TREATMENT OF CD30-POSITIVE CANCER

Abstract

Methods for treating a CD30-positive cancer in a subject are disclosed, wherein the methods comprise administering a lymphodepleting chemotherapy and CD30-specific chimeric antigen receptor (CAR)-expressing cells.

Claims

1. A method of treating a CD30-positive cancer in a subject, comprising: (i) administering a lymphodepleting chemotherapy to the subject, and (ii) subsequently administering CD30-specific chimeric antigen receptor (CAR)-expressing T cells to the subject.

2. A population of CD30-specific chimeric antigen receptor (CAR)-expressing T cells for use in a method of treating a CD30-positive cancer, wherein the method comprises: (i) administering a lymphodepleting chemotherapy to the subject, and (ii) subsequently administering CD30-specific CAR-T cells to the subject.

3. Use of a population of CD30-specific chimeric antigen receptor (CAR)-expressing T cells in the manufacture of a medicament for use in a method of treating a CD30-positive cancer, wherein the method comprises: (i) administering a lymphodepleting chemotherapy to the subject, and (ii) subsequently administering CD30-specific CAR-T cells to the subject.

4. The method according to claim 1, the population for use according to claim 2, or the use according to claim 3, wherein administering a lymphodepleting chemotherapy to the subject comprises administering fludarabine and bendamustine.

5. The method, the population for use or the use according to any one of claims 1 to 4, wherein the method comprises administering fludarabine at a dose of 15 to 60 mg/m.sup.2 per day, for 2 to 6 consecutive days.

6. The method, the population for use or the use according to any one of claims 1 to 5, wherein the method comprises administering fludarabine at a dose of 30 mg/m.sup.2 per day, for 3 consecutive days.

7. The method, the population for use or the use according to any one of claims 1 to 6, wherein the method comprises administering bendamustine at a dose of 35 to 140 mg/m.sup.2 per day, for 2 to 6 consecutive days.

8. The method, the population for use or the use according to any one of claims 1 to 7, wherein the method comprises administering bendamustine at a dose of 70 mg/m.sup.2 per day, for 3 consecutive days.

9. The method, the population for use or the use according to any one of claims 1 to 8, wherein the method comprises administering 5×10.sup.7 CD30-specific CAR-expressing T cells/m.sup.2 to 1×10.sup.9 CD30-specific CAR-expressing T cells/m.sup.2 to the subject.

10. The method, the population for use or the use according to any one of claims 1 to 9, wherein the method comprises administering 1×10.sup.8 CD30-specific CAR-expressing T cells/m.sup.2 to 6×10.sup.8 CD30-specific CAR-expressing T cells/m.sup.2 to the subject.

11. The method, the population for use or the use according to any one of claims 1 to 10, wherein the method comprises: (i) administering fludarabine at a dose of 30 mg/m.sup.2 per day and bendamustine at a dose of 70 mg/m.sup.2 per day to a subject for 3 consecutive days, and (ii) subsequently administering CD30-specific CAR-expressing T cells to the subject at a dose of 2×10.sup.8 CD30-specific CAR-expressing T cells/m.sup.2 to 6×10.sup.8 CD30-specific CAR-expressing T cells/m.sup.2.

12. The method, the population for use or the use according to any one of claims 1 to 11, wherein the CD30-positive cancer is selected from: a hematological cancer, a solid cancer, a hematopoietic malignancy, Hodgkin's lymphoma, anaplastic large cell lymphoma, peripheral T cell lymphoma, peripheral T cell lymphoma not otherwise specified, T cell leukemia, T cell lymphoma, cutaneous T cell lymphoma, NK-T cell lymphoma, extranodal NK-T cell lymphoma, non-Hodgkin's lymphoma, B cell non-Hodgkin's lymphoma, diffuse large B cell lymphoma, diffuse large B cell lymphoma not otherwise specified, EBV-positive B cell lymphoma, EBV-positive diffuse large B cell lymphoma, primary mediastinal B cell lymphoma, advanced systemic mastocytosis, a germ cell tumor and testicular embryonal carcinoma.

13. The method, the population for use or the use according to any one of claims 1 to 12, wherein the CD30-positive cancer is selected from: Hodgkin's lymphoma, non-Hodgkin's lymphoma, anaplastic large cell lymphoma, peripheral T cell lymphoma not otherwise specified, extranodal NK-T cell lymphoma, diffuse large B cell lymphoma not otherwise specified and primary mediastinal large B-cell lymphoma.

14. The method, the population for use or the use according to any one of claims 1 to 13, wherein the subject has previously failed therapy for the CD30-positive cancer.

15. The method, the population for use or the use according to any one of claims 1 to 14, wherein the CD30-positive cancer is a relapsed or refractory CD30-positive cancer.

16. The method, the population for use or the use according to any one of claims 1 to 15, wherein CD30-specific CAR-expressing T cells comprise a CAR comprising: (i) an antigen-binding domain which binds specifically to CD30, (ii) a transmembrane domain, and (iii) a signalling domain, wherein the signalling domain comprises: (a) an amino acid sequence derived from the intracellular domain of CD28, and (b) an amino acid sequence comprising an immunoreceptor tyrosine-based activation motif (ITAM).

17. The method, the population for use or the use according to claim 16, wherein the signalling domain comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:26.

18. The method, the population for use or the use according to claim 16 or claim 17 wherein the transmembrane domain is derived from the transmembrane domain of CD28.

19. The method, the population for use or the use according to any one of claims 16 to 18, wherein the transmembrane domain comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:20.

20. The method, the population for use or the use according to any one of claims 16 to 19, wherein the antigen-binding domain comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:14, and an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:15.

21. The method, the population for use or the use according to any one of claims 16 to 20, wherein the antigen-binding domain comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:18.

22. The method, the population for use or the use according to any one of claims 16 to 21, wherein the signalling domain comprises: (a) an amino acid sequence derived from the intracellular domain of CD3.

23. The method, the population for use or the use according to any one of claims 16 to 22, wherein the signalling domain comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:25.

24. The method, the population for use or the use according to any one of claims 16 to 23, wherein the CAR additionally comprises a hinge region provided between the antigen-binding domain and the transmembrane domain.

25. The method, the population for use or the use according to any one of claims 16 to 24, wherein the hinge region comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:33.

26. The method, the population for use or the use according to any one of claims 16 to 25, wherein the CAR comprises an amino acid sequence having at least 80% amino acid sequence identity to SEQ ID NO:35 or 36.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0352] Embodiments and experiments illustrating the principles of the invention will now be discussed with reference to the accompanying figures.

[0353] FIG. 1. Swimmer plot showing duration of Response of Classical HL Patients from LCCC 1532-ATL with Measurable Disease at the Time of Treatment (Data cutoff: 14 Feb. 2020).

[0354] FIG. 2. Kaplan-Meier plot showing PFS of Classical HL Patients from LCCC 1532-ATL. The progression free survival of evaluable classical HL patients from LCCC 1532-ATL treated with bendamustine and fludarabine (n=16) is shown.

[0355] FIG. 3. Schematic overview of TESSCAR001 (NCT04268706) study design. Primary Endpoints: ORR as assessed by an Independent Radiology Review Committee; Secondary Endpoints: Safety, ORR as assessed by Investigator, DOR, PFS, OS, HRQoL; Exploratory Endpoints: Expansion and persistence of CD30.CAR-T cells in blood; immunogenicity; cytokine profiling; immunological parameters; tumor marker and ctDNA; Safety Assessments: AE and concomitant medication collection; Long-term follow-up: Additional safety monitoring×15 years. *2nd infusion of CD30.CAR-T cell: at the discretion of Principal Investigator and after discussion with the Sponsor or designee, a 2nd CD30.CAR-T infusion may be administrated if the patient has CR, PR or SD at the first response assessment and subsequently has disease progression at least 3 months post CD30.CAR-T infusion. Abbreviations: CR, complete response; DOR, duration of response; HRQoL, health related quality of life; OS, overall survival; ORR, objective response rate; PFS, progression-free survival; PD, progressive disease; PR, partial response; SD, stable disease.

[0356] FIG. 4. Schematic overview of TESSCAR001 (NCT04268706) and TESSCAR002 study procedures. *Reimaging scan can be done within 1 week prior to LD; **The washout period will be based on the type of therapy used; #LD and CD30.CAR-T-association safety evaluation to be performed from Treatment Phase until LTFU Phase. T Tumor response assessments. Abbreviations: LTFU, long term follow up; ICF, informed consent form; LD, lymphodepletion chemotherapy; EOT, end of treatment; EOS, end of study; M, month; YR, year.

[0357] FIG. 5. Schematic overview of TESSCAR002 study design. $ Subjects may be replaced if they are unable to complete follow-up through the DLT period for any reason other than a DLT. § A minimum of 3 patients is required to complete the DLT assessment period before a decision can be taken to escalate the dose for the next patient. At each new dose level, 1st and 2nd patient will be dosed 4 weeks apart; if no DLT detected from the 1st patient, the 2nd and 3rd patient can be dosed concurrently. Dose-escalation/de-escalation to be performed until at least 6 patients have been evaluated at the highest dose with acceptable safety (i.e. ‘acceptable safety’ is when the BOIN recommends to either stay at same dose level or escalate), or until a total of approximately 21 patients have been dosed. #Dose will be de-escalated to DL0 1×10.sup.8/m2 CD30.CAR-T cells in the event of 1 DLT in 1 patient, or 1 DLT among 2 patients, or 2 DLTs among 3 patients at DL1. Abbreviations: DOR, Duration of response; ORR, Objective response rate; OS, Overall survival; PFS, Progression free survival; PK, Pharmacokinetics; RP2D, Recommended Phase 2 dose.

EXAMPLES

[0358] In the following Examples, the inventors describe treatment of CD30-positive cancer using methods employing lymphodepleting chemotherapy and adoptive transfer of CD30-specific CAR-expressing T cells.

Example 1: Study LCCC 1532-ATL (NCT02690545): Phase 1/2 Study of the Administration of T Lymphocytes Expressing the CD30 Chimeric Antigen Receptor (CAR) for Relapsed or Refractory CD30+ Hodgkin Lymphoma and CD30+ Non-Hodgkin Lymphoma

[0359] 1.1 Overview

[0360] A Phase 1/2 study in patients with relapsed or refractory Hodgkin Lymphoma was conducted at the University of North Carolina (UNC, Chapel Hill, N.C.) in protocol NCT02690545.

[0361] Patients received autologous CD30.CAR-T manufactured in Good Manufacturing Practice (GMP)-compliant facilities, using a clinical grade gamma-retroviral vector and following a Standard Operating Procedure (SOP).

[0362] Patients with relapsed/refractory CD30.sup.+ lymphomas who progressed after at least 2 lines of therapy were eligible for enrollment. BV treatment was allowed, but not within 4 weeks of the scheduled infusion of CD30.CAR-T cells. Documented CD30 expression by immunohistochemistry based on the institutional hematopathology standard was required, but there was no specific cutoff for the percentage of CD30.sup.+ tumor cells.

[0363] Patients with CD30.sup.+ T cell or anaplastic large cell lymphoma as well as CD30.sup.+ B-cell lymphoma were also enrolled, but the present Example only reports the outcome of 34 patients with HL (26 patients).

[0364] Bridging chemotherapy, at the discretion of the treating physician, was allowed before lymphodepletion. The conditioning regimen consisted of bendamustine at a dose of 90 mg/m.sup.2/day for 2 days for the first cohort of 8 patients, and bendamustine 70 mg/m.sup.2/day and fludarabine 30 mg/m.sup.2/day for 3 days for the subsequent 18 patients enrolled in the second cohort.

[0365] Infusion of CD30.CAR-T cells occurred 2-5 days after lymphodepletion. Patients received either 1×10.sup.8 CAR-T cells/m.sup.2 or 2×10.sup.8 CAR-T cells/m.sup.2. An expansion cohort of patients received the highest dose level of 2×10.sup.8 CAR-T cells/m.sup.2. A second infusion of CD30.CAR-T cells at the highest dose level was allowed in patients who had stable disease or partial response after the first treatment.

[0366] The primary objective of the Phase 1 portion of the study was to establish a safe dose of CD30.CAR-T cells to infuse after lymphodepletion. Secondary endpoints included ORR, duration of response (DOR), overall survival (OS), and measurement of the expansion and persistence of CD30.CAR-T in the peripheral blood after infusion. Data were analyzed separately in patients who received non-fludarabine-based lymphodepletion and those who received regimens containing fludarabine.

[0367] Response was assessed at 6-8 weeks after CD30.CAR-T cell infusion using the Lugano criteria (Cheson et al., J Clin Oncol (2014) 32: 3059-3068). Response rate was estimated only in patients who had active disease at the time of lymphodepletion.

[0368] 1.2 Patient Demographics

[0369] Between September 2016 and 14 Feb. 2020, 29 patients with HL were enrolled in LCCC 1532-ATL study and 26 received CD30.CAR-T cells. As of 14 Feb. 2020 data cutoff, there were 3 patients enrolled at LCCC 1532-ATL study who did not receive treatment. Of the 3 patients, 1 elected not to proceed with the clinical trial, 1 had no active disease at the time of collection, 1 failed CAR-T cell manufacturing due to intensive prior autologous stem cell transplant (ASCT), allogeneic stem cell transplant (alloSCT) and multiple donor lymphocyte infusions.

TABLE-US-00002 TABLE 1 Classical HL Patient Characteristics from LCCC 1532-ATL: Benda Flu/Benda Characteristics n = 8* (%) n = 18 (%) HL Subtype: n (%) NS 6 (75) 11 (61) MC 2 (25) 2 (11) NOS 0 5 (28) Stage at I-II 1 (13) 7 (39) diagnosis: n (%) III-IV 7 (88) 11 (61) Age (years), median (range) 49 (23-67) 31.5 (15-45) Male sex: n (%) 5 (63) 13 (72) Prior Therapies, median (range) 7.5 (5-17) 8 (3-23) Bridging Therapy: n (%) 8 (100) 10 (56) Prior BV: n (%) 8 (100) 17 (94) Prior CPI: n (%) 7 (88) 13 (72) Prior ASCT: n (%) 7 (88) 14 (78) Prior alloSCT: n (%) 2 (25) 8 (44) CAR-T cells/m.sup.2: 2 × 10.sup.7 0 0 n (%) 1 × 10.sup.8 3 (38) 1 (6) 2 × 10.sup.8 5 (63) 17 (94) alloSCT = allogeneic stem cell transplant; ASCT = autologous stem cell transplant; Benda = bendamustine; BV = brentuximab vedotin; CPI = checkpoint inhibitor; Flu = fludarabine; HL = Hodgkin lymphoma; MC = mixed cellularity; NOS = not otherwise specified; NS = nodular sclerosis.

[0370] 1.3 Efficacy *One patient who received two treatments (one at UNC and the second at BCM 2 years later) was considered.

[0371] Eighteen patients received bendamustine and fludarabine in LCCC 1532-ATL study. Of these, 2 patients were in CR at the time of infusion, maintained CR, and were not included in the efficacy analysis.

TABLE-US-00003 TABLE 2 Clinical Responses of Classical HL Patients from LCCC 1532- ATL with Measurable Disease at the Time of Treatment Bendamustine* Flu/Benda** n = 5 (%) n = 16 (%) ORR: n (%) CR + PR 0 (0%) 13 (81.3%) RR: n (%) CR 0 (0%) 12 (75.0%) PR 0 (0%) 1 (6.3%) SD 1 (20%) 1 (6.3%) PD 4 (80%) 2 (12.5%) CR: complete response; ORR objective response rate; PR: partial response; RR: response rate; SD: stable disease, Benda: bendamustine: Flu: fludarabine *3 patients were in CR prior to LD due to bridging therapy and are not included in the analysis. **2 patients were in CR prior to LD due to bridging therapy and are not included in the analysis.

TABLE-US-00004 TABLE 3 Characteristics, Clinical Responses and Safety Data of Classical HL Patients Under Age 18 from LCCC 1532-ATL No. of CD30. Subject Prior LD CAR-T Lymphodepletion CAR-T cell ID Age Sex Therapies Regimen Dose Response related AE related AE UNC 33 15 Female 4 Flu/ 1 × 10.sup.8/m.sup.2 CR Anemia (G1) Alanine Benda Anoexia (G1) aminotransferase Aminotransferase Elevated (G1) Elevated (G1) Cough (G1) Hyponatremia (G1) Hypophosphatemia (G1) Lymphopenia (G4) Neutropenia (G2) Thombocytopenia (G1) Vomiting (G1) Leukopenia (G2)

[0372] In patients treated with lymphodepletion chemotherapy of bendamustine and fludarabine, the ORR was 81%, with CR reported in 12 patients (75%), PR in 1 patient (6%), SD in 1 patient (6%) and PD in 2 patients (13%) (Table 2). Among them, one patient under age 18 was treated, an achieved CR (Table 3).

[0373] Eight patients received lymphodepletion using bendamustine alone prior to CD30.CAR-T cell infusion. Of these 8 patients, 3 patients were in CR prior to lymphodepletion due to bridging therapy and were not included in the analysis (Error! Reference source not found. Table 2). None of the 5 patients with active disease showed objective clinical responses when treated (Table 2).

[0374] The duration of response (DOR) patients with measurable disease at the time of treatment is shown in FIG. 1. Among the 12 patients with CR after 1.sup.st infusion 6 patients had responses ongoing.

[0375] The 1-year PFS rate of 16 patients treated with bendamustine and fludarabine regimen was 57% (95% Cl: 28%-78%), with median PFS of 13.0 months (95% Cl: 5.4-NE)—see FIG. 2.

[0376] 1.4 Safety

[0377] No dose limiting toxicities associated with CD30.CAR-T cell infusion was observed. The patient under age 18 treated did not experience CRS or other CAR-T cell related toxicity, except grade 1 ALT (Table 3).

[0378] Cytokine release syndrome (CRS) was observed in 3 patients (24%) (Table 4). All CRS events were grade 1 and resolved spontaneously with no requirement for tocilizumab or steroid administration. Cytokines associated with the occurrence of CRS, such as IL-6 and IL1Rα, and C-reactive protein (CRP) were elevated in the plasma of patients developing clinical signs of CRS. Neurotoxicity was not observed.

[0379] Six patients developed a non-pruritic, non-tender, maculopapular skin rash. None of the patients required specific treatment for the rash as it resolved spontaneously within 3-7 days.

TABLE-US-00005 TABLE 4 Grade 3 or Higher Adverse Events and Adverse Events of Special Interest of Classical HL Patients from LCCC 1532-ATL Adverse Events Benda Flu/Benda (of Special Interests or >Grade 3): n (%) (n = 8) (n = 18) Lymphopenia 8 (100) 18 (100) Leukopenia 3 (38) 8 (44) Neutropenia 2 (25) 7 (39) Thrombocytopenia 1 (13) 7 (39) Anemia 0 3 (17) Hypoalbuminemia 0 0 Hyponatremia 0 0 Dyspnea 0 0 Rash 0 0 Headache 0 0 Pharyngitis 0 1 (6) Lung Infection 0 1 (6) Cytokine Release Syndrome (all Grade 1) 1 (13) 2 (11) Grade 3/4 Neutropenia at day 28 0 2 (11) Grade 3/4 Thrombocytopenia at day 28 0 6 (33) Grade 3/4 Anemia at day 28 0 0 Prolonged Grade 3/4 Neutropenia 0 0 (at month 3)* Prolonged Grade 3/4 0 3 (17) Thrombocytopenia (at month 3)* Prolonged Grade 3/4 Anemia (at month 3)* 0 1 (6) Rash (Any Grade) 2 (25) 4 (22) Benda = bendamustine; Flu = fludarabine; LD = lymphodepletion; *3 patients did not have data at 3 months as they withdrew from study.

[0380] There were Grade 3 or higher toxicities reported during the first 6 weeks, and most of them were hematologic and consistent with toxicities caused by the conditioning chemotherapy. Grade 3 or higher toxicities included lymphopenia, leukopenia, neutropenia, thrombocytopenia, anemia, hypoalbuminemia, hyponatremia, dyspnea, pharyngitis, lung infection and headache (Table 4). Grade 3/4 neutropenia (ANC <1.0/mL) that had not resolved by Day 28 day occurred in 2 patients. However, both patients resolved their Grade 3/4 neutropenia by Day 90. Six patients had Grade 3/4 thrombocytopenia (platelets <50,000/mL) that had not resolved by Day 28. Three patients had Grade 3/4 thrombocytopenia at Month 3 (Table 4).

[0381] Incidence of Grade 3 or higher AEs in patients treated with bendamustine and fludarabine was low (Table 4), which included leukopenia (44%), neutropenia (39%), hypoalbuminemia (0%), hyponatremia (0%), dyspnea, pharyngitis, lung infection and headache (0%). One patient experienced Grade 3 acute kidney injury and hypotension after receiving the first dose of fludarabine and bendamustine, but prior to CD30.CAR-T cell infusion. This patient did not complete the scheduled lymphodepletion regimen, but their symptoms subsequently resolved and they received the scheduled dose of CD30.CAR-T cells.

[0382] 1.5 Conclusions

[0383] Patients receiving lymphodepletion using bendamustine and fludarabine followed by CD30.CAR-T cell infusion display improved overall response and complete response rates as compared to response rates reported for Patients receiving lymphodepletion using bendamustine and fludarabine followed by CD30.CAR-T cell infusion in Ramos et al., Biol Blood Marrow Transplant 25 (2019) S7-S75, Abstract 79.

[0384] Ramos et al. reports preliminary data for NCT02917083 (RELY-30), wherein 10 relapsed/refractory HL patients received cyclophosphamide 500 mg/m.sup.2 and fludarabine 30 mg/m.sup.2 daily for 3 days, and were subsequently administered CD30.CAR T cells. Out of 9 patients evaluated at 6 weeks after infusion, 6 had CR, and 3 patients had disease progression (i.e. ORR=66.6%, CR=66.6%). Three of the 9 patients progressed (PD=33.3%).

[0385] By contrast, in the present Example, 13 out of 16 patients that received bendamustine 70 mg/m.sup.2 and fludarabine 30 mg/m.sup.2 daily for 3 days and were subsequently administered CD30.CAR T cells displayed a clinical response (i.e. CR+PR; ORR=81.3%), and 12 of these patients displayed a complete response (CR=75%). Only two of the 16 patients progressed (PD=12.5%).

[0386] The results identify benda/flu treatment as a promising conditioning lymphodepleting therapy for use in methods of treatment employing CD30.CAR-T cells.

Example 2: Study TESSCAR001 (NCT04268706): A Phase 2. Multicenter, Open Label, Single Arm Study Designed to Evaluate the Efficacy and Safety of CD30-Directed Genetically Modified Autologous T-Cells (CD30.CAR-T) in Adult and Pediatric Patients with Relapsed or Refractory CD30-Positive cHL

[0387] 2.1 Study Rationale

[0388] As the outcomes for patients with relapsed or refractory cHL who have failed standard available therapy are poor, new treatment approaches are urgently needed for this population. Example demonstrates that CD30.CAR-T cell therapy is well tolerated, and that such cells have significant clinical activity in heavily pretreated patients with relapsed or refractory cHL. Higher rates of clinical response were found to be correlated with the use of lymphodepleting chemotherapy relative to response rates reported by Ramos et al., J Clin Invest. (2017) 127(9):3462-3471 for therapy with CD30.CAR-T cells without lymphodepleting chemotherapy (NCT01316146). The favourable toxicity profile and encouraging antitumor activity of autologous CD30.CAR-T demonstrated in Example 1 provides strong support for further investigation in patients with relapsed or refractory cHL.

[0389] 2.2 Objectives and Endpoints

[0390] 2.2.1 Primary Objectives

[0391] The anti-tumor effect of autologous CD30.CAR-T are evaluated using ORR (as assessed by an Independent Radiology Review Committee (IRRC) per the Revised Criteria for Response Assessment: The Lugano Classification (Cheson et al., J Clin Oncol (2014) 32: 3059-3068)).

[0392] The ORR is defined as the proportion of patients with a Best Overall Response (BOR) of CR or PR. The BOR is defined as the best disease response recorded from CD30.CAR-T infusion until progressive disease, or start of new anti-cancer therapy, whichever comes first.

[0393] 2.2.2 Secondary Objectives

[0394] The following are assessed: [0395] Safety of autologous CD30.CAR-T [0396] Additional antitumor effect of autologous CD30.CAR-T including: [0397] Objective Response Rate (ORR) as assessed by the Investigator per the Revised Criteria for Response Assessment: The Lugano Classification (Cheson et al., J Clin Oncol (2014) 32: 3059-3068) [0398] Duration of Response (DOR) [0399] Progression Free Survival (PFS) [0400] Overall Survival (OS) [0401] Health Related Quality of Life (HRQoL) assessments

[0402] All secondary efficacy endpoints are analyzed in the Full Analysis Set (FAS) unless specified otherwise.

[0403] 2.3 Study Population

[0404] The study population includes adults and pediatric patients aged 12 to 75 years with relapsed or refractory CD30 positive cHL who have failed at least 3 prior lines of therapy, including: [0405] Chemotherapy [0406] *BV and/or [0407] A *PD-1 inhibitor *Unless BV or PD-1 inhibitor is contraindicated.

[0408] Patients may have previously received an autologous and/or allogeneic stem cell transplant.

[0409] 2.4 Study Design

[0410] The study is a Phase 2, open-label, multicenter study designed to evaluate the efficacy and safety of CD30-directed genetically modified autologous T-cells (CD30.CAR-T) in adult and pediatric patients with relapsed or refractory CD30-positive cHL.

[0411] Approximately 82 patients are enrolled (to yield 66 evaluable adult patients), at approximately 30 investigative sites in the United States, Canada and European Union. Approximately 5 pediatric patients ≥12 years of age are enrolled and analyzed separately from the adult population.

[0412] An overview of the study design and study procedures is provided in FIGS. 3 and 4.

[0413] 2.5 Study Treatment

[0414] 2.5.1 Lymphodepletion Chemotherapy

[0415] Patient will undergo LD chemotherapy with fludarabine (30 mg/m.sup.2/day) (IV infusion over 30 minutes) and bendamustine (70 mg/m.sup.2/day) (IV infusion over 30 minutes) given via IV infusion for 3 consecutive days, starting 5 days prior to CD30.CAR-T infusion (i.e. Day −5).

[0416] 2.5.1 CD30.CAR-T Infusion

[0417] CD30.CAR-T cells will be administered on Day 0 as a single IV infusion at a dose of 2×10.sup.8 cells/m.sup.2. In patients that weigh less than 50 kg, a weight-based dosing of 2.0 to 5.0×10.sup.6 cells per kg body weight will be used.

Example 3: Study TESSCAR002: A Phase 1, Study of CD30-Directed Genetically Modified Autologous T-Cells (CD30.CART) in Patients with Relapsed or Refractory CD30 Positive Non-Hodgkin Lymphoma

[0418] 3.1 Study Rationale

[0419] CD30 is a validated target for CD30-positive hematologic malignancies. This is further supported by the approval of anti-CD30 antibody drug conjugate, brentuximab vedotin, for classical HL and some NHL subtypes including systemic and cutaneous ALCL, PTCL and mycosis fungoides.

[0420] CD30 is widely expressed in NHL subtype tumor cells, with expression ranged from 100% in ALCL (Gottesman, Pathology and Laboratory Medicine International (2016) 8: 27-36) to 19% in PMBCL (Hoeller et al., Histopathology (2010) 56: 217-228) tumor cells. The high expression of CD30 positivity in NHL cells further support the potential targeting of NHL malignancies with CD30 antigens.

[0421] The standard of care therapy for NHL malignancies across all subtypes is not curative. Treatment outcomes for patients with relapsed or refractory NHL who fail standard available therapy are poor. New treatment strategies for this patient population are urgently needed to meet unmet clinical needs.

[0422] The favorable toxicity profile and encouraging antitumor activity of autologous CD30.CAR-T demonstrated in Example 1 provide strong rationale to expand the study to NHL patients to fulfill important unmet clinical needs.

[0423] 3.2 Objectives and Endpoints

[0424] 3.2.1 Primary Objectives

[0425] To evaluate the safety and dose-limiting toxicities (DLTs) of autologous CD30.CAR-T and determine the recommended Phase 2 dose (RP2D).

[0426] 3.2.2 Secondary Objectives

[0427] To evaluate the preliminary antitumor effects and pharmacokinetics of autologous CD30.CAR-T by assessing [0428] Objective response rate (ORR) [0429] Pharmacokinetics of autologous CD30.CAR-T cells in the blood. [0430] Duration of response (DoR) [0431] Progression free survival (PFS) [0432] Overall Survival (OS)

[0433] 3.2.3 Exploratory Objectives

[0434] To assess: [0435] Immunogenicity against anti-CD30 scFv (single-chain variable fragment) following CD30.CAR-T infusion in blood [0436] Cytokine profiling in blood [0437] Immunological parameters in blood [0438] Tumor markers in blood and tumor tissue [0439] Circulating tumor DNA (ctDNA) in blood

[0440] 3.3 Study Population

[0441] The study population includes adult patients 18-75 years of age with relapsed or refractory CD30 positive NHL, who have failed standard of care therapy; patients may or may not have received an autologous or allogenic HSCT.

[0442] The CD30-positive NHL subtypes to be evaluated include: [0443] ALCL [0444] PTCL-NOS [0445] ENKTCL, nasal type [0446] DLBCL-NOS [0447] PMBCL

[0448] Prospective approval of protocol deviations from recruitment and enrollment criteria, also known as protocol waivers or exemptions, is not permitted.

[0449] 3.4 Study Design

[0450] This is a phase 1, open-label, multi-center dose-escalation study designed to evaluate the safety and preliminary antitumor effects of CD30-directed genetically modified autologous T-cells (CD30.CAR-T) in adult patients with relapsed or refractory CD30-positive NHL and to determine the R2PD. Patients who have failed standard therapy and may or may not have received an autologous or allogenic HSCT will be enrolled. Up to 4 study centers will participate in the study.

[0451] The CD30-positive NHL subtypes to be evaluated include: [0452] ALCL [0453] PTCL-NOS [0454] ENKTCL, nasal type [0455] DLBCL-NOS [0456] PMBCL

[0457] Overall, this study enrolls approximately 12-21 patients.

[0458] A fresh tumor biopsy is required at the time of screening to determine CD30 positivity and confirm study eligibility. If a biopsy is medically contraindicated and an archival tumor biopsy from a recent treatment that documents CD30 positivity is available (including the original diagnostic biopsy if that is the only available tissue), may be accepted after discussion with Sponsor or designee. CD30-positivity are confirmed locally for eligibility. CD30 positivity must be confirmed prior to blood collection for CD30.CAR-T cells manufacturing.

[0459] After blood collection and during the production of CD30.CAR-T, patients are allowed to receive bridging therapy as per investigator choice. If bridging therapy is administered, washout is required after the completion of bridging therapy, to ensure adequate recovery from toxicity(ies) before initiation of the first dose of lymphodepletion chemotherapy. The washout period is based on the type of agent used as bridging therapy.

[0460] All patients go through a baseline assessment for safety which is performed within 72 hours prior to lymphodepletion chemotherapy. Imaging scans for tumor assessment can be performed within 7 days prior to lymphodepletion chemotherapy. Patients must have active disease [PR, SD or progressive disease (PD)] documented by imaging prior to starting lymphodepletion chemotherapy and CD30.CAR-T infusion. Patients with evidence of a CR following bridging therapy may not be treated with CD30.CAR-T until documented progression/active disease.

[0461] Lymphodepletion chemotherapy and CD30.CAR-T infusion can be administered on an outpatient basis. Patients may also be hospitalized for the administration of lymphodepletion chemotherapy and CD30.CAR-T infusion per institutional guidance.

[0462] Lymphodepletion chemotherapy—consisting of fludarabine (30 mg/m.sup.2/day) and bendamustine (70 mg/m.sup.2/day)—is administered for 3 consecutive days starting on Day −5 to Day −3, prior to autologous CD30.CAR-T infusion on Day 0.

[0463] Prior to CD30.CAR-T infusion, all patients are premedicated with a H1 antagonist (e.g. diphenhydramine, hydroxyzine,) up to 1 mg/kg IV (max 50 mg) and acetaminophen 10 mg/kg per oral (max 650 mg). Alternative antihistamines, anti-inflammatories and anti-emetics may be prescribed per institutional guidance. The use of steroids such as glucocorticoids must be avoided.

[0464] CD30.CAR-T cells are administered on Day 0 as a single IV infusion. Subjects return daily to the treatment facility for safety monitoring for 14 days post-infusion, and then weekly thereafter at weeks 3, 4 and 6 post-infusion.

[0465] Patients are monitored for DLT assessment from after CD30.CAR-T infusion through Day 28. Any of the toxicities which are assessed as possibly related, probably related, or definitely related to autologous CD30.CAR-T cells are considered as a DLT. The DLT assessment period commences from the start of autologous CD30.CAR-T infusion and continue for 28 days following the infusion of CD30.CAR-T cells. DLTs which occur during the DLT assessment period is used in decisions regarding dose-escalation. The RP2D is established during the study.

[0466] All patients are evaluated for safety following lymphodepletion chemotherapy to CD30.CAR-T infusion through Day 42 (EOT) at pre-determined timepoints during the Treatment Phase. Post CD30.CAR-T infusion, patients are required to stay in the treating facility for an additional 4 hours, then return daily for 14 days (excluding weekends), followed by a weekly visit on Day 21 and Day 28 for safety monitoring and then on Day 42 for EOT visit. Per institutional guidance, patients may remain hospitalized post-CD30.CAR-T infusion for toxicity monitoring. The extended hospitalization is not considered an AE.

[0467] Imaging scans are reviewed locally by the investigator as assessed by the Revised Lugano Response Criteria (Cheson et al., J Clin Oncol (2014) 32: 3059-3068).

[0468] Patients undergo exploratory biomarker assessments to characterize the pharmacokinetic properties as well as the immunogenicity and immunological properties of autologous CD30.CAR-T. Details on prioritization of blood samples collected for analysis are included in the Laboratory Manual. Blood and tissue biopsy samples are collected, if clinically feasible as assessed by the investigator, at various timepoints before and during study treatment. These data are used to analyze further the mechanisms that govern therapeutic outcome as well as to identify potential biomarkers that correlate with therapeutic efficacy and/or failure.

[0469] After the EOT visit at Day 42, patients enter the Post-Treatment Follow-Up Phase which ends at M12. Follow-up for safety, efficacy and survival continue every 3 months. After that, patients enter into long term follow-up (LTFU); follow-up for efficacy continue every 3 months until M24; follow up for safety and survival continue every 6 months from M13 through M60 (Year 5) and annually thereafter for up to 15 years until disease progression, death, withdrawal of consent, start of new anti-cancer therapy or termination of study.

[0470] An overview of the study design and study procedures is provided in FIGS. 4 and 5.

[0471] 3.5 Study Treatment

[0472] 3.5.1 Lymphodepletion Chemotherapy

[0473] Patient will undergo LD chemotherapy with fludarabine (30 mg/m.sup.2/day) (IV infusion over 30 minutes) and bendamustine (70 mg/m.sup.2/day) given via IV infusion for 3 consecutive days, starting 5 days prior to CD30.CAR-T infusion (i.e. Day −5).

[0474] 3.5.2 CD30.CAR-T Infusion

[0475] CD30.CAR-T cells will be administered on Day 0 as a single IV infusion at: [0476] DL1: 2×10.sup.8 cells/m.sup.2 [0477] DL2: 4×10.sup.8 cells/m.sup.2 [0478] DL3: 6×10.sup.8 cells/m.sup.2