AU6347199A

AU6347199A - Use of antifungal agents for treating scleroses

Info

Publication number: AU6347199A
Application number: AU63471/99A
Authority: AU
Inventors: Roumen Antonov
Original assignee: CANDIDA MEDICAL BV
Current assignee: CANDIDA MEDICAL BV
Priority date: 1998-10-27
Filing date: 1999-10-27
Publication date: 2000-05-15
Also published as: CA2348160A1; AU1036999A; EP1124560A1; WO2000024403A1; BR9914792A; JP2002528064A; WO2000024402A1; IL142641A0; CN1328466A

Description

WO 00/24403 PCT/FR99/02617 USE OF ANTIFUNGAL AGENTS IN THE TREATMENT OF SCLEROSES Field of the Invention The present invention has for its object the use of antifungal agents active on fungi of the genus Candida, and as the case may be on those of the genus Rodhotorula, for 5 the preparation of medications adapted for the prevention or treatment of tissue scleroses, and other derived pathologies, in humans or animals, connected to the presence in the human or animal organism of fungi derived from those mentioned above. 10 Summary of the Invention This invention results from the discovery by the inventor of the fact that an antifungal agent active on Candida, known by the common International name (CIN) Nystatin, permits resorbing scleroses of the tissues of the 15 organism, when it is used, on the one hand at a dosage substantially less than its usual dosage in the field of treatment of candidiases, and, on the other hand, for a much longer treatment duration than the usual duration of treatment of candidiases. 20 This discovery is the result of many years of 1 WO 00/24403 PCT/FR99/02617 observation of the effects of Nystatin in vivo for long periods of treatment carried out by the inventor on himself. In his autobiography published in Bulgaria April 27, 1998 ("Chronicle of a Dance with the Devil", by Roumen 5 Antonov, Pygmalion Press ISBN954-8336-39-1), the inventor has set forth his tests carried out on himself, and several other volunteers, with a medication said to be active against Candida albicans, without at the same time identifying this medication. 10 Having discovered that this medication, which in reality was Nystatin, had an effect of resorption of the tissue necroses or scleroses originating from old scars, the inventor hypothesized that these tissue scleroses could result from the development of Candida albicans in regions of 15 the organism where the dead cells and tissue debris accumulate. Thus the inventor sought the presence of Candida albicans in atheroma placque in vitro. The experiment consisted in cutting out atheroma placque and treating it 20 with a liquid local antifungal agent active against Candida albicans. There was then noted in a reproducible manner the decomposition in vitro of the atheroma placque after addition of the antifungal agent. Moreover, the culturing of atheroma placque permit 25 ted observing a structure of Y form which, although 2 WO 00/24403 PCT/FR99/02617 relatively atypical, could suggest the presence of Candida albicans within this atheroma placque. However, this simple observation did not permit by itself concluding with certainty the presence or absence of 5 candida albicans in atheroma placque. Such a confirmation would require the performance of supplemental experiments. Recent experiments on cultures in Sabouraud medium and of specific coloration, particularly by fluorescence, of microorganisms present in atheroma placque, have permitted 10 the inventor to determine unambiguously the presence of a fungus associated with fungi of the genus Candida in atheroma placque, and other sclerotic tissues, although present as filaments having dimensions never described until now for fungi of the genus Candida. Moreover, the inventor has thus 15 been able to show that the above-mentioned fungus was some times accompanied by a fungus of the genus Rodhotorula, this latter being itself for the first time described by the inventor in filamentary form. Moreover, in vivo studies carried out for a long 20 time by the inventor, have permitted him to determine the dosages of antifungal agents necessary for the prevention or treatment of pathologies, particularly various scleroses, connected to the development of these fungi in the organism. 25 Objects of the Invention The invention has for its object a fungus derived 3 WO 00/24403 PCT/FR99/02617 particularly by mutation, from a fungus of the genus Candida, such as Candida lipolitica, or Jarowia lipolitica, or derived from other microorganisms or fungi, particularly other species of lipolytic fungi, said fungus being capable of 5 developing in a complete sexual phase in filamentary form in the human or animal organism, particularly in the vascular system, the different organisms, as well as in the skin. The above-mentioned fungus is characterized also in that it is capable, after having been removed from a host, 10 particularly from human atheroma placque, of developing in vitro in a nutrient medium, such as Sabouraud medium, in the form of filamentary colonies of whitish color, and of a substrate mycelium having filaments easily visible in the nutrient medium. 15 It can also be present in the form colonies of whitish yeast of a fatty appearance relative to the nutrient medium, and corresponding to the imperfect fungus in asexual phase. The fungus derived from the genus Candida mentioned 20 above, according to the invention, is moreover characterized in that it comprises filaments whose diameter varies between about 4 and about 8pi, and can reach more than 30pt in a nutrient medium in vivo, or in the organism. The above-mentioned fungus of the invention is also 25 characterized in that it develops in anaerobic medium, in 4 WO 00/24403 PCT/FR99/02617 The invention also relates to a fungus as defined above, deposited in the Collection Nationale de Culture des Micro-organismes (CNCM) of the Pasteur Institute at Paris, under the number 1-2336, October 15, 1999. 5 The above-mentioned fungi, and particularly those derived from the genus Candida as defined above according to the invention, are moreover characterized in that they can develop in the human or animal organism, in association or not with a fungus of the genus Rodhotorula, or a derivative 10 of this latter, particularly in the presence of Rodhotorula rubra or Rodhotorula glutinis or other subspecies of Rodhotorula, this fungus of the genus Rodhotorula being as the case may be in filamentary form corresponding to a sexual form of these fungi in a complete phase, and this, as was 15 already mentioned above, more particularly in the blood vessels (such as the veins, capillaries, arteries, arterials, coronaries, etc.), or other human tissue, the above-mentioned fungi being adapted to be extracted from this organism. The invention also has for its object the use of 20 antifungal agents active on fungi of the genus Candida, particularly on Candida albicans, Candida lipolitica, or Jarowia lipolitica, and, as the case may be, on fungi of the genus Rodhotorula, such as Rodhotorula rubra or Rodhotorula glutinis, for the preparation of a medication adapted for the 25 prevention or treatment of pathologies connected with the 5 WO 00/24403 PCT/FR99/02617 presence in the human or animal organism of an above mentioned fungus, and more particularly of a fungus derived from the genus Candida as defined above, particularly a fungus derived from Candida lipolitica, or other 5 microorganism, present in the atypical filamentary structure mentioned above, as the case may be in association with an above-mentioned fungus of the genus Rodhotorula and/or in association with yeasts of the Candida lipolitica type. The invention more particularly has for its object 10 the use of antifungal agents such as defined above and hereafter, for the preparation of a medication adapted for the prevention or treatment of tissue scleroses or necroses, or of pathologies resulting from these scleroses or necroses, and being connected to the presence in the human or animal 15 organism of fungi as described above, and more particularly of fungus derived from the genus Candida as defined above, as the case may be in association with an above-mentioned fungus of the genus Rodhotorula. The invention has more particularly for its object 20 the use of the above-identified antifungal agents as described above, for the preparation of a medication adapted for the prevention or treatment of pathologies connected with the development of scleroses or necroses in the organism, namely a medication adapted to prevent or resorb scleroses or 25 necroses resulting from the deposit and accumulation of cellular debris in tissues, as well as holding them in the 6 WO 00/24403 PCT/FR99/02617 form of a mass by the fungi as described above, and more particularly by the fungi associated with those of the genus Candida as defined above, as the case may be in association with an above-mentioned fungus of the genus Rodhotorula, 5 developing in sclerotic or necrotic structures. The invention also relates to the use of antifungal agents as defined above and hereafter, for the preparation of a medication adapted to prevent or treat atherosclerosis connected with the presence in the blood vessels of the human 10 or animal organism, of fungi as described above, and more particularly a fungus derived from the genus Candida as defined above, as the case may be in association with an above-mentioned fungus of the genus Rodhotorula, as well as the different pathologies associated with the above-mentioned 15 atherosclerosis, such as myocardial infarction, phlebitis, and cerebral vascular thromboses. The invention also has for its object the use of antifungal agents as defined above and hereafter, for the preparation of a medication adapted for the prevention or 20 treatment of scleroses or necroses of the skin, such as eczema, psoriasis, erythmas, ichthyoses, connected with the presence in the human or animal organism, of fungi as de scribed above, and more particularly a fungus derived from the genus Candida as defined above, as the case may be in 25 association with an above-mentioned fungus of the genus Rodhotorula, or else of a medication adapted for the cleaning 7 WO 00/24403 PCT/FR99/02617 of necrotic scars and wounds and the elimination of gangrene particularly in the case of ulcers (for example of the legs) of venous origin, bed sores, traumatic wounds, chronic, ulcerated or atonic wounds, superficial and deep burns, 5 cutaneous necroses of traumatic origin, in which the above mentioned fungus or fungi are found. The invention also relates to the use of antifungal agents as defined above and hereafter, for the preparation of a medication adapted for the prevention or treatment of 10 scleroses or necroses of the joints, particularly in the case of arthroses, connected to the presence in the human or animal organism of fungi such as described above, and more particularly of fungus derived from the genus Candida as defined above, as the case may be in association with an 15 above-mentioned fungus of the genus Rodhotorula. The invention also has for its object the use of antifungal agents as defined above and hereafter, for the preparation of a medication adapted for the prevention or treatment of pulmonary pathologies of sclerotic origin, and 20 more particularly of asthma or spasmodic coughing, connected with the presence in the human or animal organism or fungi as described above, and more particularly of a fungus derived from the genus Candida as defined above, as the case may be in association with an above-mentioned fungus of the genus 25 Rodhotorula. 8 WO 00/24403 PCT/FR99/02617 Detailed Description of the Invention By antifungal agents active on fungi of the genus Candida, and on those of the genus Rodhotorula, is meant particularly any compound active on all species of the genus 5 Candida and Rodhotorula, particularly any compound active on Candida albicans, Candida lipolitica, or Jarowia lipolitica, and any compound active on Rodhotorula rubra, or any species or subspecies derived, particularly by mutation, from the above-mentioned fungi. 10 Among the fungi adapted to be used in the scope of the present invention, can be cited: - Flucytosine (CIN), - antifungal contact antibiotics of the family of polyenes, 15 - antifungal agents of the group of imidazoles, such as: - Miconazole (CIN), . Ketoconazole (CIN), . Fluconazole (CIN) (bistriazolated antifungal 20 agent). The invention has more particularly for its object the above-mentioned use of antifungal contact antibiotics of the family of polyenes, and more particularly those extracted from cultures of bacteria of the genus Streptomyces, such as: 25 . Amphotericin B (CIN) extracted from a culture of Streptomyces nodosus, 9 WO 00/24403 PCT/FR99/02617 . Nystatin (CIN) extracted from a culture of Streptomyces noursei, or any derivative of these antifungal agents, such as those adapted to be obtained by chemical synthesis. 5 By way of illustration, the polyenes adapted to be used in the scope of the present invention are the following: - tetraenes of the following formulae 1: nystatin of formula la 10 15.. . pimarycine of formula lb in which R = CH 3 and arenomycine in which R = CH 2

-CH

2

-CH

2

-CH

3 20 25 10 WO 00/24403 PCT/FR99/02617 rimocide of the formula 1c 5 10 - - 04 10* . tetrine A of formula ld in which R = H, and 15 tetrine B in which R = OH 20 014 25 11 WO 00/24403 PCT/FR99/02617 - pentaenes of following formulae 2: . philipine of formula 2a in which R = H, and pentamycine in which R OH 5 10 e~~~~~~. .......-........... ,..-:....... 10 mycotizine A of formula 2b in which R = H, 15 and mycotizine B of formula 2b in which R - CH 3 20 12 WO 00/24403 PCT/FR99/02617 eirotitzine A of formula 2c in which R = CH,, and eirotitzine B of formula 2c in which R = H C - - 10 the compound of the following formula 2d 15 - hexaenes of the following formulae 3: . dermostatine A of formula 3 in which R = CH 3 , 20 and dermostatine B of formula 3 in which R = CH 2

-CH

3 ..- 4 14 01H - 40 H .C0 N 25 13 WO 00/24403 PCT/FR99/02617 - heptaenes of the following formulae 4: amphotericin B of formula 4a 5 - - I - - COCK 10 candidine of formula 4b 15 - 20 Preferably, the antifungal agent used in the scope of the present invention is Nystatin. Also preferably, the dosages of the antifungal agents used in the scope of the present invention are about 2 to about 100 times, particularly about 5 to about 50 times, 25 less than the usual dosages of these same antifungal agents (namely to the usual doses of about 10 tablets of 500,000 IU 14 WO 00/24403 PCT/FR99/02617 per day for 10 days) in the scope of the treatment of candidiases, particularly buccopharyngal, intestinal or vaginal candidiases. Moreover, the duration of treatment is much longer, 5 particularly at least about 5 times longer, in the case of the present invention, than the case of the treatment of the mentioned candidiases, and is more particularly at least about 2 months, particularly at least about 6 months. By way of illustration, the dosages used in the 10 case of the use of antifungal agents, and more particularly Nystatin, within the scope of the present invention, are the following: - less than about 100,000 units per day for an adult, particularly in persons between about 20 years old and 15 about 30 to 40 years old, or - between about 25,000 and about 50,000 units per day in persons less than about 20 years old, and in children, - between about 25,000 and about 125,000 units per week, preferably about 25,000 units per week, in persons 20 older than 40, for an unlimited treatment period. The dosages indicated above are given for one or several administrations per day during the treatment, or, preferably, for one or several dosages per day spaced by intervals of about 3 to about 7 days during the duration of 25 the treatment. At the dosage levels indicated above, the duration 15 WO 00/24403 PCT/FR99/02617 of treatment is about at least 6 months for children and persons less than about 20 years old, up to one or several years for persons more than 20 years old. It should be noted that after such periods of 5 treatment, the immune system becomes capable of taking over against the development of scleroses or necroses due to the development of fungi as described above, and more particularly of the above-mentioned fungi derived from the genus Candida, by an immunization process, and this over a 10 period comprised between about 3 years and about 5 years. A treatment with antifungal agents mentioned above can then stop. In the case in which this immunization disappears, the treatment with the help of antifungal agents mentioned 15 above could then be resumed, preferably with dosages about twice as little as those indicated above, and for a duration about twice as short. The dosages and durations of treatment indicated above permit decreasing significantly the quantities of fungi 20 as described above, and more particularly of the above mentioned fungi derived from the genus Candida, as well as, as the case may be, those of the genus Rodhotorula, in the organism. The invention also has for its object the use of 25 the above-identified antifungal agents as described above, at dosages permitting stopping the growth of said fungi in the 16 WO 00/24403 PCT/FR99/02617 organism. In this case, the dosage level used in the scope of the use of antifungal agents, and more particularly of Nystatin, is preferably about 25,000 units per week (one or several administrations), in persons more than 40 years old, 5 for an unlimited duration of treatment. The invention also has for its object methods of treating the above-mentioned pathologies, by administration to patients needing treatment, of the above-mentioned antifungal agents, particularly at the doses indicated above, 10 and for the above-mentioned times. Preferably, the above-identified antifungal agents are used, in the scope of the present invention, for the preparation of medications adapted to be administered by the oral route, particularly in the form of tablets or drinkable 15 suspensions, or by the injection route. The invention will be further illustrated with the help of the detailed description which follows, of the identification of fungi as described above in biopsies of human origin. 20 1. Patients There is used for research biopsies of vessels, carried out on the occasion of cardiovascular operations practiced on 69 patients, from two different hospitals. The sick persons were distributed in the following 25 manner: 50 men and 19 women, ages 32 to 81. The materials gathered during biopsies were 137 in 17 WO 00/24403 PCT/FR99/02617 number, divided according to their type between: aorta 35; saphenic vein 30; carotid artery 17; mammary artery 39; pulmonary artery, radial artery, etc. 6. 4 patients underwent 4 biopsies, 19 patients 3 5 biopsies, 13 patients 2 biopsies and 33 patients a single biopsy. 2. Methodology The biological materials were gathered under optimum possible conditions of sterility guaranteed by the 10 hospitals during surgical operations. In the course of operation, the biopsies were placed in sterile containers and transported immediately to the laboratory. In the laboratory, although the strictest conditions of sterility prevailed and the air was controlled, the biological 15 materials were cut up in smaller pieces for the work to be done on them and particularly the preparation of slices for direct observation, seeded with nutrient medium and the preservation by freezing for ultimate research. For the different biological materials, a maximum 20 of sections were sought so as to increase the number of seeded sections. A control of the air was carried out by the sedimentation method on a solid nutrient medium in the working area and during all the working period. 2.a. Biological materials prepared for direct 25 observation by microscope (scrapings) The scrapings were prepared in the following 18 WO 00/24403 PCT/FR99/02617 manner: a small piece of biological material was placed between two microscope slides. It was left to dry and the slides were then fixed with the use of heat. From each biopsy were prepared scrapings colored by Gram Modification 5 of Bbzuc: coloration with crystal violet and with carbon nitrate, lugol; decoloration with acetone-alcohol and obtaining contrast with safranine. The scrapings thus colored were observed with an immersion microscope with 100 power objective. Another scraping is prepared for 10 observation under fluorescence. 2.b. Seeding Microbiological seeding takes place in a solid nutrient Sabouraud medium, while seeking to obtain maximum contact of the biological material with the nutrient medium. 15 To do this, the solid material is cut up and the biological material is seeded into the cut-up material obtained. The nutrient medium contains: 10 g Peptone Difco 20 g Glucose Difco 20 15 g Agar Difco distilled water to 1000 ml. The medium is sterilized for 10 minutes at 1210 C. Once seeding is effectuated, the Petri dishes are wrapped with adhesive tape so as to decrease the risk of 25 contamination during the culture time. The culture of the material takes place at 280 C for 30 days carrying out daily 19 WO 00/24403 PCT/FR99/02617 controls without unwrapping the adhesive tape from the Petri dishes. 2.c. Subculturing the pure colonies obtained When a growth is transferred from the plate to the 5 medium, subculturing of the obtained colony takes place while respecting conditions of sterility set forth above, and once! more a scraping is prepared with Gram coloration to observe the colony in the microscope. 2.d. Identification 10 To identify the fungal colonies obtained, there is used API 32C of Bio-Merieux, which is used for yeasts, as well as methods of macroscopic and microscopic observation used for the morphological identification of other types of mycelium. 15 In the course of 137 biopsies, 535 materials were cut up and seeded primarily in a solid nutrient medium. 3. Results 3.a. Microscopic results In the course of 137 biopsies, 115 scrapings were 20 carried out for direct observation during primary cutting up of the biological pieces for their seeding. There is thus observed in the microscope yeasts and/or filaments in 87% of the scrapings. 3.b. Isolated fungal colonies 25 In a large part of the seeded biological materials and in variable periods of time, there are observed growths 20 WO 00/24403 PCT/FR99/02617 of fungal colonies of which the major part is a filamentary fungal colony of whitish color and with substrate micelia with filaments easily visible in the nutrient medium. These filaments directly leave the biological material whilst 5 making the concentric shapes easily visible. The time necessary for the definite appearance of a fungal colony is between the 4th and the 20th day after primary seeding, with two culminating points: the 6th and the 13th day. From September 21 to October 6, in 50 10 patients, 21 had biopsies with positive results (development of the colony) namely 42%. It is interesting to note that the percentage of development of the colony is 75% for patients who had 4 biopsies, 70% for those who had 3 biopsies and that this percentage descends to 29-26% for those who had 15 had 1 or 2 biopsies. Per biopsy, the percentage of positive results is 28%. The second type of fungal colonies found as a function of its proportion in the biological materials is Rhodotorula (Rubra, Glutinis). It grows directly from the 20 biological material whilst remaining in the medium. The identification of this growth was carried out by API 32C and it was seen that it is Rhodotorula Rubra and Rhodotorula Glutinis. The cases of Rhodotorula were distributed equally between the patients in the two hospitals. Rhodotorula grows 25 more rapidly, between 3 and 7 days from seeding. In most cases, it grows from the biopsy of a patient, then or later 21 WO 00/24403 PCT/FR99/02617 also will grow the whitish filamentary colony, and represents 12% of the patients. The third fungal colony that has a function of its quantity in the biopsies is Candida Lipolitica (in 2 pa 5 tients). In all cases, it was isolated beside the whitish filamentary fungal colony. During all the time of the research, in 3 of the 535 seedings that were carried out, there was seen growth of the colonies of a different type which, as a function of the 10 place where they were grown in Petri dishes, have been judged to be contaminants because they had no direct relation with the biological material. In none of the Petri dishes with a nutrient medium and controlled as to air, were found yeasts or contaminants. 22

Claims

1. Fungus derived, particularly by mutation, from a fungus of the genus Candida, such as Candida lipolitica, or Jarowia lipolitica, said fungus being capable of developing in a complete sexual phase in filamentary form in the human or animal organism, particularly in the vascular system, the different organs, as well as in the skin.

2. Fungus according to claim 1, which comprises filaments whose diameter varies between 4 and 8p, and can reach more than 30s1 in nutrient medium in vitro or in the organism.

3. Fungus according to claim 1, which develops in anaerobic medium.

4. Fungus according to claim 1, deposited at the Collection Nationale de Culture des Micro-organismes (CNCM) of the Pasteur Institute in Paris under No. 1-2336, October 15, 1999.

5. Fungus according to claim 1, which can develop in the human or animal organism, in association with a fungus 23 WO 00/24403 PCT/FR99/02617 of the genus Rodhotorula, or with a derivative of this latter, particularly in the presence of Rodhotorula rubra or Rodhotorula glutinis.

6. Use of antifungal agents active on fungi of the genus Candida, and, as the case may be, on fungi of the genus Rodhotorula, for the preparation of a medication adapted for the prevention or treatment of pathologies connected with the presence in the human or animal organism of a fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula.

7. Use of antifungal agents according to claim 6, for the preparation of a medication adapted for the prevention or treatment of tissue scleroses or necroses, or of pathologies resulting from these scleroses or necroses, and being connected to the presence in the human or animal organism of a fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula.

8. Use of antifungal agents according to claim 6, for the preparation of a medication adapted for the prevention or treatment: - of atherosclerosis connected through the presence in the blood vessels of the human or animal organism, of a fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula, as well as the 24 WO 00/24403 PCT/FR99/02617 different pathologies associated with the above-mentioned atherosclerosis, such as myocardial infarction, phlebitis, and cerebral vascular thromboses, - of scleroses or necroses of the skin, such as eczema, psoriasis, erythemas, ichthyoses, connected with the presence in the human or animal organism of a fungus defined in one of claims 1 to 5, as the case may be in associated with a fungus of the genus Rodhotorula, or else a medication adapted for the cleansing of necrotic scars and wounds and the elimination of gangrene particularly in the case of ulcers of venous origin, of bed sores, of traumatic, chronic, ulcerated or atonal wounds, or superficial and profound burns, of cutaneous necroses of traumatic origin, in which is found the fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula, - of scleroses or necroses of the joints, particu larly in the case of arthrosis, connected to the presence in the human or animal organism of a fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula, - of pulmonary pathologies of sclerotic origin, and more particularly of asthma or spasmodic coughing, connected to the presence in the human or animal organism of a fungus defined in claim 1, as the case may be in association with a fungus of the genus Rodhotorula. 25 WO 00/24403 PCT/FR99/02617

9. Use according to claim 6 of antifungal agents selected from the group consisting of - Flucytosin (CIN), - antibiotic antifungal contact agents of the family of polyenes, - antifungal agents of the group of imidazoles, such as: - Miconazole (CIN), - Ketoconazole (CIN), and - Fluconazole (CIN) (bistriazolated antifungal agent).

10. Use according to claim 6 of antifungal contact antibiotics of the family of polyenes, and more particularly those extracted from cultures of bacteria of the genus Streptomyces, such as: . Amphotericin B (CIN) extracted from the culture of Streptomyces nodosus, . Nystatin (CIN) extracted from a culture of Streptomyces noursei.

11. Use according to claim 6 of Nystatin.

12. Use according to claim 6, of antifungal agents, and more particularly Nystatin, at the following dosage levels: - less than about 100,000 units per day for an 26 WO 00/24403 PCT/FR99/02617 adult, particularly in persons between about 20 and about 30 to 40 years of age, or - between about 25,000 and about 50,000 units per day in persons less than 20 years old, and in children, or - between about 25,000 and abut 125,000 units per week, preferably about 25,000 units per week, in persons more than 40 years old, for an unlimited treatment duration.

13. Use according to claim 12, in one or several administrations per day for the duration of treatment, or preferably in one or several administrations per day spaced by intervals of about 3 to about 7 days for the duration of treatment, the duration of treatment being about at least 6 months for children and persons of less than about 25 years old, up to one or several years for persons more than 20 years old. 27