Oncolytic virotherapy is cancer treatment using a native or reprogrammed virus that has the potential to targeting and killing cancerous cell. Taking advantage of the OncoVirapy™ platform, Creative Biolabs provides customized, standardized, and reliable and high-quality oncolytic virus therapy development services for clients globally.

1. Oncolytic virotherapy is cancer treatment using a native or
reprogrammed virus that has the potential to targeting and
killing cancerous cell. Taking advantage of the OncoVirapy™
platform, Creative Biolabs provides customized, standardized,
and reliable and high-quality oncolytic virus therapy
development services for clients globally.
ONCOLYTIC VIRUS
THERAPY DEVELOPMENT

2. About
Creative Biolabs
The Best
Solutions
for Immunotherapy
Since our inception in 2004, our research and service capacity has
expanded to the entire new drug discovery and development pipeline,
including early discovery, preclinical testing, process development, cGMP
manufacturing, phase I-III clinical trails. As a global company, we have
more than 200 talented and well-trained scientists located in different
continents working closely with partners from the entire world to develop
and produce medicines of tomorrow. Facing the increasing challenges
from cancer treatment, our scientists establish systematic immunotherapy
development platforms to meet the need of our customers worldwide.
Our featured platforms include CellRapeutics™ for TCR and CAR
development, and OncoVirapy™ for oncolytic virus development.

3. ONCOLYTIC
VIROTHERAPY
Oncolytic Virus
Oncolytic viruses represent a new class
of therapeutic agents that promote anti-
tumor responses through a dual
mechanism of action that is dependent on
selective tumor cell killing and the
induction of systemic
anti-tumor immunity.

4. In vivo validation studies services for engineered oncolytic virus.
include but not limited to construction of xenograft/syngeneic
animal models, immunogenicity tests, biodistribution/PK tests,
efficacy, and toxicology studies
OncoVirapy™
Platform
Taking advantage of the OncoVirapy™
platform, Creative Biolabs provides
customized, standardized, and reliable
and high-quality oncolytic virus therapy
development services for clients
globally. Various types of oncolytic
viruses engineering systems have
been established in-house to facilitate
oncolytic virus development with less
time and reduced budget. GMP-
compliant manufacture of oncolytic
virus is also included in
our services.
Diverse engineering methods to make full use
of natural properties of various viruses, and
already developed therapeutic methods.
A variety of prove-of-concept in vitro assays
service for engineered oncolytic virus to test the
virus functional activity.
oncolytic virotherapy develp[ment service for various
type of tumor, including melanoma, genitourinary
malignancies, hepatocellular carcinoma, breast
cancer etc.

5. Oncolytic Virotherapy Development Process
Basic Cancer
Biology Study
Abnormal signaling
pathway study
Target identification
Malignant cell type
Tissue specificity
Oncolytic Virus
Vector Construction
Virus species
In vitro Validation
Replication Capacity
In vivo Validation
Non-tumor bearing
animal model
Pharmacology
Virus tropism
Tissue selectivity
Therapeutic
Strategy
Monotherapy
Combination
therapay
Arming with
therapeutic gene
Safety issue
Delivery Expression of
transgene
Function of transgene
Tumor cell selectivity
Tumor lysis capacity
Genetic stability
Infectivity
Tumor-bearing
xenograft/syngeneic
animal model
Biodistribution
Toxicology, safety
www.creative-biolabs.com
Antitumor response

6. Basic Study
Cancer Biology
Cancer is a group of diseases involving
abnormal cell growth with the potential to
invade or spread to other parts of the body.
To our knowledge, both immunologic and
cancer biologic approaches have to be
combined to enhance the efficacy of
anticancer therapy, and this will always be
a challenge for the next generation of
cancer immunotherapy. Only on the basis
of molecular mechanisms and other
proved evidence that underline the
pathogenesis of specific type of cancer,
can a better therapeutic strategy be
developed.

7. Immunotherapy in cancer is rapidly
evolving, with various treatments
being investigated for their potential
to provide long-term survival across
a broad range of tumor types, and
for their synergistic activity when
combined with other treatment
modalities. It is important now to
determine how to advance
this field and the way to use
these new immunotherapies
most effectively to achieve
the best patient outcomes.
Therapeutic Strategy
Reversing the inhibition of adaptive immunity (blocking T cell
checkpoint pathways)
CTLA-4 inhibition, PD-1/PD-L1 inhibition, LAG-3 inhibition, TIM-3 inhibition
Switching on adaptive immunity (promoting T cell co-stimulatory receptor
signaling using agonist antibodies)
Enhancing OX40 signaling, Enhancing CD27 signaling, CD40 activation,
Promoting CD137 signaling
Expanding adaptive immunity
Adoptive cell therapy
Improving the function of innate immune cells
Stimulating dendritic cell, Manipulating NK-cell inhibitory receptors
Activating the immune system (potentiating immune-cell effector
function)
Oncolytic virus, Therapeutic vaccines

8. Oncolytic Virus Vector Construction
Viruses used as oncolytic vector
Nucleic acid
Capsid
symmetry
Envelope Family Virus Cell receptor
DNA
Icosahedral
Naked Adenoviridae Adenovirus CAR
Enveloped Herpesviridae HSV-1
HVEM/nectin 1/
nectin 2
Complex Enveloped Poxviridae Vaccina virus Unknow
RNA
Icosahedral Naked
Reoviridae Reovirus Unknow
Picornaviridae Poliovirus
CAR/ICAM-1/DA
F
Helical Enveloped
Rhabdoviridae VSV LDLR
Paramxyoviridae Measles virus SLAM/CD46

9. Oncolytic Virus Vector Construction
As we move in the direction of intentionally using virus infections to mediate tumor destruction, it is apparent that a targeted
virus with exquisite tumor specificity will be superior to its non-targeted counterpart, allowing for the administration of higher
tumor destructive doses without toxicity to normal tissues. Cancer cells distinguish themselves from their normal counterparts
by alterations in cell physiology such as self sufficiency in growth signals, insensitivity to growth inhibition signals, evasion of
apoptosis, limitless replication potential, sustained angiogenesis and tissue invasion and metastasis. These alterations make
these a generous host for viruses and hence these properties can be utilized for selective replication of oncolytic viruses in
cancer cells.
Targeting Virus Tropism
Natural tropism
Deletion of viral
genes required
for virus
replication in
normal cells
Engineered
with protease
target
sequence
Ablation of
unwanted
tropism
(MicroRNA
targeting)
Engineered
with
tissue/tumor
specific
promoter
Using cellular
vehicle
Designing strategy of modification of oncolytic virus tropism

10. Oncolytic Virus Vector Construction
Basically oncolytic virus exert antitumor activity by direct lysis of cancer cell and activation of immune system. Although
OVs can extensively transfect and kill tumor cell, a percentage of tumor cell remains uninfected. Bystander killing can be
achieved, both locally at the site of a spreading infection and systemically at uninfected tumor sites, by genetically arming
the virus using one of several possible approaches.
Arming strategy
Immune-stimulatory
cytokines & receptors
Immune checkpoint
inhibitors
T cell Bispecific
Antibody
Prodrug
convertases
Short hairpin RNA

11. Oncolytic Virus Vector Construction Safety
Unwanted normal tissue pathology
Studies of the mutation rates of viral polymerases, the generation
of quasispecies, the evolution of viral populations and the
evolution between dominant subspecies within a virus population
are therefore of great interest and relevance to the OV field.
It is important that precautions for infectious
material and biological safety, and biosafety
guidelines be followed when administering
oncolytic virus. Hence, the issue of safety
worth being handled in advance, even at the
stage of oncolytic vector construction.
Careful steps must be taken to avoid the
creation of OVs that might evolve to become
serious pathogens. For the problem of
undesirable transmission, contingency plans
to terminate the spread and/or transmission
of an infection can increase clinical
confidence in viral therapy.
Unwanted transmission
The consequences of transmission to others might not be well
understood and precautions should be taken to minimize
exposure of healthcare providers, family members and other
patient contacts. Non-clinical viral shedding studies can be useful
in preparing for clinical studies and evaluating detection methods
Toxicity
The toxicology assessment of an oncolytic virus should be
comprehensive enough to identify, characterize and quantify
potential local and systemic toxicities following administration.

12. Validation of
Oncolytic vector
Plaque purification
assay
Identity assay
Replication assay
Tumor lytic assay
Transgene
expression assay
Transgene
functional assay
Immunogenicity
Test
Efficacy Study Toxicology Study
Biodistribution/PK
Test
In vivoIn vitro

13. Considerations
in Validation
Evaluation of Selectivity
For in vitro validation studies, prior to using animal models, experiments conducted to
characterize selectivity in normal and tumor cells should address selective gene expression,
cytotoxicity and viral replication. To this end, candidate oncolytic viruses should be assayed
in vitro for lysis and/or replication on tumor/permissive or nonpermissive cell lines.

14. Considerations
in Validation
Function of Transgene
If the oncolytic virus contains a transgene, it is important that the animal species be
pharmacologically responsive to the expressed protein. If the expressed transgene is inactive in
the animal species, oncolytic virus can be engineered to express the analogous species-specific
transgene and used in non-clinical studies to assess both activity and safety.

15. Considerations
in Validation
Selection of animal model
Selection of the animal model should take into consideration the purpose of the study as well
as the viral tropism, infectivity, replication ability, cytopathic potential and anti-tumor effect of
the oncolytic virus.
Ideally, a xenograft or syngeneic model should
represent the tumor biology and pathology of the
target clinical population to the clinical outcome.
Non-tumor-bearing, can be
used to evaluate the safety
of the OV.

16. Considerations
in Validation
Pharmacology & Biodistribution
Biodistribution studies in animals
address oncolytic virus dissemination
to target and non-target organs. OV
dissemination can be detected using an
assay for nucleic acid sequence. Also,
sensitive assays such as quantitative
polymerase chain reaction (QPCR) can
be applied to test the presence of
oncolytic virus sequences in animal
organs and tissues
It is critical to assess the bioactivity and
pharmacologic profile of the oncolytic
virus to understand the ability of the OV
to induce the desirable anti-tumor effect.
The experiments designed should help
to define a pharmacologically active
dose range, with establishment of an
optimal dose and a minimally effective
dose, and also to determine a
potentially optimal route for virus
product administration.

17. Featured Products
By the OncovirapyTM platform, Creative Biolabs is now able to provide
products of various oncolytic virus vector for our customers globally. To be
specific, our oncolytic vectors are designed on the basis of human adenovirus,
which has been widely used as a viral vector for gene therapy studies.
Moreover, most of the oncolytic adenovirus designed and produced with
arming of different kinds of transgene, such as for antibody, bispecific
antibody, immune modulator, miRNA and tumor-associated antigen. We
believe that our designed ready-to-build oncolytic vectors will facilitate the
research in cancer immunotherapy for scientists all over the world.
ITR E1A E1B pIX L1 L2 L3 L4 E3 L5 ITR
E2B E2A E4

18. Summary of oncolytic virus development
on OncoVirapy™ Platform
A. Basic cancer biology research
Cancer Type Malignant Cell Type
Abnormal signaling
pathway
B. Oncoytic Virus Vector Design
Tissue
selectivity
Virus
Species
Natural/Modified
Tropism
Therapeutic Gene
Arming
Safety issue
Neutralization issue
C. Therapeutic Strategy
Development
Monotherapy
Combination Therapy
D. Validation Studies
Replication
Capacity
Tumor Lysis
Capacity
Tumor cell
selectivity
Function of
Transgene
Selection and Construction
of Animal model
Pharmacology Biodistribution
Toxicology Safety
Antitumor/Host immunity Response
In vitro
In vivo
E. Oncolytic virus
Manufacture

19. The Best Solutions
for Immunotherapy
from Creative Biolabs
45-1 Ramsey Road
Shirley, NY 11967, USA
Tel: 1-631-871-5806
Fax: 1-631-207-8356
Email: inquiry@creative-biolabs.com