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1. (WO1987003202) FRACTION OBTAINED FROM LISTERIA MONOCYTOGENES HAVING THERAPEUTICAL ACTIVITY, PROCESS FOR ITS PREPARATION AND PHARMACEUTICAL COMPOSITIONS CONTAINING IT
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CLAIMS
1. Active fraction, obtained from a culture of Listeria monocytogenes strain L 79, by means of dolipidatlon of the bacteria, their disintegration, precipitation by means of an agent selected among ammonium sulphate and phosphate buffer, dialysis against distilled water, lyophilization of the insoluble fraction, solubilization in the presence of a compound selected among urea and sodium acid carbonate and lastly chromatographic purification.
2. Active fraction according to claim 1, characterized by having the following analitical characteristics:
- protein content according to Lowry: 0.75 ± 0.1 mg/mg of active fraction;
- moisture content: up to 10%;
- ashes: 0.05-0.08 mg/mg of dry weight of fraction;
- neutral sugars (anthrone method): 0.5-2% of neutral sugars/mg of active fraction
- hexosamine (Elson-Morgan method): 0.6-1% of hexosamine/mg of active fraction;
- molecular weight: between 290,000 and 330,000 Daltons, with an average value of 300,000
3. Active fraction according to claim 2, characterized by having the following aminoacidic composition, referred to a 24 hours hydrolisate:


4. Active fraction according to any of the claims 1 to 3, characterized by being capable of inhibittng the growth of tumoral masses and the forming of metastasis.
5. Active fraction according to any of the claims 1 to 3, characterized by possessing immunomobulating activity deriving from the stimulation of the phagocytary activity, by activating the microbicidal activity of the macrophages and by restoring of the antiviral activity of the macrophages.
6. Active fraction according to any of the claims 1 to 3, characterized by being capable of inducing the production of interferon.

7, Active fraction according to any of the claims 1 to 3, characterized by being capable of potentiating the antihody production.
9. A process for the preparation of the active fraction according to the preeceding claims, characterized by the steps of:
- cultivating Listeria monocytogenes, strain L 79, in a culture medium;

- collecting the resulting bacteria;
- delipidating the bacteria;
- disintegrating the bacteria;
- precipitating an insoluble fraction with an agent selected among ammonium sulphate and phosphate buffer;
- dializing against distilled water;
- lyophilizing the insoluble fraction;
- solubilizing the lyophilisate in the presence of an agent selected among urea and sodium acid carbonate, thus giving a fraction soluble in aqueous medium;
- purifying by chromatography.
9. A process according to claim 8, characterized in that said Listeria monocytogenes strain L 79 has the following characteristics:
- short Gram-positive rod, asporigen, aerobial or microaerobial; at 20 to 25ºC, seldom at 37ºC, shows a fully peculiar tumultuous rotatory mobility;
catalase +
metachromatic granules
haemolysis + ( beta)
- acid production from
glucose +


maltose +
mannitol
solicine +
saccharose
threllose + xylose -dulcitol - UVP +
- eaculin hydrolysis +
- nitrate reduction - - gelatin liquefaction - - urease - - Simmons citrate - - phenylalanine deaminase - - carrier of LM 79 prophage
Io. A process according to claim 8, characterized in that said delipidation is carried out in a Soxhlet apparatus, by alternated treatments with a) ether-ethanol (1:1) for 12 hours
b) chloroform for 12 hours
c) chloroform in admixture with methanol (2:1) for 12 hours;
d) acetone for three consecutive times, each having a duration of 12 hours.
11. A process according to claim 8, characterized in that said disintegration is carried out in a mechanical homogeneizer with simultaneous cooling.
12. A process according to claim 1, characterized in that the suspension of disintegrated bacteria is centrifugated to remove the intact bacteria.

13. A process according to claim 12, characterized in that the supernatant layer of the centrifuglng step is heated to 80°C and adjusted to a 40% saturation concentration by addition of a saturated solution of ammonium sulphate, and i3 maintained for a night at 4º C , the precipitate being thereafter recovered by centrifugation.
14. A process according to claim 13, characterized in that instead of ammonium sulphate a 1.25 M buffer of mono- and bi-potasaium phosphate is used.
15. A process according to the claims 13 or 14, characterized in that the recovered precipitate is suspended in distilled water and depurated from the ammonium sulphate or phosphate buffer, a fraction insoluble in aqueous medium being obtained.
16. A process according to claim 15, characterized in that said depuration is carried out by dialysis against distilled water.
17. A process according to claim 15, characterized in that said insoluble fraction is lyophilized.
18. A process according to claims 8 and 17, characterized in that said lyophilizate is solubilized in the presence of 6M urea or of sodium acid carbonate, a dialysis against distilled water being thereafter carried out in the case of solubilization in the presence of urea.
19. A process according to claims 8 and 18, characterized in that said solubilized fraction io purified by column chromatography.
20. Pharmaceutical composition characterized by containing, as the active ingredient, the active fraction according to claims 1 to 3.
21. Pharmaceutical composition according to claim 20, characterized by being in form suitable for the administration by intramuscular and intravenous route.
22. Pharmaceutical composition according to claim 20, characterized by containing 50-400 ɣ of said active fraction.
23. Pharmaceutical composition according to each of the claims 20 to 23, useful in the treatment of the congenic forms.
24. Pharmaceutical composition according to each of the claims 20 to 23, useful in the treatment of infections pathologic states of mierobial and viral origin.
25. Pharmaceutical composition according to each of the claims 20 to 23, useful in the treatment of the pathological states wherein the im-munodefensive system is affected.