Therapeutic Pipeline and Programs

Native high-density lipoproteins (HDLs) play roles in cholesterol metabolism, nucleic acid delivery, and inflammation. By mimicking features of native HDLs, Zylem has developed a proprietary platform of technologies that are strategically tuned to enable novel therapeutic functions while maintaining active targeting to the high-affinity ligand for native HDL called scavenger receptor type B-1 (SR-B1). Target disease types include: oncology, cardiovascular disease, topical ocular therapy (such as diabetic dry-eye disease and injury), psoriasis and atopic dermatitis.

Oncology

ZYL formulations for oncology target a multi-functional receptor (SR-B1) to simultaneously engage multiple hallmark mechanisms of cancer cell death.

SR-B1 is a cancer target. It is over-expressed in cancer cells versus normal cells and high expressing tumors correlate to worse outcomes. Tumor cells are dependent on native HDL binding SR-B1 to uptake cholesterol for survival – Increasing cholesterol removal leads to cancer cell death without affecting normal cells.

ZYL for oncology therapeutically drives natural cancer checkpoints into action:

1. Enhances immune destruction

2. Reduces tumor-promoting inflammation

3. Induces cell death

This induces a metabolic checkmate.

Our preclinical data in show multiple opportunities in in oncology:

Solid Tumor:

• Clear cell Ovarian

• Prostate

• Melanoma

• Radiosensitizer in breast and prostate

Hematological:

• Diffuse large B cell lymphoma (DLBCL)

• Chronic lymphocytic leukemia (CLL)

• Refractory/Resistant Acute Myeloid Leukemia

These cancers are the targets of our clinical strategy. Phase I clinical trial recruitment begins in the spring of 2025.

Mechanism of Action

ZYL simultaneously targets three hallmarks of cancer. This results in a metabolic checkmate for oncologic cells, while leaving healthy cells unscathed.

ZYL potently binds SR-B1 and effectively outcompetes native HDL. ZYL reduces cancer cell cholesterol, increases membrane oxidation (while simultaneously decreasing cellular antioxidant capability by down-regulating GPX4), and inhibits survival signaling. This metabolic checkmate is highly effective at abolishing cancer cells.

Cardiovascular Disease

Atherosclerotic cardiovascular disease (ASCVD) is a disease of non-functional HDL. Native High-density lipoproteins (HDL) sequester and transport cholesterol, provide anti-inflammatory and anti-oxidative effects, and reduce the risk of cardiovascular disease.

Non-functional HDL results in cardiovascular disease. CVD patients lack sufficient amounts of functional HDL. ZYL-CVD is engineered to support each protective function of native HDL, and reverse and inhibit CVD.

• Reverse Cholesterol transport

• Cholesterol efflux capacity

• Vasodilatory activity

• Antithrombotic activity

• Antioxidant activity

• Anti-inflammatory activity

ZYL-CVD Replaces Functionally Inactive HDLs as a Biomimetic to Inhibit and Reverse CVD.

• Enhanced Cholesterol Efflux

• Reduces Inflammation

• Increased Antioxidant Activity

• Eliminates Plaque Formation

Heart disease remains the most common cause of death in the world. Current treatments for cardiovascular disease leave undressed residual disease. ZYL-CVD specifically targets fat accumulating cells and reverses atherosclerotic plaques. And ZYL-CVD reduces inflammation/ oxidation. ZYL-CVD directly addresses the root causes of CVD.

ZYL-CVD addresses the failures of past treatment modalities, by being the first and only mature spherical mimic of native HDL. This breakthrough in engineering stems from building an HDL mimic using an organic polymer scaffold, whereupon the targeting protein apoA-I assembles and enables the self-assembling of HDL identifying lipids. Due to the unique spherical nature, ZYL-CVD has shown profound reduction of plaque formation in the arterial wall in preclinical disease models.

Phase I clinical trial for ZYL-CVD begins recruitment in 2025.

Partnership Opportunities

Topical Ocular Therapy - Preclinical

Native high-density lipoproteins (HDL) play a critical role in maintaining epi- and endo-thelial cell homeostasis and potently reduce inflammation. The high-affinity receptor for mature HDLs, SR-B1, is expressed by corneal epithelial cells, limbal epithelia cells, and stromal keratocytes. Topically applied first generation ZYL-003 demonstrated penetration into basal corneal epithelial cells and stromal keratocytes. In preclinical obesity models, where wound healing response is impaired, topically applied ZYL-003 significantly enhanced wound closure. Up-regulation of the Akt pathway by ZYL-003 led to re-epithelialization via increased F-actin and phosphorylation of EphA2-S897. Additional studies found ZYL-003 to attenuate initial stages of inflammation resulting from chemical burns and other stresses. Further iterations of ZYL-003 (i.e. lipids and core modifications) have led to the development of ZYL-004 and ZYL-005 that incorporate small molecules, like vitamin D and nucleic acids, respectively.

Zylem’s technology can be developed into topical eye drops to treat chronic inflammatory eye conditions, like dry eye. Also, there are several other conditions of the anterior eye treatable with ZYL eye drop formulations, such as diseases or eye injuries from chemical or thermal injury, long-term contact lends wear, severe chronic rosacea, Stevens-Johnson syndrome, atopic keratoconjunctivitis, bacterial keratitis, and post-surgical treatment to enhance wound healing.

Topical Skin Therapy - Preclinical

The skin is a highly controlled and dynamic biological barrier that protects the human body from external insults, stressors, and pathogens. The differentiation state of keratinocytes in the stratum corneum is critical for generating and preserving skin integrity via maintenance of extracellular ceramides, cholesterol, free fatty acids and other lipids. In the process of aging, skin barrier function becomes impaired by poor keratinocyte differentiation causing a lack of extracellular lipid maintenance, skin thinning, and poor barrier function. SR-B1 expressed on keratinocytes and Zylem has developed ZYL-006 therapy to promote keratinocytes differentiation. ZYL-006 binding to SR-B1 on keratinocytes enhances the expression of ABCA1, which leads to keratinocyte differentiation and extracellular lipid production.

ZYL-006 is also being explored as therapy for inflammatory skin conditions. SR-B1 is a shared receptor on immune cells (e.g. macrophages and dendritic cells) that reside in the skin. In conditions that arise from inflammation the immune cells drive a cytokine cascade that causes irritation, plaque formation, and redness of the epidermal layer. ZYL-006 binding to SR-B1 reduces proinflammatory cytokine release, thus preventing hyperactive keratinocyte activation leading to skin irritation and inflammation.

The Zylem team is currently formulating ZYL-006 for topical treatment for aging skin. Early-stage development of ZYL-006 has demonstrated success in improving skin thickening, barrier function, and lipid homeostasis, all of which are critical for optimal skin health. Further, ZYL-006 is currently being explored in applications where inflammation is a critical component of the disease process, such as in psoriasis and dermatitis. Additional topical ZYL-constructs are underway to explore lipid replenishment and vitamin D delivery to preserve the integrity of the skin barrier.

Antiviral Therapy - Preclinical

A novel therapeutic was developed by modifying the lipid composition of ZYL-001 to arrive at ZYL-002. This drug actively targets SR-B1 on the surface of airway epithelial cells, liver cells, and immune cells. The compound significantly inhibits viral entry into host cells and reduces inflammation. For SARS-CoV-2, ZYL-002 is being developed as an inhaled therapy to prevent and treat early-stage viral infection, and as an injectable drug to treat later stages of infection with visceral organ involvement.

Upon binding SR-B1, ZYL-002 depletes and disrupts cell membrane cholesterol in lipid raft micro-domains and prevents viral entry. Our preclinical data show up to 80% blockade of SARS-CoV-2 pseudovirus entry in human cell models. In addition, ZYL-002 binds SR-B1 on innate immune cells depleting cell membrane cholesterol and reducing NF-kB-mediated cytokine expression. Our data and that of others suggest that SR-B1 is a co-receptor for SARS-CoV-2 cell entry and that cholesterol is a critical membrane component for targeting and inhibition of viral cell entry.

Beyond, SARS-CoV-2 viral infection, ZYL-002 is applicable to other upper respiratory viruses and as a therapy to treat acute systemic inflammation, such as gram-negative sepsis. ZYL-002 is in early preclinical stage development with Zylem seeking partnerships for IND-enabling studies and clinical development.