Personalized KSX01-TCRT in Patients With Advanced Solid Tumors

Inclusion Criteria:

• Screening period 1- inclusion criteria:

Patients should complete all examinations for screening period 1 within 28 days after signing the informed consent form. Only those who meet the inclusion criteria for this stage can collect fresh tumor tissue, 2-3 archived pathological tissue white slides, and peripheral blood for TCR sequence screening and HLA typing testing.

1. Volunteer to participate in clinical research; Fully understand this study andvoluntarily sign an informed consent form; Willing to follow and capable ofcompleting all testing procedures.

2. Age range from 18 to 70 years old (including boundary values).

3. Solid tumors that have been confirmed by histological or cytological evaluation asincurable or metastatic, and have failed standard treatment or currently have noavailable standard treatment.

4. Expected survival time>6 months.

5. ECOG score 0 or 1.

6. Having sufficient organ function, defined as follows:

6.1) Hematology: 6.1.1) Hemoglobin 90 g/L (no blood transfusion received within 14days prior to examination); 6.1.2) Absolute value of neutrophils 1.5 109/L (did notreceive granulocyte colony stimulating factor treatment within 14 days prior toexamination); 6.1.3) Platelet count is 100 109/L in the absence of obvious liverlesions (primary or metastatic) (platelet transfusion not received within 14 daysbefore examination), or 75 109/L in the presence of liver lesions (platelettransfusion not received within 14 days before examination); 6.1.4) Absolutelymphocyte count (ALC) 0.7 109/L; 6.2) Liver function: 6.2.1) Total bilirubin (TBIL) ≤ 1.5 in the absence of obvious liver lesions (primary or metastatic) × Upper limitof normal (ULN), subjects with liver lesions or Gilbert disease ≤ 3 × ULN; 6.2.2)Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤ 2.5 × ULN (liver metastasis or liver cancer subjects can be ≤ 5 × ULN); Alkaline phosphatase (ALP) ≤ 2.5 × ULN (bone metastasis subject, ALP ≤ 5) × ULN); 6.3) Renal function:Creatinine clearance rate ≥ 60 mL/min (Cockcroft Fault formula: [140 age] × Weight [kg] × [0.85, female only]/(72 × Creatinine (mg/dl); Subjects with a creatinine clearance rate of<60 mL/min but ≥ 50 mL/min can also beenrolled if all of the following conditions are met: Serum creatinine and blood urea nitrogen (BUN) are within the normal range of theresearch center No clinical evidence showing chronic renal dysfunction (such asacidosis or electrolyte disorders) The urine routine and urine output are within thenormal range of the research center Note: It is not recommended to use IL-2 duringthe treatment period for subjects with a baseline creatinine clearance rate of<60mL/min.

7. The patient's HLA-I class molecule IHC expression is positive.

8. Patients with tumor lesions that can be collected and can screen out TCR sequencesthat can be used as drugs can enter the study. If the patient has obtainedpersonalized TCR sequences using previously collected and archived tumor tissue inother studies, they can directly enter screening period 2, but the collection timeof the archived tissue should be within one year before signing the informed consentfor this study.

9. The patient agrees to receive peripheral monocyte collection after all tests inscreening period 1 meet the standards.

• Screening period 2- inclusion criteria:

After receiving confirmation notification of TCR sequence locking from the partner, or if the patient has obtained their personalized TCR sequence in other studies, they can be arranged to undergo various inclusion evaluations in screening period 2.

The organ function and key examination items of the patient at this stage should not have significant changes compared to the examination results in screening period 1. If the patient's examination results during screening period 2 exceed the following criteria, peripheral monocyte collection should not be performed until the abnormal items return to normal range.

Screening period 2- Routine inclusion criteria

1. Confirmed screening and locking of tumor specific TCR sequences from the patient'sown body. For patients who have obtained TCR sequences through other researchprojects, they should sign an informed consent form for this research project beforeentering screening period 2.

2. Expected survival time>6 months.

3. ECOG score 0 or 1.

4. Having sufficient organ function, defined as follows:

4.1) Hematology: 4.1.1) Hemoglobin 90 g/L (no blood transfusion received within 14 days prior to examination); 4.1.2) Absolute value of neutrophils is 1.0 109/L (did not receive granulocyte colony stimulating factor treatment within 14 days before the examination); 4.1.3) Platelet count is 75 109/L in the absence of obvious liver lesions (primary or metastatic) (platelet transfusion was not received within 14 days before the examination); 4.1.4) Absolute lymphocyte count (ALC) 0.7 109/L; 4.2) Liver function: 4.2.1) Total bilirubin (TBIL) ≤ 1.5 in the absence of obvious liver lesions (primary or metastatic) × Upper limit of normal (ULN), subjects with liver lesions or Gilbert disease ≤ 3 × ULN; 4.2.2) Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) ≤ 2.5 × ULN (liver metastasis or liver cancer subjects can be ≤ 5 × ULN); Alkaline phosphatase (ALP) ≤ 2.5 × ULN (bone metastasis subject, ALP ≤ 5) × ULN); 4.3) Renal function: Creatinine clearance rate ≥ 60 mL/min (Cockcroft Fault formula: [140 age] × Weight [kg] × [0.85, female only]/(72 × Creatinine (mg/dl);

Subjects with a creatinine clearance rate of<60 mL/min but ≥ 50 mL/min can also be enrolled if all of the following conditions are met:

Serum creatinine and blood urea nitrogen (BUN) are within the normal range of the research center No acidosis or electrolyte disorders The urine routine and urine output are within the normal range of the research center Note: It is not recommended to use IL-2 during the treatment period for subjects with a baseline creatinine clearance rate of<60 mL/min.

4.4) The patient naturally breathes (without assisted oxygen supply) with a basal blood oxygen saturation of>92%.

Agree to accept peripheral monocyte collection. 6) Women of childbearing age who have the ability to conceive have a negative blood pregnancy test within 7 days before the first cell infusion (non fertility: surgical sterilization or at least 2 years after menopause), and the subjects of childbearing age use medically recognized contraceptive measures from the start of research treatment (chemotherapy) to 5 months after the last cell infusion, and no eggs have been retrieved during this period.

7. Male participants are willing to take medically approved contraceptive measureswithin 5 months after signing the informed consent form and the last cell infusion,and do not donate sperm during this period.

8. According to the iRECIST standard, there is at least one measurable lesion present.The following situations require approval from the researcher:

• During the dose increasing stage, subjects only have evaluable lesions but canevaluate their efficacy through their serum tumor markers;

• The subjects currently do not have measurable lesions, but the researchers havedetermined that they may develop or become measurable lesions within one month.

9. The toxicity and adverse reactions caused by frontline treatment should berestored to Grade 1 (excluding clinically insignificant toxicity, such as hairloss caused by chemotherapy).

Exclusion Criteria:

• Screening period 1- Exclusion criteria:

Subjects who meet any of the following criteria shall not participate in this clinical study:

1. The patient has received systemic chemotherapy on line 3 or above: Patients can be enrolled during the first line systemic chemotherapy, during thesecond line systemic chemotherapy period, or after the end of the second linesystemic chemotherapy (enrollment time is screening period 2 and meets the standardsand receives clearance chemotherapy), but they cannot receive the third linesystemic chemotherapy before enrollment:

• The second and third line systemic chemotherapy is defined as a systematicchemotherapy regimen after the progression of frontline treatment;

• For patients who complete neoadjuvant chemotherapy in the early stage of theirdisease, their neoadjuvant chemotherapy regimen can include up to two types ofchemotherapy;

• For patients who meet all other inclusion criteria but have a large number offrontline treatment lines, they can also be included after evaluation andconfirmation by the researchers. Frontline treatment plans targeting biological agents (such as immune checkpointinhibitors), small molecule targeted drugs, etc., are not considered as exclusioncriteria.

2. Within two years before signing the informed consent, the patient had a medicalhistory of other malignant tumors, except for non melanoma skin cancer, some cancersin situ (such as cervical cancer, bladder cancer, breast cancer), or low-riskprostate cancer.

3. Clinically confirmed liver diseases, including active hepatitis virus infection,alcoholic hepatitis, other types of hepatitis, cirrhosis, and hereditary liverdiseases; Among them, the subject's

• Hepatitis B surface antigen (HBsAg) and/or hepatitis B core antibody (HBcAb)are positive, and the hepatitis B virus DNA (HBV DNA) in the peripheral bloodis higher than the lower detection limit of the research center;

• HCV-Ab positive and HCV-RNA above the lower detection limit of the researchcenter Patients who can effectively control HBV-DNA and/or HCV-RNA aftertreatment can also be included in the group after being evaluated and approvedby the researcher. HBV-DNA positive subjects should receive hepatitis Btreatment after signing the informed consent form and continue to receiveKSX01-TCRT infusion for at least 6 months. Such subjects should also monitortheir HBV-DNA and hepatitis B related antigen antibodies on Day28 and at eachfollow-up visit point.

4. History of myocardial infarction, history of cardiac bypass surgery, unstableangina, active atrial fibrillation requiring treatment, symptomatic sinusbradycardia (heart rate<50 beats/min), or other clinically significant heartdiseases within 6 months prior to signing the informed consent form.

5. Tumor lesions invading the heart or large blood vessels. The patient has permanentpercutaneous nephrostomy, catheterization, bile duct, and other indwelling tubes,except for those that the researchers believe can be removed before gonorrheaclearance.

7. Primary immune deficiency. 8) HIV positive; Active HBV or HCV infection. 9) Receivedallogeneic stem cell transplantation within 6 months before signing the informedconsent form.

8. Prior to signing the informed consent form, CAR T cell therapy or other geneticallymodified T cell therapy other than this research technique was received.

9. Known allergies to dimethyl sulfoxide (DMSO) or any other cellular formulationcomponents and potential therapeutic drugs used during treatment (such ascyclophosphamide, fludarabine, and tolumab).

10. History of autoimmune diseases, except for the following:

-A history of hypothyroidism and the use of stable thyroid hormone replacement therapy;

-Patient has controllable type 1 diabetes 13) The patient's illness or condition results in their lack of understanding, participation, and/or adherence to this study plan.

14. Any other disease that researchers believe will impair the subject's tolerance tothe treatment regimen or significantly increase the risk of complications.

15. Known history of alcohol abuse, psychotropic substance abuse, or drug use. 16) Havea clear history of neurological or mental disorders in the past, such as epilepsy,dementia, schizophrenia, etc.

16. According to the judgment of the researchers, the underlying condition of thesubjects may increase their risk of receiving investigational drug treatment, or maycause confusion in the interpretation of toxic reactions and adverse events that mayoccur.

Other researchers believe that it is not suitable to participate in this study.

• Screening Period 2- Exclusion Criteria

Subjects who meet any of the following criteria shall not undergo peripheral monocyte collection:

1. Before monotherapy, the subject's anti-tumor treatment was not fully eluted (2 weeksor 5 half-lives, whichever is shorter):

-Except for the following situations: Gonadotropin (GnRH) agonists or antagonistsused for the treatment of prostate cancer Hormone replacement therapy or oralcontraceptives.

2. Known primary central nervous system (CNS) malignant tumors or symptomatic CNSmetastases. If the patient is diagnosed with a central nervous system disease and meets thefollowing conditions with the consent of the main researcher, it is not consideredan exclusion item:

• There are measurable or evaluable lesions outside the CNS;

• No history of intracranial or spinal cord bleeding;

• There is no need for ongoing or planned corticosteroid treatment within 14 daysprior to enrollment;

• Receiving stable doses of anticonvulsants;

• No stereotactic radiation or whole brain radiation therapy was received within 14 days prior to enrollment.

3. Women during pregnancy or lactation.

4. Receiving or planning to receive systemic corticosteroid therapy (>5 mg ofprednisone or equivalent treatment medication per day) or immunosuppressive drugswithin 7 days prior to monotherapy, except for the following:

• Intranasal, inhalation, topical steroids, or local administration (such asintra articular injection);

• Physiological doses of systemic steroids as an alternative therapy (such asphysiological corticosteroid replacement therapy for adrenal or pituitarydysfunction);

• Steroids are used as prophylactic drugs for hypersensitivity reactions (such ascomputed tomography [CT] prophylactic drugs).

5. The first second forced expiratory volume (FEV1)/forced vital capacity (FVC)<70%during lung function examination indicates abnormal lung function.

6. According to the judgment of the researchers, the underlying condition of thesubjects may increase their risk of receiving investigational drug treatment, or maycause confusion in the interpretation of toxic reactions and adverse events that mayoccur.

7. There are severe infections within 2 weeks before single collection. Major surgery (excluding diagnostic surgery) is performed within 4 weeks prior to singlecollection, or is expected to be performed during the study period. Subjects can beenrolled in planned or ongoing minor surgical procedures, such as establishingvenous channels.