an exploratory clinical trial of autologous humanized anti-cluster of differentiation antigen 19/20(cd19/cd20) dual specific car-t cells injection

Conditions: Diffuse Large B Cell Lymphoma

Summary: 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.

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.

Eligibility criteria:
inclusion criteria: 1. 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; 2. subjects aged 18 years or older with relapsed or refractory dlbcl (including primary mediastinal large b-cell lymphoma and transformed follicular lymphoma), 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. 3. 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. 4. confirmation for either cd19 or cd20 positivity using immunohistochemistry or flow cytometry(accepting the previous results from the a third-level grade a hospitals before the collection of peripheral blood mononuclear cells or peripheral blood. for cd20 positive only, the investigator needs to determine whether the treatment benefit); 5. according to the preliminary assessment of hodgkin's lymphoma and non-hodgkin's lymphoma, staging and response assessment recommendations (2014 version), there is at least one measurable lesion at baseline; 6. if the subject has received a single target in the past, such as cd19-car cell therapy, it must be confirmed that the disease has progressed or relapsed after treatment and is at least 1 month from the planned single collection period 7. life expectancy ≥12 weeks; 8. eastern cooperative oncology group (ecog) performance status that is either 0 or 1 at screening; 9. 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；\[egfr=186×（age）-0.203×scr-1.154（mg/dl）， female ×0.742 on the basis of the calculation results\]； * liver function defined as: * alanine aminotransferase (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; 10. hemodynamically stable and left ventricle ejection fraction (lvef) ≥ 45% confirmed by echocardiogram or multigated radionuclide angiography (muga); 11. 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; 12. 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/m2/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); 13. 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; 14. 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 polymerase chain reaction(pcr) on two consecutive tests. exclusion criteria: subjects who meet any of the following criteria will not be enrolled: 1. subjects who have received any cd19/cd20 dual-target cell therapy products before signing the informed consent form; 2. subjects with detectable cerebrospinal fluid malignant cells or brain metastases, or with a history of central nervous system (cns) lymphoma or primary cns lymphoma; 3. 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; 4. subjects who have previously received allogeneic hematopoietic stem cell transplantation (hsct); or suitable and consenting to autologous hematopoietic stem cell transplantation (asct); 5. chemotherapy other than lymphodepleting chemotherapy within 2 weeks of a-02 infusion; 6. investigational medicinal product within the last 30 days prior to sign the informed consent form; 7. 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); 8. subjects positive for hiv antibody or treponema pallidum antibody; 9. uncontrolled acute life threatening bacterial, viral or fungal infection (e.g. blood culture positive ≤ 72 hours prior to a-02 infusion); 10. unstable angina and/or myocardial infarction within 6 months prior to sign the informed consent form; 11. 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; 12. pregnant or nursing women (women of childbearing age were tested positive for pregnancy during screening period); 13. subjects with active neurological auto immune or inflammatory disorders (e.g. guillain barre syndrome, amyotrophic lateral sclerosis); 14. other conditions that the investigator thinks he/she should not be included in this clinical trial, such as poor compliance.