Download Cancer - IIPRD

DELHI . MUMBAI . BANGALORE . PUNE . INDORE
BLOOD-BASED EPIGENETIC BIO MARKER
OF HUMAN CANCER
IDENTIFYING PARTNERS/LICENSEES
BACKGROUND: MAGNITUDE OF CANCER: GLOBAL
 Global cancer burden – 32.6 mill.
 Incidence/year – 14.1 mill. (7.4 mill. ♂ + 6.7 mill. ♀) cancer patients
 Leading cause of death
 8.2 mill. (13% of all human deaths/1 death every 5 sec) per year
 ~ 65% of cancer-related death in low and middle income countries
 African Region (5.4%), Eastern Mediterranean Region (3.7%), South East Asia
Region (12.8%), and Western Pacific Region (29.7%)
 Lack of access to basic health care – Diagnosis as well as Treatment
 Low level of awareness in population
 Current status
 70 % of 65+ year-old have >2 chronic diseases, including cancer (BMC
Geriatrics, 2011)
 Current WHO projection (World Cancer Report)
 26.4 mill. patients/17 mill. Deaths (1 death every 2 sec) in 2030
 Current GLOBAL projection for 2050
 Population of 80+ year-old to triple (Nature, 2014)
BACKGROUND: MAGNITUDE OF CANCER: INDIA
 Av. life expectancy: 64 (♂) – 67 (♀) years
 2.5 mill. cancer cases (27 national registries)
  1.75 mill. (70%) in advance stage(s)
  0.55 mill. deaths every year (based on 2010 figure published in Lancet 2012)
 Current estimate: ~ 1 mill. new cases/year
 Projection 2020: ~ 1.15 mill. new cases/year
 Not a ‘notifiable’ disease in India
 Tobacco-related cancer – major cancer in India
 Cause of ≥ 50% deaths: 42% men + 18% women
 Majority of death (71%) in age group 30-70 years
 Major cancer sites in men: lip, pharynx, stomach and lungs
 Major cancer sites in women: cervix, stomach and breast
 Only 25 Regional Cancer Centers in India
 One RCC serving  45 mill. people
VISION STATEMENT - 1
 In the existing modality of cancer therapy, no convenient bio-marker is
available to monitor progress of therapy and regression
No mid-term dose/course correction possible
 Typical therapeutic protocol
Start a treatment course with a therapeutic agent
Await patient response/’feel good nod’
Continue till the end
Discontinue, if adverse
Try some other therapeutic agent
 Average dose/course
No individualized treatment possible
VISION STATEMENT - 2
 “Cancer is one of the most curable chronic diseases of humans” – Dr.
Vincent T. deVita [Former Director, NCI and Yale Cancer Center (USA)]
Condition: Timely detection
  40 % of cancers can be prevented
Cancers of lungs, cervix, head & neck, GIT, etc.
 Combating cancer by early detection
 35% enhanced clinical efficacy with existing therapeutic practice(s)
 Significant mitigation of human suffering by ‘cancer screening’
Screened population  Declining trend of cancer
Cancer statistics of USA for 2013 shows this
Incidence of cervical cancer in India
VISION STATEMENT - 3
“Prevention
is absolutely critical and it’s been somewhat
neglected”
Dr. Chris Wild, Director, International Agency for
Research on Cancer (IARC): BBC News/4th Feb. 2014
Need a ‘good’ cancer bio-marker
INTRODUCTION TO TECHNOLOGY
 Research spanning over two and a half decades has identified and
characterized a novel and ideal biomarker of human cancer, which meets
all three characteristics formulated by the international Predictive Safety
Testing Consortium (PSTC) comprising industries, non-profit
organizations and regulators (including FDA and EMEA/EMA).
 A non-radioactive, highly sensitive, reproducible assay to quantitatively
estimate the bio-marker using a minimally invasive quick process that
does not require hospitalization has been developed.
 The design of a complete and easy-to-use prototype for this test has
already been developed in a diagnostic kit format that is highly reliable
and portable to remote locations without compromising efficacy.
 Phase-I clinical study undertaken in patients with 18 different cancers
(grouped in head & neck, breast and cervical cancer categories) has
provided statistically significant verification of the ‘proof of concept’.
NEED FOR TECHNOLOGY
 Inadequacy of current therapeutic interventions for individualized mid-
course corrections of ongoing therapy (change of drug, dose, strategy,
etc.) reflects a serious gap in increasing clinical efficacy based on realtime quantitative biochemical or pathological tests.
 The novel diagnostic kit developed fills this conspicuous gap as it can
reliably and reproducibly detect multiple types of cancer using a
minimally invasive OPD procedure requiring no hospitalization within a
short time of less than 60 min and provide quantitative measurements of
treatment efficacy and cancer retardation.
 An ideal bio-marker of cancer can potentially be utilized for predictive
bio-monitoring of cancer therapy to achieve higher clinical efficacy as
well as for screening of populations for early detection of cancer.
 The clinical efficacy of cancer therapy may be enhanced by 35 to 40% by
monitoring the therapeutic progression using this sensitive and
predictive biochemical test, presented in the format of a diagnostic kit,
as proposed here.
NEED FOR TECHNOLOGY
 Feedback from a large number of clinicians suggested that the existing
clinical efficacy of cancer therapy can be enhanced greatly by:
 (a) Making mid-course quantitative assessment of therapeutic
effectiveness a reality, thus empowering the physician to take a decision
based on quantitative information for required change of the ongoing
therapeutic modality/strategy opening a corridor for individualized
therapeutic course;
 (b) Using a quick, reliable and predictive test that is minimally invasive,
does not require hospitalization and can be carried out in remote
locations with high success rates; and
 (c) Early start of therapy due to early detection of cancer by using a
screening test that is not limited to a specific tissue or type of cancer.
LIMITATIONS IN EXISTING TECHNOLOGIES
 To the best of our knowledge, there is no other diagnostic kit or
procedure for assessment of therapeutic status of cancer today that can
match the projected position of technology being offered in terms of its
spectrum of detection, sensitivity of the assay/test and ease of
application (non-invasiveness, non-radioactive, non-hazardous nature of
the components, quick result, no hospitalization, etc.).
 The other bio-markers of cancer in use today are typically specific to a
particular cancer or tissue type, the assays being time consuming and
usually requiring hospitalization.
 The scope of accessibility to these tests is expensive and limited only to
select segments of populations with close proximity to large cities.
KEY ADVANTAGES
 The novel technology has the potential to become a convenient general
biomarker of cancer enabling reliable screening of remote populations
for early detection of cancer without portability issues – an area that is
currently void of any option, thus, expanding the umbrella for marketing
of this diagnostic kit
 Armed with quantitative information derived from this quick but
sensitive test, it would become possible for the clinical oncologist to
make necessary mid-term correction in therapy for clinical gains and
make early detection of cancer in much larger segments of population a
reality.
THE BIO-MARKER MOLECULE
 ADP-ribose polymer (ARP) or poly-ADP-ribose (PAR)
 Epigenetic in nature!
 Highly heterogeneous polymer
Poly(ribose-nucleotide)
Molecular arrangement
Branched or unbranched
Different size
Multiple architecture
Multiple location
 Multiple medical applications
 Biomarker of human cancer
 Biomonitor of cancer therapeutics
 Predictive diagnosis
METABOLIC PROCESS
 Post-translation modification of proteins
 Enzyme catalyzed reaction
 Fully reversible
 Preferred target – histone proteins
 Heterogeneous, branched/unbranched polymer of ‘ADP-ribose’
units of variable lengths
Target amino acid residues: Arg & Lys
2 –ve charges/monomer added on target proteins
DNA-protein charge interaction(s) altered
Growing physical bulk of the polymer
 Conformational and functional status of chromosome strongly
influenced
SUMMARY OF TECHNOLOGY WHEN TESTED
ON MICE
 ADP-ribose polymer: indicator of carcinogenesis
Initiation phase
Promotion phase
Progression phase
 Carcinogenesis induction
DEN, DMN, Arecoline, Betel nut
Ascites
 Target as well as non-target tissue/proteins
Liver, Spleen cells, BMC, other tissues
Total cellular proteins
Histone proteins, HMG proteins
 Blood lymphocytes mirror the status
PROOF OF CONCEPT
Phase 1 study: Human Cancers
 Cancers under study
 Breast
 Cervix
 Head & Neck (14 different cancers/sites)
n = 111
n = 22
n = 24
n = 65
 Advance stage cancers
 ADP-ribose polymer quantified in peripheral blood lymphocytes
(PBL)
 2 ml blood collected
 PBL isolated, homogenized, blotted and immunoprobed
 Controls: no known history of cancer
 Mainly univ. students (volunteers)
 Relatives
 Males and females
n = 68
PROOF OF CONCEPT
Breast and Cervical Cancers:
Total cellular ARP in cancer of
breast
compared to control [Number of individuals:
Controls: All = 68, females = 27, breast
cancer patients = 22].
Category/Type/
Site of cancer
Number
of cases
PAR Level
Mean ± SD
Cancer of breast
22
7594 ± 852
Control (♀ only)
27
14125 ± 1431
Control (all: ♂ + ♀)
68
14573 ± 1452
Total cellular ARP in cancer of cervix
compared to control [Number of individuals:
Controls: All = 68 & females = 27, cervix
cancer = 24].
Category/Type/
Site of cancer
Number
of cases
PAR Level
Mean ± SD
Cancer of cervix
24
8139 ± 799.1
<0.0001
Control (♀ only)
27
14125 ± 1431
<0.0001
<0.0001
Control (all; ♂+♀)
68
14573 ± 1452
<0.0001
P
P
PROOF OF CONCEPT
Head and Neck Cancer:
Category/Type/Site of
cancer
Total cellular ARP in cancers of head and neck
compared to control
[Number of individuals:
Controls = 68, Ca Esophagus = 19, Alveolo = 6,
Pyriform sinus = 4, Nasopharynx = 10, Larynx = 1,
Tongue = 4, Tonsil = 5, Buccal mucosa = 3, Pharynx =
3, Lip = 3, Nasal cavity = 2, Oral cavity = 2, Epiglottis =
4, Vocal cord = 1].
PAR Level (Mean
± SD)
P
Control
Ca. H&N (All)
Ca. Oesophagus
Ca. Alveolo
Ca. Pyriform sinus
Ca. Nasopharynx
Ca. Larynx
Ca. Tongue
Ca. Tonsil
14573 ±1452
8059 ± 304
8007 ± 463
8173 ± 651
8196 ± 258
7890 ± 833
7537
8340 ± 885
7879 ± 1081
<0.0001
<0.0001
<0.0001
<0.0001
<0.0001
NA
0.002
<0.0001
Ca. Buccal mucosa
8172 ± 327
<0.0001
Ca.
Ca.
Ca.
Ca.
Ca.
Ca.
8177 ± 261
7801 ±906
8554 ± 440
7681
8553 ± 440
7873
<0.0001
0.0066
NA
NA
<0.0001
NA
Pharynx
Lip
Nasal cavity
Oral cavity
Epiglottis
Vocal chord
PROOF OF CONCEPT
All Cancers- Age and Gender Matched
Gender
Control
Cases
Male subjects (n)
17
23
ARP level (Mean ± SD)
14803 ± 1154
7874 ± 657
Female subjects (n)
11
33
ARP level (Mean ± SD)
14543 ± 1595
8159 ± 760
P
<0.0001
<0.0001
PROOF OF CONCEPT
2nd level R&D – Direct ‘Immuno-probe assay’
Man
Mouse spleen cell
Blood from finger tip
Heterogeneous ADP-ribose polymer (ARP)
Lysis of blood
Purification & characterization
Adsorption of lysed blood on NCM
Raise Polyclonal anti-APR in rabbit
Wash
Purification of antiserum (PAb)
ELISA based DIRECT immunoprobing using polyclonal
anti-PAR~HRP/India ink stain for total protein
Lyophilize PAb
PAb conjugated with HRP/ALP
PAb~HRP/ALP solution
Ag (PAR)
on target
Polyclonal
Ab~HRP
PAR of total
cellular proteins
in blood
Color
development
Quantification
using a digital
densitometer
THE KIT IN MAKING
ADP-Ribose
polymer mediated
Rapid and Easy test
for human Cancer
(‘ARREST – C’)
THE KIT IN MAKING
0.75 ml
Volumes
0.3 ml
Cut-out
0.5 ml
Lysis
Buffer
Blood
Sample
Wash
buffer
0.3 ml
Ab-HRP
0.5 ml
Wash
buffer
0.3 ml
0.5 ml
Colour
Developer
Water
Lysis
Buffer
CONCLUSIONS
 ADP-ribose polymer: a potential biomarker of human cancer
 Useful in prognosis & diagnosis
 Biomonitor of therapeutic progression
 Potentially usable for screening population(s)
 Multi-center human study is required
 Phase II clinical study to be initiated
 The novel immunoprobe assay of PAR to be packaged in a kit
PATENT/IP STATUS
 Application Number: 1791/KOL/2008
 Application Status: Patent Pending
 Priority Date: 23/10./2008
CONTACT DETAILS
DELHI . MUMBAI . BANGALORE . PUNE . INDORE
Contact Details
Noida (NCR) Office
E-13, UPSIDC Site-IV, Behind Grand Venice, Greater Noida, 201308
Contact Person: Tarun Khurana
Contact No.: +91-120-2342010-11/9810617992
E-Mail: [email protected], [email protected]
Website: www.iiprd.com | www.khuranaandkhurana.com