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WO1996000579A1 - Extraits cellulaires aqueux et suspensions aqueuses de fractions de membranes cellulaires provenant de mycobacteries - Google Patents

Extraits cellulaires aqueux et suspensions aqueuses de fractions de membranes cellulaires provenant de mycobacteries Download PDF

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Publication number
WO1996000579A1
WO1996000579A1 PCT/EP1995/002524 EP9502524W WO9600579A1 WO 1996000579 A1 WO1996000579 A1 WO 1996000579A1 EP 9502524 W EP9502524 W EP 9502524W WO 9600579 A1 WO9600579 A1 WO 9600579A1
Authority
WO
WIPO (PCT)
Prior art keywords
mycobacteria
aqueous
cell
cell wall
cells
Prior art date
Application number
PCT/EP1995/002524
Other languages
German (de)
English (en)
Inventor
Hans-Georg Laves
Anne Thomsen
Original Assignee
Laves-Arzneimittel Gmbh
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Laves-Arzneimittel Gmbh filed Critical Laves-Arzneimittel Gmbh
Priority to EP95924332A priority Critical patent/EP0804211A1/fr
Priority to KR1019960707482A priority patent/KR970703777A/ko
Priority to PL95317892A priority patent/PL317892A1/xx
Priority to JP8502826A priority patent/JP3031718B2/ja
Priority to AU28884/95A priority patent/AU2888495A/en
Publication of WO1996000579A1 publication Critical patent/WO1996000579A1/fr
Priority to NO965593A priority patent/NO965593L/no

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A61P29/02—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID] without antiinflammatory effect
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/08—Antiallergic agents

Definitions

  • the invention relates to aqueous extracts and aqueous suspensions of Zeil wall fractions from mycobacteria which are generally not human-pathogenic.
  • Mykobacterium tuberculosis in the variety hominis and Mykobacterium leprae are human pathogenic, but there are also a number of mycobacteria, which are usually only animal pathogenic; for example, Mycobacterium tuberculosis var. bovis is pathogenic for many mammals and Mycobacterium avium is a pathogen for birds.
  • Mycobacterium tuberculosis var. bovis is pathogenic for many mammals
  • Mycobacterium avium is a pathogen for birds.
  • tubercle bacteria which penetrate into a host organism which has already been in contact with mycobacteria in the sense of an infection which has already passed through trigger an allergy, that is to say an immunologically caused specifically altered reactivity.
  • the state of allergy to tubercle bacteria is checked by intracutaneous injection of tuberculin; Tuberculin consists of dissolved decay products from tubercle bacteria.
  • the tuberculin sample can therefore be used to determine whether a host organism is allergic to tubercle bacilli or not. An allergic organism reacts much faster to a natural infection and is therefore more likely to contain it than a non-allergic organism. This fact is used to correct a malfunction or a delayed response of the immune system by an unspecific modulation of the immune functions.
  • cytokines such as IL-1 and IL-6 play a central role in the regulation, proliferation and differentiation of immune cells.
  • An unspecific modulation of the immune function essentially arises from the activation of monocytes or macrophages, which then release cytokines to an increased extent.
  • Interleukin research is still at the experimental stage, but it is now known that interleukin 1 causes an increase in PHA-induced lymphocyte proliferation, fibroplast proliferation and an increase in the activity of natural killer cells in the presence of IFN.
  • interleukin 6 which also causes an increase in the activity of natural killer cells and a T cell proliferation.
  • the interleukins are therefore used in immunotherapy for the treatment of tumors and for the substitution of T cell defects.
  • Interleukin 2 is used successfully, for example, in metastatic renal cancer and the cytokines interferon- ⁇ and interferon- ⁇ 2B have been successful in rheumatoid arthritis and hairy cell leukemia. Recently, the granulocyte colony-stimulating factor G-CSF has also been used in the clinic for leukopenia in patients with previous myelosuppressive chemotherapy. In addition to the isolated cytokines, substances are also used therapeutically which stimulate or modulate the immune system non-specifically by activating monocytes or macrophages, which then release cytokines such as IL-1 and IL-6.
  • aqueous extracts or aqueous suspensions of cell wall fractions of mycobacteria which are generally not human-pathogenic are therefore proposed with the features of claims 1 and 2.
  • Mycobacteria which are generally not human pathogenic, can be used, inter alia, as M. tuberculosis var. Bovis, M. avium, M. kansasii, M. marinum, M. scrofulaceum, M. intercellulare, M. xenopi, M. fortuitum and M. chelonae ssp.chelone.
  • These strains are known as such and can be obtained, for example, from the comprehensive London strain collection of the Center for Public Health Laboratory, National Collection of Type Cultures. This institute also provides information about the culture conditions of the respective species or subspecies.
  • the extracts or suspensions prepared according to the invention have proven to be effective immunomodulators in the first clinical trials and can be used, for example, as supportive therapy for infectious diseases, neoplastic diseases, bronchial asthma, allergies of unknown origin and rheumatoid arthritis.
  • aqueous extract or the cell wall fraction For the preparation of the aqueous extract or the cell wall fraction, 250 ml of liquid medium from 5 g / liter sodium chloride, 14 g / liter peptone, 7 g / liter Na2HP0 4 x 2H 2 0, 2 g / liter KH2PO4, 3% glycerol (85% ) inoculated with a bacterial loop with a mycobacterial culture and incubated for 4 weeks at 35 ° C. 4.5 g of moist bacteria are filtered through a sterile suction filter with a glass frit and then triturated homogeneously. After adding 100 ml of sterilized isotonic sodium chloride solution, the bacterial count is 9.7 x 10 ⁇ CFU per ml.
  • the resuspended bacterial cells are centrifuged at 12,000 RPM for 20 minutes.
  • the pellet is mixed dry with glass beads, again taken up in sodium chloride solution and broken with a Vibrogen cell mill at 4 ° C. for 20 minutes. After suction filtering over a glass frit and adding 3 drops of an anti-foaming agent, centrifugation is carried out again (18,000 RPM, 20 minutes). The supernatant obtained is used to produce the mycobacteria extract and the pellet to produce the cell wall fraction.
  • Example 2 To obtain the cell wall fraction, the pellet is resuspended in 27 ml isotonic sodium chloride solution.
  • Example 2 To obtain the cell wall fraction, the pellet is resuspended in 27 ml isotonic sodium chloride solution.
  • Cytokines are vital messenger substances that are found between cells, e.g. mediate the cells of the immune system.
  • IL-1 and IL-6 are pleiotropic cytokines that are produced by different cells and play a central role in immune defense and hematopoiesis. They cause proliferation and differentiation of T and B cells (immunomodulating effect), the regulation of acute phase proteins by the liver in the course of an inflammatory process (anti-inflammatory effect) and the stimulation of colony-forming cells for megakaryocytes and megakaryocytes themselves (regulation of hematopoiesis ). IL-1 also induces the synthesis of other lymphokines such as IL-4, IL-5 and IL-6.
  • Heparinized blood from healthy donors was isolated by gradient centrifugation over Ficoll, washed several times and incubated in serum-free RPMI 1640 (1) with L-glutamine and antibiotics. The various stimuli were incubated with the cells for 24 hours for cytokine production (2). The supernatants were removed, provided with protective protein, stored at -20 ° C. and then examined in the corresponding cytokine test.
  • Endothelial and smooth muscle cells were isolated from pieces of the saphenous vein that could not be used in bypass surgery. EC were isolated by collagenase treatment (3) and SMC by a balancing technique (4). The cells were incubated in M199, ECGF and heparin or DMEM with 10% FCS, L-glutamine and antibiotics (5, 6, 7). The various stimuli were incubated with the cells for 24 hours for cytokine production. The supernatants were removed, provided with protective protein, stored at -20 ° C. and then examined in the corresponding cytokine test. 7TD1 assass for the detection of IL-6:
  • the above-mentioned supernatants of mono-cellular and vascular cells were diluted in 96-well culture plates in 8 to 12 steps. 2,000 cells / well of the IL-6-dependent murine B cell line 7TD1 (8) were added to these dilution series, similarly as described for B9 cells. The cultures were incubated for 72 hours, radioactive thymidine was added for a further 6 hours and the proliferation of the cells was determined by measuring the built-in radioactivity. The activity of the samples was determined by comparing the samples with a defined standard using probit analysis (9).
  • Fibroblasts were cultivated in 96 well plates. After 24 hours, the medium was changed and samples were added to the fibroblast cultures in dilution series. After 72 hours, the cultures were provided with radioactive thymidine and, after a further 24 hours, evaluated as described for 7TD1 cells.
  • the extract induces IL-1 (pg / ml) as follows:
  • the extract induces IL-6 (pg / ml) depending on the dose as follows:
  • the cell wall fraction induced dose-dependent IL-1 (pg / ml) as follows:
  • the cell wall fraction induced dose-dependent IL-6 (pg / ml) as follows:
  • T-cell growth factor parameters for production and a quantitative microassay for activity. J. Immunol. 120: 2027-2032.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
  • Immunology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Rheumatology (AREA)
  • Pain & Pain Management (AREA)
  • Pulmonology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

L'invention a pour objet des extraits aqueux provenant de mycobactéries généralement non pathogènes pour l'homme, caractérisés en ce que les extraits fournissent, dans l'essai à la cytokine, conforme à j.Immunol. 120:2027-2032, pour une variation de concentration de 1 à 4 à 1 à 16, un accroissement de la teneur en cytokines IL-1 et IL-6 dans un rapport, respectivement, de 1:6 et 1:1,25.
PCT/EP1995/002524 1994-06-30 1995-06-29 Extraits cellulaires aqueux et suspensions aqueuses de fractions de membranes cellulaires provenant de mycobacteries WO1996000579A1 (fr)

Priority Applications (6)

Application Number Priority Date Filing Date Title
EP95924332A EP0804211A1 (fr) 1994-06-30 1995-06-29 Extraits cellulaires aqueux et suspensions aqueuses de fractions de membranes cellulaires provenant de mycobacteries
KR1019960707482A KR970703777A (ko) 1994-06-30 1995-06-29 마이코박테리아의 세포벽 분획으로 구성된 수성 현탁액 및 수성 세포 추출물(aqueous cell extracts and aqueous suspensions of cell wall fractions from mycobacteria)
PL95317892A PL317892A1 (en) 1994-06-30 1995-06-29 Aqueous cellular extracts and suspensions from a fraction of mycobacteria cell membrane
JP8502826A JP3031718B2 (ja) 1994-06-30 1995-06-29 マイコバクテリアの細胞壁画分からの水性細胞抽出物および水性懸濁液
AU28884/95A AU2888495A (en) 1994-06-30 1995-06-29 Aqueous cell extracts and aqueous suspensions of cell wall fractions from mycobacteria
NO965593A NO965593L (no) 1994-06-30 1996-12-27 Vandige celleekstrakter og vandige suspensjoner av celleveggfraksjoner fra mykobakterier

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
DE4422859A DE4422859C2 (de) 1994-06-30 1994-06-30 Wäßrige Zellextrakte aus Mykobacterien
DEP4422859.7 1994-06-30

Publications (1)

Publication Number Publication Date
WO1996000579A1 true WO1996000579A1 (fr) 1996-01-11

Family

ID=6521865

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/EP1995/002524 WO1996000579A1 (fr) 1994-06-30 1995-06-29 Extraits cellulaires aqueux et suspensions aqueuses de fractions de membranes cellulaires provenant de mycobacteries

Country Status (9)

Country Link
EP (1) EP0804211A1 (fr)
JP (1) JP3031718B2 (fr)
AU (1) AU2888495A (fr)
CA (1) CA2194138A1 (fr)
CZ (1) CZ285731B6 (fr)
DE (1) DE4422859C2 (fr)
NO (1) NO965593L (fr)
PL (1) PL317892A1 (fr)
WO (1) WO1996000579A1 (fr)

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998020900A1 (fr) * 1996-11-13 1998-05-22 Centenary Institute Of Cancer Medicine And Cell Bi Compositions de paroi cellulaire de mycobacteries
US6013485A (en) * 1996-05-07 2000-01-11 Thymopharma, Ag Low-molecular weight active ingredient extract from yeasts and method for producing it
AU726734B2 (en) * 1996-11-13 2000-11-16 Centenary Institute Of Cancer Medicine & Cell Biology Mycobacterium cell wall compositions
WO2001048154A1 (fr) * 1999-12-28 2001-07-05 Toyoshima, Kumao Agent de maturation pour cellules dendritiques immatures
WO2001049319A1 (fr) * 1999-12-30 2001-07-12 Erika Mutius Von Composition de prevention et de traitement des troubles allergiques
WO2003075827A3 (fr) * 2002-03-13 2003-12-24 Bakulesh Mafatlal Khamar Utilisation de mycobacterium w pour le traitement de l'asthme (maladie pulmonaire obstructive)
US6896887B2 (en) 2001-06-11 2005-05-24 Applied Nanosystems B.V. Bacterial ghosts provided with antigens
DE202007003266U1 (de) 2007-03-02 2008-07-17 Bufe, Albrecht, Prof. Dr. Med. Pharmazeutische Zusammensetzung zum Schutz vor Allergien und entzündlichen Erkrankungen
US9375444B2 (en) 2008-08-16 2016-06-28 Protectimmun Gmbh Composition for prevention and treatment of allergic and/or inflammatory diseases
US9950017B2 (en) 2007-03-02 2018-04-24 Forschungszentrum Borstel Pharmaceutical composition for protection from allergies and inflammatory disorders
US20220047630A1 (en) * 2018-12-14 2022-02-17 Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) Barn dust extract for the prevention and treatment of diseases

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002066055A1 (fr) * 2001-02-20 2002-08-29 Maruho Co., Ltd. Nouvelle utilisation en medecine d'un antigene alpha ou d'un gene d'antigene alpha
EP1637147B1 (fr) * 2004-09-18 2008-12-10 Protectimmun GmbH Extrait de poussières d'étable pour la protection des allergies
CN103354833B (zh) * 2010-10-13 2016-11-16 特莱斯塔治疗知识产权公司 细菌核糖核酸细胞壁组合物及其制备和使用方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3172815A (en) * 1962-04-24 1965-03-09 Warner Lambert Pharmaceutical Method for preparing reticulo-endo-thelial system stimulant and stimulant thereof
GB2015876A (en) * 1978-03-10 1979-09-19 Mitsui Toatsu Chemicals Bacterial extracts of mycobacterium microorganisms
WO1987002249A1 (fr) * 1985-10-08 1987-04-23 Ragland William L Agent immunotherapeutique antiviral et sa preparation

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3172815A (en) * 1962-04-24 1965-03-09 Warner Lambert Pharmaceutical Method for preparing reticulo-endo-thelial system stimulant and stimulant thereof
GB2015876A (en) * 1978-03-10 1979-09-19 Mitsui Toatsu Chemicals Bacterial extracts of mycobacterium microorganisms
WO1987002249A1 (fr) * 1985-10-08 1987-04-23 Ragland William L Agent immunotherapeutique antiviral et sa preparation

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BARNES, P.F. ET AL: "Cytokine production induced by Mycobacterium tuberculosis Lipoarabinomannan", JOURNAL OF IMMUNOLOGY, vol. 149, no. 2, BALTIMORE US, pages 541 - 547 *
STEVEN GILLIS ET AL: "T cell growth factor: Parameters of production and a quantitative microassay for activity", JOURNAL OF IMMUNOLOGY, vol. 120, no. 6, BALTIMORE US, pages 2027 - 2032 *

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6013485A (en) * 1996-05-07 2000-01-11 Thymopharma, Ag Low-molecular weight active ingredient extract from yeasts and method for producing it
WO1998020900A1 (fr) * 1996-11-13 1998-05-22 Centenary Institute Of Cancer Medicine And Cell Bi Compositions de paroi cellulaire de mycobacteries
AU726734B2 (en) * 1996-11-13 2000-11-16 Centenary Institute Of Cancer Medicine & Cell Biology Mycobacterium cell wall compositions
US7083798B1 (en) 1996-11-13 2006-08-01 Centenary Institute Of Cancer Medicine And Cell Biology Method of immunomodulatory treatment of insulin dependent diabetes mellitus using mycobacterial cell wall compositions
WO2001048154A1 (fr) * 1999-12-28 2001-07-05 Toyoshima, Kumao Agent de maturation pour cellules dendritiques immatures
WO2001049319A1 (fr) * 1999-12-30 2001-07-12 Erika Mutius Von Composition de prevention et de traitement des troubles allergiques
US6896887B2 (en) 2001-06-11 2005-05-24 Applied Nanosystems B.V. Bacterial ghosts provided with antigens
US7067639B2 (en) 2001-06-11 2006-06-27 Applied Nanosystems B.V. Method to provide bacterial ghosts provided with antigens
US7541039B2 (en) 2001-06-11 2009-06-02 Applied Nanosystems, B.V. Immunization with bacterial ghost-based vaccines
US7858357B2 (en) 2001-06-11 2010-12-28 Applied Nanosystems B.V. Immunization with bacterial ghost-based vaccines
WO2003075827A3 (fr) * 2002-03-13 2003-12-24 Bakulesh Mafatlal Khamar Utilisation de mycobacterium w pour le traitement de l'asthme (maladie pulmonaire obstructive)
DE202007003266U1 (de) 2007-03-02 2008-07-17 Bufe, Albrecht, Prof. Dr. Med. Pharmazeutische Zusammensetzung zum Schutz vor Allergien und entzündlichen Erkrankungen
US9950017B2 (en) 2007-03-02 2018-04-24 Forschungszentrum Borstel Pharmaceutical composition for protection from allergies and inflammatory disorders
US9375444B2 (en) 2008-08-16 2016-06-28 Protectimmun Gmbh Composition for prevention and treatment of allergic and/or inflammatory diseases
US20220047630A1 (en) * 2018-12-14 2022-02-17 Helmholtz Zentrum München - Deutsches Forschungszentrum für Gesundheit und Umwelt (GmbH) Barn dust extract for the prevention and treatment of diseases

Also Published As

Publication number Publication date
AU2888495A (en) 1996-01-25
EP0804211A1 (fr) 1997-11-05
CZ285731B6 (cs) 1999-10-13
CZ375396A3 (en) 1997-06-11
NO965593D0 (no) 1996-12-27
NO965593L (no) 1997-02-26
DE4422859C2 (de) 2000-10-05
JPH09507857A (ja) 1997-08-12
DE4422859A1 (de) 1996-01-11
JP3031718B2 (ja) 2000-04-10
CA2194138A1 (fr) 1996-01-11
PL317892A1 (en) 1997-04-28

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