I'm going to share with you about our new research regarding antibody therapeutic targeting the connection hemichannels which can help improvement of the recovery from the spinal cord injury. As we know spinal cord injury is a very serious global health problem. Patients typically lose their neuronal function and also at a great risk for neuropathic complications which actually counts for the major post-traumatic loss of neurological function. As we know the part of the post-injury is related to a neuroinflammatory process which involved the activation of astrocytes and the formation of the GLIA score so which result in the formation of the impermeable barrier for regeneration of axons. That's why the therapeutic goals including limiting the size of lesions and also the axon loss which involved the approach to target the astrocytes. Therefore can result in the neurological function recovery. The molecules we focus on is a connexin 43 and this connexin 43 can form is a family of the connexin family which form the hexamers on the plasma membrane. So this typically the connexin hemichannel also called the connexons from one cell they can dock with another cell to form entire channel called the gap junction channels. And these channels allow the communication between the two adjacent cells and then they permit the exchange of the small molecules between two adjacent cells. So the recently mouse study suggests in addition to form gap junctions connexin are able to form independent hemichannels that means they not docked with another cell to form gap junction channels. So typically these hemichannels are formed in certain type of the cells such as astrocytes in the bone and astrocytes in central nerve system. So we developed the monoclonal antibody called ALMB0166 which can only target connexin 43 hemichannels but have no effect on gap junctions. And this if we apply this antibody to the in vivo we found that they have very limited effect on tissues. So then next I'm going to show this 0166 antibody can prevent the neuroinflammatory reaction. So connexin 43 hemichannel as I mentioned expressed in the astrocytes of the spinal cord and this hemichannel can open by traumatic injury. So as illustrated by this diagram you can see after the injury the hemichannel are open and this opening of the hemichannel can release the neuroinflammatory factors such as ATP, glutamate and potassium. And this can lead to the secondary injury due to the increased inflammatory reaction in the spinal cord. However if we use this 0166 antibody and then they can block the connexin 43 hemichannel they can prevent the release of this anti-pro-inflammatory factors then they can cause reduce of neuroinflammation and also leading to the reduction of the scar inflammation. So first we're looking at the properties of this antibody by looking at the binding to connexin 43. So we found this antibody can bind to connexin 43 in the high affinity. So this is a surface plasma residence experiments. So you can see there clearly you can have the high binding affinity the disassociation constant is about a 2.8 nanomolar. And then we also looking at a binding specificity in primary osteoastrocytes. So we use the both the right primary astrocytes isolating from the spinal cord and also the monkey brain astrocytes. So in both cases we can see the EC50 around the 30 microgram per ml. So then we also further looking at this antibody on the inhibition of the hemichannels. So you can see there is a dose dependent inhibition of the hemichannel opening and the EC50 is also comparable between rat and the monkey astrocytes. So here I want to mention the activation of the 43 hemichannel is induced by cytokines such as interleukin 1 beta. So then we further looking at the 016 antibody binds to connexin 43 in the murine spinal cord after spinal cord injury. So you can see after spinal cord injury the blood and spinal cord barrier is leaky. So then we can detect the antibody getting into the spinal cord. So this is detected by the anti-human secondary antibody. You can see that this antibody was detected around the spinal cord injury site. And then we also looking at the hemichannel opening in the mouse spinal cord in situ. And so then after we're looking at the hemichannel opening after spinal cord injury for half hour and also four hours and eight hours. So you can see after half hour and four hours there is significant suppression of the opening of hemichannels. At eight hours the effect is much less. Then we're looking at the secondary damage after spinal cord injury by looking at the lesion size as well as osteogliosis. So then we found with a treatment on 0166 they can reduce the lesion size after 30 minutes but have no effect after 24 hours. Also the osteogliosis size is also greatly reduced as well as the intensity of the gliosis. But after 24 hours there's no significant reduction. So then we further looking at a locomotor function of the spinal cord injury. We found that there is a dose-dependent effect with the increase of concentration of the 0166 antibody. There's an improvement of the locomotor function determined by hyaline activity using the BMS score. Then we further looking at the time-dependent study on the recovery. We found after spinal cord after 30 minutes, four hours, and eight hours they still shows the recovery with most effect after 30 minutes and four hours. So this means this treatment is very important for the acute spinal cord injury. So then with development of this antibody we also conducted R&D enabled the study toxicity studies using the mice as well as monkeys. We also demonstrated the safety profile. So in conclusion we found 0166 antibody can inhibit activation of the hemichannels in both cultured primary osteocytes as well as spinal cord astrocytes in situ. We also found a single administration waiting eight hours improved the locomotor function and recovery up to eight weeks post the injury in spinal cord injury model. Also the treatment of 0166 decreased gliosis and vision size and increased neuronal survival. And the series of the pharmacology, toxicology, and TCR and other assessment demonstrate the safety profile of 0166 antibody. Therefore ALMB 0166 is a potent first-in-class antibody therapeutic for treatment of the acute spinal cord injury. A phase one clinical trial in healthy subjects is currently underway. Thank you.

ALMB0166

acute spinal cord injury

connexin 43 hemichannels

inflammation

locomotor function