Safety of Autologous Humanized Anti-CD19 and Anti-CD20 Dual Specific CAR-T Cells in Adult Patients With Diffuse Large B-cell Lymphoma

Study Overview
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Brief Summary
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This is a single-arm, open-label, dose escalation, phase I study, aiming to evaluate the safety and efficacy of Autologous Humanized Anti-CD19 and Anti-CD20 Dual Specific Chimeric Antigen Receptor (CAR) T-cells in patient with relapsed or refractory diffuse B cell lymphoma.

Detailed Description
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CD19 CAR-T cell therapy has made breakthroughs in the treatment of B cell lymphoma and leukemia, but 30% of patients still have antigen escape, which may be related to variants in tumor cells and the expansion of CD19-negative tumor cells after treatment with CD19 CAR-T cells. CD19/CD20 bispecific CAR-T cell targeting multiple antigens can attack tumor cells while overcoming tumor antigen escape caused by a single target, maximizing efficacy and duration of treatment, and can also solve the problem of uneven distribution or low expression of single target on the tumor surface.

Official Title
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A Single-arm, Open-label, Dose Escalation Study to Explore Safety, Efficacy and Pharmacokinetics of Autologous Humanized Anti-CD19 and Anti-CD20 Dual Specific CAR-T Cells in Adult Patients With Relapsed or Refractory Diffuse Large B Cell Lymphoma

Conditions
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Relapsed or Refractory DLBCL Patients With Either CD19 or CD20 Positive

Intervention / Treatment
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* Biological: Autologous humanized anti-CD19 and anti-CD20 dual specific CAR-T Cells


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: The subject or her/his legally guardian(s) must sign the informed consent form approved by the Institutional Ethics Committee (IEC) prior to any screening procedures; Subjects aged 18 years or older with relapsed or refractory DLBCL (primary mediastinal large B-cell lymphoma and transformed follicular lymphoma included), of which refractory is defined as: Have no response to the recent treatment including: The best response to the treatment regimen is progressive disease (PD) ,or stable disease (SD) which maintained less than 6 months after the last treatment, or not suitable for autologous hematopoietic stem cell transplantation (ASCT), or ASCT refractory, including: progressive disease after ASCT or relapse within 12 months (relapse must be confirmed by biopsy), or If remedial treatment is given after ASCT, the subject must have no response or relapse after the last treatment. Subjects who have previously received ≥2 lines treatment, and at least including: Anti-CD20 monoclonal antibody(rituximab), unless the CD20 negative; A chemotherapy regimen containing anthracyclines; The DLBCL patients who transformed from follicular lymphoma must have previously received chemotherapy for follicular lymphoma and have developed chemotherapy-refractory diseases after transform to DLBCL. Confirmation for either CD19 or CD20 positivity using immunohistochemistry or flow cytometry; According to the initial evaluation, staging and response assessment of Hodgkin's and non-Hodgkin's lymphoma -the Lugano Classification (2014), there is at least one measurable lesion at baseline; Life expectancy ≥12 weeks; Eastern Cooperative Oncology Group (ECOG) performance status that is either 0 or 1 at screening; Adequate organ function: Renal function defined as: A serum creatinine of ≤1.5 × Upper Limit of Normal (ULN), or; Estimated Glomerular Filtration Rate (eGFR) ≥60 ml/min/1.73m2； Liver function defined as: ALT≤ 5 × Upper Limit of Normal (ULN) for age, and; Total bilirubin ≤ 2.0 mg/dl with the exception of patients with Gilbert-Meulengracht syndrome; patients with Gilbert-Meulengracht syndrome may be included if their total bilirubin is ≤ 3.0 × ULN and direct bilirubin ≤ 1.5 × ULN. Must have a minimum level of pulmonary reserve defined as ≤ Grade 1 dyspnea and blood oxygen saturation > 91% on room air; Hemodynamically stable and Left Ventricle Ejection Fraction (LVEF) ≥ 45% confirmed by echocardiogram or Multigated Radionuclide Angiography (MUGA); Adequate bone marrow reserve without transfusions defined as: Absolute neutrophil count (ANC) >1×10^9 /L； Absolute lymphocyte count (ALC) ≥0.3×10^9 /L； Platelets ≥50×10^9 /L; Hemoglobin > 8.0 g/dl; Must have an apheresis product of non-mobilized cells or peripheral blood harvested cells accepted for manufacturing Subjects who use the following drugs should meet the following criteria: Steroids: Therapeutic doses of steroids must be stopped 2 weeks prior to A-02 infusion. However, the following physiological replacement doses of steroids are allowed: < 6 - 12 mg/m^2/day hydrocortisone or equivalent; Immunosuppression: Any immunosuppressive medication must be stopped ≥ 4 weeks prior to sign the informed consent form; Anti-proliferative therapy other than pretreatment chemotherapy within 2 weeks of A-02 infusion; CD20 antibody-related treatment must be discontinued within 4 weeks of A-02 infusion or 5 half-lives (whichever is longer); CNS disease prophylaxis must be stopped > 1 week prior to A-02 infusion (e.g. intrathecal methotrexate); The investigator judged that the subject recovered from the toxicity of the previous anti-tumor treatment to grade 1 or below (except for special grade 2 or below toxicity that cannot be recovered in a short period of time, such as hair loss), suitable for pretreatment. Chemotherapy and treatment of CAR-T cells; Women of child-bearing potential and all male subjects must agree to use highly effective methods of contraception for at least 12 months following A-02 infusion and until CAR-T cells are no longer present by PCR on two consecutive tests. Exclusion Criteria: Prior treatment with any cell therapy before signing the informed consent form, including CAR-T therapy; Subjects with detectable cerebrospinal fluid malignant cells or brain metastases, or with a history of central nervous system (CNS) lymphoma or primary CNS lymphoma; Subjects with testicular invasion, including those who have had testicular resection; Subjects with current or previous history of central nervous system disease, such as seizures, cerebral ischemia/hemorrhage, dementia, cerebellar disease, or any autoimmune disease involving the central nervous system; Subjects who have previously received allogeneic hematopoietic stem cell transplantation (HSCT); or suitable and consenting to Autologous hematopoietic stem cell transplantation (ASCT); Chemotherapy other than lymphodepleting chemotherapy within 2 weeks of A-02 infusion; Patients on oral anticoagulation therapy within 1 week of A-02 infusion; Prior radiation therapy within 2 weeks of A-02 infusion; Investigational medicinal product within the last 30 days prior to sign the informed consent form; Subjects with active hepatitis B（defined as hepatitis B surface antigen positive, or hepatitis B core antibody positive with hepatitis B virus DNA detection value > 1000 copies/ml）or hepatitis C (HCV RNA positive) Subjects positive for HIV antibody or treponema pallidum antibody; Uncontrolled acute life threatening bacterial, viral or fungal infection (e.g. blood culture positive ≤ 72 hours prior to A-02 infusion) Unstable angina and/or myocardial infarction within 6 months prior to sign the informed consent form; Previous or concurrent malignancy with the following exceptions: Adequately treated basal cell or squamous cell carcinoma (adequate wound healing is required prior to sign the informed consent form); In situ carcinoma of the cervix or breast, treated curatively and without evidence of recurrence for at least 3 years prior to sign the informed consent form; A primary malignancy which has been completely resected and in complete remission for ≥ 5 years; Pregnant or nursing women (women of childbearing age were tested positive for pregnancy during screening period); Cardiac arrhythmia not controlled with medical management; Subjects with active neurological auto immune or inflammatory disorders (e.g. Guillain Barre Syndrome, Amyotrophic Lateral Sclerosis); Other conditions that the investigator thinks he/she should not be included in this clinical trial, such as poor compliance.

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Treatment
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Humanized anti-CD19 and anti-CD20 dual specific CAR-T cells<br> | Biological: Autologous humanized anti-CD19 and anti-CD20 dual specific CAR-T Cells<br>* Humanized anti-CD19 and CD20 bispecific autologous CAR-T cells injection: the first dose is 1.0×106 /kg, the second dose is 3.0×106 /kg, and the third dose is 8.0×106 /kg. Patients will receive lymphodepleting chemotherapy at least 1 week before CAR-T cell infusion.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The types and Incidence of adverse events |  | Up to 12 months | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Overall response rate | including CR and PR | Up to 12 months | 
| Progression-free survival (PFS) |  | Up to 12 months | 
| Response duration |  | Up to 12 months |