A Phase Ⅰb Study on Autologous GC101 TIL Injection for the Treatment of Advanced Melanoma (MIZAR-002)

Inclusion Criteria:

1. Signed the informed consent form (ICF) and able to comply with the visits andrelated procedures specified in the protocol;

2. Aged ≥18 years and ≤75 years, regardless of gender;

3. Patients with unresectable advanced, recurrent or metastatic melanoma (excludinguveal melanoma) who have failed standard treatment with PD-1 antibodies, etc. (ifBRAF mutation is carried, BRAF and MEK inhibitor treatment failure);

4. TILs can be isolated from a surgically resectable tumor region: the tissue volumemust be >150mm3, and the lesion has not received local treatment (such asradiotherapy, radiofrequency ablation, oncolytic virus, etc.) or progressed afterlocal treatment;

5. There are still at least 1 measurable lesion (according to RECIST1.1 criteria [seeAppendix 4]) even after TIL sampling and resection of surgically resectable tissue;

6. ECOG performance status 0-1;

7. Expected survival time >3 months;

8. With sufficient hematology and end-organ function as defined by the followinglaboratory test results, the test results must be completed and issued within 7 daysbefore tumor tissue collection:

• White Blood Cell (WBC)≥2.5×10^9/L#

• Absolute Lymphocyte Count (ANC)≥1.5×10^9/L;

• Absolute Lymphocyte Count(ALC)≥0.7×10^9/L;

• Platelet≥100×10^9/L#

• International Normalized Ratio#INR#≤1.5×ULN;

• Activated Partial Thromboplastin Time#APTT#≤1.5×ULN;

• Serum Creatinine (Scr)≤1.5mg/dL (or 132.6μmol/L) or Creatinine

• Clearance≥60mL/min

• Urinalysis: urine protein less than 2+, or 24-hour urine protein <1g;

• Alanine aminotransferase(AST/SGOT) ≤3×ULN;

• Alanine aminotransferase (ALT/SGPT) ≤3×ULN;

• Total Bilirubin(TBIL)≤1.5×ULN#

9. • Premenopausal women who have not undergone sterilization surgery must agree to useeffective contraception measures from the start of study treatment (preconditioning)to one year after cell infusion, and the serum pregnancy test during the screeningperiod must be negative; *Men who have not undergone sterilization surgery mustagree to use effective contraception measures from the start of study treatment (preconditioning) until one year after cell infusion;

10. No absolute or relative contraindications for surgery;

11. Any melanoma treatment methods, including radiotherapy, chemotherapy, endocrinetherapy, targeted therapy, immunotherapy, tumor embolization, or traditional Chinesemedicine/herbal medicine treatment with anti-tumor indications, must be stopped 28days before infusion. If a small molecular targeted drug was used in the previoustreatment, the withdrawal time can be shortened to 5 half-lives of the drug used;

12. Good compliance and able to adhere to the study visit plan and other agreementrequirements.

Exclusion Criteria:

1. Participation in a clinical trial of another drug or biologic therapy or receipt ofa comparable cellular therapy within 28 days prior to infusion;

2. Combination of 2 or more malignant tumors, except: Eradicated malignant tumors thathave been inactive for ≥5 years prior to study entry and are at minimal risk ofrecurrence; adequately treated non-melanoma skin cancer or malignant nevus offreckle-like nevus without evidence of disease recurrence; adequately treatedcarcinoma in situ without evidence of disease recurrence;

3. Has received live attenuated vaccination after signing informed consent or isscheduled to receive it during the study;

4. Has not recovered from a prior procedure or treatment-related adverse reaction to ≤grade 1 nci ctcae 5.0 (except for toxicities such as alopecia, etc., which in thejudgment of the investigator pose no safety risk);

5. Known history of allergy to streptomycin, ciprofloxacin, or micafungin or allergy toany component of the infused product formulation;

6. Uncontrolled co-morbidities including, but not limited to, uncontrolled arterialhypertension (systolic blood pressure ≥160 mmhg and/or diastolic blood pressure ≥100mmhg) even with standardized treatment or any unstable cardiovascular diseaseincluding transient ischemic attack, cerebrovascular accident, myocardialinfarction, unstable angina pectoris within 6 months prior to enrollment; new yorkheart association ( nyha class iii or iv congestive heart failure with an ejectionfraction <50%; or severe cardiac rhythm or conduction abnormalities, such asventricular arrhythmias, degree ii-iii atrioventricular block, etc., requiringclinical intervention; ecg results showing clinically significant abnormalities or aqtcf ≥450ms (if the first test is abnormal, it may be retested at least 5 minutesapart twice and the combined result/mean value to determine eligibility) ;

7. Patients with esophageal or gastric varices that require immediate intervention (e.g., taping or sclerotherapy) or are considered to be at high risk for bleedingbased on the opinion of the investigator or consultation with a gastroenterologistor hepatologist, have evidence of portal hypertension (including splenomegalydetected on imaging), or have a prior history of variceal bleeding must haveundergone endoscopic evaluation within 3 months prior to enrollment;

8. Uncontrolled metabolic disorders, such as diabetes mellitus known to beuncontrolled, or other non-malignant organ or systemic diseases or secondaryreactions to cancer, and which can lead to higher medical risk and/or uncertainty insurvival evaluation;

9. Hepatic encephalopathy, hepatorenal syndrome or child-pugh class b or more severecirrhosis, liver failure;

10. Comorbidity with other serious organic or psychiatric disease;

11. Have an active systemic infection requiring treatment with positive blood culturesor imaging evidence of infection, including but not limited to active tuberculosis;

12. Be hiv-positive, have a positive serologic test for syphilis, or have clinicallyactive hepatitis a, b, or c, including viral carriers: Hepatitis b, excluding thosewho are HBsAg-positive; hepatitis c, excluding those who are HCVAb-positive;

13. Who have used, or in the judgment of the investigator have a co-morbid conditionrequiring the use of glucocorticosteroids or other immunosuppressive medicationsduring the trial within 4 weeks prior to pretreatment, excluding topicalglucocorticosteroids by nasal spray, inhalation, or other routes or physiologicdoses of systemic glucocorticosteroids (i.e., no more than 10 mg/day of prednisoneor equivalent dose of other glucocorticosteroids), or who have an active autoimmunedisease ( eczema, vitiligo, psoriasis, alopecia areata, or grave's disease that doesnot require systemic treatment within the last 2 years, other autoimmune diseasesthat are not expected to recur, and hypothyroidism requiring only thyroid hormonereplacement therapy, and subjects with type i diabetes mellitus requiring onlyinsulin replacement therapy may be enrolled);

14. Any nci ctcae5.0 immune-related adverse effect (irae) grade ≥ 3 during any priorperiod of immunotherapy receipt;

15. History of organ allograft, allogeneic stem cell transplantation and renalreplacement therapy;

16. Pulmonary fibrosis, interstitial lung disease (both past history and current), andacute lung disease;

17. Clinically uncontrollable third space effusions, such as pleural and abdominaleffusions that cannot be controlled by drainage or other means prior to enrollment;

18. Patients with known molluscum contagiosum metastases;

19. Patients with clinically symptomatic central nervous system metastases (e.g.,cerebral edema, need for hormonal intervention, or progression of brain metastases).Patients with prior treatment for brain metastases, such as clinical stability (mri)that has been maintained for at least 2 months and who have discontinued systemichormone therapy (dose >10 mg/day prednisone or other equipotent hormone) for >4weeks may be included;

20. Women who are pregnant or breastfeeding;

21. History of allogeneic t-cell and nk-cell therapy;

22. If the investigator believes that other circumstances are not suitable forenrollment.