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New Strategy New Strategy of of

BiodefenseBiodefenseGeorge YeGeorge Ye President & CSOPresident & CSO

http://www.yesbiotech.comemail: info@yesbiotech.com

2355 Derry Road East, Unit 23Mississauga, Ontario, Canada, L5S 1V6

While the public health philosophy of the 20th Century – emphasizing prevention – is ideal for addressing natural disease outbreaks, it is not sufficient to confront 21st Century bio-threats by the deliberate release of viruses, bacteria, fungi or toxins from living organisms.

• Disease outbreaks, whether natural or deliberate, respect no geographic or political borders.

Bio-target

• Target Microbial• Target Cell• Target Gene• Target Protein• Target Molecule• Target Target “Cytokine “Cytokine

Storm”Storm”

• External microbes is our worst enemy during an outbreak of influenza or bronchitis, however, our own immune system is potentially more lethal.

• Our body detects foreign microorganisms indicating an infection, it might respond by over-protecting the site of infection.

Cytokine Storm• Cytokine storm occurs when the

body’s immune system overreacts to an intruder, such as a virus, by producing high levels of cytokines.

• Cytokine storm can occur in a number of infectious and non-infectious diseases including graft versus host disease (GVHD), adult respiratory distress syndrome (ARDS), sepsis, avian influenza, H1N1 flu, smallpox, and systemic inflammatory response syndrome (SIRS).

H1N1 (swine) flu, with 44,287 confirmed cases worldwide as of June 20, and with 180 deaths globally, remains high on the WHO’s radar screen.

H1N1 (swine) flu

H1N1 viruses have pandemic potential and that historical evidence supports the possibility that healthy young adults may be especially susceptible to more severe infection and poor outcomes due to the ability of a strong immune system to initiate a cytokine storm.

The relative youth of swine flu casualties bears a disquieting similarity to deaths from the infamous 1918 influenza A H1N1 pandemic. Nearly half of those approximately 50 million casualties were young adults 20 to 40 years old.

H1N1 in 1918

Of course, flu vaccines are usually effective at preventing the flu during its peak season. But they are no guarantee, especially when flu strains mutate (antigenic shift) after the vaccine has been manufactured.

Waiting for Flu Vaccine?

H1N1 and other harmful infectious diseases’ lethal mechanism may be the “cytokine storm”!

Tackling the harmful “Cytokine Storm” has been a new strategy of Biodefense.

Studies on bio-target and building an Antibody Based Defense Strategy to build improved and comprehensive biodefense.

• External Immediate Immunity to Microbial agents——

Passive Antibody Administration

• Internal Neutralizing Inflammatory factors —— Anti-Chemokine Therapy

http://www.yesbiotech.comemail: info@yesbiotech.com

2355 Derry Road East, Unit 23Mississauga, Ontario, Canada, L5S 1V6

Passive Antibody Administration – Immediate Immunity Against High-Priority Agents and Diseases:• Anthrax, including multi-drug resistant anthrax• Burkholderia bacteria• Clostridium botulinum toxins• Plague• Smallpox• Typhus• Tularemia• Various viral hemorrhagic fevers• Hand, Foot and Mouth Disease (HFMD)• Harmful Flu

HFMD

A-H1N1

• The only available countermeasure that can provide immediate immunity against a biological agent is passive antibody, while vaccines require time to induce protective immunity and depend on the host’s ability to mount an immune response.

Advantages of an Antibody-Based Defense Strategy

• Antibody-based defense were used historically for treatment of anthrax, tularemia, plague, rabies, snake envenomation, spider bites and digoxin toxicity. The First used in the late 19th, and more than 100 years of experience.

• The antibody immunization in defense provides a state of immediate immunity that can last for weeks and possibly month.

• Antibodies are natural products with minimal toxicity, provided that they contain no aggregates and have no reactivity with host tissues.

• Oral administration can be useful against certain gastrointestinal agents.

Advantages of an Antibody-Based Defense Strategy

• Ig preparations can theoretically be administered intramuscularly hence, generating antibody preparations with high specific activity suitable for a small volume administration into one of the large muscles of the arm or leg may be possible.

• Humanized monoclonal antibody cocktails (mixing MAbs) can lead to bind multiple epitopes in the target pathogen and against antigenic variation of pathogens.

• Polyclonal humanized antibodies confer broader biological activity to multiple antigenic targets. Polyclonals have the highest probability of retaining activity in the event of antigen multation.

Chemokines – The “Prime Culprit” of Cytokine Storm

• Chemokines are inflammatory proteins acting via G-protein coupled Chemokine receptors that trigger different signaling pathways.

• Monocyte Chemoattractant Protein-1 (MCP -1) and Interleukin-8 (IL-8) are two major respective members of the CC and C-C Chemokine subfamily.

• The roles of MCP-1 and IL-8 are emerging in regulating the recruitment of inflammatory cells into tissue during inflammation.

MCP-1 and IL-8

• MCP-1 and IL-8 may represent targets for diagnostic procedures and therapeutic intervention, and may be useful as a prognostic factor in the both infectious or noninfectious inflammatory diseases including gram-negative and positive infections, listeriosis, mycobacterial infections, lyme arthritis, pneumonia, fungal infections, HIV, leishmaniasis, and sepsis.

Chemokines and Diseases

Some diseases found to be medicated by Chemokines

Asthmatic reaction MCP-1, MIP-1αand RANTES

Acute pulmonary disease (SARS, ARDS)

IL-8, MCP-1, ENA78 and RANTES

Endotoxemia and Sepsis IL-8, MCP-1, MIP-1αand RANTES

Eczema and Psoriasis IL-8

Rheumatoid arthritis IL-8, MCP-1, ENA78, MIP-1α

Osteoarthritis MIP-1β

Immune complex glomerulonephritis

IL-8, MCP-1

Wound healing site MCP-1, IP-10

Cytokine Storm in SARS

Normals

(15 cases)

SARS

(210 cases)

Acute Phase Recovery Phase

MCP-1

(pg/ml) 436.2±21.5 1241.2±245.6 565.7±79.1

IL-8

(pg/ml) 0 149.7±98.1 0.6±0.4

Control mediumAnti-MCP-1 10 µg/ml

Human IL-8 (ng/ml)

0

2

4

6

8

10

12

0.1 0.5 1 5 10 100

012345678

1 10 50 100 500

Huamn MCP-1 (ng/ml)

Control mediumAnti-IL8 10 μg/ml

A. Monocyte chemotaxis

B. CXCR2B/293 cells

**

*

*

*

*

The effect of antibodies on MCP-1 and IL-8 mediated cell chemotaxis

*: significantly reduced cell migration induced my chemokines-treated with antibodies (37oC, 60 min)

Anogen-Yes Biotech has developed humanized chimeric antibodies to neutralize MCP-1 and IL-8.

• OA+LPS (“two-hit”) caused severe hypoxemia in Rabbit.

• Group A received OA (0.08ml/kg) through femoral vein (first hit), LPS (0.5mg/kg) four hours later (second hit).

• Group B received a single i.v. injection of the monoclonal anti-IL-8 antibody.

• Group C received saline for control.

Anti-IL-8 Antibody in Treatment of Rabbit ALI/ARDS Model:

PaO2 Comparison between Three Groups

0

5

10

15

0hr 4. 5hr 5hr 5. 5hr

hr

kpa

AB

C

Neutrophil Count and IL-8 Concentration in BALF

Group A Group B Group C

Neutrophil count（ 107/L） 32.46±7.30* 19.04±4.57*# 3.96±0.77

IL-8 concentration（ pg/ml）

758.38±301.92* 131.54±40.56*# 14.91±6.21

*p<0.05 vs. group C, # p<0.05 vs. group A

Histopathology Comparison

2a

Staining with hematoxylin and eosin, lung tissues were observed 10×20.

2a: the animal received two-hit, 2b: the animals were pretreated with anti-IL-8 antibody, 2c: the control animal.

2b 2c

While external agents are the root causes of complications, internal responses to these agents trigger deadly results. Thus, it is necessary to emphasize both external agents and internal responses in disease and illness prevention. By targeting internal reactions, it is possible to reduce the likelihood of fatal complications associated with external agents.

Key Point of the New Strategy:

Thank you

http://www.yesbiotech.comemail: info@yesbiotech.com2355 Derry Road East, Unit 23Mississauga, Ontario, Canada, L5S 1V6