Anti-CD19-CAR-T Cells in Subjects with Relapsed/Refractory B Cell Malignancies

Inclusion Criteria:

Subjects must meet all of the following criteria for inclusion in the study:

1. Patient able to provide written informed consent; parent or guardian of minorpatient able to provide written informed consent; ability and willingness to adhereto the study visit schedule and all protocol requirements.

2. 14 - 70 years old.

3. Relapsed/refractory disease after standard treatment (includingallogeneic/autologous hematopoietic stem cell transplantation) and not eligible forother treatment options such as a second hematopoietic stem cell transplant. A. Relapsed/refractory B-cell acute lymphoblastic leukemia (ALL) is defined as oneof the following:

• Primary refractory disease.

• Relapsed within 12 months after first remission.

• Relapsed or refractory after two or more lines of systemic therapy.

• Relapsed or refractory post allogeneic SCT. i. more than 100 days from thetransplantation at the time of enrollment. ii. not receiving immunosuppressivedrugs for 4 weeks prior to enrollment (≤5 mg prednisone or equivalent isallowed). B. Ph+ B-cell ALL are eligible if they are intolerant or ineligible for tyrosinekinase inhibitor (TKI) therapy or have relapse/refractory disease after at least twodifferent TKI treatments. C. Relapsed/refractory B-cell non-Hodgkin lymphoma (NHL) is defined as one of thefollowing:

1. No response to first-line therapy (primary refractory disease, excludingsubjects intolerant to first-line therapy):

• PD is the best response after first-line therapy.
• Best response after at least 4 cycles of first-line therapy (e.g., 4cycles of R-CHOP) is stable disease (SD), with rapid progression in 6months.

2. No response to second or subsequent lines of therapy:

• PD is the best response to last regimen.
• Best response after at least 2 cycles of the last-line therapy is SD, withrapid progression in 6 months.

3. Refractory post-autologous stem cell transplant (ASCT):

• Disease progression or relapse within 12 months (relapse must bebiopsy-proven).
• If salvage therapy is given post-ASCT, the subject must have had noresponse to or relapsed after the last line of therapy
• Relapsed or refractory after two or more lines of systemic therapy.

4. Bone marrow with ≥ 5% lymphoblasts by morphologic assessment at screening

5. Relapsed/refractory NHL as one of the following subtypes: A. DLBCL-NOS B. Primary mediastinal large B-cell lymphoma (PMBCL) C. Transformedfollicular lymphoma (TFL) following prior chemotherapy for follicular lymphoma andsubsequent transformation to DLBCL with refractory disease. D. Mantle cell lymphoma E. High-grade B-cell lymphoma F. CLL/SLL

6. ECOG performance status ≤2.

7. Life expectancy ≥ 12 weeks.

8. Adequate venous access (for apheresis) and no other contraindications for blood cellseparation.

9. Subjects must meet the following laboratory criteria at screening, and they shouldnot have received any growth factors within the 7 days prior to the hematologicassessment: A. Absolute neutrophil count ≥1.0×10^9/L. For subjects with ALL, specific criteriawill be determined by the investigator. B. Hemoglobin ≥60 g/L (without RBC transfusion within 14 days). C. Platelets ≥50×10^9/L. For subjects with ALL, specific criteria will be determined by theinvestigator. D. Absolute lymphocyte count (ALC) ≥0.5×10^9/L. If the total lymphocyte count isinsufficient with a high proportion of T cells, the investigator may discuss withthe sponsor. E. Total bilirubin <1.5×ULN; if liver involved, total bilirubin <3.0×ULN is allowed. F. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤2.5×ULN; ifliver involved, ALT/AST ≤5×ULN is allowed. G. Creatinine <1.5×ULN and estimated creatinine clearance ≥60 mL/min.

10. Cardiac ejection fraction (EF) ≥45% with no clinically significant findings onelectrocardiogram.

11. Baseline oxygen saturation >92% on room air.

12. Women of childbearing potential must have a negative serum or urine pregnancy test (women who have undergone surgical sterilization or have been postmenopausal for atleast 2 years are not considered of childbearing potential).

Exclusion Criteria:

If patients meet any of the following conditions, they cannot participate in this trial:

1. Central nervous system (CNS) involvement in ALL and clinically significantneurological changes (CNS-2 and CNS-3):

2. CNS-3, defined as detectable tumor cells in cerebrospinal fluid (CSF) with ≥ 5white blood cells (WBCs) /mm3.

3. CNS-2, defined as detectable tumor cells in CSF with <5 WBCs /mm3. Note:Subjects with CNS-1 (no detectable tumor cells in CSF) or CNS-2 withoutevidence of clinically significant neurological changes are eligible for thisstudy.

4. CNS lymphoma confirmed by MRI; active CNS DLBL unless CNS involvement has beeneffectively treated (i.e., asymptomatic) and a local treatment interval of > 4 weeksprior to enrollment.

5. Active CNS diseases such as epilepsy, cerebrovascular ischemia/hemorrhage, dementia,cerebellar disorders, or any autoimmune diseases involving the central nervoussystem.

6. Any malignancies other than CD19+ malignancies.

7. Clinically significant heart disease or arrhythmias not controlled by medication.

8. Ongoing or suspected fungal, bacterial, viral, or other infections that areuncontrolled or require intravenous antibiotic therapy; simple urinary tractinfections and simple bacterial pharyngitis are allowed.

9. Hepatitis B (positive for hepatitis B surface antigen and hepatitis B DNA >1000copies/mL) and hepatitis C (positive for hepatitis C antibodies); syphilis, humanimmunodeficiency virus (HIV) infection.

10. Presence of any indwelling or drainage catheter (e.g., percutaneous nephrostomy,indwelling Foley catheter, biliary drainage catheter, pleural/peritoneal/pericardialdrainage catheter); the use of dedicated central venous access devices such asPort-A-Cath® or Hickman® catheters is allowed.

11. Prior use of the following:

12. CD19-targeted therapy.

13. Chlorambucil or bendamustine within 3 months before enrollment, orPEG-asparaginase within 3 weeks before enrollment.

14. live vaccines within 4 weeks before enrollment.

15. Donor lymphocyte infusions (DLI) within 4 weeks before enrollment.

16. Medications for graft-versus-host disease (GVHD) treatment within 4 weeksbefore enrollment, such as calcineurin inhibitors, methotrexate, mycophenolate,rapamycin, or siltuximab, or use of immunosuppressive antibodies (anti-CD20,anti-tumor necrosis factor, anti-interleukin 6, or anti-interleukin 6 receptor)within 4 weeks before enrollment.

17. Immunostimulatory or immunosuppressive therapy within 4 weeks beforeenrollment, including interferon-α, interferon-β, IL-2, lenalidomide,efalizumab, alemtuzumab, tocilizumab, cyclosporine, or thalidomide.

18. Anti-PD-1/anti-PD-L1 therapy within 4 weeks before enrollment.

19. Systemic cytotoxic chemotherapy within 14 days before enrollment, includingdaily or weekly low-dose maintenance chemotherapy (e.g., cyclophosphamide,ifosfamide, bendamustine, chlorambucil, melphalan, or vincristine).

20. Long-acting growth factors (e.g., pegfilgrastim) within 14 days beforeapheresis or short-acting growth factors within 5 days before apheresis ormobilization agents (e.g., filgrastim/pegfilgrastim, plerixafor) within 5 daysbefore apheresis.

21. Radiation therapy within 14 days before enrollment.

22. Pharmacological doses of corticosteroids (>5 mg/day prednisone or equivalent)or other immunosuppressive drugs within 7 days before enrollment.

23. Venetoclax (BCL-2 inhibitor) within 4 days before apheresis.

24. Short-acting targeted therapy (e.g., tyrosine kinase inhibitors) within 72hours before apheresis.

25. Idelalisib (oral PI3Kδ inhibitor) within 2 days before apheresis.

26. Lenalidomide within 1 day before enrollment.

27. ≥ Grade 2 graft-versus-host disease (GVHD) per the CIBMTR grading system orrequiring systemic corticosteroid treatment exceeding physiological doses.

28. A history of autoimmune diseases in the past 2 years, such as Crohn's disease,rheumatoid arthritis, systemic lupus erythematosus, leading to end-organ damage orrequiring systemic immunosuppression or disease-modifying agents.

29. A history of heart attack, cardiac catheterization or stent implantation, unstableangina, or other clinically significant heart diseases within 12 months beforeenrollment.

30. A history of genetic syndrome with bone marrow failure, such as Fanconi anemia,Kostmann syndrome, Schwachman-Diamond syndrome, etc.

31. Symptomatic deep vein thrombosis or pulmonary embolism requiring systemicanticoagulation within 6 months before enrollment. Subjects need to be onprophylactic anticoagulation.

32. A history of or currently having other malignant tumors (excluding skin basal cellcarcinoma, breast/cervical carcinoma in situ, and other malignant tumors that havenot been treated in the past five years but are effectively controlled).

33. Use of other investigational medicinal products within 30 days before screening.

34. Pregnant, planning to become pregnant, or breastfeeding in reproductive-aged women.Women who have undergone surgical sterilization or have been postmenopausal for atleast 2 years are not considered to be of reproductive potential.

35. Male and female subjects unwilling to practice contraception from the time ofagreeing to treatment until 12 months after completion of conditioning chemotherapyor CAR-T infusion.

36. Any past medical history that may interfere with the safety of the study treatmentor the evaluation of efficacy.

37. Based on the investigator's judgment, subjects are unlikely to complete all thestudy visits or procedures required by the protocol.

38. Previous use of any CAR-T cell product or other gene-modified T cell therapy.