EP3609477A1 - Microcapsules de pectine, son procédé de fabrication et ses utilisations - Google Patents
Microcapsules de pectine, son procédé de fabrication et ses utilisationsInfo
- Publication number
- EP3609477A1 EP3609477A1 EP18719973.2A EP18719973A EP3609477A1 EP 3609477 A1 EP3609477 A1 EP 3609477A1 EP 18719973 A EP18719973 A EP 18719973A EP 3609477 A1 EP3609477 A1 EP 3609477A1
- Authority
- EP
- European Patent Office
- Prior art keywords
- microcapsules
- pectin
- probiotics
- probiotic
- bacteria
- Prior art date
- Legal status (The legal status 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 status listed.)
- Pending
Links
- 239000003094 microcapsule Substances 0.000 title claims abstract description 163
- 239000001814 pectin Substances 0.000 title claims abstract description 118
- 229920001277 pectin Polymers 0.000 title claims abstract description 115
- 235000010987 pectin Nutrition 0.000 title claims abstract description 115
- 238000000034 method Methods 0.000 title claims abstract description 38
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 18
- 239000006041 probiotic Substances 0.000 claims abstract description 124
- 235000018291 probiotics Nutrition 0.000 claims abstract description 124
- 239000000203 mixture Substances 0.000 claims abstract description 48
- 238000009472 formulation Methods 0.000 claims abstract description 26
- 241000894006 Bacteria Species 0.000 claims description 97
- 230000000529 probiotic effect Effects 0.000 claims description 61
- 239000000243 solution Substances 0.000 claims description 40
- 238000004132 cross linking Methods 0.000 claims description 20
- 239000008240 homogeneous mixture Substances 0.000 claims description 20
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 18
- 239000001963 growth medium Substances 0.000 claims description 17
- 230000008569 process Effects 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 238000005538 encapsulation Methods 0.000 claims description 12
- 210000000936 intestine Anatomy 0.000 claims description 12
- 150000008442 polyphenolic compounds Chemical class 0.000 claims description 12
- 235000013824 polyphenols Nutrition 0.000 claims description 12
- 238000007710 freezing Methods 0.000 claims description 10
- 230000008014 freezing Effects 0.000 claims description 10
- 238000011065 in-situ storage Methods 0.000 claims description 9
- 241001465754 Metazoa Species 0.000 claims description 8
- 239000013543 active substance Substances 0.000 claims description 8
- 230000032050 esterification Effects 0.000 claims description 8
- 238000005886 esterification reaction Methods 0.000 claims description 8
- 244000005709 gut microbiome Species 0.000 claims description 8
- 239000007924 injection Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- 235000013406 prebiotics Nutrition 0.000 claims description 7
- 210000002784 stomach Anatomy 0.000 claims description 7
- 235000013343 vitamin Nutrition 0.000 claims description 7
- 239000011782 vitamin Substances 0.000 claims description 7
- 229940088594 vitamin Drugs 0.000 claims description 7
- 229930003231 vitamin Natural products 0.000 claims description 7
- 230000009435 amidation Effects 0.000 claims description 6
- 238000007112 amidation reaction Methods 0.000 claims description 6
- -1 Ca 2+ Chemical class 0.000 claims description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 5
- 239000003795 chemical substances by application Substances 0.000 claims description 5
- 230000001717 pathogenic effect Effects 0.000 claims description 5
- 239000000725 suspension Substances 0.000 claims description 5
- 208000027244 Dysbiosis Diseases 0.000 claims description 4
- 150000001768 cations Chemical class 0.000 claims description 4
- 230000018044 dehydration Effects 0.000 claims description 4
- 238000006297 dehydration reaction Methods 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 230000007140 dysbiosis Effects 0.000 claims description 4
- 239000000839 emulsion Substances 0.000 claims description 4
- 230000028993 immune response Effects 0.000 claims description 4
- 230000035755 proliferation Effects 0.000 claims description 4
- PYMYPHUHKUWMLA-UHFFFAOYSA-N 2,3,4,5-tetrahydroxypentanal Chemical compound OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 claims description 3
- UCKMPCXJQFINFW-UHFFFAOYSA-N Sulphide Chemical compound [S-2] UCKMPCXJQFINFW-UHFFFAOYSA-N 0.000 claims description 3
- 235000019824 amidated pectin Nutrition 0.000 claims description 3
- 230000035800 maturation Effects 0.000 claims description 3
- 239000000304 virulence factor Substances 0.000 claims description 3
- 230000007923 virulence factor Effects 0.000 claims description 3
- 150000003722 vitamin derivatives Chemical class 0.000 claims description 3
- 244000052769 pathogen Species 0.000 claims description 2
- 239000002775 capsule Substances 0.000 abstract description 4
- 241000699670 Mus sp. Species 0.000 description 17
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 description 17
- 238000004108 freeze drying Methods 0.000 description 16
- 244000199866 Lactobacillus casei Species 0.000 description 15
- 239000000499 gel Substances 0.000 description 15
- 230000001580 bacterial effect Effects 0.000 description 14
- 229920003045 dextran sodium sulfate Polymers 0.000 description 13
- 238000012360 testing method Methods 0.000 description 13
- 210000001035 gastrointestinal tract Anatomy 0.000 description 12
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 11
- 229940072056 alginate Drugs 0.000 description 11
- 235000010443 alginic acid Nutrition 0.000 description 11
- 229920000615 alginic acid Polymers 0.000 description 11
- 239000002609 medium Substances 0.000 description 11
- 230000035882 stress Effects 0.000 description 11
- 230000000968 intestinal effect Effects 0.000 description 10
- 210000001072 colon Anatomy 0.000 description 9
- 230000002496 gastric effect Effects 0.000 description 9
- 229920001282 polysaccharide Polymers 0.000 description 9
- 230000004083 survival effect Effects 0.000 description 9
- 239000011575 calcium Substances 0.000 description 8
- 210000002919 epithelial cell Anatomy 0.000 description 8
- 150000004676 glycans Chemical class 0.000 description 8
- 239000005017 polysaccharide Substances 0.000 description 8
- 229920001817 Agar Polymers 0.000 description 7
- 241000186660 Lactobacillus Species 0.000 description 7
- 239000008272 agar Substances 0.000 description 7
- 229940039696 lactobacillus Drugs 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 229910052500 inorganic mineral Inorganic materials 0.000 description 6
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 6
- 239000011707 mineral Substances 0.000 description 6
- 235000010755 mineral Nutrition 0.000 description 6
- 241000282414 Homo sapiens Species 0.000 description 5
- 238000001035 drying Methods 0.000 description 5
- 238000000684 flow cytometry Methods 0.000 description 5
- 230000037406 food intake Effects 0.000 description 5
- 238000001879 gelation Methods 0.000 description 5
- 239000011159 matrix material Substances 0.000 description 5
- 229910021642 ultra pure water Inorganic materials 0.000 description 5
- 239000012498 ultrapure water Substances 0.000 description 5
- 230000035899 viability Effects 0.000 description 5
- 241000186000 Bifidobacterium Species 0.000 description 4
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 4
- 229920001661 Chitosan Polymers 0.000 description 4
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 4
- 241000917009 Lactobacillus rhamnosus GG Species 0.000 description 4
- AEMOLEFTQBMNLQ-BKBMJHBISA-N alpha-D-galacturonic acid Chemical group O[C@H]1O[C@H](C(O)=O)[C@H](O)[C@H](O)[C@H]1O AEMOLEFTQBMNLQ-BKBMJHBISA-N 0.000 description 4
- 229910052791 calcium Inorganic materials 0.000 description 4
- 238000004624 confocal microscopy Methods 0.000 description 4
- 210000003608 fece Anatomy 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 229940059406 lactobacillus rhamnosus gg Drugs 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 230000011987 methylation Effects 0.000 description 4
- 238000007069 methylation reaction Methods 0.000 description 4
- 239000011859 microparticle Substances 0.000 description 4
- 239000006872 mrs medium Substances 0.000 description 4
- 239000002504 physiological saline solution Substances 0.000 description 4
- 238000002360 preparation method Methods 0.000 description 4
- 108090000790 Enzymes Proteins 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical group Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 3
- 229920001202 Inulin Polymers 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 150000007513 acids Chemical class 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 230000005684 electric field Effects 0.000 description 3
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 3
- 230000007366 host health Effects 0.000 description 3
- 229940029339 inulin Drugs 0.000 description 3
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 235000016709 nutrition Nutrition 0.000 description 3
- 210000001819 pancreatic juice Anatomy 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 239000001509 sodium citrate Substances 0.000 description 3
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 235000000346 sugar Nutrition 0.000 description 3
- 230000004580 weight loss Effects 0.000 description 3
- 239000011701 zinc Substances 0.000 description 3
- GHOKWGTUZJEAQD-ZETCQYMHSA-N (D)-(+)-Pantothenic acid Chemical compound OCC(C)(C)[C@@H](O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-ZETCQYMHSA-N 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000186016 Bifidobacterium bifidum Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- 240000008886 Ceratonia siliqua Species 0.000 description 2
- 235000013912 Ceratonia siliqua Nutrition 0.000 description 2
- 241000194033 Enterococcus Species 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 235000013958 Lactobacillus casei Nutrition 0.000 description 2
- 241001616242 Lactobacillus paracasei ATCC 334 Species 0.000 description 2
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 2
- 241000186429 Propionibacterium Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241000194017 Streptococcus Species 0.000 description 2
- 230000003110 anti-inflammatory effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000032770 biofilm formation Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- 239000000679 carrageenan Substances 0.000 description 2
- 235000010418 carrageenan Nutrition 0.000 description 2
- 229920001525 carrageenan Polymers 0.000 description 2
- 229940113118 carrageenan Drugs 0.000 description 2
- 239000002577 cryoprotective agent Substances 0.000 description 2
- RMRCNWBMXRMIRW-BYFNXCQMSA-M cyanocobalamin Chemical compound N#C[Co+]N([C@]1([H])[C@H](CC(N)=O)[C@]\2(CCC(=O)NC[C@H](C)OP(O)(=O)OC3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)C)C/2=C(C)\C([C@H](C/2(C)C)CCC(N)=O)=N\C\2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O RMRCNWBMXRMIRW-BYFNXCQMSA-M 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 235000015872 dietary supplement Nutrition 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 230000006353 environmental stress Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000003301 hydrolyzing effect Effects 0.000 description 2
- 230000036737 immune function Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 150000002500 ions Chemical class 0.000 description 2
- 229910052742 iron Inorganic materials 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 229940017800 lactobacillus casei Drugs 0.000 description 2
- 239000011777 magnesium Substances 0.000 description 2
- 229910052749 magnesium Inorganic materials 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000008520 organization Effects 0.000 description 2
- 229920000642 polymer Polymers 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- LXNHXLLTXMVWPM-UHFFFAOYSA-N pyridoxine Chemical compound CC1=NC=C(CO)C(CO)=C1O LXNHXLLTXMVWPM-UHFFFAOYSA-N 0.000 description 2
- 238000004626 scanning electron microscopy Methods 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 239000001974 tryptic soy broth Substances 0.000 description 2
- 108010050327 trypticase-soy broth Proteins 0.000 description 2
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 2
- UHVMMEOXYDMDKI-JKYCWFKZSA-L zinc;1-(5-cyanopyridin-2-yl)-3-[(1s,2s)-2-(6-fluoro-2-hydroxy-3-propanoylphenyl)cyclopropyl]urea;diacetate Chemical compound [Zn+2].CC([O-])=O.CC([O-])=O.CCC(=O)C1=CC=C(F)C([C@H]2[C@H](C2)NC(=O)NC=2N=CC(=CC=2)C#N)=C1O UHVMMEOXYDMDKI-JKYCWFKZSA-L 0.000 description 2
- BZTDTCNHAFUJOG-UHFFFAOYSA-N 6-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=CC=C(C(=O)O)C=C21 BZTDTCNHAFUJOG-UHFFFAOYSA-N 0.000 description 1
- AUNGANRZJHBGPY-MBNYWOFBSA-N 7,8-dimethyl-10-[(2R,3R,4S)-2,3,4,5-tetrahydroxypentyl]benzo[g]pteridine-2,4-dione Chemical compound OC[C@H](O)[C@H](O)[C@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-MBNYWOFBSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- PLXMOAALOJOTIY-FPTXNFDTSA-N Aesculin Natural products OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)[C@H](O)[C@H]1Oc2cc3C=CC(=O)Oc3cc2O PLXMOAALOJOTIY-FPTXNFDTSA-N 0.000 description 1
- 244000028821 Annona squamosa Species 0.000 description 1
- 235000005274 Annona squamosa Nutrition 0.000 description 1
- 241000271566 Aves Species 0.000 description 1
- 241000219310 Beta vulgaris subsp. vulgaris Species 0.000 description 1
- 241001655328 Bifidobacteriales Species 0.000 description 1
- 241000186018 Bifidobacterium adolescentis Species 0.000 description 1
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 description 1
- 241000186011 Bifidobacterium catenulatum Species 0.000 description 1
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 1
- 241001134772 Bifidobacterium pseudocatenulatum Species 0.000 description 1
- 206010065687 Bone loss Diseases 0.000 description 1
- GHOKWGTUZJEAQD-UHFFFAOYSA-N Chick antidermatitis factor Natural products OCC(C)(C)C(O)C(=O)NCCC(O)=O GHOKWGTUZJEAQD-UHFFFAOYSA-N 0.000 description 1
- WNBCMONIPIJTSB-BGNCJLHMSA-N Cichoriin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1)c1c(O)cc2c(OC(=O)C=C2)c1 WNBCMONIPIJTSB-BGNCJLHMSA-N 0.000 description 1
- 235000005979 Citrus limon Nutrition 0.000 description 1
- 244000131522 Citrus pyriformis Species 0.000 description 1
- 241001112695 Clostridiales Species 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical class OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- IAJILQKETJEXLJ-UHFFFAOYSA-N Galacturonsaeure Natural products O=CC(O)C(O)C(O)C(O)C(O)=O IAJILQKETJEXLJ-UHFFFAOYSA-N 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 240000001929 Lactobacillus brevis Species 0.000 description 1
- 241000186679 Lactobacillus buchneri Species 0.000 description 1
- 241000218492 Lactobacillus crispatus Species 0.000 description 1
- 241000186673 Lactobacillus delbrueckii Species 0.000 description 1
- 241000186840 Lactobacillus fermentum Species 0.000 description 1
- 241000186606 Lactobacillus gasseri Species 0.000 description 1
- 241000186605 Lactobacillus paracasei Species 0.000 description 1
- 240000006024 Lactobacillus plantarum Species 0.000 description 1
- 241000186604 Lactobacillus reuteri Species 0.000 description 1
- 241000186869 Lactobacillus salivarius Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000235070 Saccharomyces Species 0.000 description 1
- 235000021536 Sugar beet Nutrition 0.000 description 1
- JZRWCGZRTZMZEH-UHFFFAOYSA-N Thiamine Natural products CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N JZRWCGZRTZMZEH-UHFFFAOYSA-N 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000006518 acidic stress Effects 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- IAJILQKETJEXLJ-RSJOWCBRSA-N aldehydo-D-galacturonic acid Chemical group O=C[C@H](O)[C@@H](O)[C@@H](O)[C@H](O)C(O)=O IAJILQKETJEXLJ-RSJOWCBRSA-N 0.000 description 1
- 125000003368 amide group Chemical group 0.000 description 1
- 239000003708 ampul Substances 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 235000013361 beverage Nutrition 0.000 description 1
- 229940002008 bifidobacterium bifidum Drugs 0.000 description 1
- 210000000941 bile Anatomy 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007963 capsule composition Substances 0.000 description 1
- 150000001732 carboxylic acid derivatives Chemical group 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 230000001332 colony forming effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 235000015140 cultured milk Nutrition 0.000 description 1
- 229960002104 cyanocobalamin Drugs 0.000 description 1
- 235000000639 cyanocobalamin Nutrition 0.000 description 1
- 239000011666 cyanocobalamin Substances 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 238000010520 demethylation reaction Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 210000001842 enterocyte Anatomy 0.000 description 1
- 229940093496 esculin Drugs 0.000 description 1
- AWRMZKLXZLNBBK-UHFFFAOYSA-N esculin Natural products OC1OC(COc2cc3C=CC(=O)Oc3cc2O)C(O)C(O)C1O AWRMZKLXZLNBBK-UHFFFAOYSA-N 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- WQMLFJWIKARBFW-BKKMTDGVSA-N evomonoside Chemical compound O[C@@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@H]1O[C@@H]1C[C@@H](CC[C@H]2[C@]3(CC[C@@H]([C@@]3(C)CC[C@H]32)C=2COC(=O)C=2)O)[C@]3(C)CC1 WQMLFJWIKARBFW-BKKMTDGVSA-N 0.000 description 1
- 229920000912 exopolymer Polymers 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 235000021001 fermented dairy product Nutrition 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000012388 gravitational sedimentation Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- WHWDWIHXSPCOKZ-UHFFFAOYSA-N hexahydrofarnesyl acetone Natural products CC(C)CCCC(C)CCCC(C)CCCC(C)=O WHWDWIHXSPCOKZ-UHFFFAOYSA-N 0.000 description 1
- 238000000265 homogenisation Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002519 immonomodulatory effect Effects 0.000 description 1
- 239000007943 implant Substances 0.000 description 1
- 238000002513 implantation Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 238000001802 infusion Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 210000002490 intestinal epithelial cell Anatomy 0.000 description 1
- 230000004609 intestinal homeostasis Effects 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000006193 liquid solution Substances 0.000 description 1
- 230000028744 lysogeny Effects 0.000 description 1
- WPBNNNQJVZRUHP-UHFFFAOYSA-L manganese(2+);methyl n-[[2-(methoxycarbonylcarbamothioylamino)phenyl]carbamothioyl]carbamate;n-[2-(sulfidocarbothioylamino)ethyl]carbamodithioate Chemical compound [Mn+2].[S-]C(=S)NCCNC([S-])=S.COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC WPBNNNQJVZRUHP-UHFFFAOYSA-L 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 244000005706 microflora Species 0.000 description 1
- 239000004005 microsphere Substances 0.000 description 1
- 210000000110 microvilli Anatomy 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 229960003512 nicotinic acid Drugs 0.000 description 1
- 235000001968 nicotinic acid Nutrition 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 235000019520 non-alcoholic beverage Nutrition 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 229940055726 pantothenic acid Drugs 0.000 description 1
- 235000019161 pantothenic acid Nutrition 0.000 description 1
- 239000011713 pantothenic acid Substances 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- 108010009004 proteose-peptone Proteins 0.000 description 1
- 235000008160 pyridoxine Nutrition 0.000 description 1
- 239000011677 pyridoxine Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 229960002181 saccharomyces boulardii Drugs 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000007901 soft capsule Substances 0.000 description 1
- 239000008174 sterile solution Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 235000019157 thiamine Nutrition 0.000 description 1
- KYMBYSLLVAOCFI-UHFFFAOYSA-N thiamine Chemical compound CC1=C(CCO)SCN1CC1=CN=C(C)N=C1N KYMBYSLLVAOCFI-UHFFFAOYSA-N 0.000 description 1
- 229960003495 thiamine Drugs 0.000 description 1
- 239000011721 thiamine Substances 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 235000015193 tomato juice Nutrition 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-O vancomycin(1+) Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C([O-])=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)[NH2+]C)[C@H]1C[C@](C)([NH3+])[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-O 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 229940011671 vitamin b6 Drugs 0.000 description 1
- 239000011240 wet gel Substances 0.000 description 1
- 229920001285 xanthan gum Polymers 0.000 description 1
- 239000000230 xanthan gum Substances 0.000 description 1
- 235000010493 xanthan gum Nutrition 0.000 description 1
- 229940082509 xanthan gum Drugs 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5036—Polysaccharides, e.g. gums, alginate; Cyclodextrin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1232—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt in powdered, granulated or dried solid form
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/065—Microorganisms
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
-
- 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
- A61K35/741—Probiotics
- A61K35/742—Spore-forming bacteria, e.g. Bacillus coagulans, Bacillus subtilis, clostridium or Lactobacillus sporogenes
-
- 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
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- 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
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/062—Ascomycota
- A61K36/064—Saccharomycetales, e.g. baker's yeast
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5089—Processes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P33/00—Antiparasitic agents
- A61P33/02—Antiprotozoals, e.g. for leishmaniasis, trichomoniasis, toxoplasmosis
- A61P33/04—Amoebicides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
- B01J13/00—Colloid chemistry, e.g. the production of colloidal materials or their solutions, not otherwise provided for; Making microcapsules or microballoons
- B01J13/02—Making microcapsules or microballoons
- B01J13/06—Making microcapsules or microballoons by phase separation
- B01J13/14—Polymerisation; cross-linking
Definitions
- the present invention relates to the field of probiotics.
- the present invention relates to pectin microcapsules comprising biofilm probiotics, its method of manufacture and uses thereof.
- the gastrointestinal tract of man has 10 14 micro-organisms in cohabitation, most of which are in the colon with a concentration around 10 11 -10 12 bacteria / mL. Thanks to molecular and culture techniques in constant evolution, to date more than 1000 bacterial species have been identified as resident of the human gastrointestinal tract and an individual would host at least 160 different bacterial species.
- This intestinal population known as the gut microbiota, plays an important role in host physiology (digestion and assimilation of nutrients, protection against colonization of pathogens and modulation of immune responses).
- Dysbiosis of the intestinal microbiota corresponding to an imbalance in its composition could be associated with many human pathologies, such as obesity, Crohn's disease, or ulcerative colitis.
- the ingestion of probiotics can modify the composition of the intestinal microbiota and thus allow to restore its equilibrium.
- the microorganisms most used as probiotics are bacteria belonging to the genera Lactobacillus and Bifidobacterium, natural hosts of the gut microbiota of humans.
- the effectiveness of the probiotics is strain-specific, and each strain can contribute to the health of the host through different mechanisms. Among these mechanisms, the modulation of immune functions at the intestinal level is sought for probiotics in relation to their anti-inflammatory capacity.
- probiotics face a number of environmental stresses before arriving at their site of action, such as gastric acidity, the presence of hydrolytic enzymes or bile salts produced by the intestine. hail.
- probiotics must achieve this in sufficient numbers. In particular, they must be able to withstand gastric acidity and pancreatic juice during their passage through the stomach to be released alive in the intestines. In fact, about 90% of the probiotics ingested are destroyed in the face of these physicochemical stress conditions (acidic pH and high ionic strength).
- a first solution Provided by the state of the art consists in formulating the probiotics with a large bacterial load. Indeed, the amount of live probiotics during ingestion must be sufficient for them to persist in the gastrointestinal tract.
- a second solution is to encapsulate the probiotic bacteria in a matrix, usually alginate, and / or to adapt the probiotic bacteria to the stress conditions encountered in the gastrointestinal tract before possibly encapsulating them.
- alginate or carrageenan microcapsules comprising probiotics in biofilm form do not make it possible to improve their viability in a simulated gastric liquid compared to microcapsules comprising planktonic probiotics.
- This publication concludes that chitosan-coated alginate capsules represent the most appropriate probiotic delivery system due to their release profile and viability during storage. It also indicates that there is a need in the state of the art for providing new capsules that incorporate additional protective materials to protect probiotics from thermal and acid exposures.
- microcapsules although satisfactory, can be improved to allow, for example, optimal adhesion of probiotics in the intestines.
- the culture conditions of the probiotics and the microcapsule / capsule formulation methods remain to be defined and optimized, in particular to improve the survival of the probiotic bacteria, as well as their functionality in the intestine.
- the object of the present invention is thus to provide a new microcapsule avoiding at least partly the aforementioned drawbacks and in particular to improve the survival of probiotic bacteria in the gastrointestinal tract, while being simple to implement.
- an object of the invention is to provide microcapsules for improving the survival of probiotic bacteria in the stomach, to vectorize them into the colon and to release them in a physiological state favorable to their implantation and to their probiotic activity.
- the present invention relates to microcapsules intended to protect probiotics (bacteria or yeasts, for example), comprising an envelope and a core in which said probiotics are dispersed in the form of biofilms distributed in distinct clusters, characterized in that said envelope and said heart are composed of pectin.
- probiotics bacteria or yeasts, for example
- probiotics are defined as living microorganisms (yeasts or bacteria) exerting a beneficial action on the health of the host which ingests them by improving the balance of the intestinal flora, beyond the effects traditional nutritional products. This definition has been approved by the United Nations Food and Agriculture Organization (UNAA) and the World Health Organization (WHO).
- UNAA United Nations Food and Agriculture Organization
- WHO World Health Organization
- the following characteristics may be used alone or in any technically possible combination: the biofilms derived from probiotics are formed in situ;
- the concentration of probiotics in microcapsules varies from 8 to 1 log CFU / g, preferably varies from 9.5 to 10.5 log CFU / g, and typically is 10 log CFU / g;
- said envelope and the core composing the microcapsules are of amidated pectin and methylated having:
- DA degree of amidation
- a degree of esterification ranging from 5 to 50%, preferably from 20 to 30%;
- the probiotics are chosen from: a probiotic bacterium, such as Lactobacillus, Bifidobacterium, Enterococcus, Propionibacterium, Bacillus and Streptococcus, etc. or a yeast, such as a yeast of the genus Saccharomyces cerevisiae, Saccharomyces. boulardi, etc., or a mixture thereof;
- a probiotic bacterium such as Lactobacillus, Bifidobacterium, Enterococcus, Propionibacterium, Bacillus and Streptococcus, etc.
- a yeast such as a yeast of the genus Saccharomyces cerevisiae, Saccharomyces. boulardi, etc., or a mixture thereof;
- said heart comprises another non-probiotic active substance (ie different from a probiotic bacterium or a yeast), such as a polyphenol, a vitamin, a prebiotic, such as inulin, fructo-oligosaccharides (FOS ), etc., or a mixture thereof;
- microcapsules are in dehydrated (for example freeze-dried) or frozen form, so as to optimize their preservation.
- the present invention also relates to a probiotic formulation, characterized in that it comprises at least the microcapsules as described above.
- Another subject of the present invention relates to a method of manufacturing microcapsules as described above, comprising the following steps:
- microcapsules comprising: a pectin shell and a pectin ring forming core, wherein the probiotics are immobilized;
- step (iii) culturing the microcapsules obtained at the end of step (ii) in a culture medium, so as to form in situ within the microcapsules biofilms distributed in distinct clusters from the immobilized probiotics;
- step (iv) optionally, the dehydration (such as lyophilization) or the freezing of the microcapsules obtained at the end of step (iii).
- the homogeneous mixture of step (i) comprises a pectin content, by weight, relative to the total volume of the solution, ranging from 2% (w / v) to 10% (w / v), preferably from 3% (w / v) to 8% (w / v) and typically 3.5% (w / v) to 5% (w / v);
- the mixture of step (i) comprises a probiotic concentration ranging from 10 5 CFU / ml to 10 9 CFU / ml, preferably from 10 6 CFU / ml to 10 8 CFU / ml and typically of the order of 10 7 CFU / mL;
- the encapsulation step (ii) comprises a first substep of injection (iia) by means making it possible to drip the mixture of step (i) into the solution of crosslinking stirred, so that each drop forms a microcapsule when it comes into contact with the crosslinking solution;
- the encapsulation step (ii) comprises a second sub-step called crosslinking of the microcapsules (iib) comprising the maturation of the microcapsules obtained during the injection sub-step (iia) for a period of from minus 3 minutes, preferably from 5 to 60 minutes, and in particular from 10 to 25 minutes;
- the crosslinking solution is composed of divalent cation, such as Ca 2+ , Zn 2+ , Ba 2+ , Fe 2+ in the form of chloride, sulphide, acetate, or a mixture thereof, and is in particular Ca 2+ in the form of chloride;
- step (i) another non-probiotic active substance, such as polyphenol, vitamins, minerals, a prebiotic, etc. is added to the homogeneous mixture.
- another non-probiotic active substance such as polyphenol, vitamins, minerals, a prebiotic, etc.
- the present invention also relates to the use of the microcapsules as described above, or of the abovementioned probiotic formulation comprising said microcapsules, or microcapsules obtained according to the above-mentioned method, for protecting the probiotics during passage in the stomach and for deliver them in an active form to the intestine in an animal.
- animal refers to mammals, birds, fish, insects, etc., as well as humans.
- the present invention also relates to the microcapsules described above, or the abovementioned probiotic formulation or microcapsules obtained according to the process described above, for its use as a medicament.
- the invention also relates to microcapsules described above, or to the abovementioned probiotic formulation or microcapsules obtained according to the method described above for use in animals in order to:
- FIG. 2 is an observation in confocal microscopy: (a) an overview (730 ⁇ scale) representing 12 pectin microcapsules according to example 1 of the invention which were incubated in an MRS culture medium in which, within each microcapsule, the various distinct clusters of biofilm comprising the probiotics L. casei ATCC334 are distinguished, (b) an enlarged view (scale 150 ⁇ ) of the microcapsules of FIG. 2 (a) in which one can see 4 microcapsules inside which are arranged the distinct clusters of biofilm included in the pectin network; and (c) an even more enlarged view (scale 5 ⁇ ) of the interior of a microcapsule of Figure 2 (b) and showing two clusters of biofilm;
- FIG. 3 is a cryo-scanning electron microscopy observation: (a) an overview (2.0 mm scale) of several pectin microcapsules according to example 1 of the invention which have been incubated in a MRS culture medium; (b) an enlarged view (100 ⁇ scale) of the inside of a microcapsule of FIG. 3 (a) in which distinct clusters of biofilm (including L. casei ATCC334 bacteria) distributed in the network can be distinguished; pectin; (c) an enlarged view (scale 40.0 ⁇ ) of FIG.
- FIG. 4 is a graph showing the weight loss of mice over time with DSS treatment (from day 5 to day 11): the condition "physiological water + DSS” corresponds to the mice that have received the DSS treatment and fed with physiological water; the "formulation + DSS” condition corresponds to the mice treated with DSS and gavaged with a formulation comprising the microcapsules according to the invention (pectin microparticles containing L. casei probiotic biofilms);
- FIG. 5 and FIG. 6 are graphs representing the quantification over time of the bacteria of the genus Lactobacillus spp (FIG. 5) and L. casei (FIG. 6) in the faeces of the group of mice which have received the formulation according to said invention. as shown in Figure 4 (pectin microparticles containing L. casei probiotic biofilms).
- the Applicant has focused on the development of new microcapsules including probiotics able to withstand gastric acidity and that of pancreatic juice during their passage in the stomach to release viable probiotics in the intestines where they have particular beneficial action on the health of the host that ingests them.
- microcapsules comprising a pectin shell and a pectin-forming core in which probiotics in the form of a biofilm are immobilized make it possible to obtain this technical effect.
- microcapsules pectin could increase the survival of bacteria or probiotic yeasts at different levels of the gastrointestinal tract and release them in viable and active form in the intestine of the 'host.
- the microcapsules according to the invention after ingestion, thus make it possible to resist the gastric acidity, to that of the pancreatic juice, as well as to the hydrolytic enzymes and to implant themselves, moreover in the intestine.
- the simple use of pectin among all the existing polysaccharides makes it possible to form microcapsules having a pectin shell and a core composed of a pectin network having a similar resistance, or even an improvement in gastric stress compared to Microcapsules of the prior art for which it is necessary to apply at least two layers of polysaccharide (alginate / chitosan) to protect bacteria or probiotic yeasts.
- pectin is furthermore not suggested in the prior art insofar as the three-dimensional network formed by pectin in the presence of calcium ions is relatively heterogeneous with respect to the alginate network. This is due to the presence of ester and amide groups and the presence of branched zones in pectin (Assifaoui et al., Soft Matter 2015). However, surprisingly, this heterogeneity of structure is responsible for the development of bacteria and probiotic yeasts in biofilms. Thus, unlike alginate, which has a linear structure, the particular pectin gel network unexpectedly allows for better growth of probiotic bacteria and yeasts within that network.
- the present invention firstly relates to microcapsules intended to protect probiotics, comprising an envelope and a heart in which said probiotics are dispersed in the form of biofilms distributed in distinct clusters, characterized in that said envelope and said core are composed of pectin.
- Biofilm refers to structured communities of bacteria or yeast enclosed in a self-producing polymer matrix that adheres to a living or inert surface (Costerton et al., 1999).
- biofilms derived from probiotics are formed in situ.
- probiotics and in particular the probiotic bacteria of the genus Lactobacillus, grown in biofilm, are more resistant to stress mimicking the conditions encountered in the gastrointestinal tract and moreover exhibit an activity. increased anti-inflammatory.
- the biofilm formation of bacteria and probiotic yeasts combined with the use of pectin, makes it possible to form microcapsules with good resistance to environmental stresses encountered between ingestion and the site of action (colon).
- the biofilm is also preserved until the place of delivery of the bacteria.
- the microcapsules according to the invention are capable of releasing at the level of the colon the bacteria with a biofilm phenotype, that is to say having, on the one hand, adhesion properties which are much greater than the planktonic cells and on the other hand on the other hand, increased probiotic activity (immunomodulation).
- the enzyme that degrades pectin is naturally present in the colon. It is therefore a targeted delivery of bacteria in a selected compartment of the intestine, the colon.
- the probiotic bacteria and yeast suitable for the present invention are thus able to form a biofilm and can be chosen from: Lactobacillus, Bifidobacterium, Enterococcus, Propionibacterium, Bacillus and Streptococcus or a mixture thereof.
- the probiotic bacteria may be chosen from: acidophilus L. crispatus L. gasseri L. delbrueckii L. salivarius L. casei L. paracasei L. plantarum L. rhamnosus L. reuteri L. brevis, L. buchneri, L. fermentum, B. adolescentis, B. angulation, B. bifidum, B. brief, B. catenulatum, B. infantis, B. lactis, B. longum, B. pseudocatenulatum, S. thermophilic, or a mixture thereof, preferably the probiotic bacteria are L. casei and L. rhamnosus, or a mixture thereof.
- probiotic yeasts that are suitable for the present invention can be chosen from: Saccharomyces cerevisiae, Saccharomyces boulardii, etc. or a mixture thereof.
- the concentration of probiotics in microcapsules is very high. It varies, for example, from 8 to 1 log CFU / g, preferably from 9.5 to 10.5 log CFU / g, and is typically 10 log CFU / g.
- other non-probiotic active substances namely different from a bacterium or a yeast, can be included in the core of the microcapsules. In particular, they are trapped in the pectin network.
- These other active substances may be chosen in particular from: a polyphenol, a vitamin (thiamine (B1), riboflavin (B2), niacin (B3), pantothenic acid (B5), pyridoxine (B6), folic acid (B9) and cyanocobalamin (B12), a mineral (magnesium, calcium, iron, etc.), a prebiotic, such as inulin, FOS, etc., or a mixture thereof.
- thiamine B1
- riboflavin B2
- niacin B3
- pantothenic acid B5
- pyridoxine B6
- folic acid B9
- cyanocobalamin B12
- a mineral magnesium, calcium, iron, etc.
- prebiotic such as inulin, FOS, etc., or a mixture thereof.
- polyphenols of plant origin are very reactive molecules which, according to in vitro and in vivo studies carried out in mice, can have a strong impact on the signaling of the intestinal cells, but also on the bacteria of the gut microbiota.
- the microcapsules according to the invention advantageously contain a high amount of vitamins and minerals that will be easily assimilated by the body.
- the "prebiotics” are short chain oligosaccharides or polysaccharides consisting of approximately two to twenty units of sugar. They escape digestion in the small intestine and are potential substrates for hydrolysis and fermentation by intestinal bacteria.
- the envelope and the core of the microcapsules according to the invention are pectin.
- the microcapsules according to the invention comprise or may consist of an envelope and a heart in which said probiotics are dispersed in the form of biofilms distributed in distinct clusters, characterized in that said envelope and said core are composed of pectin.
- the microcapsules are only composed of pectin to form the envelope and the heart, that is to say that the microcapsules do not include other polysaccharides to form their heart and their envelope.
- the microcapsules according to the invention do not comprise a coating, ie they are not covered by another protective material, such as a polysaccharide, such as chitosan.
- pectin is a polysaccharide of vegetable origin characterized by an ⁇ -D-galacturonic acid backbone and small amounts of ⁇ -L-rhamnose more or less branched.
- it comprises a sequence of two majority structures: a homogalacturonic main chain (or “smooth zone”, called HG) and a rhamnogalacturonic chain (or “spiky zone”, called RG).
- Homogalacturonans are the main chain that makes up pectins (generally representing more than 60% of pectins). These are ⁇ -D-galacturonic acid polymers bound in (1 ⁇ 4). The length of these chains can range from 70 to 100 galacturonic acid residues in lemon, sugar beet or apple, that is to say having molecular weights of about 12 to 20 kilodalton (kDa). .
- the carboxylic function of ⁇ -D-galacturonic acids may be in acid form, or ionized by different cations including calcium, or esterified with methanol.
- the galacturonic acids can be acetylated at 0-2 and / or O-3.
- the pectins are characterized by a degree of methylation (DM) and a degree of acetylation (DAc) which correspond to the ratio esterified galacturonic acids (respectively methylated or acetylated) on the total galacturonic acids.
- HM highly methylated pectins
- the degree of esterification of pectins has an impact on the flexibility of the molecule: the lower the degree of esterification, the more rigid the pectin. He also has a strong impact on their gelation properties.
- the carboxylic acid function of ⁇ -D-galacturonic acids may, during an industrial demethylation treatment in an ammoniacal medium, be amidated.
- the pectin is amidated and has a degree of DA amidation.
- said envelope and the core comprising the microcapsules are amidated pectin having a degree of amidation (DA) ranging from 3 to 30%, preferably from 20 to 30% and typically of the order of 24%.
- DA degree of amidation
- the degrees of amidation (DA) and esterification (DE) are determined by the titration method (Food Chemical Codex, 1981) (Washington, DC: National Academy). of Sciences.
- the microcapsules according to the invention have a mean diameter ranging from 100 ⁇ to 5,000 ⁇ , preferably ranging from 250 ⁇ to 1,000 ⁇ and in particular ranging from 400 ⁇ to 800 ⁇ .
- the microcapsules are in dehydrated form (for example freeze-dried), or frozen, so as to optimize their preservation.
- the present invention also relates to a probiotic formulation, in particular for animals, characterized in that it comprises at least the microcapsules as described above.
- the probiotic formulation may be in various galenical forms, preferably for oral administration, such as a capsule, a soft capsule, a tablet, a beverage, an ampoule, a powder or any other ingestible dosage form. by a host or allowing the preparation of drinks or nutritional dishes.
- the drinks or nutritional dishes may be fermented milk, an ice dairy product, a cereal bar, a non-alcoholic beverage, dietary supplements, a nutritional supplement, croquettes and treats for pets, etc.
- the present invention also relates to a method of manufacturing microcapsules as described above, comprising the following steps:
- microcapsules comprising: a pectin shell and a pectin network core, wherein the probiotics are immobilized;
- step (iii) culturing the microcapsules obtained at the end of step (ii) in a culture medium, so as to form in situ within the microcapsules biofilms distributed in distinct clusters from the immobilized probiotics;
- step (iv) optionally, the dehydration (such as lyophilization) or the freezing of the microcapsules obtained at the end of step (iii).
- a solution or an oil / water pectin emulsion is prepared on the one hand and a suspension of probiotics on the other hand; then the two preparations are mixed until homogenization and the homogeneous mixture is obtained.
- the pectin may also be amidated and has a degree of amidation (DA) ranging from 3 to 30%, preferably from 20 to 30% and typically of the order of 24%.
- the homogeneous mixture comprises a pectin content, by weight, relative to the total volume of the mixture, ranging from 2% (w / v) to 10% (w / v), preferably 3% (m / v) at 8% (w / v) and typically 3.5% (w / v) to 5% (w / v).
- the homogeneous mixture of step (i) comprises a probiotic concentration ranging from 10 5 CFU / mL to 10 9 CFU / mL, preferably from 10 6 CFU / mL to 10 8 CFU / mL. and typically of the order of 10 7 CFU / mL.
- step (i) it is possible to add to the homogeneous mixture of step (i), another non-probiotic active substance, such as polyphenol, vitamins, minerals, a prebiotic, or a mixture thereof.
- this other active substance represents, by weight, relative to the total mass of the homogeneous mixture of from 0.1 to 30%, preferably from 0.1 to 20% and typically from 5% to 15%.
- this encapsulation step (ii) comprises a first injection sub-step (iia) by means making it possible to drop the homogeneous mixture of step (i) into the stirred crosslinking solution, so that each drop forms a microcapsule when it comes into contact with the crosslinking solution.
- this injection step (iia) can be performed by means of a hand shower.
- the cutting of the drops is done by gravity and allows in particular to obtain microcapsules having a mean diameter ranging from 1000 to 5000 ⁇ .
- This step (iia) can also be performed by means of an encapsulator.
- the cutting of the drops may be, according to one embodiment, be performed by applying an electric field.
- the applied electric field ranging from 0.1 kv to 10 kv, preferably from 2 kv to 10 kv and typically from 5 kv to 8 kv. This technique generally makes it possible to obtain microcapsules having a mean diameter ranging from 400 to 1000 ⁇ .
- the cutting of the drops can be performed by vibration. This technique generally makes it possible to obtain microcapsules having a mean diameter ranging from 100 to 1000 ⁇ .
- the drip rate generally varies from 20 ⁇ s to 300 ⁇ -ds, preferably ranges from 40 ⁇ s to 100 ⁇ s, and typically ranges from 40 ⁇ s to 80 ⁇ s. Us; while the injection height of the homogeneous mixture of step (i) in the crosslinking solution varies from 1 cm to 15 cm, preferably ranges from 2 cm to 10 cm and typically ranges from 2 cm to 6 cm.
- the crosslinking solution is composed of divalent cation, such as Ca 2+ , Zn 2+ , Ba 2+ , Fe 2+ , for example in the form of chloride, sulphide or acetate, or is composed of one of their mixtures.
- the crosslinking solution is composed of Ca 2+ ions in chloride form.
- the divalent ion concentration of the crosslinking solution ranges from 25 to 750 mM, preferably from 200 to 750 mM, and in particular from 500 to 750 mM.
- the encapsulation step (ii) comprises in particular a second sub-step called crosslinking microcapsules (iib).
- This second substep comprises the maturation of the microcapsules obtained during the injection sub-step (iia) for a period of at least 8 minutes, preferably ranging from 0 to 60 minutes, and in particular ranging from at 25 minutes.
- this substep of crosslinking (iib) is carried out at a temperature below 40 ° C, preferably below 30 ° C and in particular ranging from 4 ° C to 25 ° C.
- the microcapsules thus comprise a pectin envelope and a heart formed of a pectin network and in which the probiotics are immobilized.
- the microcapsules obtained according to the process of the invention have a mean diameter ranging from 100 ⁇ to 5000 ⁇ , preferably ranging from 250 ⁇ to 1,000 ⁇ and in particular ranging from 400 ⁇ to 800 ⁇ .
- step (iii) of culturing the microcapsules obtained at the end of step (ii) are preferably recovered, generally by gravitational sedimentation, and then rinsed, generally with distilled water. Then, the step of culturing the microcapsules (iii) in a culture medium, so as to form in situ within the microcapsules biofilms in separate clusters from immobilized probiotics.
- the culture medium during this culturing step (iii) is chosen from: MRS (deMan, Rogosa, Sharpe), AOAC (Association of Official Analytical Chemists), LB (Lysogeny Broth), TSB (Tryptic Soy Broth), TPY (Tryptone Phytone Yeast), BSM (Bifidus Selective Medium), Enterococcosel Broth (Bile Esculin Azide Broth), Nutrient Broth # 4, CA SO Broth (Casein Peptone Soybean), AC Broth (AH Culture Broth) ), Reinforced Clostridial Medium, BHI (Brain Heart Infusion), Bifidobacterium Broth, Tomato Juice Broth, or a mixture thereof, and is preferably selected from: MRS (deMan, Rogosa, Sharpe), AOAC (Association of Official Analytical Chemists) ).
- the pH of the culture medium varies from 3 to 8, preferably from 4 to 7 and typically from 5 to 7.
- the temperature of the culture medium varies, for example, from 20 to 40 ° C, preferably from 25 ° C to 37 ° C and generally from 25 to 30 ° C.
- This step is generally carried out for a duration greater than or equal to 12 hours, preferably ranging from 12 to 72 hours and generally from 20 to 48 hours.
- microcapsules obtained at the end of step (iii) are optionally frozen or dehydrated.
- This freeze-drying step comprises, for example, the freezing of the microcapsules to about -80 ° C., for example in 10 mL vials, ie about 1 g of samples per vial, with a ramp of 8 ° C./min. .
- cryo-protectants glycerol type, sugars, antioxidants, etc.
- the samples are then lyophilized, for example 20 hours at -55 ° C. with a pressure of 0.05 mbar, applying 5 steps (-40, 10, 0, 20 and 30 ° C.). After lyophilization, the samples are stored for example at room temperature.
- the freezing step is carried out by freezing the microcapsules, for example in 10 mL vials, ie about 1 g of samples per vial.
- cryo-protectants glycerol type, sugars, antioxidants, etc.
- the freezing temperatures are for example between -20 ° C and -80 ° C, and the temperature decrease is for example with a ramp of 8 ° C / min.
- the present invention relates to the use of the microcapsules as described above, or the abovementioned probiotic formulation comprising said microcapsules, or microcapsules obtained according to the aforementioned method, for protecting the probiotics during passage in the stomach and delivering them in an active form to the gut in an animal.
- the present invention relates to microcapsules described above, or the abovementioned probiotic formulation or microcapsules obtained according to the process described above, for its use as a medicament.
- the invention also relates to the microcapsules described above, or the aforementioned probiotic formulation or microcapsules obtained according to the method described above, for use in animals to:
- a Nisco Var 1 type encapsulator was used (parameters: 8 kV, tip 0.7 mm) with a syringe pump (parameters: flow 98.4 ⁇ / h, volume 0.20 mL).
- the homogeneous mixture obtained above is introduced into a 20 ml syringe before being injected into the encapsulator using the syringe pump.
- the 8 kV electric field will force the formation of microdroplets that will fall into a stirred CaCl 2 crosslinking bath, each of which will form a microcapsule.
- the average diameter of the wet microcapsules thus produced is of the order of 1000 ⁇ .
- the gelation is carried out under a sterile atmosphere for 20 minutes by contact of the pectin solution with the sterile solution of CaCl 2 at 0.75 M.
- microcapsules comprising: a pectin shell and a heart forming a pectin network, in which the probiotics are immobilized.
- MRS medium pH 5.8 (Conda) or AOAC medium pH 6.8 (Difco).
- the pectin microcapsules containing the bacterial biofilms are washed three times in ultra pure water (15 mL).
- the bacteria are liberated from the pectin gels.
- the pectin gels (1 batch of microcapsules of 4 mL of pectin) are broken up using a buffer solution (50 mL) of 0.1 M sodium citrate, which allows to release and suspend in solution the bacteria.
- the re-suspended bacteria are then counted according to two methods: the method of the units forming the colonies or flow cytometry. * Colonies Forms Method
- the solution of re-suspended bacteria is diluted in a cascade of 10 "1 to 10 " 5 with physiological saline and 3 drops of 10 ⁇ are deposited at 10 "5 to 10 ° dilutions on an MRS medium agar at pH 5, 8.
- the petri dish is then incubated for 48 hours at 28 ° C. This method makes it possible to measure the bacterial cultivability.
- the solution of re-suspended bacteria is first washed: centrifugation of 1 ml at 10 000 g and recovery of the bacterial cells in 1 ml of 1 ⁇ filtered PBS. Then this bacterial suspension is diluted to be from 10 5/10 6 bacteria / mL. Finally, the bacteria are labeled with cFDA (carboxy-Fluorescein DiAcetate) and with ⁇ (Propidium lodide). The labeled bacterial solutions are then analyzed by flow cytometry (BD Acuri, C6). The cFDA labeling makes it possible to measure the bacterial viability while the marquage labeling makes it possible to measure the membrane integrity.
- cFDA carboxy-Fluorescein DiAcetate
- ⁇ Propidium lodide
- the protocol consists in introducing a solution of pectin at 4% (m / v) in a petri dish before covering it with a dialysis membrane (Thermofisher, Snakeskin Dialysis Tubing, 10 kDa) .
- the pectin solution is obtained by dissolving powdered pectin, sterile in autoclaved ultra-pure water and in a sterile atmosphere so as to obtain a pectin content of 4% (w / v).
- This box is then introduced upside down (membrane down) for 20 minutes in a bath of CaCl 2 .
- the calcium ions will diffuse via the dialysis membrane into the pectin solution to allow gelation of the pectin.
- the gel is then dried to form a film.
- the Applicant also tested under the same experimental conditions and successfully another strain Lactobacillus rhamnosus GG in the MRS culture medium for 24 hours. Indeed, biofilms were formed on a flat film of pectin, as well as inside the microcapsules of pectin.
- Drying tests were conducted using freeze-drying, a method widely used in the probiotic and ferment industry, but which has high mortality rates.
- freeze-drying protocol is as follows:
- the samples are then lyophilized for 20 hours at -55 ° C. with a pressure of 0.05 mbar, applying 5 steps (-40, 10, 0, 20 and 30 ° C.);
- the counts were carried out on agar culture medium and in flow cytometry.
- the biomass obtained is 9.7 Log CFU / g (Cheow, Kiew, and Hadinoto 2014) and 9.38 Log CFU / g for Bifidobacterium bifidum in alginate microspheres (Châvarri et al., 2010).
- results without optimization of the survival of the bacteria after freeze-drying for the "24 h biofilm AOAC" condition are 10.24 log CFU / g and 9.86 for the "24 hr biofilm in MRS" condition.
- EXAMPLE 3 ACID STRESS RESISTANCE TEST OF THE MICROCAPSULES OF EXAMPLE 1 IN COMPARISON WITH PLANCTONIC PROBIOTIC BACTERIA
- planktonic bacteria or microcapsules (0.8 g) were introduced in a known quantity, ie 8.5 log of CFU, in 5 ml of gastric solution at 37 ° C. After 2 hours of incubation, the planktonic bacteria, the biofilm bacteria in the pectin microcapsules and the bacteria re-released in the medium were counted on MRS agar according to the UFC method (previously described). Beforehand, the microcapsules were recovered and disintegrated in 10 ml of sodium citrate (0.1 M) in order to suspend the bacteria in solution.
- probiotic biofilm bacteria in the pectin microcapsules according to the invention have a relatively low mortality compared to planktonic bacteria namely on average 1, 15 log CFU loss.
- EXAMPLE 4 TEST OF THE ADHESION PROPERTIES OF THE MICROCAPSULES OF EXAMPLE 1 IN COMPARISON WITH PLANCTONIC PROBIOTIC BACTERIA Test 1
- Lactobacillus casei bacteria was studied in vitro in a model of intestinal epithelial cells of the Caco-2 line.
- the cells are cultured in microplate (24 wells) at the concentration of 10 5 cells per well and maintained for 15 days to obtain a differentiated cell mat (with a brush border).
- the biofilm Lactobacillus casei bacteria released from the pectin microcapsules according to the invention are brought into contact with the epithelial cells for 1 h 30 at 37 ° C., at a known concentration (100 times more bacteria than epithelial cells (ME). 100), or 10 times more bacteria than epithelial cells (MOI 10)). After this incubation time, the epithelial cells are washed and the bacteria adhered to the epithelial cells are counted on MRS agar plates.
- probiotic bacteria of Lactobacillus casei in planktonic form originated from a 1% revitalized culture for 24 h in MRS culture medium at pH 5.8 from a 24 hour cryotube output culture in MRS pH 5.8. These bacteria are contacted with epithelial cells as previously described.
- the adhesion rate of planktonic bacteria Lactobacillus casei is similar to that already described in the literature namely 0.68% at MOI 100.
- the adhesion rate increased considerably, namely to 20%.
- the biofilm bacteria in the pectin microcapsules thus showed a similar adhesion capacity to bacteria in planktonic form.
- the freeze-drying of biofilm bacteria in pectin microcapsules has increased their adhesion, and initial results suggest that pectin plays a role in promoting adhesion to the intestinal mucosa.
- planktonic bacteria have an adhesion capacity to epithelial cells increased by pectin. Indeed planktonic bacteria without pectin have an average adhesion rate of 3.32% and in contact with pectin (4 mg) this rate increases to 44.22%.
- EXAMPLE 5 IN VIVO TEST WITH DSS (DEXTRAN SODIUM SULFATE)
- mice Male C57BL mice from Charles River Laboratories were divided into two groups:
- mice received 2% (w / v) DSS (Dextran Sodium Sulfate) treatment in drinking water from day 5 to day 11 of the experiment. DSS caused inflammation in the intestines of the mice.
- DSS Extran Sodium Sulfate
- mice of each group were weighed to observe the evolution of the weight of the mice as a function of time ( Figure 4).
- mice were recovered in order to quantify the bacteria of the Lactobacillus genus on the one hand ( Figure 5) and the L. casei species on the other hand ( Figure 6).
- 140 mg of faeces from each group of mice were diluted in physiological saline, and disintegrated with stirring with glass beads, giving a liquid solution (in triplicate).
- the solution containing the bacteria was counted according to the method of colony-forming units: the solution of re- suspended bacteria was diluted in cascade io th with physiological water and dilutions were plated on MRS agar medium at pH 6 containing 20 mg / ml of vancomycin (medium allowing the growth of bacteria of the genus Lactobacillus) and on an M-RTLV medium agar at pH 6 (medium allowing the growth of bacteria of the L. casei species). The agar plates were then incubated for 48 h at 37 ° C. This method measures bacterial cultivability. Result
- mice which have received the pectin microparticles comprising the probiotic bacteria in biofilm form according to the invention have an improved state of health compared to the other mice which have received physiological saline.
- the DSS caused in the different groups of mice tested intestinal inflammation (diarrhea, weight loss, etc.).
- the weight loss was lower in the group having received the formulation according to the invention as this is illustrated in Figure 4.
- microcapsules according to the invention allows the vectorization and release of probiotics in viable form and allows to retain the functionality of probiotic bacteria.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Veterinary Medicine (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Organic Chemistry (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Botany (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Zoology (AREA)
- Nutrition Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Inorganic Chemistry (AREA)
- Dispersion Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Medicinal Preparation (AREA)
Abstract
Description
Claims
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
FR1753248A FR3065162B1 (fr) | 2017-04-13 | 2017-04-13 | Microcapsules de pectine, son procede de fabrication et ses utilisations |
PCT/FR2018/050927 WO2018189490A1 (fr) | 2017-04-13 | 2018-04-12 | Microcapsules de pectine, son procédé de fabrication et ses utilisations |
Publications (1)
Publication Number | Publication Date |
---|---|
EP3609477A1 true EP3609477A1 (fr) | 2020-02-19 |
Family
ID=59297029
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
EP18719973.2A Pending EP3609477A1 (fr) | 2017-04-13 | 2018-04-12 | Microcapsules de pectine, son procédé de fabrication et ses utilisations |
Country Status (4)
Country | Link |
---|---|
US (1) | US20200155470A1 (fr) |
EP (1) | EP3609477A1 (fr) |
FR (1) | FR3065162B1 (fr) |
WO (1) | WO2018189490A1 (fr) |
Families Citing this family (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2020508696A (ja) | 2017-01-31 | 2020-03-26 | カンザス ステート ユニバーシティー リサーチ ファウンデーション | 微生物細胞、それを生成する方法、およびその使用 |
EP3697220A4 (fr) | 2017-10-20 | 2021-08-11 | MS Biotech, Inc. | Procédés de production de matieres végétales ensilées à l'aide de megasphaera elsdenii |
WO2023117364A1 (fr) * | 2021-12-01 | 2023-06-29 | Vilnius University | Microcapsule et procédés de fabrication et d'utilisation de celle-ci |
CN115193350B (zh) * | 2022-07-18 | 2023-07-14 | 齐鲁工业大学 | 乳杆菌在低pH值果汁中微胶囊化包封方法 |
TW202416850A (zh) * | 2022-09-01 | 2024-05-01 | 紐西蘭商恆天然合作集團有限公司 | 包含生物活性劑的顆粒 |
CN116508994A (zh) * | 2023-05-06 | 2023-08-01 | 浙江大学 | 一种益生菌rg-i果胶微胶囊及其制备方法 |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
BE1019142A3 (fr) * | 2011-01-21 | 2012-03-06 | Vesale Pharma S A | Substance probiotique microencapsulee. |
CN105919970A (zh) * | 2016-06-02 | 2016-09-07 | 天津欣益源科技发展有限公司 | 一种用于提高益生菌生物活性的包埋微胶囊 |
-
2017
- 2017-04-13 FR FR1753248A patent/FR3065162B1/fr active Active
-
2018
- 2018-04-12 WO PCT/FR2018/050927 patent/WO2018189490A1/fr unknown
- 2018-04-12 US US16/604,591 patent/US20200155470A1/en not_active Abandoned
- 2018-04-12 EP EP18719973.2A patent/EP3609477A1/fr active Pending
Also Published As
Publication number | Publication date |
---|---|
FR3065162B1 (fr) | 2019-06-28 |
WO2018189490A1 (fr) | 2018-10-18 |
US20200155470A1 (en) | 2020-05-21 |
FR3065162A1 (fr) | 2018-10-19 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
FR3065162B1 (fr) | Microcapsules de pectine, son procede de fabrication et ses utilisations | |
Silva et al. | Symbiotic microencapsulation to enhance Lactobacillus acidophilus survival | |
Dafe et al. | Development of novel carboxymethyl cellulose/k-carrageenan blends as an enteric delivery vehicle for probiotic bacteria | |
KR101918089B1 (ko) | 장내 생존율이 증대된 유산균의 코팅 방법 | |
US10117884B2 (en) | Processing of natural polysaccharides by selected non-pathogenic microorganisms and methods of making and using the same | |
Islam et al. | Microencapsulation of live probiotic bacteria | |
Anal et al. | Recent advances in microencapsulation of probiotics for industrial applications and targeted delivery | |
Barajas-Álvarez et al. | Microencapsulation of Lactobacillus rhamnosus HN001 by spray drying and its evaluation under gastrointestinal and storage conditions | |
CA2825473C (fr) | Protection de cellules microbiennes contre la degradation acide | |
Haghshenas et al. | Effect of addition of inulin and fenugreek on the survival of microencapsulated Enterococcus durans 39C in alginate-psyllium polymeric blends in simulated digestive system and yogurt | |
Khorasani et al. | Starch-and carboxymethylcellulose-coated bacterial nanocellulose-pectin bionanocomposite as novel protective prebiotic matrices | |
Chaikham et al. | Influence of encapsulated probiotics combined with pressurized longan juice on colon microflora and their metabolic activities on the exposure to simulated dynamic gastrointestinal tract | |
BE1024197B1 (fr) | Procédé d'enrobage de microorganismes, poudre desdits microorganismes enrobés obtenue et composition pharmaceutique, nutraceutique, cosmétique, alimentaire ou sanitaire la comprenant. | |
Torp et al. | Optimizing oral delivery of next generation probiotics | |
CHACKOSHIAN et al. | Improvement of probiotic survival in fruit juice and under gastrointestinal conditions using pectin-nanochitin-nanolignocellulose as a novel prebiotic gastrointestinal-resistant matrix | |
He et al. | Encapsulation of Lactobacillus in low-methoxyl pectin-based microcapsules stimulates biofilm formation: enhanced resistances to heat shock and simulated gastrointestinal digestion | |
Mudgil et al. | Fortification of Chami (traditional soft cheese) with probiotic-loaded protein and starch microparticles: Characterization, bioactive properties, and storage stability | |
Chotiko et al. | Three protective agents for pectin-rice bran capsules for encapsulating Lactobacillus plantarum | |
Lee et al. | Effect of pectic oligosaccharide on probiotic survival and physicochemical properties of hydrogel beads for synbiotic encapsulation of Lactobacillus bulgaricus | |
Srivastava et al. | Enhanced encapsulation efficiency and controlled release of co-encapsulated Bacillus coagulans spores and vitamin B9 in gellan/κ-carrageenan/chitosan tri-composite hydrogel | |
Soto et al. | Recent developments on wall materials for the microencapsulation of probiotics: a review | |
Chen et al. | Enhance the resistance of probiotics by microencapsulation and biofilm construction based on rhamnogalacturonan I rich pectin | |
Srisuk et al. | Characteristics co-encapsulation of Lactobacillus acidophilus with Dictyophora indusiata. | |
Evivie | Preliminary studies on pharmaceutical microencapsulation for synbiotic application | |
Sekhavatizadeh et al. | Evaluation of Physicochemical Properties of Lactobacillus acidophilus ATCC 4356 Cells Encapsulated with Sodium Alginate and Balangu (Lallemantia royleana) Seed Mucilage |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: UNKNOWN |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: THE INTERNATIONAL PUBLICATION HAS BEEN MADE |
|
PUAI | Public reference made under article 153(3) epc to a published international application that has entered the european phase |
Free format text: ORIGINAL CODE: 0009012 |
|
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: REQUEST FOR EXAMINATION WAS MADE |
|
17P | Request for examination filed |
Effective date: 20191010 |
|
AK | Designated contracting states |
Kind code of ref document: A1 Designated state(s): AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR |
|
AX | Request for extension of the european patent |
Extension state: BA ME |
|
DAV | Request for validation of the european patent (deleted) | ||
DAX | Request for extension of the european patent (deleted) | ||
STAA | Information on the status of an ep patent application or granted ep patent |
Free format text: STATUS: EXAMINATION IS IN PROGRESS |
|
17Q | First examination report despatched |
Effective date: 20220811 |