Bioray Bio Announces Breakthrough in Non-Viral CAR-T Therapy with First-Ever PD1-Targeted Integration Showing Complete Responses in Relapsed/Refractory Lymphoma Patients

Shanghai, September 25, 2020 /PRNewswire/ -- On September 25, 2020, BRL Medicine Inc. (“BRL Medicine”), a company focused on the research and development of gene therapy and cell-based drugs, announced for the first time the breakthrough results of its collaborative clinical trial with the First Affiliated Hospital, Zhejiang University School of Medicine, titled “Clinical Trial of PD-1 Knockout, Non-Viral Site-Specific Integration CD19-CAR-T Cell Therapy for Relapsed/Refractory Non-Hodgkin Lymphoma.”This is the world's first use of gene-editing technology inCAR-T therapy with site-specific integration at the PD-1 locus is also the world’s first clinical trial of non-viral, site-specifically integrated CAR-T cell therapy for lymphoma.Meanwhile,The latest research findings were published on the preprint platform medRxiv on September 23, 2020., jointly completed by East China Normal University, the First Affiliated Hospital of Zhejiang University School of Medicine, and BRL Medicine Inc.

PD1-Site-Integrated CD19-CAR-T is developed by BRL Medicine using its proprietary Quikin CAR-T™ platform technology, a CAR-T product prepared in a single step without the use of viral vectors. This product integrates PD-1 immune checkpoint inhibition with CAR-T tumor-killing functionality, achieving the combined therapeutic effect of PD-1 immunotherapy and CAR-T therapy.

2Case patients undergoing treatment3Complete Remission After Months

This clinical study planned to enroll 15 patients. Among the 4 patients currently evaluable, all achieved partial response (PR) at 1 month. Of the 2 patients who have reached the 3-month evaluation time point, both achieved complete response (CR).

The first patient to achieve complete remission was diagnosed in 2018 with stage IVBE diffuse large B-cell lymphoma. Despite multiple courses of chemotherapy and radiotherapy, the disease remained poorly controlled. Pre-enrollment imaging revealed a lesion measuring 3.6 × 3.5 cm in the mesenteric space along the course of the small intestine in the left lower abdomen, with abnormally increased radioactive uptake. In May 2020, the patient was enrolled and received an infusion of PD-1 knockout, non-viral, site-specifically integrated CD19 CAR-T cells. Twenty-eight days post-treatment, PET-CT imaging showed a significant reduction in FDG metabolism compared to baseline, indicating a partial response (PR). Ninety days after treatment, follow-up PET-CT imaging demonstrated complete disappearance of the lesion with no increased FDG metabolism, achieving a complete response (CR).

No drug-related adverse events of grade 3 or higher occurred during the entire CAR-T treatment process.including cytokine release syndrome and neurotoxicity. Following infusion, CAR-T cells exhibited robust expansion and sustained persistence in vivo,D90Flow cytometry analysis revealed that CAR-T cells in peripheral blood remained at a certain proportion. Currently, two patients have recovered and been discharged, and they are still under long-term follow-up.

The Principal Investigator of this clinical study (PI) Professor Huang He, President of the First Affiliated Hospital, Zhejiang University School of Medicine, stated:“Diffuse large B-cell lymphoma is the most common type of non-Hodgkin lymphoma, and there is currently a lack of effective treatments for patients with refractory or relapsed disease. This year, we initiated clinical research on non-viral, site-specific integration CAR-T therapy based on gene-editing technology. We are very pleased to observe that patients rapidly achieved complete remission following treatment. We anticipate that this novel CAR-T technology will provide more convenient, safer, and definitively effective long-term therapeutic outcomes for patients with refractory or relapsed disease.”

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BRL Medicine’s non-viral site-specific integration CAR-T technology (Quikin CART™) utilizes CRISPR/Cas9 gene-editing to insert CAR elements into specific genomic loci without viral vectors, achieving simultaneous gene knockout and stable CAR integration in a single step. This technology is part of BRL Medicine’s pipeline of second-generation CAR-T products.

PD-L1/PD-1 is a critical immune checkpoint that inhibits T-cell function. Currently, inhibitors targeting PD-L1/PD-1 have demonstrated favorable efficacy in various types of malignant tumors, and numerous studies have reported that PD-1 knockout can effectively enhance the function of CAR-T cells. Non-Hodgkin lymphoma (NHL) is a hematologic malignancy originating in lymphoid tissue, accounting for 80%–90% of all lymphomas. Although patients often achieve disease remission after initial treatment, relapse frequently occurs subsequently. While CAR-T cell products have been approved for the clinical treatment of relapsed/refractory non-Hodgkin lymphoma, overall efficacy remains limited. However, studies suggest that inhibition of the PD-1 pathway may yield improved clinical outcomes.

Therefore, BRL Medicine utilized the Quikin CART™ platform to develop non-viral CD19-CART cells with site-specific integration and PD-1 knockout, for the clinical treatment of relapsed or refractory non-Hodgkin lymphoma. This product offers two major advantages: uniform and stable CAR expression and PD-1 gene knockout, effectively combining the strengths of PD-1 inhibitors and CART cells. In multiple clinical trials conducted to date, this CART product has demonstrated excellent safety and efficacy.

Traditional CAR-T products are primarily manufactured using viral vectors, which impose stringent requirements on viral vector production processes. This significantly increases the manufacturing cost and complexity of CAR-T cells, hindering their large-scale clinical application. Furthermore, these therapies are prohibitively expensive; for instance, Novartis’s Kymriah is priced at $475,000, and Kite’s Yescarta at $373,000. Additionally, because viral vectors integrate CAR elements into the cell genome via random insertion, they may alter normal gene expression, posing a potential safety risk of oncogenesis. The Quikin CART™ technology eliminates the need for viral vectors in cell preparation, substantially reducing the production costs of CAR-T products while avoiding the oncogenic risks associated with random insertion.

Quikin CART™ technology enables simultaneous regulation of endogenous T-cell genes and sustained CAR expression in a single step. Compared to other CAR-T technologies, it offers advantages such as a simplified process, fewer production steps, shorter preparation time, and high product uniformity. This technology platform can be utilized for the preparation of multiple immune checkpoint knockout-enhanced CAR-T cells, facilitate the rapid production of universal CAR-T cells, and support the development of safely regulated CAR-T products with dynamic control, thereby providing robust technical support for the diverse engineering of CAR-T cells in the future.

Professor Liu Mingyao, Chief Scientist at BRL Medicine and Professor at the School of Life Sciences, East China Normal University, stated:“Compared with traditional autologous CAR-T technology, Quikin CAR-T™ technology enables one-step, site-specific integration of CAR elements into the genome and regulatory intervention of endogenous T-cell genes without using viruses. This means that the manufacturing process, production steps, and preparation time for CAR-T cells are significantly simplified and shortened, thereby greatly reducing the production cost of CAR-T cells, minimizing the tumorigenic risk caused by random viral insertion, and improving the uniformity of CAR-T products. Current results show that our non-viral, PD-1 site-specifically integrated CD19 CAR-T cells have enormous therapeutic potential, demonstrating excellent safety and response rates in clinical trials.”

Principal Investigator of the Project and First Author of the Paper, BRL Medicine InnovationDr. Zhang Jiqin, Associate Researcher at the School of Life Sciences, East China Normal University, and R&D Director of CAR-T, said：“In recent years, the continuous maturation and development of CRISPR/Cas9 gene-editing technology have inspired us to explore its application in CAR-T therapy. Through an in-depth analysis of the limitations associated with existing CAR-T technologies, we recognized that leveraging gene-editing tools to generate non-viral, site-specifically integrated CAR-T cells represents a highly promising direction. After extensive methodological trials and optimization of conditions, we have established the robust Quikin CAR-T™ technology platform. This virus-free platform enables the one-step preparation of CAR-T cells, offering numerous advantages unmatched by current CAR-T technologies. We look forward to leveraging this platform to develop more successful CAR-T products and achieve greater breakthroughs in clinical treatment.”

In addition toClinical studies on non-viral, site-specific integration of PD1-targeted CD19-CAR-T cells are currently underway. BRL Medicine is also developing other non-viral, site-specifically integrated CAR-T products targeting solid tumors, aiming to achieve greater breakthroughs in CAR-T therapy.