Gram positive bacteria preparations for the treatment of diseases comprising an immune dysregulation

A technology for autoimmune diseases and preparations, applied in metabolic diseases, skin diseases, sensory diseases, etc., can solve problems such as local ulcers, impossible identification of thermally unstable components, and inability to be used for immunosuppression

Inactive Publication Date: 2012-04-04
INST PASTEUR +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, although live BCG vaccines are useful for the prevention of tuberculosis (JB Milstein et al., WHO Bull.OMS, 1990, 68, 93-108), they have some disadvantages: first, they cannot be used for immunosuppression due to their residual virulence Second, intradermal BCG vaccination causes local reactions proportional to the total amount of bacteria used, and can cause local ulcers, or more severe reactions (abscesses) in case of accidental subcutaneous injection. )
However, it is not possible to identify heat-labile components of heat-inactivated formulations by heat treatment at 120°C for 10 to 30 minutes

Method used

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  • Gram positive bacteria preparations for the treatment of diseases comprising an immune dysregulation
  • Gram positive bacteria preparations for the treatment of diseases comprising an immune dysregulation
  • Gram positive bacteria preparations for the treatment of diseases comprising an immune dysregulation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0133] Example 1: Preparation of deep freeze-dried inactivated mycobacteria

[0134] 1) Materials and methods

[0135] Mycobacterium bovis BCG cells (BCG Pasteur vaccine strain 1173P2, deposited in the French Collection of Microorganisms (CNCM) on April 24, 1978, 25 rue du Docteur Roux, 75724 Paris Cedex 15 (France), deposit number n° I-059 (M. Gheorghiu et al., 1983, Bull. Inst. Pasteur, 81, 281-288) was grown in sterile Sauton medium (HOOCCH (NH 2 O)CH 2 CONH 2 H 2 O (Asparagine), 4g / l; C 6 H 8 O 7 ; H 2 O (citric acid), 2g / l; K 2 HPO 4 (Dipotassium hydrogen phosphate), 0.5g / l; MgSO 4 -H 2 O (magnesium sulfate), 0.50g / l; iron III citrate, 0.05g / l; glycerin, 60ml; zinc sulfate solution (0.155g zinc sulfate dissolved in 10ml pyrogen-free water), 240μl / l, pH 7 ). More specifically, the 1173P2 strain was grown in a 250ml spherical culture flask containing 130ml of sterile Sauton medium and cultured at 37°C for 14 days, which is equivalent to the culture reaching the end of the logar...

Embodiment 2

[0150] Example 2: Preparation of deep freeze-dried inactivated mycobacterial components

[0151] 1) Delipidated fraction (Component A)

[0152] About 10mg / ml of 10 10 Deeply freeze-dried inactivated BCG cells suspended in boric acid buffer (Na 2 B 4 O 7 10H 2 O 0.363%; H 3 BO 3 0.525%; NaCl 0.619% and Tween 200.0005%, dissolved in distilled water, pH 8), centrifuge at 12000g for 10 minutes, discard the supernatant. The precipitate was resuspended in 1ml of chloroform / methanol (9 / 1) and reacted at room temperature for 24h to extract lipids. Centrifuge at 12000g for 10 min to remove chloroform / methanol. The lipid-free precipitate from the chloroform / methanol extract was dried in vacuo.

[0153] 2) Deglycosylated fraction (Component B)

[0154] About 10mg / ml of 10 10 Deeply freeze-dried inactivated BCG cells suspended in boric acid buffer (Na 2 B 4 O 7 10H 2 O 0.363%; H 3 BO 3 0.525%; NaCl 0.619% and Tween 200.0005% were dissolved in distilled water, pH 8), centrifuged at 12000g fo...

Embodiment 3

[0171] Example 3: The protective effect of deep freeze-dried inactivated mycobacteria on asthma in a mouse model

[0172] 1) Materials and methods

[0173] a) Animals

[0174] Male mature (6 to 7 weeks old) BP2 (H-2 q ) The mice were from Centre d'élevage R. Janvier (Le Genest, Saint Isle, France) and were raised in an animal breeding facility free of specific pathogens.

[0175] b) Mouse model of asthma-like immunogen-specific disease

[0176] On day 0 (D 0 ) And the 7th day (D 7 ), with 1 μg ovalbumin (OVA; ICN Laboratories) and 1.6 mg aluminum hydroxide adjuvant (dissolved in saline, final volume 0.4 ml), male mature BP2 mice were immunized subcutaneously (neck dorsal side). An intranasal challenge experiment was performed with 10 μg OVA dissolved in 50 μl saline on day 96-98 to induce allergic reactions. Alternatively, a water-soluble ray-grass pollen extract is used as the immunogen, and the immunization is added the same as OVA, but the amount of the extract is 10 μg.

[0177] c)...

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Abstract

Compositions comprising components prepared from Gram positive bacteria such as Gram positive facultative intracellular bacteria, for example mycobacteria, including extended freeze-dried killed Gram positive bacteria, their method of preparation and their use in humans and animals, for the prevention and / or the treatment of disorders comprising an immune dysregulation such as cancer, autoimmune diseases, allergy and tuberculosis.

Description

Technical field [0001] The present invention relates to a composition comprising ingredients prepared from Gram-positive bacteria (such as Gram-positive facultative intracellular bacteria, such as mycobacteria), said composition being used for the treatment of humans and animals including immune disorders disease. The present invention also relates to the formulation of said ingredients and compositions. Background technique [0002] Diseases that include immune disorders (such as Th1-Th2 imbalance) include various cancers, autoimmune diseases (such as multiple sclerosis, rheumatoid arthritis, Crohn disease and diabetes), and allergic diseases (such as asthma) , Allergic rhinitis, conjunctivitis and atopic dermatitis). [0003] E.g: [0004] Allergic asthma is a common disease that involves allergen-induced airway inflammation; T lymphocytes reach the respiratory tract and activate in response to inhaled allergens, resulting in inflammatory leukocytes, eosinophils, and sometimes n...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/04A61K35/74A61P35/00A61P37/00A61P31/06A61K39/02A61K39/05A61P1/00A61P3/10A61P11/06A61P17/00A61P25/00A61P27/02A61P29/00A61P37/02A61P37/06A61P37/08
CPCC12N1/20A61K35/74A61K45/06A61P1/00A61P1/04A61P11/02A61P11/06A61P17/00A61P19/02A61P25/00A61P27/02A61P27/14A61P29/00A61P31/00A61P31/06A61P35/00A61P37/00A61P37/02A61P37/06A61P37/08A61P3/10A61K39/04
Inventor 玛丽-安妮·拿获里米舍利娜·拉格朗迪吉勒·马沙尔贝尔纳多·鲍里斯·瓦尔加夫季格让·勒福尔费利克斯·罗曼乔治·海基梅因菲利普·佩尔特
Owner INST PASTEUR
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