GB2240922A - Bacterial lysate for the treatment of dermatitis - Google Patents

Abstract

A bacterial lysate derived from at least one of the following strains: Haemophilus influenzae, serotype B NCTC 8467 Diplococcus pneumoniae, serotypes 1, 2, 3 and 47 NCTC 7465, 7466, 7978 and 10319 Klebsiella pneumoniae NCTC 204 and 5056 Klebsiella ozaenae NCTC 5050 Staphylococcus aureus I-049, I-050, I-051, I-052, I-053, and I-054 Streptococcus viridans I-046, I-047 and I-048 Treptococcus pyogenes, group A NCTC 8191 Branhamella catarrhalis (formerly Neisseria catarrhalis) I-045, NCTC 3622 and 3625 is used to manufacture a drug for treating atopic dermatitis.

Description

DRUG FOR THE TREATMENT OF DERMATITIS The present invention is related to a drug for the treatement of atopic dermatitis and particularly the new use of a known bacterial lysate for the manufacture of a drug destined for the treatement of the said disease.

It is already known from the Swiss patent No. CH 633.188 held by the same patentee, about an immunotherapeutic drug against respiratory tract infections which contains as active principle a bacterial lysate derived from at least one of the following strains: - Staphylococcus aureus I-049, I-050, I-051, I-052, I-053 and I-054 - Streptococcus viridarts I-046, I-047 and I-048 - Neisseria catarrhalis It0/25 and from at least one clf the following strains:: - Hemophilus influenzae serotype b NCTC 8467 - Diplococcus pneumoniae serotype 1, 2, 3 and 47 NCTC 7465, 7466, 7978 and 10319 - Kiebsiella pneumoniae NCTC 204 and 5056 - Klebsiella ozaenae NCTC 5050 - Streptococcus pyogenes group A NCTC 8191 - Neisseria cararrhalis NCTC 3622 and 3625 The NCTC strains are catalogued by the National Collection of Type Cultures, 175 Colindale Avenue, London NW9 5HT, Great Britain, and are accessible to the public, while the I strains were registered on March 14, 1978 by the holder of the above-mentioned patent with the National Collection of Microorganism Cultures, Pasteur Institute, 28, rue du Dr Roux, F-75724 Paris, France.All these strains are from germs responsible for r respiratory tract infections.

The bacterial lysate covered by the above-mentioned Swiss patent has demonstrated, in a totally unexpected mamner, a remarkable effect on atopic dermatitis which is a chronic, hereditarily determined disease.

Recent studies showed that atopic dermatitis would be linked to a complex immunodeficiency characterized especially by weak reactions of cell-mediated delayed hypersensitivity ( weak skin reaction) and an increase in Immunoglobulln E (IgE) synthesis. Moreover, the gastrointestinal tract may serve as an entrance door for the allergens responsible for the disease.

Consequently, the present invention covers the use of a bacterial lysate as defined in Claim I for the manufacture of a drug destined to treat atopic dermatitis.

Preferably, the bacterial lysate is derived from all the strains mentioned in Claim I and comes in a concentrated and lyophilized form.

The drug is produced preferably in the form of capsules for oral administration. Each capsule contains 7 mg of lyophilized bacterial lysates.

Described below, as an example, are the preparation of bacterial lysates as well as a galenic form of the drug and the results of clinical trials.

Preparation of Bacterial Lysates A basic sterile standard culture medium can be prepared for all bacterial strains. It contains the following substances in the amounts indicated for a final volume of one liter: Sodium chloride 2,5 g Sodium monohydrogenophosphate 2 g Sodium acetate 0,5 g Aneurine 0,003 g Nicotinic acid 0,003 g Sodium lactate (70% solution) 2 ml Ammonium lactate (50% solution) 2 ml Meat extract 22,5 g Yeast extract 7,5 g Glucose 3g The different strains are cultured separately in optimized conditions in a liquid medium. The basic culture medium is preferably enriched to 2% with the yeast extract and to 0,5% with the Fildes extract for culturing the Hemophilus influenza strain.Also preferably, the basic culture is enriched to 0,5% with the horse serum and to 0,3% with glucose for culturing the Diplococcus pneumoniae strains.

Finally, the same basic medium is preferably enriched to 0,3 % with glucose for culturing the other strains, i.e., Streptococcus viridans and pyogenes, Klebsiella ozaenae and pneumoniae, Staphylococcus aureus and Branhamella catarrhalis.

These separate cultures are grown in a fermentor at 370C and at a pH of 7. After being harvested and counted, the germs are centrifuged and suspended in a saline solution separately for each bacterial strain. The various bacterial suspensions are then subjected to alkaline lysis in standardized conditions of pH (9-10) and of temperature (20-40 C), while the whole process is monitored using a microscope.

The lysate volume coming from each bacterial strain is then adjusted according to the number of germs and the final volume of concentrate.

The various lysates are then mixed in appropriate proportions and centrifuged. The supernatant is diluted in a sallne solution to a final volume calculated for each production batch. The concentrate is then purified by in-depth filtration and then sterilized on a 0,2 ,u membrane.

The product is then tested chemically and microbiologically. It can be stored at 40C.

Preparation of Capsules Before preparing the chosen galenic form, one proceeds, in a known manner, to neutralize the concentrate of bacterial lysates and then to lyophilize this concentrate in order to obtain a lyophilizate which is harvested aseptically, filtered and vacuum-stored in a suitable container. The product thus obtained and called "standardized lysate" contains, for 40 mg, preferably 7 mg of lyophilized bacterial lysates which will constitute the active principle of the future drug. This standardized lyophilizate undergoes chemical and microbiological quality control.

Next, capsules of the drug for oral administration are prepared also in a known manner. Each capsule contains 40 mg of standardized lyophilizate. Capsules, of Capsugel type for example, are filled with a mixture of the lyophilizate and appropriate excipients easily selected by the man skilled in the art.

Toxicity studies Acute toxicity studies in mice at doses of up to 5,000 mg/kg of body weight for determining the lethal dose 50 (L.D. 50) showed that the drug has no toxic effect. Moreover, macro- and microscopic examinations revealed no change in the animal organs.

Clinical studies Clinical studies were performed with the drug thus prepared to determine its effects on atopic dermatitis. Twenty-six adult patients with long-lasting atopic dermatitis were treated with the drug for five months. The drug was administered at the dose of one capsule (7 mg of bacterial lysate) per day for one month and on ten consecutive days of each of the third, fourth and fifth months of the study. A year earlier, 15 of these patients had already received the drug at the same dose for five months.

Special attention was paid to the severity of itching and dermatitis with "lichenification" which was evaluated monthly on the following scale: 0 = absent, 1 = mild, 2 = moderate, 3 = severe and 4 = very severe.

A clinical improvement was considered when the severity of the symptoms (itching and dermatitis) and the use of conventional treatments diminished by more than 50 & Serum IgA, IgG and IgM levels (measured by radial immunodiffusion) and the total serum IgE level (measured by radioimmunoassays) were determined before the study and after one and five months of treatment respectively.

Cell-mediated skin tests (CMI Multitest) were also conducted. A clinical improvement was observed in 57,7% of the 26 patients treated during the five months of the study. If one considers only the 15 patients who were treated for two five-month periods, an improvement was noted in 73,3% of cases.

Serum IgA, IgG, IgM and IgE levels in the 26 patients showed a statistically significant decline after one and five months of treatment (Table I) .

In the 15 patients treated with the drug for two periods, a statistically significant decline of IgE level was observed at the end of the first period and became more pronounced during the second period (Table 2).

The skin tests (CMI Multitest) showed in the 26 patients a weak response before treatment (reflecting a cellular immunodeficiency), i.e., 7,21 + 1,5 mm, which increased after five months of treatment to reach normal levels, 1. e., 22,25 + 2,7 mm (the normal mean values of skin induration varying between 18 and 28 mm).

The results of the tests performed thus show that the drug covered by the present invention has a remarkable effect on atopic dermatitis.

Table I Mean serum IgA, IgG, IgM and IgE levels in 26 patients with atopic dermatitis treated for one five-month period Parameter Initial levels after 1 month p = after 5 months p IgA (IU/ml) 332,62+ 17,29 313,23+ 18,42 0,077 306,15+ 16,36 0,012 IgG (IU/ml) 1844,23+ 68,18 1687,31+ 59,62 0,001 1672,69+ 60,72 0,004 IgM (IU/ml) 228,84+ 12,74 182,31* 8,22 0,0001 195,73* 11,86 0,002 IgE (IU/ml) 1935,54468,65 1553,08368,49 0,007 1286,46281,38 0,012 Table II Mean serum IgE level in 15 patients with atopic dermatitis treated for two five-month periods.

Mean IgE levels (IU/ml) Study period Initial levels After one month After 5 months First 4593,6ffi1089,6 4315,41014,8* 3304,1+ 819,5* Second 2961,7+ 681,6 2329,7+ 530,9*** 1888,6+ 384,33###### * p = 0,038 ** p - 0,007 #### p = 0,008 ##### p = 0,014 Summary The present invention relates to the use of a bacterial lysate in a concentrated and lyophilized form derived from one of the following strains: Haemophilus influenzae (serotype b), Diplococcus pneumoniae (serotypes 1, 2, 3 and 47), Klebsiella pneumoniae, Klebsiella ozaenae, Staphylococcus aureus, Streptococcus viridans, Streptococcus pyogenes (serotype A), Branhamella catarrhalls (formerly Neisseria catarrhalls) for the manufacture of a drug designed to treat atopic dermatitis.

Claims (4)

1. Use of a bacterial lysate derived from at least one of the following strains: Haemophilus influenzae, serotype b NCTC 8467 Diplococcus pneumoniae, serotypes 1, 2, 3 and 47 NCTC 7465, 7466, 7978 and 10319 Klebsiella pneumoniae NCTC 204 and 5056 Klebsiella ozaenne NCTC 5050 Staphylococcus aureus It049, I-050, I-051, I-052, I-053, and I-054 Streptococcus viridans I-046, I-047 and I-048 Streptococcus pyogenes, group A NCTC 8191 Branhamella catarrhalis (formerly Neisserla catarrhalis) I-045, NCTC 3622 and 3625 for the manufacture of a drug destined to treat atopic dermatitis.

2. Use according to Claim 1., characterized by the fact that the bacterial lysate is derived from all the strains mentioned In Claim I.

3. Use according to Claim 1 or 2., characterized by the fact that the bacterial lysate is in a concentrated and lyophilized form.

4. Use according to one of Clams 1 to 3., characterized by the fact that the drug comes in the form of capsules for oral administration, each capsule containing 7 mg of bacterial lysate for a daily dose.