CT011 Autologous CAR-T Cells in Patients With Hepatocellular Carcinoma at Risk of Recurrence After Surgical Resection

Inclusion Criteria:

To be included in the trial, participants must meet all of the following criteria:

1. Volunteer to participate in the clinical trial; fully understand and are informed ofthis trial and sign the informed consent form; Willing to follow and able tocomplete all trial procedures;

2. Age 18-75 years, inclusive, male or female;

3. Initially diagnosed with CNLC stage IIIa HCC with any of the following vasculartumor thrombi and absence of atrial tumor thrombi on preoperative imaging:

• Portal vein tumor thrombus (PVTT);

• Hepatic vein tumor thrombus (HVTT);

• Inferior vena cava tumor thrombus (IVCTT);

4. Has undergone surgical resection:

• Pathological evaluation of surgical resection specimen with negative margins;

• Preoperative conversion/neoadjuvant therapy and/or postoperative therapy areallowed;

5. The participant has recovered from liver resection and postoperative progressiveincrease in AFP level(including: a. AFP increase of at least 20% in any 3 monthsafter surgery; or b. AFP increase of ≥ 10% in any 2 consecutive tests after surgery)with a potential tendency to recurrence as assessed by the investigator.

6. Tumor tissue samples positive for GPC3 by immunohistochemistry (IHC) (stainingintensity ≥ 1 +, percentage of stained tumor cells ≥ 10%);

7. Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1 (within 7days prior to apheresis);

8. Child-Pugh score ≤ 7 points (within 7 days prior to apheresis);

9. Estimated survival > 12 weeks;

10. Adequate venous access for apheresis;

11. Laboratory test results within 7 days prior to apheresis should meet the followingcriteria (if the laboratory test results do not meet the following criteria, arepeat test within 1 week is allowed; if the laboratory test results still do notmeet the criteria, it will be considered a screening failure):

• Hematology (without transfusion, platelet transfusion, colony-stimulatingfactor and other supportive treatment within 7 days before detection, exceptrecombinant human erythropoietin): neutrophil count (ANC) ≥ 1.5 × 109/L,lymphocyte count (LY) ≥ 0.5 × 109/L, platelet count (PLT) ≥ 75 × 109/L,hemoglobin (Hb) ≥ 9.0 g/dL; The results of hematology within 24 hours beforeapheresis shall also meet this standard;

• Blood chemistry: serum creatinine ≤ 1. 5 × upper limit of normal (ULN),endogenous creatinine clearance ≥ 40 mL/min (using Cockcroft-Gault formula),alanine aminotransferase (ALT) ≤ 5×ULN, aspartate aminotransferase (AST) ≤ 5 ×ULN, alkaline phosphatase (ALP) ≤ 5 × ULN, total bilirubin (TBil) ≤ 3 × ULN,serum albumin (ALB) ≥ 28 g/L, and serum lipase and amylase ≤ 2 × ULN;

• Prothrombin time (PT) prolongation ≤ 4 s;

12. Female participants of childbearing potential must have a negative serum pregnancytest at screening and agree to remain abstinent or use highly effective and reliablecontraception (< 1% failure rate per year) during the treatment period and for atleast 1 year after the last dose of trial treatment, during which time they must notdonate eggs:

• Women of childbearing potential are defined as women who have not reachedpost-menarche but have not reached a post-menopausal state (no menses for ≥ 12consecutive months, with no cause other than menopause) and who have not beenpermanently infertile due to surgery (removal of ovaries, fallopian tubes,and/or uterus) or other reasons (e.g. M ü llerian agenesis, etc.) as determinedby the investigator;

• Contraceptive methods with an annual failure rate of < 1% include: bilateraltubal ligation, male sterilization, approved hormonal contraceptives,hormone-releasing intrauterine devices, copper-containing intrauterine devices;Periodic abstinence (e.g., calendar, ovulation, symptothermal, or postovulationrhythm methods) and withdrawal are not adequate methods of contraception.

13. Male participants who are sexually active with a female of childbearing potential,who have not had a vasectomy, must agree to remain abstinent or to use contraception (e.g., condoms in combination with other contraceptive measures to achieve an annualfailure rate of < 1%, see inclusion criterion # 12) during the treatment period andfor 1 year after the last dose of study treatment and refrain from donating spermduring this period.

Exclusion Criteria:

Participants were not included in the trial if they met any of the following criteria:

1. Known fibrolamellar HCC, sarcomatoid HCC, or mixedhepatocellular-cholangiocarcinoma;

2. Intrahepatic recurrence or extrahepatic metastasis, or residual hepatocellularcarcinoma detected before apheresis (imaging evidence according to RECIST v1.1);

3. More than 2 years since surgical resection;

4. Pregnant or lactating females;

5. Positive test results for any of the following: human immunodeficiency virus (HIV)antibody, Treponema pallidum antibody, hepatitis C virus (HCV) ribonucleic acid (RNA), hepatitis B surface antigen (HBsAg) and/or hepatitis B core antibody (HBcAb)positive and hepatitis B virus deoxyribonucleic acid [HBV DNA] ≥ 1000 IU/mL (HBsAg-positive or HBcAb-positive participants must receive antiviral therapy),cytomegalovirus (CMV) DNA, Epstein-Barr virus (EBV) DNA;

6. Any uncontrolled active infection, including but not limited to active tuberculosis,infectious diseases requiring systemic treatment, etc.; Patients who use drugs toprevent infection can be enrolled at the discretion of the investigator;

7. Subjects with clinically significant abnormal thyroid function (freetriiodothyronine [FT3], free thyroxine [FT4] and serum thyroid stimulating hormone [TSH] for serum thyroid hormones, and total thyroxine [TT4] and totaltriiodothyronine [TT3] for serum thyroid hormones if necessary) judged by theinvestigator and not suitable for entry into the trial after assessment; Patientswith stable thyroid function after treatment can be considered for inclusion;

8. Previous or current hepatic encephalopathy;

9. Presence of clinically significant massive abdominal/pleural effusion, defined as:positive signs of pleural/peritoneal effusion on physical examination orpleural/peritoneal effusion requiring intervention (e.g., paracentesis or drugtherapy) for control;

10. Toxicities caused by previous treatment have not recovered to Common TerminologyCriteria for Adverse Events (CTCAE) v5.0 ≤ Grade 1, except for alopecia,pigmentation, other laboratory abnormalities that do not affect the tolerability ofthe participants as judged by the investigator;

11. Received anti-tumor treatment for the disease under study within 2 weeks prior toapheresis, including but not limited to surgery, systemic drug therapy (or within 5half-lives of the drug, whichever is shorter), radiotherapy, interventional therapy,etc.;

12. Received immunotherapy including anti-PD-1/PD-L1, anti-CTLA-4, or any otherinvestigational therapy within 4 weeks (or within 5 half-lives of the drug,whichever is shorter) prior to apheresis;

13. Previously received any cell therapy (including CAR-T cells, TCR-T cells, TILs,etc.);

14. Received systemic glucocorticoid therapy within 7 days prior to apheresis; Patientswith recent or current use of inhaled or topical corticosteroids and physiologicdose replacement therapy may be enrolled;

15. Vaccination with live or live attenuated vaccines within 4 weeks prior to apheresisor planned during the trial;

16. Known active autoimmune disease, including but not limited to rheumatoid arthritis,systemic lupus erythematosus, autoimmune hepatitis, interstitial lung disease,inflammatory bowel disease, antiphospholipid syndrome, Wegener's granulomatosis,Sjogren's syndrome, multiple sclerosis, glomerulonephritis, etc.; Or otherparticipants who require chronic use of immunosuppressive therapy;

17. Participants with a history of organ transplantation, allogeneic hematopoietic stemcell transplantation or awaiting organ transplantation;

18. Previous allergies to immunotherapy, tocilizumab, cyclophosphamide or fludarabineand other related drugs, previous history of severe allergies, or known allergies tocomponents of CT011 cell infusion preparation (such as albumin or dimethylsulfoxide, etc.);

19. Presence of central nervous system metastasis or related symptoms, or clinicallysignificant central nervous system disease or abnormal neurological examinationresults or psychiatric disease;

20. Need for long-term anticoagulation/thrombolysis/antiplatelet therapy (such aswarfarin, heparin, rivaroxaban, aspirin, dipyridamole, clopidogrel, etc.); Patientsreceiving prophylactic anticoagulation to maintain patency of venous access devicesmay be included in this trial;

21. Major surgical procedure or significant trauma within 4 weeks prior to apheresis, oranticipation of the need for major surgery during the trial;

22. Pre-apheresis oxygen saturation ≤ 95% (under room air, the finger pulse oxygen testis accepted);

23. Any other disease, metabolic disorder, sign, or test result that contraindicates theuse of the trial drug, affects the interpretation of the results, or places theparticipant at high risk of treatment complications, which, as assessed by theinvestigator, would not be appropriate for participation in the trial, including butnot limited to: poorly controlled diabetes mellitus (treated glycosylated hemoglobin [HbA1c] > 8%), poorly controlled hypertension (blood pressure > 160 mmHg/100 mmHg),hypotension requiring vasopressor medication, uncontrolled congestive heart failure (New York Heart Association [NYHA] Class III-IV), Left ventricular ejection fraction [LVEF] < 50%, myocardial infarction within the past 6 months, arrhythmia not wellcontrolled by drug therapy, unstable angina pectoris, or other serious heartdisease, pulmonary embolism, chronic obstructive pulmonary disease, interstitiallung disease or clinically significant pulmonary function test abnormality as judgedby the investigator; Presence of currently gastrointestinal obstruction oractive/unstable gastrointestinal ulcer, gastrointestinal bleeding within the past 3months or with bleeding risk (e.g., esophageal and gastric varices caused by portalhypertension or bleeding tendency; patients with liver cirrhosis are recommended toundergo additional gastroscopy during the screening period according to the judgmentof the investigator to determine the condition of varices); The participant is in asevere inflammatory state overall (e.g., increased neutrophil count and/orC-reactive protein, fever ≥ 38 ℃ unrelated to the underlying disease);

24. Presence of other incurable malignancies within the past 5 years or at the sametime, except for appropriately treated cervical carcinoma in situ, skin basal cellcarcinoma and other malignancies with very low risk of metastasis/death;

25. Inability or unwillingness of the participant to comply with the requirements of thetrial protocol as assessed by the investigator.