PHARMACEUTICAL COMPOSITIONS COMPRISING NATURAL HUMAN alpha -INTERFERON

Abstract: Use of natural human alpha -interferon for the preparation of a medicament in liquid form to be administered through peroral route at dosages comprised between 100 UI and 500 UI/day, for therapy of viral infections, in particular viral hepatitis, neoplasia and immune diseases in humans and animals.

Description: PHARMACEUTICAL COMPOSITIONS COMPRISING NATURAL HUMAN O.-INTERFERON The invention concerns pharmaceutical compositions for a peroral administration comprising natural human α- mterfercn isolated from lympnoblastoid or leukoc t c cells. In particular compositions are useful for therapy of viral infections, m particular viral nepatitis, neoplasia and immunodeficiency syndromes. The mterferon efficient dosages are clearly lower than dosages utilized for parenteral administration. α-, β-, γ-mterferons are usually administered by infection and are used for therapy, α-mterferon is the most largely utilized mterferon (1) . In an updated study of medicaments for either acute or chronic viral hepatitis tnerapy (2), only α-mterferon is widely accepted as single therapeutic agent. "Viral hepatitis" means at least five different pathologies, having different agents, namely A, B, C, D, E. The therapeutic trend is to treat said pathologies with α-mterferon, with dosages according to the kind of hepatitis, to the overall status of tne subject and to other variable factors. In general, further to the mterferon treatment an almost normalisation clinical and biochemical parameters is achieved for cnronic hepatitis (B, C, D) . The mterferon activity on acute hepatitis has not been focused yet, though for hepatitis C, a therapeutic treatment with α-mterferon lowers the chromcition rate of the disease. Therapeutic cycles indicate the day alternate administration through subcutaneous route of recombinant α-interferon (r α-IFN) at dosages of app. 5.000.000 UI, that m special cases can be up to 9.000.000 Ul/day. The lengtn of therapeutic cycles is cf from six months up to one year (nine months average) . In many cases, unαes reα side effects interfere with the course of therapeutic treatment. In fact some patients, particular tnose at an advanced stage of disease or with severe physiologic damages, do not tolerate the tnerapy and tnerefore the treatment snculd be interrupted. Claimed side effects are: fever, nausea, vomit, tiredness, algia and depression. Moreover the therapeutic cost are quite relevant ooth due to tne high amount of active principle (more than 8.000 new cases each year Italy and 300.000 world-wide 1 and to the necessity of hospitalisation just in consideration of said side effects further to the parenteral administration (day πospitai or outpatients' department) . Finally, as far as cnronic active viral hepatitis the only alternative to the terferon treatment is represented by liver transplant. The clinical trend is to increase -the posology dosage and the length of therapeutic cycle (3), but clinical data show (4') : severe side effects; low acceptance by the patient; high therapeutic costs. Garcia et al. (5) report that the estimate for eacn cured patient is between 700.000 and 2.000.000 English pounds Capri S. (6) report that the cost of eacn mterferon therapeutic treatment is of Lit. 70.000.000/subject. It is therefore evident that tne actual composition of mterferon for therapeutic treatment of hepatitis is not optimal. Moreover clinical resul t s show a better tnerapeut c efficacy m patients which ~ e not the ma target for therapy, namely: young subjects, subjects with a disease at an initial stage, subjects infected with genotipic virus 2 or 3, low viremia subjects. On the contrary a less therapeutic efficacy can be found in those subjects which really need tne therapeutic treatment (subjects poco respondent), as subjects affected by an aggressive form (active cnronic hepatitis) , long length diseases affected subjects, over 50 suojects. Thus patients that really need an immediate mterferon treatment are tnose that have a lower chance of success (7) . The autncrs of the instant invention have found a pharmaceutical composition comprising natural human α- mterferon from either ly pnoblastoid or leukocitic cells to be administered through peroral route, with dosages clearly lower than those used for parenteral administration. The composition maintains as unaltered chemical-physical, biological and pharmacological characteristics of the active principle, having a therapeutic effect substantially analogous to the compositions of prior art but overcoming disadvantages thereof. The composition is preferably in a liquid form with a concentration of 100 to 500 Ul/ml, preferably approx. 150 Ul/ml, most preferably mono-cosage units, most preferably of appr. 1 ml. The composition acts by activating the defence mechanisms against viral infections, tumour growth and stimulates an immune response. The utilisation of natural mterferon was chosen for the setter chances of therapeutic success with respects to recombinant mterferon, obtained by cloning of a single subtype. Though leukocitic and lympnoblastoid interferons exert the same therapeutic properties, the former can be advantageously produced. As a matter of fact it is obtainable by stabilised cell lines, without the need of blood donors. Processes for purifying interferons are known to those skilled in the art, and for example are shown m US Patent 4,732,683; in Cantell K. and Hirvonen S. Texas Reports on Biology and Medicine, Vol. 35, p.138, 1977; m Zoon K.C. et ai. Science 207, p. 527, 1980. The peroral route is generally mucn more accepted b subjects, makes easier nosology schemes and dosages, lowers to stops the antigenic risk, induces the transmission and amplification signal mecnanism, with a irato therapeutic effect, with dosages 100 times lower than known formulations for parenteral administrations. The low dosage annuls the risk of toxic effects; allows a better availability of medicine to satisfy an increasing request and a drastic lowering of therapeutic costs. The preferred formulation m dosage units of small volumes (1 ml) to drink allows an immediate availability cf the active principle, a good standard of cleanliness from the monodosage primary container; the certainty of the taken dosage; the taking of the active principle to be immediately adsorbed by the oro-pharyngeal mucosa, easily preventing the deglutition, an ease and safe way of administration for all of patients, as opposite to lozenges or tablets formulations that snould be kept in the mouth till to full dissolution, with high chances of swallowing. Moreover the composition of the invention is conveniently used for home therapies or on the job place, as precautionary measure for the prophylaxis of viral pathologies, and to control chronic diseases which need of long therapeutic cycles (even yearly) and often recurrent. The composition can be used also in association with other drugs to get synergism and optimize therapeutic schemes. The following clinical studies show the therapeutic effect. A comparison of the electrophoretic protein pattern and of the concentration of IgG, IgA, IgM, before the beginning of the peroral therapy with natural human α-mterferon of hepatitis or other pathologies affected subjects, before and after two weeks of therapeutic treatment, allows to foreseen quail-quantitatively the subject response. Subjects wnicn respond to tne therapy witn 450UI/dιe dosages show a decrease of α2- and β-globulms, of IgGs, of the IgG/IgA ratio, tcgetner to an increase of IgA and IgM concentrations, have a good chance of eliminate the HBVe antigen and to seroconvert, namely to confer a stable remission of the pathology. On the other hand subjects which respond to the same therapy with a decrease of albumin serum concentration, of IgGs, IgAs, IgMs, together to an increase of αl-globul fractions, snould seronvert with longer times. Moreover subjects that respond with an increase of IgGs, of the IgG/IgA ratio, together to a decrease of IgM and of the IgA/IgM ration, could be resistant to the therapy. The monitoring of said parameters (markers) is useful for a planning of therapeutic strategies in clinic and also for the clinical practitioner. Clinical studies on healthy subjects Table 1 shows different therapeutic scnemes . Table 1 T = background; T = Id further the first administration, T = 2d further the first administration, T = 3d further the first administration, T ; = 4d further the first administration, T = 5d further the first administration, T„ = Id after the treatment suspension, T- = 2d after the treatment suspension. The change of the induced biological response with respect to the therapeutic scheme, has been measured on samples of blood, taken at different times. In particular the activity with respect to the day dosage of active principle, to the mono- or pluπ-administration, to the length of the therapeutic cycle was measured. The analysis of data shew that natural human α- interferon from either lymphoblastoid or leukocitic cells, administered at low dosages for a peroral route, is able to modulate (according to the dosage and to the length of the therapeutic cycle) the expression of membrane antigen of healthy subject blood mononuclear cells. In particular, according to therapeutic scheme, the pharmaceutical composition seems to be able to increase both CD4 and CD8 cell population. It is also evident an increased expression of markers of cell activation, as DR antigens and interleukm 2 receptor. The therapeutic scheme with 450 U/die x 5 d (exp.b) is the one provided better results, as shown in Tables 2 and 3. In fact there is an increase ( and absolute) of CD3, CD4, DR1, CD25 lymphocytes. Said increases are, according to different cases, better evident at T 3 , T 4 , T 5 times to later decrease at T, and T- times. The same posology dosage, but with a shorter therapeutic cycle (1 day) (exp.a), interferes less evidently with the and absolute numbers of mononuclear cells the blood (Tables 4 e 5) . In fact in this experiment an increase of average percentage values but not of absolute T, CD8, and class II hystccompatibility antigen lymphocytes values, is evident at time T 3 . Other experimental conditions snow lower increases of the immune response. Therefore, natural human α-mterferon from either lymphoblastoid or leukocitic cells, administered at low dosages trougn peroral route, snows an important role m modulating the immune response, both in the phase afferent than efferent, e has a therapeutic application for the treatment of infective diseases and of other conditions of immunodeficiency. Clinical studies on hepatitis subjects Viral B Hepatitis 14 patients affected by cnronic viral 3 hepatitis, with an age comprised between 4 and 59, were used for random studies. All of subject were previously treated for different periods ranging from some montns to some years with steroids, or with steroid-azothiopurme, with no beneficial effects, neither for the clinical symptomatology nor for the biochemical parameters of the disease, which evolved,' in some cases, to nepat c cirrhosis . The therapeutic treatment of a one administration of 150U/day was initiated immediately after the suspension of tne previous treatment, and effects of said treatment were monitored by checking any alteration of the immune response; of the haematological and biochemical parameters; of serum markers of the viral infection and of the hystoche istry of hepatic bioptic samples. The time of observation varied from 15 to 32 montns and results can be summarized in the following: 1) all of patients during the first 3-6 weeks of treatment registered a transient decay of hepatic biochemical functions (i.e. a 2-3 fold increase of alanmeaminetransferase (ALT) levels) , with no clinical symptoms of disease worsening; 2 > the pnenomenon goes on for 4-6 weeks; 3) in all of treated patients an intense activation of the immune system was ooservec, even after the therapeutic treatment; 4) 7 patients eliminate KBV DNA and HBeAg from serum and stable seroconvert; 5) 1 patient nas an HBcAg increased title, more than the original value; 6) in other 9 patients said titre decreases significatively. Therefore, 50 D of patients get a stable remission of the disease. Viral C Hepatitis The tnerapeutic standard cf viral hepatitis C foresees the use of α-mterferon through parenteral route. 6 active chronic hepatitis C affected patients were subjected to therapy with peroral administration at 150U/dιe, by starting the treatment just after the suspension of the steroid therapy. The observation time (equal to the length of the treatment) resulted to be variade from 19 to 69 weeks. In general tne treatment was well tolerated and all of patients registered a significant increase of vivacity and appetite, with a oetter tolerance to pnysical exercises . No patients got a normalization of transammase levels during the observation period, but one whicn registered the biochemical and clinical remission of the disease, after the treatment susoension at tne 19th weeK due to an increasing of articular pains. Results are shown tables 2-5. BIBLIOGRAPHY 1) Howard M. et al. Le Scienze n.311 Vol LIII 72-80 (1994) . 2) Saracco G., Rizzetto M. Biomed. Pharmacother . 49 (2) ,55-57 (1995) . 3) Kasahara A.K., et al., Hepatology; 21, 291-29 " ? (1995) . 4) Paoletti A. et al . ; Clin. Ter. 146(5) , 343-349 (1995) . 5) Garcia De Ancos J.L. et al.; J. Hepatol. Il:sll-sl8 (1990) . 6) Capri S. Adis International, Milano, pp 41-49 (1994) . 7) Bianchi F.B., La rivista del medico pratico, Ott. (suppl.5) (1995) . b vs a - p<0,05 ; c vs a - p<0,01 ; e vs d p<0,01 ; f vs d = p<0,05 Student's "t" test Tab. ?- - Tab. /» - b vs a - p<0,05 ; d v p<0,05 ; f vs - p<0. l Student's "t" test Tab. £"-

Claims: CLAIMS 1. Use of natural human α-interferon for the preparation of a medicament in liquid form to be administered through peroral route at dosages comprised between 100 UI and 500 Ul/day, for therapy of viral hepatitis in humans and animals. 2. Use of natural human α-interferon for the preparation of a medicament in liquid form to be administered through peroral route at dosages comprised between 100 UI and 500 Ul/day, for therapy of neoplasia and immunologic diseases in humans and animals. 3. Use of natural human α-mterferon according to claims 1 or 2 wherein said mterferon is obtained from lymphoblastoid cell cultures. 4. Use of natural human α-interferon according to claims 1 or 2 wherein said interferon is obtained from lymphocyte cells. 5. Use of natural human α-interferon according to any of previous claims ' wherein said medicament is administered in mono dosage units of appr. 1 ml. 6. Pharmaceutical liquid composition for peroral administration comprising natural human α-interferon either from lymphoblastoid cell cultures or from lymphocyte cells at a concentration between 100 Ul/ml and 500 Ul/ml.

Inventors: BROZZO RENZO, TARRO GIULIO, BROZZO, RENZO, TARRO, GIULIO
Applicants: BROZZO RENZO, IFI ISTITUTO FARMACOTERAPICO I, TARRO GIULIO, BROZZO, RENZO, IFI ISTITUTO FARMACOTERAPICO ITALIANO S.P.A., TARRO, GIULIO
Application year: 1997
Main classification: A61K38
IPC codes: A61K9, A61K38