CZ20012544A3 - Pharmaceutical packing - Google Patents

Abstract

This invention provides a pharmaceutical pack comprising as active ingredients (1) an antiviral agent active against hepatitis B virus and (2) a vaccine for the prophylaxis and/or treatment of hepatitis B infection, the active ingredients being for simultaneous or sequential use. Preferred components are a nucleoside analogue as the antiviral agent, together with a hepatitis B virus vaccine which comprises a hepatitis B virus surface antigen.

Description

Technical field

The invention relates to the use of a nucleoside analogue effective against hepatitis B virus, HBV or other anti-viral agents against HBV, such as gamma-interferon or a nucleotide analogue and a vaccine against HBV in the treatment of hepatitis B.

BACKGROUND OF THE INVENTION

Chronic infection with hepatitis B virus, HBV, which is currently not yet effectively treated, is a major global problem. Patients who are chronic carriers of HBV and are expected to have more than 300 million worldwide are at high risk for the development of active chronic hepatitis, cirrhosis and primary liver cell carcinoma.

EP-A-388049 (Beecham Group p.l.c.) describes the use of penciclovir / famciclovir in the treatment of hepatitis B virus infection. All data on penciclovir / famciclovir refer to pharmaceutically acceptable salts, for example hydrochloride and solvates, for example hydrate.

EP-A-494119 (IAF Biochem. International Inc) discloses the use of 1,3-oxathiolane nucleoside analogues including lamivudine in the treatment of hepatitis B.

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SUMMARY OF THE INVENTION

SUMMARY OF THE INVENTION The present invention provides a pharmaceutical package comprising, as active ingredients, (1) an antiviral active against hepatitis B virus and (2) a vaccine for the prophylaxis and / or treatment of hepatitis B virus infection, wherein the active ingredients are intended for simultaneous or sequential use.

A pharmaceutical package is a package or package that may contain one or more unit doses of the active ingredients. It may be, for example, a foil of metal or plastic, such as a blister. The package may be accompanied by instructions for the administration of the active ingredients. Where the antiviral and HBV vaccine are to be administered separately, it may be a double pack.

The package according to the invention is intended for use in the therapeutic or prophylaxis of hepatitis B virus infection.

In particular, the package according to the invention is intended for the treatment of chronic hepatitis B.

The antiviral agent used in the pharmaceutical package of the invention may be a nucleoside. In another embodiment, the antiviral agent may be a nucleotide.

Suitable substances for this use are, for example, penciclovir, famciclovir, lamivudine, ganciclovir, lobucariv, adefovir, ribavirin, BMS200,475, vidarabine or ARA-AMP. Preferred nucleoside analogs are penciclovir, famciclovir and lamivudine.

Another potential antiviral agent is interferon, with alpha-interferon being particularly preferred.

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Information on the structure and potency of nucleoside analogs can be obtained in the literature, for example, in Pharmaprojects, PJB publications Limited, Richmond, Surrey, U.K. or from R & D Foc, published by IMS World Publications, 364 Euston Road, London NW1 3BL.

The data on anti-viral agents including the nucleoside analogues mentioned above and their salts also refer to solvates, for example hydrates.

Examples of pharmaceutically acceptable salts can be found in the aforementioned patent publications (Beecham Group p.l.c.) and in the references cited therein.

It will be appreciated that nucleoside or nucleotide analogues having activity against hepatitis B virus and HBV vaccine in the combination of the invention may still be administered in combination with other pharmacologically active substances, in particular other antiviral agents.

Throughout the application, the term vaccines for the prophylaxis and / or treatment of hepatitis B virus infection includes all vaccines comprising HBV antigens, such as surface antigen, core antigen and polymerase, and therapeutic vaccines are also included.

In one embodiment of the invention, the hepatitis B virus antigen is HBsAg surface antigen of hepatitis B virus. The preparation of this surface antigen is well documented and has been described, for example, in a publication

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Harford et al., Develop. Biol. Standard 54, p. 125 1983, Gregg et al., Biotechnology, 5, p. 479, 1987, and in EP-A-0 226 846, EP-A-0 299 108 and the references cited therein. files listed.

The term "hepatitis B surface antigen" or "HBsAg" refers to any antigen of this type or an immunogenic derivative thereof, particularly a fragment with the same antigenicity as the full surface antigen. It will be appreciated that in addition to the 226 amino acid residues of HBsAg S as described in Tiollais et al., Nature 317, 489, 1985 and references therein, the surface antigen described in the present application may optionally contain the entire S antigen signal sequence. or a portion thereof as described in the above references and also in EP-A-0 278 940. It may also be a surface antigen variant, for example a mutant, described in International Application WO 91/14703. According to another embodiment, the surface antigen may consist of a protein designated L * in European Patent Application No. 0 414 374, i.e. a protein whose amino acid sequence consists of part of the amino acid sequence of the large (L) protein of hepatitis B virus or its subtype, wherein the amino acid sequence of the protein consists of (a) residues 15-22, followed by residues 133-145 and then residues 175-400 of said L protein, or (b) residue 12 followed by residues 14-52, then residues 133-145 and residues 175-400 of said protein.

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The hepatitis B virus surface antigen may also be the polypeptides described in EP 0 198 474 or EP 0 304 578.

Usually, the surface antigen is in particulate form. It may consist of only protein S or may be composite particles, for example (S, L *), where L * is as defined above and S denotes the hepatitis B virus surface antigen S-protein.

The preferred hepatitis B virus antigen is the composite particles defined as S, L *.

Another preferred hepatitis B virus antigen is the 226 amino acid sequence of the HBV surface antigen in particulate form.

A vaccine of this type may advantageously contain pharmaceutically acceptable excipients. A suitable adjuvant is, for example, an aluminum salt, such as aluminum hydroxide gel (alum) or aluminum phosphate, as described in WO 93/24148, but may also be a calcium, iron or zinc salt, or an insoluble suspension of acylated tyrosine, acylated sugars, cationically or anionically derivatized polysaccharides or polyphosphazenes.

Preferably, a strong helper system is used for the hepatitis B virus antigen. In the case of the vaccine of the invention, it is preferred that the vaccine elicits an immune system response in terms of TH1. Suitable auxiliary systems are, for example, a combination of monophosphoryl lipid A, preferably 3-de-O-acylated monophosphoryl lipid A (3D-MPL). • · · · · · · · · · · · · · · · · · · · · · · · φ · · · φ · φ · φ · φ · φ · φ spolu φ spolu spolu spolu spolu spolu spolu spolu spolu spolu spolu spolu spolu spolu spolu A vaccine comprising hepatitis B surface antigen in conjunction with 3D-MPL has been described in European Patent Application 0 633 784.

A preferred system comprises a combination of monophosphoryl lipid A and a saponin derivative, especially a combination of QS21 and 3D-MPL as disclosed in International Application WO 94/00153, or may be a less reactive composition comprising QS21 in combination with cholesterol and described in International Application WO 96/33739.

Other excipients which may be used in this case include CpG-containing oligonucleotides as described in WO 96/02555 (University of Iowa).

In a preferred embodiment, the invention relates to a vaccine comprising HBV antigen and, as adjuvant, monophosphoryl lipid A or a derivative thereof.

Preferably, the vaccine further comprises a saponin, particularly a saponin QS21.

Preferably, the vaccine further comprises an oil-in-water emulsion and comprises tocopherol.

A particularly preferred combination of adjuvants comprising QS21, 3D-MPL and tocopherol in an oil-in-water emulsion is described in WO 95/17210.

The vaccine of the invention may be used for the treatment and / or prophylaxis of hepatitis B virus infection by:

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Administering an effective and safe amount of (1) an antiviral agent active against hepatitis B virus and (2) a vaccine for the prophylaxis and / or treatment of hepatitis B infection, simultaneously or sequentially in any order, to humans or other animals at risk of or suffering from this infection.

Antiviral agents such as penciclovir / famciclovir and HBV vaccine or pharmaceutically acceptable salts or esters thereof may be administered simultaneously in the form of two separate pharmaceutical formulations for simultaneous or sequential use. Usually, the active compounds will be administered separately in dosages which are usually for separate administration of these components. A vaccine or antiviral agent may be administered at the start of treatment.

The invention also relates to the use of anti-viral agents for the manufacture of a pharmaceutical composition for the treatment of patients already having a hepatitis B virus vaccine and suffering from a hepatitis B virus infection. who have already been treated with an antiviral agent and are suffering from hepatitis B virus infection. A preferred antiviral agent is a nucleoside analogue, in particular penciclovir / famciclovir or lamivudine. Preferred hepatitis B virus vaccines have been mentioned above.

A single dose of a nucleoside or nucleotide analog may be administered, for example, 1 to 4 times daily. The exact dose will depend on the route of administration and the severity of the disease being treated, and it will be understood that it will also vary according to the age and weight of the patient, in patients. With a weakened immune system, higher doses may be required.

The vaccines are usually administered in multiple doses at different intervals. They are usually 6 to 12 doses given once every two weeks or once a month.

A preferred component of a pharmaceutical pack when administered concurrently are separately stored components, for example for vaccination into each arm. However, it is also possible to provide simultaneous administration by mixing the two components prior to administration. The components may be administered enterally, for example orally or also parenterally, such as intramuscularly and particularly intravenously.

According to the invention, the antiviral compounds can be formulated into tablets prepared in a conventional manner. Oral formulations such as tablets or capsules may be prepared by conventional techniques using conventional pharmaceutically acceptable excipients such as binding agents such as pregelatinized corn starch, polyvinylpyrrolidone or hydroxypropylmethylcellulose, and fillers such as lactose, microcrystalline cellulose or hydrogen phosphate. calcium, glidants such as magnesium stearate, talc or silica, a disintegrating agent such as potato starch or sodium starch glycolate, and also wetting agents such as sodium lauryl sulfate. The tablets may be coated by known methods. Liquid compositions for oral administration may, for example, be in the form of a solution, syrup or suspension, or may be presented in a dry state to be mixed with water or other suitable vehicle just prior to use. Liquid

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The compositions may be prepared by conventional methods using conventional additives, for example, suspending agents such as sorbitol syrup, cellulose derivatives or hydrogenated edible oils, emulsifying agents such as lecithin or acacia, non-aqueous vehicles such as almond oil, esters types of oils, ethyl alcohol or fractionated vegetable oils and also preservatives, for example methyl or propyl p-hydroxybenzoates or sorbic acid. Compositions of this type may also contain buffer-type salts, flavorings, colorants and sweeteners as appropriate.

Compositions intended for oral administration may also be formulated to control the release of one or both of the active ingredients.

For parenteral administration, the compositions may be formulated in a form suitable for a single injection or continuous infusion. Injectable formulations may be presented in unit dose form, for example in syringes, ampoules or vials containing multiple doses, together with a preservative.

The active antiviral agent may be used in suspension, solution or emulsion in an oily or aqueous vehicle and may contain adjuvants such as suspending, stabilizing and / or dispersing agents. In this case, the active ingredient can also be supplied in the form of a powder to be mixed with a suitable vehicle, for example sterile pyrogen-free water, just prior to use.

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For rectal administration, the antiviral agents may be formulated as suppositories or retention enemas, and suppositories may contain a conventional suppository base, for example, cocoa butter or other glycerides.

The antiviral compositions of the present invention may be prepared in a manner conventional in the pharmaceutical industry.

For example, lamivudine (penciclovir) / famciclovir may be mixed with suitable excipients. For example, tablets may be prepared by direct compression of the resulting mixture. Capsules may be prepared by filling the mixture containing suitable excipients in gelatine capsules in an appropriate filling machine. Controlled-release oral or rectal dosage forms can also be prepared in conventional manner.

Nucleoside analogs active against hepatitis B virus can be identified by conventional procedures, for example, in vitro assays on a culture of primary duck liver cells infected with duck hepatitis B virus, DHBV. Changes in the level of preS1 and / or viral DNA in the culture to which these analogs have been added demonstrate efficacy. Analogs can also be identified by their ability to interfere with the normal acylation of synthetic peptides containing the N-terminal amino acids of DHBV or hepatitis B virus of humans, forest marmots, squirrels or other animals.

The following examples are intended to illustrate the invention.

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DETAILED DESCRIPTION OF THE INVENTION

Pharmacokinetic interactions of the vaccine containing hepatitis B / lamivudine surface antigen in dogs

Method

A vaccine of the following composition was used. The HBV surface antigen was equivalent to that used in the commercially available Engerix-B vaccine ™ vaccine (SmithKline Beecham Biologicals) except that it was a lyophilized vaccine.

Lyophilized Ag antigen:

HBsAG 100 ALIGN! pg sucrose 12, 6 mg NaCl 20, 3 mM NaH ₂ PO ₄ / Na ₂ HPO ₄ 1.35 mM Auxiliary system: oil-in-water emulsions 250 μΐ skvalen 10, 7 mg DL alpha-tocopherol 11, 9 mg polyoxyethylene sorbitan monooleate (Tween 80) 4.8 mg monophosphoryl lipid A 100 ALIGN! μς QS21 100 ALIGN! μυ Water for injection into 0.5 ml Na ₂ HPO ₄ 575 μυ KH ₂ PO ₄ 100 ALIGN! μυ

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KCI 100 pg

NaCl 4 mg pH 6.8 ± 0.2

Lamivudine (Zeffix ™, Glaxo Wellcome) was administered daily as a capsule for oral administration to three male dogs and three female dogs at 100 mg / dog / day for 6 weeks. In addition, on days 14, 28 and 42, a vaccine containing hepatitis B virus surface antigen and adjuvant was administered as described above, the vaccine was administered by intramuscular injection just prior to lamivudine administration. Blood samples were taken before administration, 0.5, 0.75, 1, 2, 4, 6, 8, 12 and 24 hours after lamivudine administration on days 7, 14, 28 and 42. Separated blood plasma was frozen at -20 ° C. ° C and then sent to Pharma Bio-Research for analysis of lamivudine plasma concentrations.

Serum samples for evaluation of surface antigen antibodies were collected on days 0, 29 and 43.

Results

Pharmacokinetics of lamivudine

Blood samples were collected on days 7, 14, 28 and 42 of a six-week toxicity test to evaluate the systemic action of lamivudine in male and female dogs by oral administration of lamivudine 100 mg / dog / day concurrently with a vaccine containing hepatitis B virus surface antigen intramuscularly on days 14, 28 and 42 immediately prior to lamivudine administration. Concentrations of lamivudine in blood plasma in samples • 9 · 9 · ♦ 99 «

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9 > 99 taken 24 hours after dosing, were measured in Pharma Bio-Research.

Peak mean lamivudine plasma concentrations were achieved 2 hours after dosing on all days on which lamivudine was administered except females on day 7, when the maximum mean plasma lamivudine concentration was reached 1 hour after dosing. At day 28, the maximum mean lamivudine concentration was lower than on days 7, 14 and 42. After reaching the maximum, the mean plasma lamivudine concentration decreased in an apparent biexponential manner.

The mean maximum lamivudine plasma concentrations, Cmax and area under the plasma lamivudine concentration curve over time up to 24 hours after administration, AUC ₂₄ on days 7, 14, 28 and 42 are summarized in the following table, with standard deviation in brackets .

Cmax (ng / ml)

Day 7 Day 14 Day 28 Den 42 male female male female male female male female 3045 4290 3176 3555 2053 2542 3277 3287 (1516) (3335) (871) (1901) (515) (1255) (567) (1256)

AUC ₂₄ (ng.h / ml)

Day 7 Day 14 Day 28 Den 42 male female male female male female male female 12541 11514 12858 13567 11629 8883 12585 11049 (2211) (4324) (3231) (5957) (2694) (2534) (1182) (4334)

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The time to peak plasma concentration T _max was typically 2 hours in individual dogs ranging from 0.75 to 4 hours and was apparently independent of HBs vaccine administration.

Plasma lamivudine concentrations could be determined quantitatively in male dogs # 71 and 73 and female dogs # 70 at all time intervals on days 7,

14, 28 and 42, therefore, these animals were used for quantification assays following administration of lamivudine throughout the experiment.

The C _max value for lamivudine was somewhat higher in females than in males. The area under the AUC24 curve was somewhat lower when systemically administered to lamivudine in canine females than in males. However, there were no statistically significant gender differences in the systemic trial, p> 0.57.

On days 14, 28 and 42, the C _max and AUC24 values for systemically administered lamivudine in dogs were generally similar to those on day 7, but the C _max values for canine females on days 14, 28 and 42 were slightly lower than on day 7. However, there was no statistically significant difference in this value for any time interval, (p> 0.08). The mean of the AUC24-based drug build-up ratio is summarized in the following table.

Build-up ratio

male female day 14 / day 7 1.0 1,2 day 28 / day 7 0.9 0.8 day 42 / day 7 1.0 1.0

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From the accumulation values, it can be concluded that there is no accumulation of lamivudine following administration of the HBs vaccine due to values approaching 1.0.

The values for terminal concentration and corresponding terminal half-lives for lamivudine on days 7, 14, 28 and 42 are shown in Tables 5 to 8. The terminal value in the range 0.3239 to 0.1364 / h corresponds to a terminal half-life of lamivudine of 2.1 to 5, 1 hour.

Serology

Method

Quantitative determination of antibodies against hepatitis B virus surface antigen was performed by ELISA using HBs (Hep 286) as envelope antigen. Antigen and antibody solutions were used per well at 100 μΐ. The antigen was diluted to a final concentration of 1 μρ / γηΐ in PBS and was adsorbed overnight at 4 ° C into a 96 well microtiter plate (Maxisorb Immuno-plate, Nunc, Denmark). The plates were then incubated for 1 hour and 30 minutes at 37 ° C with PBS containing 5% nonfat powdered milk and 0.1% Tween 20. Two-fold dilutions of serum from 1/50 or 1/200 in PBS containing PBS were added. 0.5% Gloria milk and 0.1% Tween 20 to HBs-coated plates, and the plates were incubated for 1 hour at 37 ° C. Plates were then washed 4 times with PBS with 0.1% Tween 20. Thereafter, HRPO-conjugated IgG against canine material (Rockland, USA) was added to each well at a 1:40,000 dilution in 0.5% non-fat milk powder and buffer with 0 , 1% Tween 20, after which the plates were incubated for 1 hour at φ φφφφ

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9 Φ Φ Φ · φ · ΦΦ room temperature. After washing, the plates were incubated for 10 minutes at room temperature with a solution of tetramethylbenzidine TMB (Biorad, USA) at 2-fold dilution in 0.1 M citrate buffer pH 5.8. The reaction was stopped by adding 0.5 N sulfuric acid and the plates were then read at 450/630 nm. ELISA titers were expressed as mean titers.

Results

Serological response against hepatitis B virus was measured by ELISA on days 0, 29 and 43. Mean titers are shown in the following table:

Antibodies titers against HBs surface antigen

Dog no. Day 0 Den 29 Den 43 69 25 679 7258 71 25 389 3780 73 25 705 6496 70 25 63 1027 and 72 25 176 3821 74 25 582 11482 Diameter 25 383 5321

Mean titers at different time intervals are 25 on day 0 (probably 1/2 of the first dilution), 383 on day 29 and 5321 on day 43. These values clearly demonstrate elicitation of the immune system response.

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Conclusion

From the above data, it is clear that the oral response and lamivudine systemic response rates in dogs at 100 mg / dog / day are independent of the administration of the hepatitis B virus surface antigen on days 14, 28 and 42 in a pharmacokinetic study, lasting 6 weeks. No differences in the extent or rate of systemic response to lamivudine administration between canine males and canine females could be demonstrated.

Obviously, administration of the vaccine induces the production of high levels of anti-surface antigen antibodies in the bloodstream, which at the same time means that the use of small vertebrate dogs (beagle) is suitable for this type of bioassay.

Represented by:

Claims (19)

PATENT CLAIMS A pharmaceutical package comprising (1) an antiviral agent active against hepatitis B virus and (2) a vaccine for the prophylaxis and / or treatment of hepatitis B virus infection, wherein the active ingredients are for simultaneous or sequential use administration. Pharmaceutical package according to claim 1, characterized in that it is intended for ⁽ treatment of hepatitis B virus infection) Pharmaceutical package according to claim 1, characterized in that it is intended for '1 ...... 10'_; Prevent hepatitis B virus infection Pharmaceutical package according to one of Claims 1 to 3, characterized in that it contains a nucleoside analogue as an antiviral agent. Pharmaceutical package according to claim 4, characterized in that it contains a compound selected from the group of penciclovir, famciclovir or lamivudine as the antiviral agent. Pharmaceutical package according to any one of claims 1 to 6 3, characterized in that it contains a nucleotide analogue as an antiviral agent. = Pharmaceutical package according to claim 4 or 6, characterized in that it comprises: 9 · t · 9 · 9 9 9 9 9 9 9 9 9 · 9 9 9 · · · 999 9,999 9 9,999 9 9 9 9 9 » 99 99 99 99 antiviral agent of the group ganciclovir, lobucavir, adefovir, ribavirin, MBS200,475, vidarabine or ARA-AMP. Pharmaceutical package according to one of Claims 1 to 3, characterized in that it contains alpha-interferon as the antiviral agent. Pharmaceutical package according to one of Claims 1 to 8, characterized in that it contains the hepatitis B virus surface antigen as a vaccine active against hepatitis B. Pharmaceutical package according to claim 9, characterized in that the vaccine effective against hepatitis B virus consists of an SL * antigen. Pharmaceutical package according to claim 9, characterized in that the vaccine effective against hepatitis B consists of 226 amino acid residues of antigen S. Pharmaceutical package according to any one of claims 1 to 11, characterized in that the vaccine comprises an adjuvant. Pharmaceutical package according to claim 12, characterized in that it comprises a 3D-MPL adjuvant, QS21, a mixture of QS21 and cholesterol, a CpG oligonucleotide, aluminum hydroxide, aluminum phosphate, tocopherol and an oil-in-water emulsion or a combination of two or more number of said auxiliary means. • a «« · r r a a a a a a 4 4 44 4 4 4 4 4 · <4 4 ··· 4 4 43 4 4 ·· «44443 444 4,444 44 44 44 44 14. The pharmaceutical package of claim 13 wherein the excipient is 3D-MPL, QS21, and an oil-in-water emulsion. Pharmaceutical package according to claim 14, characterized in that the oil-in-water emulsion comprises squalene, tocopherol and polyoxyethylene sorbitan monooleate (Tween 80). 16. A method of treating a patient suffering from or at risk of infection with hepatitis B virus, comprising administering simultaneously or sequentially in any order a safe and effective amount of 1) an antiviral agent active against hepatitis B virus and 2) a prophylaxis vaccine and / or treating hepatitis B virus infection Method according to claim 16, characterized in that a pharmaceutical package according to any one of claims 1 to 15 is administered. Use of an antiviral agent for the manufacture of a medicament for the treatment of patients who have already received a hepatitis B virus vaccine and who are suffering from a hepatitis B virus infection. Use of a hepatitis B virus vaccine for the manufacture of a medicament for the treatment of a patient already treated with an antiviral agent and suffering from a hepatitis B virus infection.