49.0 t _max [h] 3.73 ± 1.22 4.03 ± 0.04 - - % PTF [%] - - 164.00 ± 45.33 305.24 ± 80.46

Example 4

An in vitro assay is performed to evaluate the cytotoxic effect of silymarin, silibinin, silibinin bis (hydrogen succinate) disodium salt and succinic acid using the XTT test using the mouse cell line L929 (see DAScudiero et al., Cancer Res. 48, pp. 4827 -4833; OSWeislow et al., J. Natl. Cancer Inst., 81, 577-586; NW Rohirm et al., J. Immunol. Methods, 142).

Investigate the following concentrations at the studied positions: 9.77, 19.53, 39.06, 78.13, 156.25, 312.5, 625, 1250 μg / ml. A complete medium (RPMI 1640 containing 10 vol.% FST) is used as a negative control. The solvent control for the test position is RPMI 1640 medium containing 10 vol.% FST and 1% DMSO. The solvent control for the positive control is also RPMI 1640 medium containing 10 vol.% FST and 10.0 vol.% Deionized water. Sodium dodecyl sulfate is used as a positive control. The following concentrations are used: 3.125, 6.25, 12.5, 25, 50, 100, 125, 250 μg / ml. The incubation time is 24 hours at 37 ± 1.5 ° C.

Negative control and solvent control do not show a decrease in cell viability. Positive Control (SDS) induces a particular dose-dependent decrease in cell viability.

Toxic effects are observed after incubation with silymarin from 39.06 μg / ml and to the highest studied concentration (1250 μg / ml). The calculated value of XTT ₅₀ is 35.2 μg / ml.

Toxic effects are observed after incubation with silibinin with 78.13 μg / ml and to the highest studied concentration (1250 μg / ml). The calculated value of XTT ₅₀ is 67.5 μg / ml.

Significant cytotoxic effects are not observed after incubation with the sodium salt of silibinin bis (hydrogen succinate) up to the highest analyzed concentration (1250 μg / ml). Due to the loss of cytotoxicity, the value of XXT ₅₀ cannot be calculated.

No significant cytotoxic effects were observed after incubation with succinic acid to the highest test concentration (1250 μg / ml). Due to the loss of cytotoxicity, the value of XXT ₅₀ cannot be calculated.

Studies have shown that under these conditions, the cytotoxic potential of silymarin is approximately 100% higher than the cytotoxic potential of silybinin. Therefore, it can be expected that silibinin can be administered in higher doses relative to silymarin without inducing serious side effects.

Example 5

NS5B RNA-dependent RNA polymerase (RNA-dependent RNA polymerase - RdRp) is an important enzyme for viral replication (see S.B. Hwang et al., Virology 227, 1997, pp. 439-446). The purified compounds are then tested in a cell-free enzyme assay to detect HCV RdRp effects: silibinin A, silibinin B, isosilibinin A, isosilibinin B, silicristin, silidianin and the silibinin ester salt of silibinin-C-2 ', 3-bis (active hydrogen succinate) Legalon® SIL product ingredient).

The concentration of DMSO in all reactions was kept constant at 5%. Prepare stock solutions (100 mM) of the compounds in 100% DMSO. The target enzyme in the study is HCV NS5BΔ21 polymerase genotype J4 (1b).

11 shows data obtained for six purified constituents of silymarin (i.e., silibinin A, silibinin B, isosilibinin A, isosilibinin B, silicristinin, and silidianin). 12, respectively, shows silibinin ester data. Silibinin ester is most effective.

The value of the IC ₅₀ ester of silibinin is determined by the dose-response curve in two measurements. The determined IC ₅₀ value is 47 ± 14 μM. Curves agree on data points and IC ₅₀ values are interpolated on the obtained curves using SigmaPlot 8.0 software.

Claims (21)

1. The use of silibinin component with the General formula (I)

in which R ₁ , R ₂ , R ₃ , R ₄ and R _{5 are} independently selected from the group consisting of —H, —SO ₃ H, —PO ₃ H ₂ , —CO — C ₁ —C ₈ alkylene -OH, -CO-C ₁ -C ₈ -alkylene-CO ₂ H, -CO-C ₁ -C ₈ -alkylene-SO ₃ H, -CO-C ₁ -C ₈ -alkylene-ORO ₃ H ₂ , - CO-C ₁ -C ₈ -alkylene-PO ₃ H ₂ , - (C ₂ -C ₃ -alkylene-O) _n -H, where n = 1 to 20, -CO-C ₁ -C ₈ -alkylene- N (C ₁ -C ₃ alkyl) ₃ ⁺ X ^- , where X ^- represents a pharmaceutically acceptable anion,
or their pharmaceutically acceptable salts,
to obtain a drug designed for parenteral administration in the treatment of viral hepatitis, the drug optionally containing cyclodextrin and / or phospholipid. 2. The use according to claim 1, wherein the silibinin component is a silibinin ester. 3. The use according to claim 2, wherein the silibinin ester is silibinin-C-2 ', 3-bis (hydrogen succinate) or a physiologically acceptable salt thereof. 4. The use according to claim 1, wherein the drug is substantially free of silidianin and / or silicristin and / or isosilibinin. 5. The use according to claim 1, wherein the viral hepatitis is hepatitis B or hepatitis C. 6. The use according to claim 1, wherein the drug is processed for injection or infusion. 7. The use according to claim 1, wherein the drug is processed for intravenous administration. 8. The use according to claim 1, wherein the drug does not contain any of the constituent parts of silymarin in addition to the silibinin component. 9. The use according to claim 1, wherein the drug contains a silibinin component in a dose of at least 50 mg silibinin. 10. The use according to claim 1, wherein the drug contains additional pharmaceutical agents in addition to the silibinin component. 11. The use according to claim 10, wherein the additional drug is selected from the group consisting of arginine glutamate, cythiolone, epomediol, ornithine oxoglurate, thidiatsikarginin, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiopronin, thiopronin, , cyanocobalamin, leucine, levulose, acyclovir, idoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valacyclovir, cidofovir, penciclovir, valganciclovir, brivudine, interferon alpha, interferon beta, interferon gamma, interferon 2a, interferon alpha-2b, interferon alpha-n1, interferon beta-1a, interferon beta-1b, interferon alfacon-1, peginterferon alfa-2b, peginterferon alfa-2a and interferon gamma-1b. 12. The use according to claim 1 for reducing viral load in patients with hepatitis. 13. The use according to claim 1 for the treatment of viral hepatitis in a patient who has or has already undergone a liver transplant. 14. The use according to claim 1 for the treatment of viral hepatitis in a patient who does not respond to treatment with ribavirin / interferon. 15. The use according to claim 1 for the maintenance and / or preventive treatment of viral hepatitis with a pharmaceutical agent selected from the group consisting of arginine glutamate, silymarin, cythiolone, epomediol, ornithine oxoglurate, thidiacycarginine, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiopronin, betaine, cyanocobalamin, leucine, levulose, acyclovir, idoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valaciclovir, cidofovir, penciclovir, valganciclovir, brivudine, interferon alpha, interferon beta, interferon gamma, interferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon beta-1a, interferon beta-1b, interferon alfacon-1, peginterferon alpha-2b, peginterferon alpha-2a and interferon gamma -1b. 16. The use according to claim 1, and after treatment of viral hepatitis with the specified drug, viral hepatitis is treated with another drug. 17. The use according to claim 1, wherein the drug is created as an integral part for subsequent treatment, first the drug is administered during the first period and then another drug is administered during the second period. 18. The use of claim 17, wherein the first period is at least 2 days. 19. The use according to clause 16, wherein the other drug contains one or more pharmaceutical agents selected from the group consisting of arginine glutamate, silymarin, citiolone, epomediol, ornithine oxoglurate, thidiacycarginine, myoinositol, methionine and N-acetylmethionine, choline, ornithine aspartate, cyanidanol, thiopronin, betaine, cyanocobalamin, leucine, levulose, acyclovir, idoxuridine, vidarabine, ribavirin, ganciclovir, famciclovir, valaciclovir, cidofovir, penciclovir, valganciclovir, brivudine, interferon lfa, interferon beta, interferon gamma, interferon alpha-2a, interferon alpha-2b, interferon alpha-n1, interferon beta-1a, interferon beta-1b, interferon alfacon-1, peginterferon alpha-2b, peginterferon alpha-2a and interferon gamma -1b. 20. The use according to clause 16, and another drug is created for oral administration. 21. A kit for treating viral hepatitis, comprising at least one drug according to one of claims 1 to 11 and at least one drug according to claim 19 or 20.

Images