L-DOS47: A Pharmacological, Tumor-Targeted Alkalization Therapy

One of the most consistent hallmarks of cancer biology is the acidity of solid tumors, a characteristic that gives cancer cells a survival advantage. This acidity is not a passive byproduct of cancer metabolism, as was first noted as early as the 1920s with observations of glycolytic metabolism in tumors, but is now understood as an active driver of tumor progression, immune evasion, metastasis, and drug resistance.123
With growing recognition that the tumor microenvironment (TME) is central to therapeutic success, acidity has come into focus as a decisive factor, reshaping not only tumor growth and metastasis but also the tumor immune microenvironment (TIME), where it can dictate the success or failure of cancer immunotherapies.4

In response, a growing body of research has focused on strategies to neutralize tumor acidity and establish a TME more conducive to therapy. Approaches such as systemic buffering agents and inhibitors of acid transport have been explored, but their clinical translation has been limited by challenges in achieving effective and sustained alkalization.123
Efforts to overcome these barriers have given rise to a new generation of tumor-targeted therapies that seek to directly remodel the acidic TME. Among them is Helix BioPharma’s Tumor Defense Breaker™ L-DOS47, a first-in-class antibody-enzyme conjugate (AEC) designed to locally convert urea into ammonia, raising pH within the TME and re-sensitizing tumors to cancer immunotherapies.

Why Acidity Matters in Cancer

Most healthy tissues in the body maintain a tightly regulated extracellular pH (pHe) of 7.2 to 7.4. By contrast, solid tumors often display pHe values ranging from 6.5 to 6.9.12
This acidification arises from a phenomenon known as the Warburg Effect, in which tumors preferentially generate energy by breaking glucose into acidic lactate through glycolysis, even in the presence of oxygen and functional mitochondria.125
When coupled with dysfunctional vasculature, the result is a buildup of hydrogen ions that drives a persistently acidic environment in and around tumor cells.135

The acidic TME drives cancer progression by selecting for more aggressive, therapy-resistant clones.123
At the same time, low pH activates enzymes that degrade the extracellular matrix, breaking down the physical barriers around the tumor and allowing cancer cells to invade surrounding tissue and metastasize.125
Acidity also switches on angiogenic signals such as VEGF, which drive the formation of abnormal new blood vessels, fueling hypoxia, acid production, and tumor expansion.125
It blunts immune surveillance, co-opting immune cells to suppress the activity of cytotoxic T cells (the immune system’s frontline fighters), disrupts T cell motility and competes for glucose with tumor-infiltrating T cells, leading to their exhaustion and driving tumor progression.46
The result is a competitive advantage for the tumor and a hostile setting for therapy.1236

Beyond Buffering: Pharmacologic Approaches to Tumor Alkalization

Neutralizing tumor acidity through the oral administration of buffers like sodium bicarbonate has shown promise in preclinical models.127
Studies have demonstrated that buffering can reduce metastasis and enhance the efficacy of checkpoint inhibitors in animal models of melanoma and pancreatic cancer.127

This has driven interest in alternative alkalization strategies:

  • Proton transport inhibitors (PTIs): Agents targeting proton pumps (e.g., V-ATPase inhibitors) or transporters (e.g., Na⁺/H⁺ exchangers, monocarboxylate transporters [MCTs]) reduce extracellular acidification by blocking H⁺ efflux. While effective in preclinical studies, most PTIs have faced hurdles in systemic toxicity and poor selectivity for tumor cells, limiting clinical adoption.123
  • Nano-enabled extracellular alkalization: Nanoparticles engineered to release buffering agents (e.g., calcium carbonate, magnesium hydroxide) within the TME can locally raise pHe. These nanoplatforms also allow co-delivery of chemotherapeutics, enhancing drug uptake in an alkalized tumor milieu. However, scalability, biocompatibility, and controlled biodistribution remain challenges.7
  • Enzymatic and metabolic interventions: Inhibition of lactate dehydrogenase (LDHA) reduces lactate accumulation and indirectly alleviates extracellular acidification. While promising, systemic LDHA inhibition risks toxicity to normal tissues that also rely on glycolysis.356
  • Intracellular acidification approaches: Complementary to TME alkalization, some investigational strategies aim to selectively lower the intracellular pH (pHi) of cancer cells. This exploits the fact that tumor cells maintain an alkaline cytosol (pHi ~7.4–7.6) as an adaptive survival mechanism. Pharmacologic inhibition of proton exporters or delivery of acid-generating nanoparticles can collapse this gradient, triggering apoptosis or necrosis. This “inside-out” strategy remains experimental but highlights the broader therapeutic potential of targeting tumor pH dysregulation.37

Collectively, these findings underscore both the promise and the limitations of current alkalization therapies. Most approaches have been challenged by lack of tumor selectivity, systemic side effects, or delivery issues fueling the need for more precise solutions.1237

Why Pharmacological Alkalization Works

Neutralizing tumor acidity offers several benefits, all of which align with known hallmarks of cancer:

  • Immune restoration: Acidic conditions suppress T-cell function by impairing cytokine secretion and downregulating key surface receptors. Buffering the TME restores the ability of T cells to recognize and attack tumor cells, thereby improving immunotherapy efficacy.46
  • Reduced invasion and metastasis: An acidic pH promotes protease release and matrix remodeling, enabling tumor cells to invade nearby tissues. Alkalization inhibits this process and can reduce the likelihood of distant spread.1235
  • Overcoming drug resistance: Many chemotherapeutics are weak bases that become protonated and trapped in the extracellular space or lysosomes. A neutralized TME enhances drug penetration and intracellular retention.1237
  • Targeting metabolic vulnerabilities: Tumor cells adapted to acidic pH show specific metabolic dependencies (e.g., glutamine metabolism, lipid synthesis). By altering the pH, these vulnerabilities may be exposed and therapeutically exploited.36

A Smarter Strategy: Tumor-Targeted Alkalization

L-DOS47 is a fusion of a camelid-derived single-domain antibody (nanobody) targeting CEACAM6 (an antigen overexpressed in many solid tumors), and the enzyme urease, which converts endogenous urea into ammonia and bicarbonate.4
Once administered intravenously, L-DOS47 navigates through the bloodstream to locate and bind with CEACAM6-expressing cancer cells.4
Upon binding, the urease component becomes catalytically active within the acidic TME, locally converting urea into two molecules of ammonium and one molecule of bicarbonate.4
Recent evidence suggests that the cytotoxic effect of ammonia is highly dependent on the extracellular pH. At higher pH levels, a greater fraction of ammonia exists in a membrane-permeable form, enhancing its ability to enter cells, disrupt organelle function, and suppress tumor growth.3
In parallel, the generation of bicarbonate at the tumor site contributes to raising extracellular pHe, which not only counteracts tumor acidity but also improves therapeutic responsiveness.4
A more alkaline microenvironment increases the activity of ionizable, weak-base chemotherapeutics such as doxorubicin by facilitating their penetration into cancer cells, while also promoting infiltration and activation of immune populations including T cells, B cells, and macrophages.7

The result is a highly localized increase in pHe in the tumor microenvironment.4
By bypassing systemic buffer delivery, L-DOS47 offers an elegant and targeted way to neutralize tumor acidity at the source.4

How L-DOS47 Stands Apart

Unlike systemic buffers, PTIs, or nanoparticle approaches, L-DOS47 uniquely combines tumor antigen targeting with enzyme-driven alkalization. Its localized activity avoids systemic pH disruption, patient compliance barriers, and high-dose regimens required for oral buffers.4

CEACAM6 provides an ideal target because it is not only highly expressed in multiple aggressive tumors (e.g., NSCLC, PDAC, CRC where therapeutic options remain limited), but also implicated in immune evasion and resistance pathways. Importantly, CEACAM6 blockade itself may confer therapeutic benefit: disruption of CEACAM6–CEACAM1 interactions has been shown to enhance T cell responses, suggesting that L-DOS47 nanobody binding could offer dual benefits—enzyme delivery plus partial immune reactivation.4

Safety and tolerability have been demonstrated in early Phase I/II NSCLC trials, with encouraging signals of efficacy in chemotherapy combinations.4
L-DOS47 has already demonstrated safety and tolerability in Phase I/II trials in NSCLC, with promising signals of efficacy when combined with chemotherapy.4

A Platform for the Future

L-DOS47 is more than a single asset; it is proof of concept for pharmacological alkalization. It opens the door for a new class of therapies that don’t just attack the tumor directly but reshape its surroundings to enable better outcomes from existing treatments.

As immunotherapy and precision oncology continue to evolve, so must our approach to the tumor microenvironment. Targeting cancer’s hidden enablers like acidity represents a bold, science-driven path forward. By unlocking tumors from within, therapies like L-DOS47 hold the promise to turn resistance into response and the intractable into treatable, toward more durable, accessible, and effective cancer care.

References:

1 Hamaguchi R, Isowa M, Narui R, Morikawa H, Wada H. Clinical review of alkalization therapy in cancer treatment. Front Oncol. 2022;12:1003588.

2 Ibrahim-Hashim A, Estrella V. Acidosis and cancer: from mechanism to neutralization. Cancer Metastasis Rev. 2019;38(1-2):149–55.

3 Pillai SR, Damaghi M, Marunaka Y, Spugnini EP, Fais S, Gillies RJ. Causes, consequences, and therapy of tumors acidosis. Cancer Metastasis Rev. 2019;38(1-2):205–22.

4 Tang T, Huang X, Zhang G, et al. Advantages of targeting the tumor immune microenvironment over blocking immune checkpoint in cancer immunotherapy. Signal Transduct Target Ther. 2021;6(1):72.

5 Kato Y, Ozawa S, Miyamoto C, et al. Acidic extracellular microenvironment and cancer. Cancer Cell Int. 2013;13:89.

6 Najdi R, et al. Microenvironmental metabolism regulates anti-tumor immunity. Cancer Res. 2019;79(4):793–801.

7 Yang Y, et al. Sodium bicarbonate nanoparticles modulate the tumor pH and enhance the cellular uptake of doxorubicin. J Control Release. 2019;296:73–82.

Jacek Antas

Chief Executive Officer


Jacek Antas is a shareholder of the Company, has spent more than 25 years in the financial services industry holding various positions in sales and consulting.

Mr. Antas obtained a master’s degree from the Warsaw School of Economics and has served as a board member of various
companies throughout his career.

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James B. Murphy

Chief Financial Officer


Mr. Murphy is a certified public accountant with over thirty years of experience in finance and operations management. He is currently a consultant with Danforth Advisors LLC (“Danforth”), a leading provider of outsourced strategic and operational specialists across functions in the life sciences industry. While at Danforth, Mr. Murphy has served over fifteen private and publicly held life sciences companies as CFO and CFO Advisor, helping them secure over USD 0.5 billion in financing and successfully execute pivotal asset transactions. Mr. Murphy functions as a consultant to Helix pursuant to a consulting agreement between the Company and Danforth.

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Medical Adviser


Thomas Mehrling (PhD in Pharmacology and MD) has over 20 years’ experience in multinational Pharma companies developing novel oncology compounds from preclinical research through to registration. Prior to entering the industry, he spent 13 years as an MD at the University Hospital in Frankfurt, working on preclinical and translational projects. He served as Director of European Oncology at Mundipharma International (2003–2013), building the company’s first European oncology business from the ground up out of Cambridge, UK, and completing the clinical development, registration and launch of two major products in Europe, DepoCyte® and Levact® (Ribomustin® and Treanda®). In 2013, he led the establishment of the Mundipharma Group’s start-up, Mundipharma EDO, developing anti-cancer therapeutics for solid tumours out of Basel, Switzerland.

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Director Quality Assurance


Kim is the Director of Quality Assurance at Helix BioPharma Corp. An experienced quality assurance professional with expertise in Canadian, US, and EU regulations, she has been involved in all aspects of Phase I/II biopharmaceutical product development over the years, including regulatory submissions, QC laboratory compliance, tech transfer and third-party oversight of CMC activities, clinical QA, and bioanalytical data analysis. Kim joined Helix in 2000, transitioning into QA in 2003. She holds a B.Sc in Biochemistry and a Ph.D in Veterinary Physiological Sciences, both from the University of Saskatchewan.

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Brenda Lee

Director Clinical Operations


Brenda is the Clinical Operations Director at Helix Biopharma Corp. A clinical research operations professional with 25 years of experience managing clinical trials, ranging from early Phase I to late Phase IIIb/IV studies, she brings experience in clinical study protocol writing and development, trial start-up and vendor management, and a proven track record in planning and managing clinical trials to quality standards, timelines and budget. Brenda joined Helix Biopharma Corp. in 2018, working to advance the clinical program of L-DOS47. She holds B.Sc and M.Sc. degrees from the University of Toronto, specializing in Nutritional Sciences and Human Biology.

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Director


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Malgorzata Laube

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Jacek Antas

Chairman of the Board


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Mr. Antas obtained a master’s degree from the Warsaw School of Economics and has served as a board member of various
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Jonathan Davis

Advisor, ADC Discovery


Jonathan Davis received his Ph.D. from University of California, San Francisco, where he studied protein structure and function using NMR. After a post-doc at Harvard Medical School exploring RNA selection and structure in the labs of Jack Szostak and Gerhard Wagner, he went to work at EMD Serono, where his work involved improving antibody-based therapeutics, inventing a platform technology for generating heterodimeric Fcs as a basis for multifunctional molecules, and developing a novel scaffold based on an artificially-designed protein from David Baker’s lab. In 2008 he took a job at Bristol-Myers Squibb in Waltham/Cambridge MA, working on antibody discovery and platform development in a wide range of therapeutic areas, with a particular focus on multispecific therapeutics. He moved to Madison, WI in 2019 to take on the role of VP of Innovation and Strategy at Invenra, a biotech focused on bispecific antibodies, and where he is currently head of the Scientific Advisory Board. In early 2024 he left the corporate world to found Creative Antibodies, a consulting firm that helps guide companies to successful antibody discovery and development projects, from mAbs to multispecifics, ADCs, and other formats. Outside of science, Jonathan is a conservatory trained cellist, plays numerous other instruments, and founded the UCSF Orchestra (now Symphony Parnassus) in San Francisco, where he was Music Director for six years.

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Davide Guggi

Advisor, CMC


Davide graduated as a pharmacist and received his PhD in Pharmaceutical Technology and Biotechnology from the University of Vienna. He has over 20 years of experience in the pharmaceutical industry, principally in the field of oncology. At the beginning of his career, Davide led oncology business units and commercial departments at Mundipharma and Gilead across Austria and Eastern Europe. Since over 10 years he has been working as a CMC expert, covering operational and regulatory CMC functions on behalf of over 20 different small- and medium-sized biotech companies across the world. He has served as CMC Director and CSO/CTO for several years, developing both small molecules and biologics (mABs, Fab, ADCs and Radio-immuno-conjugates) from early discovery to NDA/BLA in the US, EU and Canada, with a focus on First-in-Human and Phase I/II studies in oncology indications.

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Tumor Defense Breaker™, L-DOS47


L‑DOS47 is a first‑in‑class, clinical-stage antibody‑enzyme conjugate designed to deliver a game-changing assist to anti-cancer immunity and today’s leading cancer immunotherapies for the treatment of prevalent, hard-to-treat solid tumors. The compound precisely targets CEACAM6, a cell-surface protein overexpressed in non‑small cell lung cancer (NSCLC) and other aggressive tumors, where it delivers an enzymatic payload that raises the extracellular pH of the acidic tumor microenvironment (TME). By neutralizing tumor acidity, L-DOS47 restores immune cell infiltration and activity, helps turn immunologically “cold” tumors “hot”, and enhances the therapeutic reach of immune checkpoint inhibitors. With patented composition-of-matter coverage through 2036 and demonstrated synergy with PD-1 inhibitor, pembrolizumab, L-DOS47 is poised to significantly increase the efficacy of immune checkpoint blockade and unlock broader and more durable responses in NSCLC and other aggressive solid tumors.

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LEUMUNA™


LEUMUNA™ is an oral immune checkpoint modulator designed to activate the donor immune system to recognize and fight relapsing leukemia in patients who have undergone allogeneic stem cell transplantation (allo-SCT). Although a life-saving procedure, up to 30% of patients who undergo allo-SCT see their cancer return, facing a median survival of just four months. LEUMUNA aims to offer these patients a new lease on life, by activating an immune cascade and inciting graft-versus-leukemia (GvL) effect, potentially offering long-term remission. Backed by strong preclinical data and a promising safety record from trials with its precursor compound, ulodesine, LEUMUNA offers a patient‑friendly, oral approach to a difficult-to-treat condition, with patent protection through 2041 and an Orphan Drug Designation granted by the US FDA.

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GEMCEDA™


GEMCEDA is a first-in-class oral prodrug of gemcitabine that opens up the possibility for convenient at-home administration, metronomic dosing and seamless integration into combination regimens with immune checkpoint inhibitors. To date, gemcitabine is only administered intravenously because oral forms have shown poor bioavailability of about 10%. GEMCEDA was developed as a prodrug to enable new uses of gemcitabine by combining it with cedazuridine, an enzyme inhibitor that helps boost its bioavailability to 90%. This remarkable innovation allows for greater flexibility in dosing schedules, fewer clinic visits, and a better quality of life, while achieving bioavailability on par with intravenous gemcitabine. Supported by a well‑established safety profile, scalable manufacturing, and patent coverage to 2043, GEMCEDA reimagines how chemotherapy can fit into patients’ lives.

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