Probiotic in Diabetes Treatment: - Effective Identification of Probiotic Bacteria Isolated from Regional Biota

Tariful IslamSenior Officer

Biotechnology Derived Product FacilityIncepta Pharmaceuticals Ltd

1st International Conference DRUG DISCOERY & DEVELOPMENT RESEARCH IN DEVELOPING COUNTRIES (DRDC)

4th October,2015 1

Elie Metchnikoff (1885 – 1916)

• First described by Metchnikoff in 1908 as “Live microbial food ingredients that alter the microflora and confer health benefit”

• “Live microorganisms which when administered in adequate amounts confer a health benefit on the host” (FAO/WHO)

Probiotics

Probiotics Research Trends

1761

91

191

453432

564

307

271

127

421

294

1997

1998

1999

2000

2001

2002

2003

2004

2005

2006

2007

2008

Number of Published Clinical Studies on Probiotic

Source: www.ncbi.nlm.nih.gov/pubmed 3

Research Publications on Probiotic

Gut functionAcute diarrheaTravelers diarrhea

AllergyAtopic dermatitisAsthma

Oral microbiologyDental caries

Colds, respiratory infections

Skin microbiology, inflammation

CNS

Metabolic syndromeDiabetes Obesity

Probiotics - Biological Swiss Army Knife

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Classification of Probiotic Bacteria

Probiotic Bacteria

Lactobacillus Bifidobacterium Enterococcus

L. casei group L. acidophilus group

L. reuteri/ L. fermentum group

L. caseiL. paracasei

L. zeaeL. rahmnosus

5Georgieva et al. (2014)

~ 200-230bp ~ 198-210bp

~1500bp ~ 2900bp ~ 100bp

16S 23S 5S

Identification of Lactobacillus paracasei

rRNA Intergenic Spacer Regions

6

M

Genomic DNA of L. paracasei

7

Identification of Lactobacillus paracasei

M M

8

Lp1

Identification of Lactobacillus paracasei

Sequencing of 16S-23S ISR rRNA Amplified Region of L. paracasei

9

Environmental Factors HFD Stress Life Style

Altered Microbiota

Increased Intestinal

Permeability

LPS

Activation of TLR-4

Cytokine Release

Interruption in Insulin

receptor’s activity

T1D/T2D

Destruction of pancreatic β- cell

Diabetes and Gut Microbiota

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Insulin Resistance in Presence of LPS

Golgi

(signal)

-P -

Obstacle in glucose

transport

Glucose

Insulin receptor

Insulin

P

Insulin binding and signal transduction

GLUT4

LPS binding with TLR-4

T2D

11

Probiotic Treatment for T2D

12Gomes et al. (2014)

GLP-1

Insulin secretion

L-cell

Butyrate

Butyrate Producing Bacteria (Probiotics)

Treatment for T1D

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Potential probiotic bacteria

Identification at molecular

level

Selection of identified

probiotic bacteria for diabetes treatment

In-vitro and In-vivo test for

effectiveness

Fermentation (R&D Scale)

Cell separation Lyophilization (R&D scale)

Formulation & Clinical trial

Final Product

Development Pathway of Probiotic Formulation for Diabetes Treatment

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Importance of Microbiota

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State of the art facility of Incepta Pharmaceuticals Ltd

Our Strength

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Bio-safety Cabinet class II,

PCR machine, Applied Biosystems, USA/Germany.

Gel Electrophoresis apparatus, BioRad, USA.

Gel Documentation System, Cell Biosystem, USA

Fermenter (Bioreactor), New Brunswick, Eppendorf, USA

Lyophilizer (Vertis, USA)

Animal House

Our Strength

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•Ahmed FE. Genetically modified probiotics in foods. Trends Biotechnol 2003;21:491–497•Alipio Z, Liao W, Roemer EJ, et al. Reversal of hyperglycemia in diabetic mouse models using induced-pluripotent stem (iPS)-derived pancreatic beta-like cells. Proc Natl Acad Sci U S A 2010;107:13426–13431 •Daniel C, Repa A, Wild C, et al. Modulation of allergic immune responses by mucosal application of recombinant lactic acid bacteria producing the major birch pollen allergen Bet v 1. Allergy 2006;61:812–819 •Ferber S, Halkin A, Cohen H, et al. Pancreatic and duodenal homeobox gene 1 induces expression of insulin genes in liver and ameliorates streptozotocin induced hyperglycemia. Nat Med 2000;6:568–572 •Georgieva . M., Andonova . L., Peikova L., Zlatkov. Al., Probiotics – health benefits, classification, quality assurance and quality control – review, PHARMACIA, vol. 61, No. 4/2014•Gomes AC, Bueno AA, Souza RGM, and Mota JF, Gut microbiota, probiotics and diabetes, Nutrition Journal, 2014, 13:60•Honi U, Sabrin F, Islam T, et al. Enzymatic activity and antibiotic resistance profile of Lactobacillus paracasei ssp. paracasei-1 isolated from regional yogurts of Bangladesh, J. Microbiol. Biotechnol. Food Sci., 2013, 3, 3, 235-239 •Islam T, Khan MI, Hossain MS, et al. Effective differentiation of Lactobacillus paracasei from Lactobacillus casei group, Open Life Sci. 2015 (Received) •Islam T, Sabrin F, Islam ME, et al. Analysis of antimicrobial activity of Lactobacillus paracasei ssp. paracasei-1 isolated from regional yogurt, Int. Res. J. Appl. Life Sci., 2012, 1, 4, 66-72 •Kuehn MJ. Genetically engineered probiotic competition. Gastroenterology 2006;130:1915–1916 •Noguchi H. Stem cells for the treatment of diabetes. Endocr J 2007;54:7–16 •Soria B, Roche E, Berná G, León-Quinto T, Reig JA, Martín F. Insulin-secreting cells derived from embryonic stem cells normalize glycemia in streptozotocin- induced diabetic mice. Diabetes 2000;49:157–162 •Suzuki A, Nakauchi H, Taniguchi H. Glucagon-like peptide 1 (1-37) converts intestinal epithelial cells into insulin-producing cells. Proc Natl Acad Sci U S A 2003;100:5034–5039 •Yadav H, Lee JH, Lloyd J, et al. Beneficial metabolic effects of a probiotic via butyrate induced GLP-1 secretion. J Biol Chem 2013 •Zalzman M, Gupta S, Giri RK, et al. Reversal of hyperglycemia in mice by using human expandable insulin-producing cells differentiated from fetal liver progenitor cells. Proc Natl Acad Sci U S A 2003;100:7253–7258

References

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Thank You

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