golden jubilee commemoration oration of insa

8/7/2019 Golden Jubilee Commemoration Oration of INSA

1/48

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


2/48

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.


3/48

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


4/48

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


5/48

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


6/48

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


7/48

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


8/48

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


9/48

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


10/48

Approved by DCGI, India and US FDA


11/48

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


12/48


13/48

Patient code: CR

Initial After 5 doses


14/48

Patient code: RPS

After 2 dosesInitial


15/48

Patient code: BWS



16/48

Patient code: CHB

Initial After 1 Year


17/48

Patient code: RAB



18/48

Patient code: SNC

Initial After 2 doses (six months)


19/48

5

4

B1-Skin

B1-Tissue

3

2

1

0


20/48

mmuno . n ec. s. , , -


21/48

(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.


22/48

Mw

Singh IG, Mukherjee R & Talwar GP. Vaccine, 9, 10 14, 1991


23/48

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


24/48

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


25/48

CHRONIC TUBERCULOSIS

43.248.6

40

50

60 Mw control

00 05.4 5.4

0

10

20

30

2 6 12


26/48

Genomic analysis of M.w. done

by a National consortium of

. ,

Tyagi and Anil Tyagi -

laboratories


27/48

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


28/48

Landscape of genome evolution across the generalist and specialistmycobacterial lineages

PloS ONE (2007), 10/e968, 1 8


29/48

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)


30/48

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.


31/48

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


32/48

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


33/48

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


34/48

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


35/48

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 /


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


36/48

Effect on disease causation and Herpes

Simplex virus-2 (HSV-2) multiplication

PreparationPreparation # Shedding/# Shedding/


# with lesions/# with lesions/


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


37/48

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


38/48

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


39/48

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.


40/48

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


41/48

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


42/48

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


43/48

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


44/48

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


45/48

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


46/48

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


47/48

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


48/48

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

golden jubilee commemoration oration of insa

Documents