Pharmaceutical compositions comprising selected lactobacillus strains

Abstract: Topical pharmaceutical compositions, suited for the use in gynecolocy and urology, comprise as active principles selected Lactobacillus strains isolated from vaginal or urologic habitat of asymptomatic patients.

Description: The present invention concerns pharmaceutical compositions suited for the treatment of vaginal and urological diseases, containing selected strains of microorganisms of the " Lactobacillus " genus. A further object of the invention is provided by said strains. The use of lactic acid bacteria in gynecology has already been proposed (FR 6836 M and US 4592748). Their use in urology has been disclosed, for instance, by: a) Andrew W. BRUCE - Gregor REID: "Intravaginal instillation of lactobacilli for prevention of recurrent urinary tract infections"; Can. J. Microbiol. 34(1988) 339-342 b) Gregor REID - Andrew W. BRUCE - Mojtaba BEHESATI: "Effect of antibiotic treatment on receptivity of uroepithelial cells to uropathogens"; ibid. 327-331 c) Gregor REID - Jacqueline A. McGROARTY - Rosanne ANGOTTI and Roger L.COOK: "Lactobacillus inhibitor production against "Escherichia coli" and coaggregation ability with uropathogens"; ibid. 344-351 It has now been found that the use of a particular strain of Lactobacillus , isolated from the vaginal habitat, taxonomically characterized and selected by means of "in vitro" activity tests, allows to obtain particularly favourable therapeutic results, thanks to: (a) its high affinity towards the vaginal epithelium allowing its implant on the vaginal mucosa both in physiological and in pathological conditions, restoring therefore the optimal microflora and pH conditions; (b) its direct action on the pathogenic microorganism, apparently not due to the production of diffusible inhibitory substances but to a coaggregation with the pathogenic microorganism. Of course, the validity of the invention should not be considered to be connected with the verification of the above suggested mechanism of action. The pharmaceutical compositions of the invention are particularly useful for the treatment of vaginal and urological affections of mycotic and bacterial origin. The strains were deposited at the Collection National de Cultures de Microorganisms (CNCM) of Institut Pasteur - Paris (France) on July, 21 , 1988; The identity and deposit numbers of the strains of the invention is hereinbelow reported. LACTOBACILLUS CASEI I-785 LACTOBACILLUS GASSERI I-786 LACTOBACILLUS FERMENTUM I-789 LACTOBACILLUS CASEI SUBSP.PSEUDOPLANTARUM I-790 LACTOBACILLUS CRISPATUS I-787 LACTOBACILLUS FERMENTUM I-788 Some information relevant to the characterisation of the strain of the invention are hereinbelow reported. Lactobacillus Casei : I-785 Guanine + cytosine content (G + C %): 39.7% Fermented sugars according to API CHL 5041 test: L-arabinose ribose galactose glucose fructose N-acetylglucosamine maltose melibiose sucrose melicitose raffinose gluconate 5-oxo-gluconate Plasmids: 1 plasmid 30M daltons Surface protein (S-layer): Molecular weight 56-66.000 daltons. Lactobacillus Gasseri : I-786 G + C %: 34.5 Fermented sugars according to API CHL 5041 test: galactose glucose fructose mannose N-acetylglucosamine amygdaline arlutine salicine cellobiose maltose sucrose trealose gentiliose D tagatose Plasmids: 1 32M daltons Surface protein: two bands with Molecular weight = 50.000 Hybridization with Lactobacillus Gasseri DSM 20243:78% Lactobacillus Fermentum : I-789 G + C %: 51.2 Fermented sugar according to HPI CHL 5041 test: L-arabinose ribose galactose glucose fructose mannose maltose lactose melibiose sucrose raffinose gluconate 5-oxo-gluconate Hybridization with Lactobacillus Fermentum ATCC 14931:100% Lactobacillus Casei supsp.Pseudoplantarum : I-790 G + C%: 46 Hybridization with ATCC 2558:92% Lactobacillus Crispatus: I-787 G + C %: 38 Hybridization with ATCC 33820:80% Lactobacillus Fermentum (LF4): I-788 G + C%: 52.2 Hybridization with ATCC 14931 The compositions of the invention comprise Lactobacillus Fermentum I-789 strain, optionally in combination with one or more strains selected in the above defined group. Particularly favourable therapeutic results were obtained with the following mixture: MIXTURE A 1 Lactobacillus Casei I-785 Lactobacillus Gasseri I-786 Lactobacillus Fermentum I-789 Lactobacillus fermentum I-789 proved to be endowed with a direct inhibitory action on some species of pathogenic fungi, particularly on Candida strains, as it has been shown by means of in vitro tests on agar plates whereupon Candida strain was contacted with Lactobacillus Fermentum I-789. An inhibitory halo of Candida was noticed and a test carried out using the sterile surnatant of the I-789 culture instead of the whole cells shows that said inhibitory action is not due to inhibitory substances released in the culture medium. For the practical use, the microorganisms of the invention are formulated in suitable administration forms such as ovules, creams, vaginal capsules, solutions for lavages, sachets and the like. Unit doses may comprise from 10³ to 10¹⁰ cells of each single strain, the preferred dosage being about 10⁶ cells per unit dose. The bacterial cultures are preferably in lyophilized forms and may be prepared according to conventional methods. For the preparation of aqueous formulations for lavages and irrigations small bottles are used, provided with resevoirs containing the lyophilized microorganisms, to be dissolved before use in a suitable liquid carrier contained in the bottles. The compositions of the invention are practically devoid of any toxicity and show no systemic absorptions so as to allow the administration also to pregnant patients and in cases of intolerability to antimycotic or antibacterial drugs. The compositions of the invention are therefore particularly useful, inter alia, for the prophylaxis of fungal infections due to antibiotic treatments and corresponding complications (formation of resistant strains, relapses etc.). The clinical experience up to now acquired shows as particularly convenient and effective a treatment schedule comprising the administration of the composition of the invention in form of capsules, creams or ovules to be administered before sleeping followed by a lavage on the subsequent morning. The following non limitative examples further illustrate the compositions of the invention. EXAMPLE 1 OVULES Active principle Mixture A 1 1 x 10⁶ cells of each strain Excipient Semi-synthetic glycerides 2800 mg EXAMPLE 2 VAGINAL CREAM (30 g tube) Active principle Lactobacillus fermentum I-789 1 x 10⁶ cells of each strain Excipient Hydrogenated lanoline 5 g Vaseline oil 5 g Dimethylpolysiloxane 10 g SiO₂ (Aerosil 200 R ) 15 g Dodecylglycol polyethylenglycol 1,5 g 1000 copolymer 6 g EXAMPLE 3 VAGINAL CAPSULES Active principle Mixture A 1 1 x 10⁶ cells of each strain Excipient Triglycerides with C₈-C₁₂ fatty acids (Migliol R ) 1500 mg Glycerine 2500 mg Titanium dioxide 10 mg EXAMPLE 4 BOTTLES WITH RESERVOIR Each bottle contains: a) RESERVOIR Lyophilized mixture A 1 1 x 10⁶ cells of each strain b) BOTTLE (10 ml) Glycerine 4,0 g Water q.s. to 10 ml

Claims: Claims for the following Contracting States : AT, BE, CH, DE, FR, GB, IT, LI, LU, NL, SE A Lactobacillus strain deposited at C.N.C.M. of Institut Pasteur on 21.7.1988 having the following identity and deposit number: Lactobacillus Fermentum I-789 Pharmaceutical compositions containing the Lactobacillus strain of claim 1. pharmaceutical compositions according to claim 1 containing a mixture of Lactobacillus strain of claim 1 and at least one Lactobacillus strain selected from - Lactobacillus Casei I-785 - Lactobacillus Gasseri I-786 - Lactobacillus Casei Subsp. Pseudoplantarum I-790 - Lactobacillus Crispatus I-787 - Lactobacillus Fermentum I-788; said strains having been deposited at C.N.C.M. of Institut Pasteur on 21.7.1988. pharmaceutical compositions according to claims 2 or 3 containing from 10³ to 10¹⁰ cells of each strain per unit dose. Pharmaceutical compositions according to claim 4 containing about 10⁶ cells of each strain per unit base. Pharmaceutical compositions according to any one of claims 1-5, containing a mixture of: - Lactobacillus Casei I-785 - Lactobacillus Gasseri I-786 - Lactobacillus Fermentum I-789 Pharmaceutical composition according to any one of the preceding claims in form of creams, ovules, vaginal capsules, solutions for lavages. Claims for the following Contracting States : ES, GR A process for the preparation of pharmaceutical compositions which comprises culturing a Lactobacillus Fermentum strain deposited at C.N.C.M. of Institut Pasteur on 21.7.1988, under deposit number I-789, optionally liophylizing the cultured cells and distributing them in suitable administration forms. A process according to claim 1 which further comprises culturing Lactobacillus strains selected from - Lactobacillus Casei I-785 - Lactobacillus Gasseri I-786 - Lactobacillus Casei Subsp. Pseudoplantarum I-790 - Lactobacillus Crispatus I-787 - Lactobacillus Fermentum I-788; said strains having been deposited at C.N.C.M. of Institut Pasteur on 21.7.1988.

Inventors: BOTTAZZI VITTORIO, DELLAGLIO FRANCO, DONDI GIANCARLA, MORELLI LORENZO, TOSI SILVANA, BOTTAZZI, VITTORIO, DELLAGLIO, FRANCO, DONDI, GIANCARLA, MORELLI, LORENZO, TOSI, SILVANA
Applicants: TOSI A FARMA SRL, DR. A. TOSI FARMACEUTICI S.R.L.
Application year: 1989
Main classification: C12N1
IPC codes: A61P13, A61P31, C12R1, C12N1, A61K9, A61P15, A61K35