Cascade Biotechnology INC.

Complement System Therapeutics

On September 27-29, 2022 in Boston, MA, Dr. Dan Benjamin will be speaking at the 6th annual Complement-based Drug Development Summit, www.complement-therapeutics.com.

The title of the talk is:Targeted Delivery Platform of sC3, a Complement Depleting Protein, Using Viral Delivery, Conjugate Guidance, & Localized Application Modalities

Topics:

• Discover how specific cell types in the retina and kidney can be targeted to express sC3

• Conjugate guidance and tissue specific local delivery are being developed to increase efficacy and decrease off target effects

• Taken together, these technologies allow Cascade to optimize treatment based on the therapeutic target

Please contact Dr. Benjamin on this website or at: daniel.benjamin@cascadebiotechnology.com

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Cascade Biotechnology is developing therapeutics that use the body’s complement system to attack diseases and disorders such as Alzheimer’s disease (AD), schizophrenia (SZ), myasthenia gravis (MG), Neuromyelitis Optica (NMO), cancer, adult macular degeneration (ADM), blood diseases such as paroxysmal nocturnal hemoglobinuria (PNH), atypical hemolytic uremic syndrome (aHUS) and others.
The complement system is an important part of the innate immune system, which helps to protect the host against infection and connects innate and adaptive immune systems.
Complement consists of more than 30 proteins that are activated through a series of proteolytic cleavages, which in turn activate proteins in the next step of the complement cascade.
There are three complement activation pathways, the classical pathway, activated by the presence of antibodies on the cell surface, the lectin pathway, activated by the recognition of certain carbohydrates on cell surfaces, and the Alternative Pathway, which is constantly activated by spontaneous conformational changes to C3, the most common complement protein.
All three activation pathways come together through C3 activation. C3 activation allows for the activation of another complement protein, C5, that initiates the terminal complement pathway. This pathway forms the macromolecular Membrane Attack Complex (MAC), which is able to destroy invading pathogens by punching holes in their cellular membrane.
While complement proteins serve an important regulatory role in inflammation, over activation of complement ultimately leads to disease states. Inhibiting overactive complement offers one strategy for managing and potentially preventing a range of disease and disorders.
See: Fritzinger DC, Benjamin DE (2016) The Complement System in Neuropathic and Postoperative Pain. Open Pain J. (2016);9:26-37.

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Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Complement resolves immunological problems, yet over-activation may lead to these diseases:

Renal: Diabetic Nephropathy, Lupus Nephritis
Neurological & autoimmune: Myasthenia Gravis (MG), Multiple Sclerosis (MS), Neuromyelitis Optica (NMO)
Retinal: Age-related macular degeneration (AMD)
Cognitive: Alzheimer’s Disease (AD), Prodromal Schizophrenia (SZ)



Disease Program	Research Project Codes	Project Phase	Status

Diabetic Nephropathy Lupus Nephritis	CB101	Development	Scale-up for IND-Enabling GLP Studies
Retinal Gene Therapy	CB201	Discovery	In Vivo Testing

Neuromyelitis Optica	CB301	Discovery	In Vitro Research
Myasthenia Gravis	CB401	Discovery	In Vitro Research

Prodromal Schizophrenia	CB701	Early Discovery	In Vitro Research
Alzheimer's disease	CB801	Early Discovery	In Vitro Research

Amyotrophic Lateral Sclerosis	CB901	Early Discovery	In Vitro Research
	*links to detailed page

At Cascade, we are developing a gene therapy approach to these common and deadly renal inflammatory diseases.

LN patients typically require long-term corticosteroids and immunosuppressants. This constitutes an unmet medical need for at least 750,000 patients in the US. Lupus patients will need up to 3 kidney transplants in their lifetime. Many cannot receive a transplant.

Statistically, 1 in 3 of the 37.3 million diabetics in the US will develop DN, a progressive and deadly disease. The cumulative death rate is nearly 20%, even with current treatments. Many are not eligible for a renal transplant, and many cannot withstand long term hemodialysis.

Cascade is developing Renalta™, a:
once in a lifetime single-dose
lentivirus delivered
complement depletion therapy
lentivirus incorporates permanently into the renal cell genome and Renalta is continuously secreted to protect the kidney

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Cognitive Disorders: Schizophrenia

Schizophrenia is a life long psychiatric disorder that often strikes in the late teens or early twenties.
There is strong genetic component that is also significantly shaped by environmental factors.
The disease often involves excessive loss of gray matter (Cannon TD, et al. (2002) Cortex mapping reveals regionally specific patterns of genetic and disease- specific gray-matter deficits in twins discordant for schizophrenia. Proceedings of the National Academy of Sciences of the United States of America. 2002; 99:3228–32332) (Cannon TD et al.,(2015) Progressive reduction in cortical thickness as psychosis develops: a multisite longitudinal neuroimaging study of youth at elevated clinical risk. Biol Psychiatry Jan 15;77(2):147-57. doi: 10.1016/j.biopsych.2014.05.023) and reduced numbers of synaptic structures on neurons, (Garey LJ, et al. Reduced dendritic spine density on cerebral cortical pyramidal neurons in schizophrenia. J Neurol Neurosurg Psychiatry. 1998; 65:446–453).
Schizophrenia is associated with decreased dendritic spine density on prefrontal cortical pyramidal neurons (Glausier JR, Lewis DA. Dendritic spine pathology in schizophrenia. Neuroscience. 2013; 251:90– 107).
Consequently cognition, memory and perception are often impaired.
Psychotic symptoms (positive which are: hallucinations, delusions, thought disorders and paranoia, even movement disorders: negative which are: flat affect, lack or loss of emotion and speech, reduced sense of pleasure, difficultly initiation, maintaining activity) of schizophrenia are treated medicinally with varying degrees of success.
The cause and mechanisms involved with schizophrenia are not well understood.
Significant effort has been put forth to understand the genetic basis of schizophrenia.
More than 100 loci in the human genome contain SNP haplotypes that associate with risk of schizophrenia (Schizophrenia Working Group of the Psychiatric Genomics Consortium. Biological insights from 108 schizophrenia-associated genetic loci. Nature. 2014; 511:421–427).
There is an association between schizophrenia and genetic markers across the Major Histocompatibility Complex (MHC) locus.
The MHC locus is known for its role in immunity, containing highly polymorphic human leukocyte antigen (HLA) genes that encode many antigen-presenting molecules. Autoimmune diseases may have genetic associations at the MHC locus that arise from alleles of HLA genes. Schizophrenia’s association to the MHC is not yet explained.
Studies have shown that strongly associated markers are near a complex, multi-allelic, and partially characterized form of genome variation that affects the C4 gene encoding complement component 4. Schizophrenia is associated to CSMD16,10, which encodes a regulator of C413. (Sekar. et al., (2016) Schizophrenia risk from complex variation of complement component 4 Nature. February 11; 530(7589): 177–183).
In addition, very recently C5 levels in CSF were abnormally high in several psychiatric disorders, including bipolar disorder and schizophrenia. (Ishii T et al. (2018) Increased cerebrospinal fluid complement C5 levels in major depressive disorder and schizophrenia.Biochem Biophys Res Commun. Mar 4;497(2):683-688).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Multiple Sclerosis-Autoimmune

High levels of complement system factors are found in blood and cerebrospinal fluid of MS patients, suggesting that the system is actively contributing to disease.
Studies have shown that increased gray matter lesions predict disease course. During brain development, the complement system is needed for synaptic pruning or elimination of excess synapses.
The complement system may also facilitate synaptic pruning in neurodegenerative diseases, but in the case of MS, the amount of synaptic pruning is incorrect.
Studies have confirmed this unregulated pruning by showing the presence of certain activated complement factors, along with a lack of molecules that normally control them.
One consequence of unregulated synaptic pruning could be lesions in brain tissue. Indeed, it has been shown that a large set of complement factors, receptors, and regulatory molecules were present in the brains of deceased MS patients. High levels of activated complement can be found in gray matter lesions.
These lesions are found in both in the surface, and in deeper brain regions. (Watkins et al. (2016) Complement is activated in progressive multiple sclerosis cortical grey matter lesions. Journal of Neuroinflammation 13:161).
The extent of nerve cell loss in a brain area was linked to the numbers of cells with complement factors in that area.
Complement is activated in the MS cortical grey matter lesions in areas of elevated numbers of complement receptor-positive microglia and suggests that complement over-activation may contribute to the worsening pathology that underlies the irreversible progression of MS.

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Neuromyelitis Optica-Autoimmune

Neuromyelitis optica (NMO) is an autoimmune inflammatory disease that selectively targets the optic nerves and spinal cord, leading to blindness and paralysis (J. Ratelade, et al. (2013) Involvement of antibody- dependent cell-mediated cytotoxicity in inflammatory demy- elination in a mouse model of neuromyelitis optica. Acta Neuropathologica, vol. 126, no. 5, pp. 699–709).
Since NMO patients often present demyelinating lesions in the central nervous system (CNS), it has long been considered a variant of multiple sclerosis (MS); however, recent data suggest that its pathogenesis may be different (M. Jurynczyk et al. (2015) Overlapping CNS inflammatory diseases: differentiating features of NMO and MS,.Journal of Neurology, Neurosurgery, and Psychiatry, vol. 86, no. 1, pp. 20–25).
The immunopathology of NMO includes restricted demyelination and inflammation of the optic nerves and several spinal segments (Li and Y. Yan. Experimental models of neuromyelitis optica: current status, challenges and future directions. Neuroscience Bulletin, vol. 31, no. 6, pp. 735–744, 2015).
Combination Treatment of C16 Peptide and Angiopoietin-1 Alleviates Neuromyelitis Optica in an Experimental Model (Mediators of Inflammation Volume 2018, Article ID 4187347).
In contrast to multiple sclerosis, autoantibodies against aquaporin-4 (AQP4) expressed on astrocytic end-feet have been exclusively detected in sera of NMOSD patients.
Several lines of evidence suggested that anti-AQP4 autoantibodies are pathogenic, but the mechanism triggering inflammation, impairment of astrocyte function, and the role of neutrophils presented in NMOSD cerebrospinal fluid remains unknown. pathogenicity of NMOSD serum, which by consecutive action of anti-AQP4 Abs, complement system, and neutrophils affected astrocyte function. Anti-AQP4 Ab binding astrocytes initiated two parallel complementary reactions.
The first one was dependent on the complement cytotoxicity via C5b-9 complex formation, and the second one on the reverse of complex formation, and the second one on the reverse of astrocyte glutamate pump into extracellular space by C5a pre-activated neutrophils. Piatek P, et al. (2018) C5a Pre-activated Neutrophils Are Critical for Autoimmune-Induced Astrocyte Dysregulation In Neuromyelitis Optica Spectrum Disorder .Front Immunol. 23 July 2018 doi: 10.3389/fimmu.2018.01694).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Myasthenia Gravis-Autoimmune

Myasthenia gravis (MG) is primarily caused by antibodies directed towards the skeletal muscle acetylcholine receptor, leading to muscle weakness.
Although these antibodies may induce compromise of neuromuscular transmission by blocking acetylcholine receptor function, the predominant mechanism of injury to the neuromuscular junction is complement-mediated lysis of the postsynaptic membrane.
Complement plays a central role in EAMG and MG pathogenesis, as well as an important role in T- and B-cell function. Complement inhibitor-based therapy has great potential to treat human MG.
Complement is important in both innate and adaptive immunity (Kemper C, Atkinson JP. T-cell regulation: with complements from innate immunity. Nat Rev Immunol. 2007;7(1):9–18) (Morgan BP and Kavanaugh D. Introduction to complement in health and disease. Semin Immunopathol. 2018 Jan;40(1):1-2).
Activation of the complement system protects the host against invading pathogens by distinct mechanisms, which include cell lysis of pathogens, opsonization with complement fragments, chemotaxis of inflammatory cells and formation of the membrane attack complex (MAC) ( Kohl J. Self, non-self, and danger: a complementary view. Adv Exp Med Biol. 2006;586:71–94).
In the adaptive immune response, complement is the effector system for the primary and secondary antibody responses of B cells (Boackle SA, Holers VM. Role of complement in the development of autoimmunity. Curr Dir Autoimmun. 2003;6:154–168.) (Blank M, Schoenfeld Y. B cell targeted therapy in autoimmunity. J Autoimmun. 2007;28(2–3):62–68).
Complement activation is regulated by a series of approximately 30 plasma and membrane proteins participating in classical, alternative and lectin pathways. The classical pathway is activated by immune complexes containing antigen and IgM or complement fixing IgG (Austen KF, Feeron DT. A molecular basis of activation of the alternative pathway of human complement. Adv Exp Med Biol. 1979;120B:3–17).
The alternative pathway is activated by foreign pathogens and polymeric IgA. (Austen KF, Feeron DT. A molecular basis of activation of the alternative pathway of human complement. Adv Exp Med Biol. 1979;120B:3–17.) (Bogers WM et al. Complement enhances the clearance of large- sized soluble IgA aggregates in rats. Eur J Immunol. 1991;21(5):1093–1099).
The lectin pathway is initiated by binding of mannose-binding lectin to microbial pathogens and may involve IgA-containing immune complexes (Roos A et al. IgA (2001) and activates the complement system via the mannan-binding lectin pathway. J Immunol. 167(5):2861–2868.) (Thiel S, et al. A second serine protease associated with mannan-binding lectin that activates complement. Nature. 1997;386(6624):506–510).
Although initiated differently, all three pathways converge at the cleavage of C3 by specific convertase enzymes followed by the generation of the MAC.
The existence of a new activation pathway by which generation of C5a through a coagulation pathway involving thrombin in the absence of C3 has been proposed (Huber-Lang Met al. Generation of C5a in the absence of C3: a new complement activation pathway. Nat Med. 2006;12(6):682–687).
Complement activity is modulated by regulatory proteins that prevent the cascade from progressing toward tissue damage as a result of inadvertent binding of activated complement components. (Kusner et al. 2008 Effect of complement and its regulation on myasthenia gravis pathogenesis Expert Rev Clin Immunol. January ; 4(1): 43–520).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Amyotrophic Lateral Sclerosis (ALS)-Autoimmune

Amyotrophic lateral sclerosis (ALS) is a fatal progressive neurodegenerative disease with no available therapy. Components of the innate immune system are activated in the spinal cord and central nervous system of ALS patients.
Studies in the SOD1G93A mouse show deposition of C1q and C3/C3b at the motor end-plate before neurological symptoms are apparent, suggesting that complement activation precedes neurodegeneration in this model.
Analyzing post-mortem tissue of ALS donors may lead to a better understanding of the complement system’s role and it’s activation at the motor end-plate in human ALS. (Bahia , Idrissi , Bosch , Ramaglia , Aronica , Baas Troost D (2016) Complement activation at the motor end-plates in amyotrophic lateral sclerosis. J Neuroinflammation. Apr 7;13(1):72. doi: 10.1186/s12974-016-0538-2).

Cascade Biotechnology INC| Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Retinal and Corneal Disease

Age-related macular degeneration

Age-related macular degeneration (AMD) is the leading cause of irreversible vision loss among the elderly, accounting for 8.7% of blindness worldwide. Globally, the total number of patients with any type of AMD is expected to increase over the next 25 years to 288 million affected individuals (Wong et al., 2014).
The disease is characterized by a gradual loss of central vision due to photoreceptor cell degeneration in the center of the retina at the back of the eye, known as the macula.
Photoreceptors are in close contact with a layer of cells called the retinal pigment epithelium (RPE). RPE cells support the function of the photoreceptors and play an important role in maintaining retinal homeostasis.
In AMD, this natural function of the RPE is disturbed, resulting in the accumulation of retinal waste products called drusen underneath the RPE.
Two forms of advanced AMD are distinguished. Neovascular AMD, is characterized by infiltration of abnormal blood vessels into the retina.
These newly formed vessels are fragile and when they break, the leakage of blood constituents in the retina leads to sudden vision loss.
The second form of advanced AMD, geographic atrophy, is the result of gradual degeneration of the RPE and photoreceptors cells.
However, no treatment is available for the remaining majority of early, intermediate or geographic atrophy AMD cases, and furthermore there are no effective means of preventing progression of early to advanced stages (Chakravarthy et al., 2010a, Jager et al., 2008).
The complement system plays a central role in the etiology of AMD2. it is known that AMD is the result of a complex interaction of environmental and genetic risk factors.
Studies into the molecular constituents of drusen had already suggested that AMD may have an immunological component. This suggestion arose after proteins involved in inflammation and/or other immune-associated responses, including components of the complement system, were found within drusen. (Hageman et al., 2001, Johnson et al., 2001, Mullins et al., 2001).Evidence for a strong genetic component in AMD arose from twin and family studies.
Twin studies showed high concordance between AMD between monozygotic pairs, even double compared to dizygotic pairs, and estimated that the heritability of AMD may be as high as 45 to 70% (Hammond et al., 2002, Meyers et al., 1995, Seddon et al., 2005).
These findings were consistent with familial aggregation analyses that observed a higher prevalence of AMD characteristics and an earlier onset of disease symptoms among relatives of patients compared to control families (Klaver et al., 1998, Seddon et al., 1997).
There is genetic evidence for a role of the complement system in AMD. (Abecasis et al., 2004, Iyengar et al., 2004, Majewski et al., 2003, Seddon et al., 2003, Weeks et al., 2004).
When the first genome-wide association study (GWAS) for AMD was performed in 2005, it identified that same genomic region, which lead to the discovery of a highly associated genetic variant in complement factor H (CFH; Tyr402His) (Klein et al., 2005).
These findings were corroborated by three additional studies (Edwards et al., 2005, Hageman et al., 2005, Haines et al., 2005).
Through genetic studies that followed over the next decade, the understanding of the genetic basis of AMD increased dramatically with the identification of disease-associated variants across several biological systems (Fritsche et al., 2013).
The genetic link between AMD and the complement genetic variants in or near complement factor I (CFI), complement component 3 (C3), complement component 2 (C2), complement component 9 (C9), complement factor B (CFB) and vitronectin (VTN) were also found to be associated with the disease (Fagerness et al., 2009, Fritsche et al., 2013, Fritsche et al., 2016, Gold et al., 2006, Maller et al., 2007, Yates et al., 2007).
In addition, a common haplotype carrying a deletion of complement factor H related genes CFHR1 and CFHR3 was found to be protective for AMD (Hughes et al., 2006).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Blood Disorders

Paroxysmal nocturnal hemoglobinuria (PNH)

When the complement system is activated, it triggers a variety of events leading to cleavage of protein C5. Once C5 is cleaved, a variety of events occur that propagate the formation of the membrane attack complex (MAC).
This MAC generates pores, or holes, in cells ultimately leading to the cellʼs demise of the system that are needed. These regulators sit on the outer membrane of cells, so the complement system recognizes that these cells are of the self.
When those regulators are missing, as is the case in PNH, this leads to the destruction of the cells that are missing protein shields. Some of those shields, (2 proteins known as CD 55 and CD 59) are anchored the cell surface by a ‘tailʼ.
We call this tail a GPI anchor – but in PNH this GPI anchor is missing because of a mutation in a gene called PIG-A. This defective gene causes the cellʼs inability to form this GPI anchor.
So the complement regulator proteins are lost because they arenʼt anchored to the cell surface.
When the complement system becomes highly active from infections, surgery, or similar events, it creates increased cell death of those cells missing this protein shield.
Because of the missing CD 59 protein [note: CD 55 is not mentioned here] on the surface of the red blood cell the membrane attack complex takes place, which makes the holes and pores on the cell surface, releasing the hemoglobin inside the cell through the holes – the hemoglobin escapes the cell walls.
This is the point where hemolysis occurs. Eventually the cell completely ruptures, releasing all the free hemoglobin intravascularly.
That has a variety of consequences, including hemolytic anemia , thrombosis because of inflammation, and kidney problems because of free hemoglobin filtering through the kidney tubes—leading to hemoglobinuria (red urine).
Continued hemoglobinuria can lead to kidney damage.(Expert Rev Hematol. 2014 October ; 7(5):583–598).
Because of the missing CD 59 protein [note: CD 55 is not mentioned here] on the surface of the red blood cell , the membrane attack complex takes place, which makes the holes and pores on the cell surface, releasing the hemoglobin inside the cell through the holes – the hemoglobin escapes the cell walls.
This is the point where hemolysis occurs. Eventually the cell completely ruptures, releasing all the free hemoglobin intravascularly.
That has a variety of consequences, including hemolytic anemia , thrombosis because of inflammation, and kidney problems because of free hemoglobin filtering through the kidney tubes—leading to hemoglobinuria (red urine).
Continued hemoglobinuria can lead to kidney damage. In its normal form, patients present with overt hemolysis and hemoglobinuria. Subclinical PNH implies that you have a PNH clone with defective hemolysis.
The ability of high-sensitivity flow cytometry to identify a small amount of PNH clones has resulted in the classification called ‘subclinical PNHʼ. So these patient may not present with hemolysis but once a PNH cline is identified, it is important to monitor the sized of the clone and understand the potential consequences of clone growth and the potential for hemolytic events to begin (Jamile Shammo, MD, FACP, FASCP Interviews with the Experts: The Complement System in PNH | Aplastic Anemi and MDS International Foundation (AA MDS) Sun, 03/31/2013 - 8G37pm. Last updated on Fri, 03/11/2016).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Coronary Disease

Cardiac and Myocardial Ischemia

Regarding inflammation as one mediator of I/R injury, experimental and clinical studies have shown that reperfusion after transient ischemia results in activation of endothelial cells, the contact and the complement system and attraction of neutrophils to the site of infarction (Wouters D et al. (2008) C1 inhibitor: just a serine protease inhibitor? New and old considerations on therapeutic applications of C1 inhibitor. Expert Opin Biol Ther 8(8):1225–40).
Complement-mediated ischemia-reperfusion injury: lessons learned from animal and clinical studies. Diepenhorst GM et al., (2009) complement-mediated ischemia-reperfusion injury: lessons learned from animal and clinical studies. Ann Surg 249(6):889–99).
The complement system is a major component of innate immunity, which is involved in both recognition and response to pathogens (Walport MJ. (2001) Complement. Second of two parts. N Engl J Med 344(15):1140–4).
It is further implicated in an increasing number of homeostatic and disease processes such as the immune complex catabolism, the clearance of dead and dying cells and the modulation of adaptive immune responses (Carroll MC. (2004)The complement system in regulation of adaptive immunity. Nat Immunol. 5(10):981–6. doi:10.1038/ni1113).
Three pathways can activate the complement cascade: the classical, the alternative, and the lectin pathway. After initiation, these three pathways converge at the level of cleavage and activation of complement component C3, which subsequently leads to the generation of the anaphylatoxins (C3a, C5a) and the membrane attack complex (MAC; C5b-9). (Panagiotou et al. 2018 The lectin pathway of Complement in Myocardial Ischemia/Reperfusion Injury—Review of Its Significance and the Potential Impact of Therapeutic Interference by C1 Esterase Inhibitor. Frontiers in Immunology | Volume 9 | Article 1151).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Renal Disease

Atypical Hemolytic Uremic Syndrome (aHUS)

The complement cascade provides an important line of defense against invasive pathogens. However, the complement system also causes kidney injury in a variety of different diseases.
Complement activation is particularly important in the development of and C3 glomerulopathy. Complement inhibition is effective for treatment of aHUS, and complement inhibitors will likely be tested in other kidney diseases in the future.
While the role of the complement system in the pathogenesis of many kidney diseases is well established, however, there is not a simple algorithm for identifying which patients should be treated with complement inhibitors or for how long complement inhibition should be continued. (Joshua M. Thurman (2015) Complement in Kidney Disease: Core Curriculum 2015 Am J Kidney Dis. 2015 Jan; 65(1): 156–168).
Complement cascade is involved in several renal diseases and in renal transplantation. The different components of the complement cascade might represent an optimal target for innovative therapies.
Study of complement involvement in renal diseases and transplantation has led to a reclassification of some renal diseases, moving from a histopathological to a physiopathological classification.
Renal diseases involve complement over activation and complement dysregulation.
Many targets such as C1, C3, C5a and C5aR are currently the subject of national or international trials.
In addition, many molecules proved to be effective in vitro or in preclinical trials and are waiting to move to human trials in the future. (Maurizio Salvadori, Giuseppina Rosso, and Elisabetta Bertoni (2015) Complement involvement in kidney diseases: From physiopathology to therapeutical targeting World J Nephrol. 2015 May 6; 4(2): 169–184).

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Our Science:Technology

**Discovery Chemistry Design and High Throughput in silico/in vitro screening**

Cascade has established a robust platform to design anti-complement drugs that target and inhibit the complement system at various levels.We rely upon a core set of chemical modifications to tailor the specificity, potency, and safety of our small molecules. Cascade benefits from over 30 years of innovation and experience in drug design and discovery. We have state of the art in silico protein modeling and drug docking technology to drive novel drug synthesis, and novel cell-based model systems to test activity and predict bioavailability.

Protein engineering and Gene Therapy

Cascade has state of the art in silico protein modeling and analysis technology to create superior biologics based on complement proteins. Our in silico platform uses advanced immunogenicity prediction using massive informational libraries of immunogenic motifs. We have a full BL2 cell culture facility to create and test our therapeutic proteins. Cascade has developed novel purified protein, cell-based and ex vivo model systems to test activity and predict bioavailability. We are using cutting edge lentivirus technology in the development of our gene therapy agents. Because the future of healthcare will likely involve multiple gene therapy interventions per individual, our lentiviral vectors provide a significant competitive advantage. Lentivirus produces minimal immune activation and allows for subsequent gene therapy treatments. A single adenovirus gene therapy would produce an immune response to subsequent adenovirus treatments, thus making it practical only to give one treatment in a lifetime.

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Staff Publications

Vogel CW, Fritzinger DC, Gorsuch WB et al. Complement depletion with humanised cobra venom factor: efficacy in preclinical models of vascular diseases. Thromb Haemost 2015;113:548-52
Vogel CW, Finnegan PW, Fritzinger DC. Humanized cobra venom factor: structure, activity, and therapeutic efficacy in preclinical disease models. Mol Immunol 2014;61:191-203
Huda R, Fritzinger DC, Finnegan PF, et al. Complement depletion with humanized Cobra Venom Factor (CVF) improves the severity of Experimental Autoimmune Myasthenia Gravis (EAMG). Mol Immunol 2011;48:1712
Fritzinger DC, Dean R, Meschter C, et al. Complement depletion with humanized cobra venom factor in a mouse model of age-related macular degeneration. Adv Exp Med Biol 2010;703:151-62
Fritzinger DC, Damaj BB, Wong K, et al. Pharmacokinetics of humanized cobra venom factor in mice. Mol Immunol 2010;47:2268-2269
Vogel C-W, Fritzinger DC. Cobra venom factor: structure, function, and humanization for therapeutic complement depletion. Toxicon 2010;56:1198-1222
Wang SY, Veeramani S, Racila E, et al. Depletion of the C3 component of complement enhances the ability of rituximab-coated target cells to activate human NK cells and improves the efficacy of monoclonal antibody therapy in an in vivo model. Blood 2009;114:5322-30
Gorsuch WB, Guikema BJ, Fritzinger DC, et al. Humanized cobra venom factor decreases myocardial ischemia-reperfusion injury. Mol Immunol 2009;47:506-10
Fritzinger DC, Hew BE, Thorne M, et al. Functional characterization of human C3/cobra venom factor hybrid proteins for therapeutic complement depletion. Dev Comp Immunol 2009;33:105–116
Fritzinger DC, Ferreira VP, Hew BE, et al. A novel concept for the treatment of paroxysmal nocturnal haemoglobinuria (PNH): complement depletion with a human C3 derivative with cobra venom factor-like activity prevents lysis of PNH erythrocytes. Mol Immunol 2008;45:4177
Fritzinger DC, Hew BE, Lee JQ, et al. Human C3/cobra venom factor hybrid proteins for therapeutic complement depletion: in vivo activity and lack of toxicity in primates. Mol Immunol 2008;45:4112
Fritzinger DC, Hew BE, Lee JQ, et al. Derivatives of human complement component C3 for therapeutic complement depletion: a novel class of therapeutic agents. Adv Exp Med Biol 2008;632:293-307
Vogel CW, Fritzinger DC. Humanized cobra venom factor: experimental therapeutics for targeted complement activation and complement depletion. Curr Pharm Des 2007;13:2916-26
Fritzinger DC. Cancer research center hotline: complement depletion: use of human C3/cobra venom factor (CVF) chimeric proteins as therapeutic agents. Hawaii Med J 2005;64:133-4, 137

Fritzinger DC, Benjamin DE. The Complement System in Neuropathic and Postoperative Pain. Open Pain J. 2016 ; 9: 26–37

Brucato FH, Benjamin DE. Synaptic Pruning in Alzheimer's Disease:Role of the Complement System. Global Journal of Medical Research F:Diseases 2020;Volume 20 Issue 6 Version1.0. ISSN: 2249-4618 & Print ISSN:0975-5888

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Funding

NIH Funding

Grant Number: 1 R43 HL139170-01A1Principal Investigator: Fritzinger, David Charles Project Title: Preparation and development of a stabilized C3 analog for the treatment of paroxysmal nocturnal hemoglobinemia Institution: CASCADE BIOTECHNOLOGY Award Issue Date: 09/21/2018

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Baerbel Rohrer, Ph.D.

Professor of OphthalmologyMedical University of South Carolina

Claes Wahlestedt, MD, Ph.D.

Professor University of Miami Miller School of Medicine

www.scripps.edu/florida/neuro/cwahlestedt.html

Anish Suri, Ph.D.

Chief Scientific Officer, Cue Biopharma

https://be.linkedin.com/in/anishsuri

Daniel E. Benjamin Ph.D.

Daniel E. Benjamin Ph.D. is the Chief Executive Officer (CEO) of our company. With his extensive knowledge and experience, he leads the organization in setting strategic goals and implementing effective business strategies. He oversees the overall operations of the company, including financial performance, resource allocation, and risk management. Under his leadership, the company has achieved significant growth and success.

Edwin Holland B.A.

Chief Operating Officer, V.P. Business Development

David Frizinger Ph.D.

Chief Biotechnology Officer

Duncan Taylor Ph.D.

Consultant to Business Development

Dalakas, M. C., Alexopoulos, H., & Spaeth, P. J. (2020). Complement in neurological disorders and emerging complement-targeted therapeutics. In Nature Reviews Neurology (Vol. 16, Issue 11, pp. 601–617). Nature Research. https://doi.org/10.1038/s41582-020-0400-0

Goetzl, E. J., Srihari, V. H., Guloksuz, S., Ferrara, M., Tek, C., & Heninger, G. R. (2020). Decreased mitochondrial electron transport proteins and increased complement mediators in plasma neural-derived exosomes of early psychosis. Translational Psychiatry 10 (1). https://doi.org/10.1038/s41398-020-01046-3

Kamitaki, N., Sekar, A., Handsaker, R. E., de Rivera, H., Tooley, K., Morris, D. L., Taylor, K. E., Whelan, C. W., Tombleson, P., Loohuis, L. M. O., Ripke, S., Neale, B. M., Corvin, A., Walters, J. T. R., Farh, K. H., Holmans, P. A., Lee, P., Bulik-Sullivan, B., Collier, D. A., … Spencer, C. C. A. (2020). Complement genes contribute sex-biased vulnerability in diverse disorders. Nature https://doi.org/10.1038/s41586-020-2277

Kopczynska, M., Zelek, W., Touchard, S., Gaughran, F., Di Forti, M., Mondelli, V., Murray, R., O’Donovan, M. C., Morgan, B. P. (2019). Complement system biomarkers in first episode psychosis. Schizophrenia Research 204,16–22. https://doi.org/10.1016/j.schres.2017.12.012

Magdalon, J., Mansur, F., Teles e Silva, A. L., de Goes, V. A., Reiner, O., Sertié, A. L. (2020). Complement System in Brain Architecture and Neurodevelopmental Disorders. In Frontiers in Neuroscience (Vol. 14). Frontiers Media S.A. https://doi.org/10.3389/fnins.2020.00023

Mongan, D., Sabherwal, S., Susai, S. R., Föcking, M., Cannon, M., Cotter, D. R. (2020). Peripheral complement proteins in schizophrenia: A systematic review and meta-analysis of serological studies. In Schizophrenia Research (Vol. 222, pp. 58–72). Elsevier B.V. https://doi.org/10.1016/j.schres.2020.05.036

Purves-Tyson, T. D., Robinson, K., Brown, A. M., Boerrigter, D., Cai, H. Q., Weissleder, C., Owens, S. J., Rothmond, D. A., & Shannon Weickert, C. (2020). Increased Macrophages and C1qA, C3, C4 Transcripts in the Midbrain of People With Schizophrenia. Frontiers in Immunology 11 https://doi.org/10.3389/fimmu.2020.02002

Sellgren, C. M., Gracias, J., Watmuff, B., Biag, J. D., Thanos, J. M., Whittredge, P. B., Fu, T., Worringer, K., Brown, H. E., Wang, J., Kaykas, A., Karmacharya, R., Goold, C. P., Sheridan, S. D. Perlis, R. H. (2019). Increased synapse elimination by microglia in schizophrenia patient-derived models of synaptic pruning. Nature Neuroscience 22 (3), 374–385. https://doi.org/10.1038/s41593-018-0334-7

Wallace, J., Lord, J., Dissing-Olesen, L., Stevens, B., Murthy, V. N. (2020). Microglial depletion disrupts normal functional development of adult-born neurons in the olfactory bulb. ELife. https://doi.org/10.7554/eLife.50531

Woo, J. J., Pouget, J. G., Zai, C. C., Kennedy, J. L. (2020). The complement system in schizophrenia: where are we now and what’s next? In Molecular Psychiatry (Vol. 25, Issue 1, pp. 114–130). Springer Nature. https://doi.org/10.1038/s41380-019-0479-0

Cascade Biotechnology INC.

Complement System Therapeutics

Announcements

Complement Therapeutics General Overview

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

The complement system is an important part of the innate immune system, which helps to protect the host against infection and connects innate and adaptive immune systems.

Complement consists of more than 30 proteins that are activated through a series of proteolytic cleavages, which in turn activate proteins in the next step of the complement cascade.

While complement proteins serve an important regulatory role in inflammation, over activation of complement ultimately leads to disease states. Inhibiting overactive complement offers one strategy for managing and potentially preventing a range of disease and disorders.

See: Fritzinger DC, Benjamin DE (2016) The Complement System in Neuropathic and Postoperative Pain. Open Pain J. (2016);9:26-37.*By using the Cascade Biotechnology, Inc. website, without changing your settings,you agree to our use of cookies to enhance your browsing experience .

Complement Therapeutics Disease Indications

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Complement resolves immunological problems, yet over-activation may lead to these diseases:

Renal: Diabetic Nephropathy, Lupus Nephritis

Neurological & autoimmune: Myasthenia Gravis (MG), Multiple Sclerosis (MS), Neuromyelitis Optica (NMO)

Retinal: Age-related macular degeneration (AMD)

Cognitive: Alzheimer’s Disease (AD), Prodromal Schizophrenia (SZ)

Pipeline

Diabetic Nephropathy (DN) and lupus nephritis (LN)

Schizophrenia

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Cognitive Disorders: Schizophrenia

Schizophrenia is a life long psychiatric disorder that often strikes in the late teens or early twenties.

There is strong genetic component that is also significantly shaped by environmental factors.

Schizophrenia is associated with decreased dendritic spine density on prefrontal cortical pyramidal neurons (Glausier JR, Lewis DA. Dendritic spine pathology in schizophrenia. Neuroscience. 2013; 251:90– 107).

Consequently cognition, memory and perception are often impaired.

The cause and mechanisms involved with schizophrenia are not well understood.

Significant effort has been put forth to understand the genetic basis of schizophrenia.

More than 100 loci in the human genome contain SNP haplotypes that associate with risk of schizophrenia (Schizophrenia Working Group of the Psychiatric Genomics Consortium. Biological insights from 108 schizophrenia-associated genetic loci. Nature. 2014; 511:421–427).

There is an association between schizophrenia and genetic markers across the Major Histocompatibility Complex (MHC) locus.

Multiple Sclerosis-Autoimmune

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Multiple Sclerosis-Autoimmune

High levels of complement system factors are found in blood and cerebrospinal fluid of MS patients, suggesting that the system is actively contributing to disease.

Studies have shown that increased gray matter lesions predict disease course. During brain development, the complement system is needed for synaptic pruning or elimination of excess synapses.

The complement system may also facilitate synaptic pruning in neurodegenerative diseases, but in the case of MS, the amount of synaptic pruning is incorrect.

Studies have confirmed this unregulated pruning by showing the presence of certain activated complement factors, along with a lack of molecules that normally control them.

These lesions are found in both in the surface, and in deeper brain regions. (Watkins et al. (2016) Complement is activated in progressive multiple sclerosis cortical grey matter lesions. Journal of Neuroinflammation 13:161).

The extent of nerve cell loss in a brain area was linked to the numbers of cells with complement factors in that area.

Complement is activated in the MS cortical grey matter lesions in areas of elevated numbers of complement receptor-positive microglia and suggests that complement over-activation may contribute to the worsening pathology that underlies the irreversible progression of MS.

Neuromyelitis Optica-Autoimmune

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Neuromyelitis Optica-Autoimmune

The immunopathology of NMO includes restricted demyelination and inflammation of the optic nerves and several spinal segments (Li and Y. Yan. Experimental models of neuromyelitis optica: current status, challenges and future directions. Neuroscience Bulletin, vol. 31, no. 6, pp. 735–744, 2015).

Combination Treatment of C16 Peptide and Angiopoietin-1 Alleviates Neuromyelitis Optica in an Experimental Model (Mediators of Inflammation Volume 2018, Article ID 4187347).

In contrast to multiple sclerosis, autoantibodies against aquaporin-4 (AQP4) expressed on astrocytic end-feet have been exclusively detected in sera of NMOSD patients.

Myasthenia Gravis-Autoimmune

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Myasthenia Gravis-Autoimmune

Myasthenia gravis (MG) is primarily caused by antibodies directed towards the skeletal muscle acetylcholine receptor, leading to muscle weakness.

Although these antibodies may induce compromise of neuromuscular transmission by blocking acetylcholine receptor function, the predominant mechanism of injury to the neuromuscular junction is complement-mediated lysis of the postsynaptic membrane.

Complement plays a central role in EAMG and MG pathogenesis, as well as an important role in T- and B-cell function. Complement inhibitor-based therapy has great potential to treat human MG.

Complement is important in both innate and adaptive immunity (Kemper C, Atkinson JP. T-cell regulation: with complements from innate immunity. Nat Rev Immunol. 2007;7(1):9–18) (Morgan BP and Kavanaugh D. Introduction to complement in health and disease. Semin Immunopathol. 2018 Jan;40(1):1-2).

Although initiated differently, all three pathways converge at the cleavage of C3 by specific convertase enzymes followed by the generation of the MAC.

The existence of a new activation pathway by which generation of C5a through a coagulation pathway involving thrombin in the absence of C3 has been proposed (Huber-Lang Met al. Generation of C5a in the absence of C3: a new complement activation pathway. Nat Med. 2006;12(6):682–687).

Amyotrophic Lateral Sclerosis (ALS) Autoimmune

Cascade Biotechnology INC | Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Neurological Diseases

Amyotrophic Lateral Sclerosis (ALS)-Autoimmune

Amyotrophic lateral sclerosis (ALS) is a fatal progressive neurodegenerative disease with no available therapy. Components of the innate immune system are activated in the spinal cord and central nervous system of ALS patients.

Studies in the SOD1G93A mouse show deposition of C1q and C3/C3b at the motor end-plate before neurological symptoms are apparent, suggesting that complement activation precedes neurodegeneration in this model.

Age-Related Macular Degeneration

Cascade Biotechnology INC| Complement Therapeutics; novel approach to CNS/PNS disease management using the innate complement system.

Retinal and Corneal Disease

Age-related macular degeneration

The disease is characterized by a gradual loss of central vision due to photoreceptor cell degeneration in the center of the retina at the back of the eye, known as the macula.

Photoreceptors are in close contact with a layer of cells called the retinal pigment epithelium (RPE). RPE cells support the function of the photoreceptors and play an important role in maintaining retinal homeostasis.

In AMD, this natural function of the RPE is disturbed, resulting in the accumulation of retinal waste products called drusen underneath the RPE.

Two forms of advanced AMD are distinguished. Neovascular AMD, is characterized by infiltration of abnormal blood vessels into the retina.

These newly formed vessels are fragile and when they break, the leakage of blood constituents in the retina leads to sudden vision loss.

The second form of advanced AMD, geographic atrophy, is the result of gradual degeneration of the RPE and photoreceptors cells.

However, no treatment is available for the remaining majority of early, intermediate or geographic atrophy AMD cases, and furthermore there are no effective means of preventing progression of early to advanced stages (Chakravarthy et al., 2010a, Jager et al., 2008).

The complement system plays a central role in the etiology of AMD2. it is known that AMD is the result of a complex interaction of environmental and genetic risk factors.

Twin studies showed high concordance between AMD between monozygotic pairs, even double compared to dizygotic pairs, and estimated that the heritability of AMD may be as high as 45 to 70% (Hammond et al., 2002, Meyers et al., 1995, Seddon et al., 2005).

These findings were consistent with familial aggregation analyses that observed a higher prevalence of AMD characteristics and an earlier onset of disease symptoms among relatives of patients compared to control families (Klaver et al., 1998, Seddon et al., 1997).

There is genetic evidence for a role of the complement system in AMD. (Abecasis et al., 2004, Iyengar et al., 2004, Majewski et al., 2003, Seddon et al., 2003, Weeks et al., 2004).

When the first genome-wide association study (GWAS) for AMD was performed in 2005, it identified that same genomic region, which lead to the discovery of a highly associated genetic variant in complement factor H (CFH; Tyr402His) (Klein et al., 2005).

These findings were corroborated by three additional studies (Edwards et al., 2005, Hageman et al., 2005, Haines et al., 2005).

See: Fritzinger DC, Benjamin DE (2016) The Complement System in Neuropathic and Postoperative Pain. Open Pain J. (2016);9:26-37.

*By using the Cascade Biotechnology, Inc. website, without changing your settings,
you agree to our use of cookies to enhance your browsing experience .