MXPA00002570A

MXPA00002570A - Use of ifn-alpha and amantadine for the treatment of chronic hepatitis c

Info

Publication number: MXPA00002570A
Application number: MXPA/A/2000/002570A
Authority: MX
Inventors: Friederike Zahm
Original assignee: F Hoffmannla Roche Ag
Priority date: 1997-09-18
Filing date: 2000-03-14
Publication date: 2001-03-05

Abstract

The present invention provides the use of IFN-a in association with Amantadine for the manufacture of medicaments for the treatment of chronic hepatitis C infections. The present invention also provides medicaments containing the IFN-a and Amantadine as a combined preparation for simultaneous, separate or sequential use in therapy of chronic hepatitis C infections. The present invention further provides a method for treating chronic hepatitis C infections in patients in need of such treating comprising administering an amount of IFN-a in association with an amount of Amantadine effective to treat hepatitis C.

Description

USE OF IFN-ALPHA AND AMANTADINE FOR THE TREATMENT OF CHRONIC HEPATITIS C Field of the Invention The present invention relates to the treatment of chronic hepatitis 0 infections, by the use of an amount of IFN-a in association with Amantadine which is effective in treating hepatitis C.

Background of the Invention Interferons (IFNs) are natural proteins that have antiviral, antiproliferative and immunoregulatory activity. Four different classes of interferons are known to exist in humans (Pest et al (1987) Ann. Rev. Biochem 5_ß 727-777 and Emanual &Pestka (1993) J. Biol. Chem. 266, 12565-12569) . The IFN-a family represents the predominant class of IFNs produced by stimulated peripheral blood leukocytes (Pestka et al., Cit., Havell et al (1975) Proc. Nati. Acad. Sci. USA 72_, 2185-2187; Cavalieri et al (1977) Proc. Nati, Acad. Sci. USA 74_ 3287-3291.}, And lymphoblastoid and myeloblastoid cell lines REF .: 32975 (Familletti et al. (1981) Antimicrob. Agents. Chemother. 5-9) The antiviral effect of IFN-a is achieved not by the direct influence of the same viruses, but by an activity in their target cells, in the sense of protection against viral infection. exert effects on cancerous tumors and can influence the immune system of the body because they inactivate, for example, macrophages and NK cells and intensify the expression of several immunologically significant constituents of the cell membrane The details of the preparation of interferon- CDNA and the direct expression of them, especially in E. Coli, have been the subject of many publications. Thus, for example, the preparation of recombinant interferons are known, for example, from Nature 295 (1982), 503-508, Nature 284 (1980), 326-320, Nature, 290 (1981), 20-26, Nucleic Acids Res. 8_ (? 980), 4057-4074, as well as European Patent Nos. 32134, 43980 and 211148.

Monotherapy with IFN-a is commonly used in the treatment of chronic hepatitis infections, but nevertheless, this treatment is not always effective. Amantadine has been proposed as a monotherapy treatment for chronic hepatitis C hepatitis (JP Smith et al, "Treatment of chronic hepatitis C with amantadine hydrochloride, Abstract of the Annual Meeting of the American Gastroenterology Association," May 1996). However, this treatment with monotherapy does not always respond with a good result for all patients. The combination therapy may therefore be more effective than any monotherapy.

Description of the Invention The present invention therefore provides the use of IFN-a in association with Amantadine for the manufacture of medicaments for the treatment of chronic hepatitis C infections. The present invention also provides medicaments containing IFN-a and Amantadine. as a combined preparation for simultaneous, separate or sequential use in therapy of chronic hepatitis C infections. In addition, the present invention provides a method for the treatment of chronic hepatitis C infections in patients in need of such treatment comprising administering an amount of IFN- a in association with an amount of Amantadine which is effective in treating chronic hepatitis C. The term "IFN-a, as used herein includes IFN-as derived from any natural material (e.g. from leukocytes, fibroblasts, lymphocytes) or from material derived from them, (for example cell lines), or from those prepared with recombinant DNA technology The details of the donation of IFN-a and the direct expression of it, especially in E coli, have been the subject of many publications. The preparation of recombinant IFN-as has been disclosed, for example by Goeddel et al. (1980) Nature 284, 316-320 and (1981), Nature 290, 20-26, and European Patent Nos. 32134, 43980 and 211148. There are many types of IFN-a such as IFN-al, IFN-a2; and in addition its subtypes including but not limited to IFN-a2A; and, IFN-a2B, and IFN-a2C and IFN-alI, (also referred to as IFN-all or? -IFN). The term "IFN-a" also includes consensus IFN-a obtainable in Amgen or mixtures of natural and / or recombinant IFN-as. IFN-as. USE of IFN-a2A is preferred. The manufacture of IFN-a2A has been described in European Patent Nos. 43980 and 211148. The IFN-a used in this invention can be conjugated to a polymer such as a polyalkylene glycol (substituted or unsubstituted), for example polyethylene glycol, for form PEG IFN-a. The conjugation can be carried out by several linkers known in the art, in particular, by means of linkers such as those described in the European patent applications, publication Nos. 0510356 and 593868 and European Patent Application No. 97108261.5. The molecular weight of the polymer, which is preferably polyethylene glycol, can be comprised between 300 to 30,000 daltons, and one or more, preferably one to three polymers can be conjugated with IFN-a. A preferred IFN-a is formed using IFN-a2A. The Amantadine, which is a tricycle [3.3.1. I3 '7] decane-1-amine, is described in the Merck Index, compound No. 373, Tenth Edition. Its manufacture was described in U.S. Patent No. 3,152,180. To carry out the invention, IFN-a and Amantadine are administered to a patient suffering from chronic hepatitis C infection, in sufficient quantities to eliminate or at least to alleviate one or more of the signs or symptoms of chronic hepatitis C including elevated ALT, positive test for anti-HCV antibodies, presence of HCV demonstrated by a positive test to HCV-RNA, clinical stigmata of chronic diseases of the liver and hepatocellular deterioration. The dosage of IFN-a to carry out the combination therapy of this invention is from about 1 to 6 million international units (Ul) administered twice or three times a week, every other day or daily. The preferred dosage for carrying out the combination therapy of this invention is approximately 3 million Ul administered three times per week.

The dosage of Amantadine to practice this invention is from about 100 to 400 mg per day, preferably 200 mg. This daily dosage can be administered once a day in a single dose or in divided doses two or three times per day. Amantadine is administered to the patient in association with IFN-a, ie the dose of IFN-a is administered during the same period of time or in periods other than the periods in which the patient receives doses of Amantadine. Currently, the IFN-a formulations are not effective when administered orally, so that the preferred method of administration of IFN-a is parenteral, preferably by subcutaneous (s) or intramuscular (s) injection (s). . Amantadine can be administered orally, in capsules or in tablet form in association with parenteral administration of IFN-a. Naturally, other types of administration of both drugs are contemplated in the form in which they are available, such as in the form of administration such as nasal spray, transdermal route, suppositories, sustained release form, etc. Any form of administration will be appropriate with the proviso that the appropriate doses are administered without destroying the active ingredient. The efficacy of the treatment can be determined by controlled clinical trials of combination therapy versus morotherapy. The efficacy of combination therapy to alleviate the signs and symptoms of chronic hepatitis C infections and the frequency and severity of side effects is compared with previous monotherapy with IFN-a, and Amantadire. Three populations suffering from chronic hepatitis C 1 infections are evaluated. Patients who were not previously treated. 2. Patients who were previously treated with IFN-a, or with any other drug and who subsequently suffered relapses. 3. Patients who did not respond to previous treatments with IFN-a, or with any other drug. The efficacy of the combination therapy will be determined by the degree to which the signs or symptoms of chronic hepatitis were alleviated. Example Antiviral Effect of Amantadine and IFN-a, 2A Against the Hepatitis C Virus in Peripheral Blood Mononuclear Cells (PBMC) of Patients with Chronic Hepatitis Mononuclear cells from patients with chronic hepatitis C (seropositive to anti- HCV and HCV RNA, with histologically proven chronic hepatitis), for the purpose of determining the presence of HCV RNA, using reverse transcription and PCR techniques with universal primers from the highly conserved 5 'noncoding region of the HCV genome (Navas et al. , J. Hepatol, 21, 182-186 (1994)). The typing and subtyping of HCV genomes was carried out by RFLP analysis of the PCR products (Navas et al, J. Clin Microbiol 21, 317-321 (1997)). For the purposes of this study, only cases infected by a single genotype were considered, in order to minimize the possible interference of multiple genotypes, in the present population, with the subtype "Ib of HCV (Pernas et al., J. Gen. Virol., 76, 415-420 (1995).) Thus, HCV RNA positive PBMC obtained from 15 patients were analyzed in vi tro to determine the effects of PBMC treatment of 10 paired healthy donors were used as controls and analyzed in a similar manner PBMCs were isolated from heparinized venous blood by Ficoll-Hypaque gradient sedimentation.The urea-PBMC were isolated, washed twice with phosphate-buffered saline, and suspended in RPMI. The viability of these cells was verified by trypan blue exclusion PBMC were cultured in duplicate at a concentration of 2 x 10 6 viable cells / ml in sets of 6 wells with cultures. of fabrics under humid atmosphere with 5% C02 for 7 days. The cultures were kept without mitogens (medium only) or stimulated with unique mitogens (Phytohemaglutinin (PHA) or Lipopolysaccharide (LPS) chronic hepatitis C infections or with PHA in addition to LPS (10 μg / ml each) (Martin et al., Cytokine 8, 313-317 (1996)). The proliferation of PBMC, and possible drug-induced cytotoxicity, were measured using non-isotopic cell proliferation and cytotoxicity assays The effect of experimental treatments with Amantadine alone, in combination with IFN-a2A and those of IFN-a2A, only, they were established through the trial. of HCV RNA in cultured mononuclear cells, compared to untreated PBMCs in patients (Martin et al. supra); the specificity controls were as previously described by Navas et al. in J. Hepatol. 21, 182-186 (1994). The treatment of mononuclear cells from healthy donors with Amantadine only, in combination with IFN-a2A, or IFN-a2A alone, served as controls. Changes in HCV RNA concentrations were measured by the AMPLICOR ™ HCV MONITOR assay (Roche Diagnostic System, Inc. Branchburg). Doses of amantadine in the physiological range of l-5μM (2μM corresponds to the level in the blood, therapeutically recommended, daily dose of the drug: 100 mg / 12 hours) did not affect the viability of the cell and had minor effects on the response to mitogens during PBMC isolated from patients with HCV and from healthy donors. The highest doses of Amantadine (50 and 500 μM) were investigated only in PBMC from healthy donors. The dose of 50μM slightly decreased the proliferation of PBMC, while the dose of 500μM showed a marked anti-proliferation effect. All PBMC cultures of patients with HCV, but not of the donors, were positive to HCV RNA with or without mitogens, evaluated by a modification of the AMPLICOR ™ HCV MONITOR assay. The Dosage of 2μM Amantadine reduced the average amount of HCV RNA (number of copies / μg RNA) in < 70% alone or in combination with lOOOUI / ml of IFN-a2A. In a single patient, different degrees of reduction of the concentration of HCV RNA in PBMC were obtained after treatment with 1, 2 and 5 μM of Amantadine. alone and in combination with 1000 'Ul / ml of IFN-a2A (Table 1). In addition, HCV RNA was negative in up to 3/15 (20%) of PBMC cultures. (Table 1). TABLE 1. Number of cases with reduction or disappearance of HCV RNA in PBMC after experimental treatments (n = 15) Amantadine IFN-a2A Reduced concentration of HCV RNA (μM) (Ul / ml) > 25: > 50% > 75% Negative 1 0 3 2 3 0 2 0 5 2 4 1 5 0 2 2 3 3 1000 • 0 2 3 2 1000 3 3 4 0 2 1000 3 1 3 3 1000 0 0 2 3 HCVARN was negative in PBMC cultures of 1/15 (7%) and 3/15 (20%) patients with doses of 2 and 5 μM of Amantadine respectively compared to 2/15 (13%) with IFN-a2A alone. With the combination of Amantadine and IFN-a2A (20%) the PBMC cultures were negative HCV RNA. The combination of 2μM of Amantadine / IFN-a2A gave better results in the disappearance of HCV RNA in individual PBMCs (up to 20% of the cases, Table 1) showed a superior effect than with the same doses of Amantadine alone.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is that which is clear from the present description of the invention. Having described the invention as above, property is claimed as contained in the following:

Claims

1. Use of IFN-a in association with Amantadine for the manufacture of medicines for the treatment of chronic hepatitis C infections

2. Use according to claim 1, wherein the amount of IFN-a approximately 1 to 6 million Ul twice or three times per week, one day yes and one day no.

3. Use according to claim 1, wherein the amount of Amantadine is from 100 to 400 mg daily, preferably 200 mg daily.

4. Use according to claims 1 wherein IFN-μa is IFN-a2A or PEG-IFN-0C2A.

5. Drugs characterized because they contain IFN-a and Amantadine as a combined preparation for simultaneous, separate or sequential use in therapy of chronic hepatitis C infections,

6. Medicaments according to claim 5, characterized in that IFN-cc is IFN-a2A.

7. Medicaments according to claim 5, characterized in that IFN-a is PEG-IFN-a.

8. Medicaments according to claim 5, characterized in that IFN-a is PEG-IFN-0C2A.

9. A method for treating chronic hepatitis C infections, characterized in that it comprises administering an amount of IFN-a in association with an amount of Amantadine that is effective in treating chronic hepatitis C.

10. The method according to claim 9, characterized in that the amount of IFN-a administered in said method is approximately 1 to 6 million Ul twice or three times per week.

The method according to claim 9, characterized in that the amount of Amantadine administered in said method is from 100 to 400 mg per day.

12. The method of any of claims 9 to 11, characterized in that the IFN-a is IFN-a2A or PEG-IFN-a2A.

13. Use of IFN-a and amantadine for the treatment of chronic hepatitis C infections • ** 'OF CHRONIC HEPATITIS C SUMMARY OF THE INVENTION The present invention provides the use of IFN-a in association with Amantadine for the manufacture of 5 medicaments for the treatment of chronic hepatitis C infections. The present invention also provides medicaments containing IFN-a and Amantadine as a combined preparation for simultaneous use, Sequential or separate in therapy of infections 10 Chronicles of Hepatitis C. The present invention further provides a method for treating chronic hepatitis C infections in patients in need of such treatment comprising administering an amount of IFN-a in association with an amount of 15 Amantadine that is effective in treating hepatitis C.