This document discusses probiotics and their potential role in treating diabetes. It begins by providing background on probiotics and how their use has increased in clinical studies over time. Probiotics have been shown to impact gut function, immunity, oral health and metabolic conditions like diabetes and obesity. The document then discusses how gut microbiota alterations from factors like diet and stress can increase intestinal permeability, activate the immune system and interrupt insulin activity, leading to diabetes. It proposes that probiotic treatment may help by producing substances like butyrate and GLP-1 that aid insulin secretion. The remainder outlines a process for developing a probiotic formulation to treat diabetes, from identifying potential probiotic bacteria to clinical trials. It emphasizes the importance of the gut

1. Probiotic in Diabetes Treatment: - Effective Identification of
Probiotic Bacteria Isolated from Regional Biota
Tariful Islam
Senior Officer
Biotechnology Derived Product Facility
Incepta Pharmaceuticals Ltd
1st
International Conference
DRUG DISCOERY & DEVELOPMENT RESEARCH IN DEVELOPING COUNTRIES (DRDC)
4th
October,2015 1

2. 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

3. Probiotics Research Trends
17
61
91
191
453
432
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

4. Gut function
Acute diarrhea
Travelers diarrhea
Allergy
Atopic dermatitis
Asthma
Oral microbiology
Dental caries
Colds, respiratory infections
Skin microbiology,
inflammation
CNS
Metabolic syndrome
Diabetes
Obesity
Probiotics - Biological Swiss Army Knife
4

5. Classification of Probiotic Bacteria
Probiotic BacteriaProbiotic Bacteria
LactobacillusLactobacillus BifidobacteriumBifidobacterium EnterococcusEnterococcus
L. casei groupL. casei group
L. acidophilus
group
L. acidophilus
group
L. reuteri/ L. fermentum
group
L. reuteri/ L. fermentum
group
L. casei
L. paracasei
L. zeae
L. rahmnosus
L. casei
L. paracasei
L. zeae
L. rahmnosus
5Georgieva et al. (2014)

6. ~ 200-230bp ~ 198-210bp
~1500bp ~ 2900bp ~ 100bp
16S16S 23S23S 5S5S
Identification of Lactobacillus paracasei
rRNA Intergenic Spacer Regions
6

7. M
Genomic DNA of L. paracasei
7
Identification of Lactobacillus paracasei

8. M M
8
Lp1
Identification of Lactobacillus paracasei

9. Sequencing of 16S-23S ISR rRNA Amplified Region of L. paracasei
9

10. Environmental
Factors
Environmental
Factors HFDHFD StressStress Life StyleLife Style
Altered
Microbiota
Altered
Microbiota
Increased
Intestinal
Permeability
Increased
Intestinal
Permeability
LPSLPS
Activation
of TLR-4
Activation
of TLR-4
Cytokine
Release
Cytokine
Release
Interruption in
Insulin
receptor’s
activity
Interruption in
Insulin
receptor’s
activity
T1D/T2
D
T1D/T2
D
Destruction of
pancreatic β- cell
Destruction of
pancreatic β- cell
Diabetes and Gut Microbiota
10

11. Insulin Resistance in Presence of LPS
Golgi
(signal)
-
P
-
Obstacle in
glucose
transport
Obstacle in
glucose
transport
GlucoseGlucose
Insulin receptor
Insulin
P
Insulin binding and
signal transduction
Insulin binding and
signal transduction
GLUT4
LPS binding with TLR-4LPS binding with TLR-4
T2DT2D
11

12. Probiotic Treatment for T2D
12Gomes et al. (2014)

13. GLP-1GLP-1
Insulin secretion
L-cellL-cell
ButyrateButyrate
Butyrate Producing
Bacteria (Probiotics)
Treatment for T1D
13

14. Potential
probiotic
bacteria
Potential
probiotic
bacteria
Identification
at molecular
level
Identification
at molecular
level
Selection of
identified
probiotic bacteria
for diabetes
treatment
Selection of
identified
probiotic bacteria
for diabetes
treatment
In-vitro and In-
vivo test for
effectiveness
In-vitro and In-
vivo test for
effectiveness
Fermentation
(R&D Scale)
Fermentation
(R&D Scale)
Cell separationCell separation
Lyophilization
(R&D scale)
Lyophilization
(R&D scale)
Formulation &
Clinical trial
Formulation &
Clinical trial
Final
Product
Final
Product
Development Pathway of Probiotic Formulation for Diabetes Treatment
14

15. Importance of Microbiota
15

16. State of the art facility of Incepta Pharmaceuticals Ltd
Our Strength
16

17. 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
17

18. •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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19. Thank You
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