golden jubilee commemoration oration of insa
TRANSCRIPT
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Immunotherapeutic vaccines and
Microbicides for reproductive health
A unique immunotherapeutic vaccineA unique immunotherapeutic vaccine
for leprosy approved by DCGI andfor leprosy approved by DCGI and
USFDA, its new found utility inUSFDA, its new found utility in
u ercu os s an cancersu ercu os s an cancers
Two microbicides for cure andTwo microbicides for cure and
prevention of reproductive tract andprevention of reproductive tract and
sexually transmitted infectionssexually transmitted infections
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Importance of Immunity inLeprosy
Most ( 99 %) do not contract the
. .
Those who do, fall in a spectrum
between polar TT to polar LLdepending on the degree of Immune
deficit.
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Nature of defect is the inability tomount CMI response of the Th1 type
to M. leprae
eliminate M. leprae
Normalcy of reactivity to other
bacteria and Microorganisms
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No medium known for culture of M.
leprae, requires a host cell obligatorily
Developed anDeveloped an inin--vitrovitro system forsystem for
MonocyteMonocyte -- derived macrophages fromderived macrophages from
peripheral bloodperipheral blood
Krishnan and Talwar (1974). IJMR; 62, 313 316
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Slow growerSlow grower division time 13 daysdivision time 13 days
M. leprae sourceM. leprae source Human lepromasHuman lepromascontaining unknown number of live andcontaining unknown number of live and
dead bacteriadead bacteria
Devised a selective approach for incorporation
of Methyl 3H-thymidine into DNA by M. leprae
without interference of host macrophages
Talwar, Krishnan and Gupta (1974). Infection and Immunity; 9, 187 191
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Mycobacterial multiplication in cultivated
macrophages derived from peripheral blood
monocytes of Leprosy patients
Patient
no
Clinical
status
CPM 3H-thymidine incorporated per 5 x 105 Phagocytic cells
Macrophages +Lymphocytes + M. leprae
Macrophages + M. leprae
1.1. LLLL 36,45836,458 45,62845,628
2.2.3.3.
4.4.
5.5.
6.6.
LLLLLLLL
TTTT
TTTT
TTTT
53,92953,92952,35452,354
6,3326,332
3232
381381
59,59659,59683,47683,476
54,96954,969
78,44778,447
26,26026,260
Talwar, Krishnan, Jha, Verma. Biochimie (1974). 56, 231 - 237
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Nature of defect is the specific inability toNature of defect is the specific inability to
mount CMI response of the TH1 type to M.mount CMI response of the TH1 type to M.
lepraeleprae
a war, r s nan, e ra, um, earson . n. xp. mmuno . ,
195 - 203
Mehra, Talwar, Balakrishnan, Bhutani (1972). Clin. Exp. Immunol. 12,205 - 213
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An Immuno-therapeutic vaccine
Developed For Multi bacillary Leprosybased on Heterologous Approach
(14 papers in Golden Jubilee issue of Leprosy In India Oct.
1978)
Strategy adopted Search for an
immunologically cross-reactivemycobacteria eliciting immune response
from not only TT but also LL patients; in-
vitro followed by in-vivo investigations
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A non-pathogenic fast growing Mycobacterium
coded as M.w, sharing antigens with M. leprae
and M. tuberculosis discovered
Key antigens heat stable; usable as autoclaved
vaccine; Simplification of toxicology studies
Preclinical toxicologyPreclinical toxicology Phase I human safetyPhase I human safety
Phase II, Phase III efficacy to field trialsPhase II, Phase III efficacy to field trialswith approvals of Drugs Controller General ofwith approvals of Drugs Controller General of
India and Ethics committeesIndia and Ethics committees
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Approved by DCGI, India and US FDA
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Mw vaccine
Combined Immunotherapy withCombined Immunotherapy withChemotherapyChemotherapy
BENEFITSBENEFITS Faster bacterial clearanceFaster bacterial clearance
Shortens recover timeShortens recover time
Clears granulomasClears granulomas
Reactions (number and severity) reducedReactions (number and severity) reduced
Lepromin conversion to positivityLepromin conversion to positivity
Effective in slow respondersEffective in slow responders
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Patient code: CR
Initial After 5 doses
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Patient code: RPS
After 2 dosesInitial
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Patient code: BWS
Initial After 4 doses
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Patient code: CHB
Initial After 1 Year
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Patient code: RAB
Initial After 4 doses
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Patient code: SNC
Initial After 2 doses (six months)
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5
4
B1-Skin
B1-Tissue
3
2
1
0
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mmuno . n ec. s. , , -
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(a) (b)
Protection test of M.wgrown in Bio-reactor
(c) (d)
Spleen Lungs
(a) & (b) Guinea pigs challenged with M.tuberculosis H 37Rv
(c) & (d) Immunized with M.w.
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Mw
Singh IG, Mukherjee R & Talwar GP. Vaccine, 9, 10 14, 1991
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OUTCOME OF THERAPY(CAT II Tuberculosis)
Mw + MDT 48/49* 97.96%
CURED
MDT alone 21/27** 77.77%
* Defaulter for 6 doses, sputum negative after intensive phase
** 2 2+ No effect of therapy
4 1+ No effect of therapy
P=0.007
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RELAPSE RATE
( 24 MTHS AFTER COMPLETION)
MDT + Mw 0/16(0.0%) 2/46(4.34%)
CATEGORY I II
MDT alone 8/33(24.24%) 11/20(55%)*
*P=0.002
9TH NORTH AMERICAN IUATALD CONFERENCE-2005
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CHRONIC TUBERCULOSIS
43.248.6
40
50
60 Mw control
00 05.4 5.4
0
10
20
30
2 6 12
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Genomic analysis of M.w. done
by a National consortium of
. ,
Tyagi and Anil Tyagi -
laboratories
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MIP ancestor of M. tuberculosis,
M. leprae and M. aviumintracellular complex (MAIC)
, ,
M.w. named as Mycobacterium
indicus pranii (MIP)Infect. Genet. Evol (2008) 8, 100-101
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Landscape of genome evolution across the generalist and specialistmycobacterial lineages
PloS ONE (2007), 10/e968, 1 8
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Ongoing therapeutic clinical trials on
Mycobacterium indicus pranii (MIP)
Phase III multicenter trials under DBT intuberculosis
Bladder Cancer(B Khamar, Ahemadabad; Pant, K G MedicalCollege, Lucknow )
Crohns disease (S. Hasnain and others)
Psoriasis (Rath & Kar, 2003. Int. J. Dermat. 42, 756-57)
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CREDITS
G. P. Talwar, A.D.Krishnan, Vijay Lakshmi
Mehra, A.S.Mustafa, Arun Fotedar,
H.K.Prasad, S.A.Zaheer, R.S.Misra, H.K.Kar,
Ashok Mukherjee, R.Walia, Pankaj Sharma,Rama Mukherjee, P.Prabhakar Reddy, Kiran
Katoch, Rajni Rani.
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Dimensions of STIs and RTIs (WHO) 340 million cases of new sexually transmitted infections occurred
in the year 1999 worldwide
New cases of various STDs each year (2006)
92 million of Chlamydia trachomatis
62 million of N. gonorrhoeae (Gonorrhea)
5.5 million of Human Papillomavirus (HPV)
5 million of Trichomoniasis
1 million of Genital Herpes
77,000 of Hepatitis B
300,000 500,000 of Candida albicans / year in the US only
300 million vaginosis / vaginitis
Over 40 million people are living with human immunodeficiency virus
(HIV) / AIDS worldwide and AIDS has assumed pandemic dimensions in
Africa & South East Asia
http://www.who.int/reproductive-health/rtis/docs/sti_factsheet
T Mi bi id
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Two Microbicides
PRANEEM Polyherbal BASANT Polyherbal1. Tablet
2. 15-30 min dispersion time
Cream / gel
Ready to use
Both have wide spectrum anti-microbial action
Inhibit N. onnorrhoeae includin Dru s resistant
isolates); E.coli, Staph. aureus Candida albicans (including Flucanozole resistant
strain) C. tropicalis, C. krusei, C. glabrata
Herpes simplex-2; Chlamydia trachomatis
Anti-HPV-16 High virucidal action on HIV-1
Talwar et al. Am Jour Reprod Immunology, 43: 144-151; 2000
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Praneem: Purified extracts of Neem leaves (Azadirachta
indica); Saponins of Sapindus mukerosii; Menthacitrata oil
BASANT: Curcumin (Diferuloyl methane); purified extractof Amla (Emblica officinalis); Saponins of S.
mukerossi; Aloe vera; Rose water
All ingredients Quality controlled
Tablets & Cream made under GMP conditions for clinical trials
In these polyherbal formulations combination ofingredients extends the range
Synergistic action, enhancing efficacy
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Lack of action of Amla on various Candidas and
Enhancement of Anti-Candida action of Saponins
and Curcumin by Combination in total formulation
BASANT
Zone of Inhibition (mm)
Amla Saponins Curcumin BASANT
1. Candida albicans
2. Candida glabrata
3. Candida krusei
4. Candida tropicalis
NZ
NZ
NZ
NZ
10
10
10
10
12
11
NZ
10
20
22
18
19
NZ = No zone of Inhibition
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Effect of Polyherbal formulation on
prevention of Chlamydia vaginal infection
PreparationPreparation # Positive on day 4/# Positive on day 4/
# Inoculated# Inoculated
# Positive on day 8 /# Positive on day 8 /
# Inoculated# Inoculated
PBS control
Polyherbal Pessary
PBS control
11/12 11/12
02/12
10/12
02/12
11/12
Investigations by K. Whalley, Johns Hopkins university
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Effect on disease causation and Herpes
Simplex virus-2 (HSV-2) multiplication
PreparationPreparation # Shedding/# Shedding/
# Inoculated# Inoculated
# with lesions/# with lesions/
# Inoculated# Inoculated
PBS control
Polyherbal Cream
PBS control
8/8 8/8
0/6
6/6
0/6
6/6
Investigations by K. Whalley, Johns Hopkins university
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Phase II Efficacy Trials in women suffering
from abnormal vaginal discharge due to RTIs
6 centers of the Indian Council
of Medical Research at :-
Postgraduate Institute Medical
Full or Partial Relief of symptoms
in 124 women [96.8 %],
No Relief in 4 women [3.1%]
,
Chandigarh; AIIMS, New Delhi;
Postgraduate Institute Kolkata;
KEM Mumbai;
KEM, Pune;
National Institute for Researchin Reproductive Health,Mumbai.
Subjects investigated = 128
x en o re e o serve y
Microbiology assays
Trichomonas 8/8 100%
vaginalis
Candida 12/13 92.3%
Bacterial
vaginosis by 24/33 72.7%
Nugent score
f
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Acceptability by both partners of
Praneem Polyherbal vaginal Tablet forpre-intercourse use (NARI trials)
20 women & their partners
,
90% participants 80% & higher acceptability score
95% liked the smell & reported that the product was
easy to use
Joglekar NS et al. Ind. J. Med. Res. 123, 547-552, 2006
Phase II comparative safety and efficacy
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Phase II comparative safety and efficacy
study of Praneem polyherbal tablet and
Betadine in symptoms of chronic abnormal
vaginal discharge in women employing WHO
syndromic approachTreatment Patients treated Cured after 2 weeks
Praneem
Betadine
50
49
46 / 50 (92%)
40 / 49 (81.6%)
Salhan et al (2007) J. Obst. Gynaecol. Res.
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A PROSPECTIVE, COMPARATIVE,
RANDOMIZED, MULTICENTRIC STUDY TO
COMPARE THE EFFICACY AND SAFETY OF
PRANEEM POLYHERBAL VAGINAL TABLET
SYMPTOMS OF ABNORMAL VAGINAL
DISCHARGE
PHASE-III TRIAL in progress in 7 centers
Licensed to Panacea Biotech
I hibiti f HIV NL4 3 li ti b
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Inhibition of HIV NL4.3 replication by
BASANTBASANT Dilution In Human CD4+ In Hela-CD4-LTR-
CEM-GFP cells gal
Percent Inhibition
1:1000 98 99
1: 10,000
1: 20,000
1: 40,000
81
51
22
64
53
32
Studies by Debashish Mitra at National Cell Science Center, Pune
INHIBITION OF HPV 16 TRANSDUCTION IN HeLa
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Inhibition of HPV16 Transduction
50
75
100
Aloe
Amlaki
INHIBITION OF HPV-16 TRANSDUCTION IN HeLa
CELLS
100 g/ml--Saponin
5 g/ml--Curcumin
300 g/ml2.5 - 6.0 g/ml3.9 g/mlAmlaki
nontoxic (10 mg/ml)420 - 640 g/ml520 g/mlAloe
toxic dose95% CI of IC50IC50Compound:
10 -1 10 0 10 1 10 2 10 3 10 4025
Saponin
Drug concentration (g/ml)
Studies by Dr. John Schiller at NCI / NIH
Clearance of HPV DNA in Praneem-treated Women
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M + - 1 2 3 4 5 6 7 8 9 10
Cases
~ 450bp
(trials at Institute of Cytology and Preventive Oncology of ICMR)
Controls
Before Praneem Treatment
PCR L1 Consensus
450b
PCR L1 Consensus
M + - 7 8 9 10
CasesControls
M + - 1 2 3 4 5 6 7 8 9 10
Controls
PCR L1 Consensus
~450bp
Cases
After One (1) Course of Praneem TreatmentAfter Two (2) Courses of
Praneem Treatment
Women in Placebo-treated Arm showed
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450bp
M + - 1 2 3
CasesControls
Women in Placebo-treated Arm showed
Persistent HPV infection
BeforeTreatment
Placebo GroupPlacebo Group
PCR L1 Consensus
M + - 1 2 3
CasesControls
PCR L1 Consensus
450bp
After
Treatment
Colposcopic Analysis of Regression of
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Colposcopic Analysis of Regression of
preneoplastic lesions in Women treated with
PraneemBefore Treatment After Treatment
Representative case showingabnormal transformation zone ischaracterized by acetowhiteepithelium (flat warts indicated by
arrows) on the posterior lip of thecervix with extensive ectopy.
Remission of acetowhite lesion(condylomatous changes) after
Praneem treatment
Cytological improvements in Pap Smears of
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Cytological improvements in Pap Smears of
Women treated with PraneemBefore Treatment After Praneem Treatment
Pap Smear Showing intermediateand parabasal cells with rounded
margins, thickening and separation
of ectoplasm, and dark pyknotic
ecentric nuclei, a hallmark feature of
HPV infection. Pap Stain, original
magnification 400X
Pap Smear Showing clearance of HPV infection and reversal of cellular
changes in intermediate and
parabasal cells characteristic of HPV
infection. Pap Stain, original
magnification 400X
SUMMARY
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SUMMARY
Have developed Two Microbicides: Praneem Tablet;
BASANT Cream
Both have wide spectrum action against RTIs & STIs
Praneem regresses AVDS in 92 - 96% women; Currently
in Phase III trials; Licensed to Panacea Biotech
Therapeutic Phase II / III trials in progress for eliminationof HPV 16/18 in women with cervical dysplasia
molecularly positive for HPV; Encouraging early results
Credits
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CreditsTalwar Research Foundation, NewDelhi
National Center for Cell Science,Pune
CONRAD, Eastern Virginia MedicalSchool, Virginia, USA
GP Talwar, Sajad A Dar, Mahendra K
Rai, Kavita Bansal
Debashish Mitra, Sujata V Kulkarni
Gustavo Doncel
NCI / NIH, Bethesda, MD, USA
Institute of Cytology & PreventiveOncology (ICMR), Noida
National Institute for Research inReproductive Health (ICMR),Mumbai
Johns Hopkins University,Baltimore, Maryland
John Schiller, Christopher B Buck
Bhudev C. Das, Alok C. Bharti,Suresh Hedau, Shirish Shukla,Showket Hussain, Uma Kailash
K V R Reddy
K Whalley