DK3030247T3 - Probiotikum til excessiv spædbarnsgråd - Google Patents

Probiotikum til excessiv spædbarnsgråd Download PDF

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DK3030247T3
DK3030247T3 DK14750457.5T DK14750457T DK3030247T3 DK 3030247 T3 DK3030247 T3 DK 3030247T3 DK 14750457 T DK14750457 T DK 14750457T DK 3030247 T3 DK3030247 T3 DK 3030247T3
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pediococcus pentosaceus
thp
cells
macrophages
bacterial composition
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Castellana Jordi Cuñé
Mallén Elisabet Lázaro
Mazo Jordi Espadaler
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Ab Biotics Sa
Venpharma Laboratorios S A
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    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
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    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
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    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/14—Prodigestives, e.g. acids, enzymes, appetite stimulants, antidyspeptics, tonics, antiflatulents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/04—Centrally acting analgesics, e.g. opioids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/20—Hypnotics; Sedatives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/22—Anxiolytics
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    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
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    • C12N1/00—Microorganisms; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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    • C12N1/205—Bacterial isolates
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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    • A23V2250/00—Food ingredients
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Claims (14)

1. Bakteriel sammensætning, der omfatter fra 104 til 1012cfu/g levedygtige Pediococcus pentosaceus-ceWer, der har evnen til at inducere produktion af interleukin-10, hvor produktionen af interleukin-10 ved THP-1-makrofager i tilstedeværelse af Pediococcus pentosaceus-ceWer udtrykt som normaliseret stigning er større end produktionen af interleukin-10 ved den negative kontrol, der er THP-1-makrofager i fravær af Pediococcus pentosaceus-ceWer, når den normaliserede stigning bestemmes ved følgende trin: (a) differentiering af THP-1-monocytter i makrofager ved dyrkning af THP-1-monocytcellelinjen opnået fra cellesamlingen hos Public Health England, katalognummer 88081201, i RPMI- (Roswell Park Memorial Institute) 1640 medium med 10 % føtalt bovint serum (FBS) og med phorbol 12-myristat 13-acetat (PMA) til en endelig koncentration på 0,16 μΜ; (b) dyrkning af THP-1-makrofagerne i RPMI 1640-medium med 10 % FBS i 24-brønds ELISA-plader til en endelig koncentration på 106makrofager/brønd; (c) inkubation i 2½ time af THP-1-makrofagerne med lipopolysaccharid (LPS) til en endelig koncentration på 10 ng/ml, og vask af THP-1-makrofagerne med D-PBS-(Dulbecco's Phosphate Buffered Saline) medium; (d) klargøring af en kultur af Pediococcus pentosaceus-ceWer ved dyrkning natten over i MRS- (Man, Rogosa and Sharpe) medium ved 37 °C i en 5 % CO2 atmosfære; (e) tilsætning til hver ELISA-brønd af 500 pi RPMI 1640-medium med 10 % FBS og en passende mængde afen opløsning af Pediococcus pentosaceus-ceWer tor at opnå et endeligt forhold på 25:1, dvs. 2,5 x 107cfu Pediococcus pentosaceus-ceWer. 106 THP-1 -makrofager; (f) inkubation af THP-1-makrofagerne med Pediococcus pentosaceus-ceWerne i 2½ time ved 37 °C eller uden Pediococcus pentosaceus-ceWerne på de samme betingelser som negativ kontrol; (g) vask af THP-1-makrofagerne med D-PBS-medium for at fjerne Pediococcus pentosaceus-ceWerne, efterfølgende tilsætning til THP-1-makrofagerne af RPMI 1640-medium med 10 % FBS tilsat 50 pg/ml gentamicin, 10 pg/ml ampicillin og 12 pg/ml chloramphenicol, inkubation ved 37 °Cved 5-7 % CO2 og udtagning afalikvoter ved 5 og 24 timer; (h) centrifugering af alikvoterne og analyse af de ovenstående væsker for interleukin-10-kvantificering ved strømningscytometri; og (i) beregning af den normaliserede stigning af interleukin-1 O-koncentration med formlen (IL1024h - IL10sh)/IL1 Osh; hvor ILIOsh og IL1024her koncentrationen af interleukin-10 i pg/ml ved henholdsvis 5 og 24 timer.
2. Bakteriel sammensætning ifølge krav 1, hvor produktionen af interleukin-10 ved THP-1-makrofagerne i tilstedeværelse af Pediococcus pentosaceus-ceWer udtrykt som normaliseret stigning er mindst 2 gange større end produktionen af interleukin-10 ved THP-1-makrofagerne i fravær af Pediococcus pentosaceus-ceWer, når den normaliserede stigning bestemmes ved trinnene (a)-(i) som defineret i krav 1.
3. Bakteriel sammensætning ifølge et hvilket som helst af kravene 1-2, hvor Pediococcus pentosaceus-ceWerne har evnen til at antagonisere grampositive og gramnegative tarmbakterier.
4. Bakteriel sammensætning ifølge krav 3, hvor de grampositive bakterier omfatter bakterier valgt fra gruppen bestående af Clostridium difficile og Enterococcus faecalis.
5. Bakteriel sammensætning ifølge krav 3, hvor de gramnegative bakterier omfatter bakterier valgt fra gruppen bestående af Escherichia coli, Enterobacter aerogenes, Klebsiella oxytoca og Bacteroides vulgatus.
6. Bakteriel sammensætning ifølge krav 3, hvor Pediococcus pentosaceus-ceWerne har evnen til at antagonisere Clostridium difficile, Enterococcus faecalis, Escherichia coli, Enterobacter aerogenes, Klebsiella oxytoca og Bacteroides vulgatus, hvor evnen til at antagonisere bestemmes ved følgende trin: (i) ensartet udstrygning af patogenstammer på plader, der indeholder Oxoid-medium, og dyrkning til konfluens i en CO2-inkubator ved passende temperaturer og % CO2 for vækst af hvert patogen; (ii) anbringelse af to cylindersektioner med en 6 mm diameter af en ensartet udsået konfluent agarplade af Pediococcus pentosaceus-ceWeme i kontakt med den udsåede plade med patogenet, konfrontering af både (a) vækstsiden af én cylindersektion med den udsåede plade med patogenet; og (b) ikke-vækstsiden af den anden cylindersektion med den udsåede plade med patogenet; og inkubation natten over ved 37 °C; (iii) måling næste dag af hæmningszonerne ved anbringelse af agarpladen over en flad lineal; og (iv) beregning af den væksthæmmende aktivitet ved fratrækning af cylinderdiameteren (CD) fra hæmningszonediameteren (IZD) målt i centimeter og dividering af denne forskel med 2 ifølge formlen GI = (IZD-CD) / 2.
7. Bakteriel sammensætning ifølge et hvilket som helst af kravene 1-6, hvor Pediococcus pentosaceus er Pediococcus pentosaceus indleveret til den spanske artskultursamling under accessionsnummeret CECT 8330.
8. Bakteriel sammensætning ifølge et hvilket som helst af kravene 1-7, der endvidere omfatter fra 104 til 1012 cfu/g celler af Bifidobacterium longum CECT 7894.
9. Bakteriel sammensætning som defineret i et hvilket som helst af kravene 1-8 til anvendelse i bedringen af excessiv gråd hos spædbørn.
10. Bakteriel sammensætning ifølge krav 9 til anvendelse i bedringen af excessiv gråd forbundet med spædbarnskolik.
11. Bakteriel sammensætning ifølge krav 9, hvor spædbørnene har en alder fra tre uger til tolv måneder.
12. Bakteriel sammensætning ifølge et hvilket som helst af kravene 1-8, der er i en form valgt fra gruppen bestående af et kosttilskud, et lægemiddel, en modermælkserstatning, et spiseligt produkt og et fødevareprodukt.
13. Bakteriel sammensætning ifølge krav 12, der er i form af et spædbarnskosttilskud i form af en olieagtig suspension.
14. Fremgangsmåde til screening og isolering af hidtil ukendte Pediococcus pentosaceus-celler, der omfatter følgende trin: (i) analyse af nye Pediococcus pentosaceus-ceWer fra en pulje af Pediococcus pentosaceus-ceWer for deres evne til at inducere produktion af interleukin-10 ved at følge trinnene som beskrevet i krav 1; og (ii) udvælgelse og isolering af de nye Pediococcus pentosaceus-ceWer fra puljen, der inducerer en produktion af interleukin-10, udtrykt som normaliseret stigning, der er større end produktionen af interleukin-10 ved den negative kontrol, når den normaliserede stigning bestemmes i henhold til trinnene ifølge krav 1.
DK14750457.5T 2013-08-09 2014-08-07 Probiotikum til excessiv spædbarnsgråd DK3030247T3 (da)

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EP13382324 2013-08-09
PCT/EP2014/066970 WO2015018883A2 (en) 2013-08-09 2014-08-07 Probiotic for infantile excessive crying

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BR112016002650A2 (pt) 2017-09-12
MX380752B (es) 2025-03-12
CN111676171B (zh) 2023-06-06
WO2015018883A3 (en) 2015-04-02
WO2015018883A2 (en) 2015-02-12
ES2683190T3 (es) 2018-09-25
US20180008652A1 (en) 2018-01-11
US20160184371A1 (en) 2016-06-30
CN105555283A (zh) 2016-05-04
TW201536302A (zh) 2015-10-01
MX2021000259A (es) 2021-03-25
KR20160046826A (ko) 2016-04-29
HUE039336T2 (hu) 2018-12-28
PL3030247T3 (pl) 2019-02-28
EA201690196A1 (ru) 2016-12-30
EP3030247B1 (en) 2018-05-16
US10265351B2 (en) 2019-04-23
CA2920461C (en) 2023-01-03
JP2016526918A (ja) 2016-09-08
JP6643988B2 (ja) 2020-02-12
AU2014304535A1 (en) 2016-03-10
CA3176938A1 (en) 2015-02-12
CN111676171A (zh) 2020-09-18
CN105555283B (zh) 2020-08-11
AU2014304535B2 (en) 2017-06-15
MX2016001635A (es) 2016-10-07
CA2920461A1 (en) 2015-02-12
EP3030247A2 (en) 2016-06-15

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