Sponsor: Philogen S.p.A. (industry)
Phase: 2
Start date: July 12, 2024
Planned enrollment: 162
L19IL2 (darleukin; bifikafusp alfa) is an investigational cancer immunotherapy studied alone and in combinations, principally in melanoma. L19IL2 is one drug; Daromun is a combination of L19IL2 and a different drug, L19TNF. Results for the combination cannot establish the effect of L19IL2 alone. (ema.europa.eu)
L19IL2 joins interleukin-2 (IL-2), an immune-stimulating cytokine, to an antibody fragment that binds the extra-domain B of fibronectin. This target is associated with newly formed tumor blood vessels; the design aims to concentrate IL-2 activity at tumors. (ema.europa.eu)
The injected-L19IL2 study reported no serious adverse events. The earlier dacarbazine-combination study described toxicity as manageable and reversible, with no treatment-related deaths. The updated Daromun trial reported no new safety signals; its findings apply to the two-drug combination, not L19IL2 alone. (pubmed.ncbi.nlm.nih.gov)
Last updated: Sep 2026
L19TNF (Fibromun; onfekafusp alfa) is an investigational cancer treatment studied alone and with other drugs, including doxorubicin for soft-tissue sarcoma. It is also a component of the melanoma combination L19-IL2/L19-TNF; results for that combination should not be attributed to L19TNF alone. (pubmed.ncbi.nlm.nih.gov)
L19TNF joins human tumor necrosis factor (TNF) to an L19 antibody fragment. L19 binds the extra-domain B of fibronectin, a marker associated with newly formed tumor blood vessels, with the aim of delivering TNF to tumor tissue. (pmc.ncbi.nlm.nih.gov)
Phase I investigators identified 13 μg/kg L19TNF plus 60 mg/m² doxorubicin as the recommended combination dose; dose-limiting toxicity occurred at higher studied doses. In the 22-patient intralesional melanoma combination study, adverse events were mostly grade 1–2, but two patients had grade 3 injection-site reactions. These findings describe different regimens and routes of administration, not a single established safety profile for L19TNF. (pubmed.ncbi.nlm.nih.gov)
Last updated: Sep 2026
Goal: Determine whether intralesional L19IL2, L19TNF, or their combination, each given with pembrolizumab, produces objective responses in melanoma that has progressed on anti-PD-1/PD-L1 therapy. Each arm’s activity will be assessed against historical outcomes for anti-PD-1 rechallenge, not a concurrent control arm.
Patients: Adults with ECOG performance status 0–1, measurable unresectable stage IIIB–IV melanoma, confirmed progression on anti-PD-1/PD-L1 therapy, and at least one injectable cutaneous, subcutaneous, or nodal metastasis. Stage IV disease involving lung, liver, bone, or brain is limited to oligometastatic disease of up to 10 lesions; active or symptomatic CNS disease and significant prior immune-related toxicity generally exclude participation.
Design: Open-label, randomized, three-arm phase 2 study using a Simon two-stage design. Participants are allocated equally among the three experimental regimens. An initial safety run-in assesses the first 12 patients in each arm during the first 21-day cycle.
Treatments: All arms receive intravenous pembrolizumab every three weeks for up to 35 cycles or approximately two years, plus four weeks of intralesional L19IL2, L19TNF, or both. Pembrolizumab is an anti-PD-1 antibody used here as systemic rechallenge. L19IL2 and L19TNF fuse IL-2 and TNF, respectively, to an antibody fragment targeting the extra-domain B of fibronectin, with the aim of concentrating cytokine activity at tumors. In a small uncontrolled intralesional melanoma study, L19IL2 produced complete responses in all treated metastases in 6 of 24 evaluable patients; this does not establish systemic efficacy with pembrolizumab. A small intralesional study of L19IL2 plus L19TNF reported responses in treated lesions in 11 of 20 evaluable patients, but could not isolate either drug’s contribution; intravenous L19TNF alone produced no objective responses among 31 evaluable patients with advanced solid tumors.
Outcomes: The primary endpoint is confirmed objective response rate by RECIST v1.1 in the intention-to-treat population, assessed for up to two years. Secondary measures include best overall response, duration of response, response by iRECIST and itRECIST, progression-free and overall survival, and pathological response in an injected lesion at week 18. Safety assessments include adverse events, immune-related events, laboratory tests, cardiac assessments, and antibodies against the investigational proteins.
Burden on patient: Moderate to high. Participation requires screening and tumor tissue sampling, repeated intralesional injections over four weeks, and intravenous pembrolizumab visits every three weeks for up to two years. Scheduled imaging extends through week 96; baseline and week-18 biopsies of an injected lesion, laboratory monitoring, and ECG and echocardiogram assessments add procedures and travel beyond routine treatment. Progression and survival follow-up can continue for approximately two and three years, respectively.
Last updated: Sep 2026
Inclusion Criteria:
1. The participant (or legally acceptable representative if applicable) provides written informed consent for the trial.
2. Be \> or equal to 18 years of age on day of signing informed consent.
3. Participant with histologically or cytologically confirmed diagnosis of unresectable metastatic melanoma at stage III B, C, D or IV M1a (AJCC 8th ed.). Patients with Stage IVM1b, M1c and M1d oligometastatic disease \[up to 10 lesions in aggregate including lung, liver, bone or brain, with or without lymph node involvement\], are eligible. However, patients with symptomatic or rapidly enlarging/bleeding brain lesions are excluded. Patients with acral lentiginous melanoma are eligible as well. Detailed information about prior therapies and burden of disease at study entry must be available.
4. Patients must have confirmed primary resistance to or acquired resistance on treatment with an anti-PD1/L1 monoclonal antibody (mAb) administered either as monotherapy, or in combination with other checkpoint inhibitors or other therapies. PD-1 treatment progression is defined by meeting all of the following criteria: a.) Has received at least 2 doses of an approved anti-PD-1/L1 mAb.
b.) Has demonstrated disease progression (PD) after PD-1/L1 as defined by RECIST v1.1. The initial evidence of PD is to be confirmed by a second assessment no less than four weeks from the date of the first documented PD, in the absence of rapid clinical progression. c.) Progressive disease has been documented within 12 weeks from the last dose of anti- PD-1/L1 mAb.
i. Progressive disease is determined according to iRECIST. ii. This determination is made by the investigator. Once PD is confirmed, the initial date of PD documentation will be considered the date of disease progression.
5. Patients harboring the BRAF mutation who received BRAF/MEK inhibition (or declined BRAF/MEK inhibitors) and received thereafter anti-PD1 therapy showing resistance to such immunotherapy are eligible to the study. Anti-PD1 immunotherapy must be the last therapy received by the patient prior to randomization.
6. Eligible subjects must have measurable disease (according to RECIST v1.1) as assessed by the local site investigator/radiology. Lesions situated in a previously irradiated area are considered measurable if progression has been demonstrated in such lesions. Eligible subjects must be a candidate for intralesional therapy with at least one injectable cutaneous, subcutaneous, or nodal metastatic melanoma lesion (≥ 5 mm in longest diameter) or with multiple injectable lesions that in aggregate have a diameter of ≥ 5 mm.
7. A female participant is eligible to participate if she is not pregnant, not breastfeeding, and at least one of the following conditions applies: a.) Not a woman of childbearing potential (WOCBP) b.) A WOCBP who agrees to follow contraceptive guidance during the treatment period and for at least 120 days after the last dose of study treatment. WOCBP must be using for the time period indicated highly effective contraception methods. WOCBP and effective contraception methods are defined by the "Recommendations for contraception and pregnancy testing in clinical trials" issued by the Head of Medicine Agencies' Clinical Trial Facilitation Group and which include, for instance, progesterone-only or combined (estrogen- and progesterone-containing) hormonal contraception associated with inhibition of ovulation, intrauterine devices, intrauterine hormone-releasing systems, bilateral tubal occlusion, vasectomized partner or sexual abstinence. Pregnancy test will be repeated at the safety visit (only WOCBP).
8. Have provided archival tumor tissue sample or newly obtained core or excisional biopsy of a tumor lesion not previously irradiated. Formalin-fixed, paraffin embedded (FFPE) tissue blocks are preferred to slides. Newly obtained biopsies are preferred to archived tissue.
Note: If submitting unstained cut slides, newly cut slides should be submitted to the testing laboratory within 14 days from the date slides are cut (details pertaining to tumor tissue submission can be found in the Procedures Manual).
9. Have an Eastern Cooperative Oncology Group (ECOG) performance status of 0 to 1.
10. Have adequate organ function as defined in the following table. Specimens must be collected within 14 days prior to the start of study treatment.
11. Be able to provide a core or excisional lymph node biopsy for biomarker analysis from an archival or newly obtained biopsy at Screening. In addition, participants may provide additional biopsy at Week 18 and at the time of discontinuation due to progression
12. Documented negative test for HIV, HBV and HCV. For HBV serology, the determination of HBsAg and anti-HBcAg Ab is required. In patients with serology documenting previous exposure to HBV (e.g. anti-HBsAg and/or anti-HBc Ab) a negative serum HBV-DNA test is also required.
13. All acute toxic effects (excluding alopecia and vitiligo) of any prior therapy must have resolved to National Cancer Institute (NCI) Common Terminology Criteria for Adverse Events (CTCAE) (v. 5.0) Grade ≤ 1 or baseline unless otherwise specified above. Note: Participants with ≤Grade β neuropathy may be eligible. Participants with endocrinerelated AEs Grade ≤β requiring treatment or hormone replacement may be eligible. Note: If the participant had major surgery, the participant must have recovered adequately from the procedure and/or any complications from the surgery prior to starting study intervention.
14. Full resolution of checkpoint blockade therapy-related adverse effects (including immunerelated adverse effects) and no treatment for these AEs for at least 4 weeks prior to the time of enrollment.
15. No history of severe immune related adverse effects from prior given immune checkpoint blockade therapy (CTCAE Grade 4; CTCAE Grade 3 requiring treatment \>4 weeks).
16. Male patients with WOCBP partners must agree to use simultaneously two acceptable methods of contraception (i.e. spermicidal gel plus condom) from the screening to three months following the last study drug administration. In addition, male participant must refrain from sperm donation during the treatment period.
17. Willingness and ability to comply with the scheduled visits, treatment plan, laboratory tests and other study procedures.
Exclusion Criteria:
Participants are excluded from the study if any of the following criteria apply:
Medical Conditions
1. Patients with more than 10 distant melanoma lesions in lung, liver, bone or brain combined. Patients with symptomatic or rapidly enlarging/bleeding brain lesions are excluded.
2. Uveal melanoma or mucosal melanoma or melanoma with unknown primary.
Pregnancy Exclusion
3. A WOCBP who has a positive urine pregnancy test (within 72 hours) prior to treatment. If the urine test is positive or cannot be confirmed as negative, a serum pregnancy test will be required.
Prior/Concomitant Therapy
4. Has received prior therapy with an anti-PD-1, anti-PD-L1, or anti PD L2 agent or with an agent directed to another stimulatory or co-inhibitory T-cell receptor (e.g., CTLA-4, OX 40, CD137), and was discontinued from that treatment due to a Grade 3 or higher irAE.
5. Has received prior systemic anti-cancer therapy including investigational agents or has used an investigational device within 4 weeks prior to the first dose of study treatment. Note: Participants who have entered the follow-up phase of an investigational study may participate as long as it has been 4 weeks after the last dose of the previous investigational agent.
6. Has received prior radiotherapy within 2 weeks of start of study treatment or have had a history of radiation pneumonitis. Note: Participants must have recovered from all radiation-related toxicities and not require corticosteroids. A 1-week washout is permitted for palliative radiation (≤β weeks of radiotherapy) to non-CNS disease.
7. Has undergone prior allogeneic hematopoietic stem cell transplantation within the last 5 years. (Participants who have had a transplant greater than 5 years ago are eligible as long as there are no symptoms of GVHD).
8. Has had an allogeneic tissue/solid organ transplant.
9. Has received live live or live attenuated vaccines within 30 days prior to the first dose of study treatment and while participating in the study. Note: Killed vaccines are allowed. Note: Any licensed COVID-19 vaccine (including for Emergency use) in a particular country is allowed in the study as long as they are mRNA vaccines, adenoviral vaccines, or inactivated vaccines. These vaccines will be treated just as any other concomitant therapy. Investigational vaccines (i.e., those not licensed or approved for Emergency Use) are not allowed.
Diagnostic assessments
10. Has a diagnosis of immunodeficiency or is receiving chronic systemic steroid therapy (in dosing exceeding 10 mg daily of prednisone equivalent) or any other form of immunosuppressive therapy within 7 days prior the first dose of study drug.
11. Has a known additional malignancy that is progressing or has required active treatment within the past two (2) years. Note: Participants with basal cell carcinoma of the skin, squamous cell carcinoma of the skin, superficial bladder tumors (Ta, Tis \& T1), second primary melanoma in situ, or carcinoma in situ, that have undergone potentially curative therapy are not excluded.
12. Has known active CNS metastases and/or carcinomatous meningitis. Participants with previously treated brain metastases may participate provided they are radiologically stable, i.e., without evidence of progression for at least 4 weeks by repeat imaging (note that the repeat imaging should be performed during study screening), clinically stable and without requirement of steroid treatment for at least 14 days prior to first dose of study treatment.
13. Has severe hypersensitivity (≥Grade γ) to pembrolizumab and/or any of its excipients, or to (immuno)cytokines IL2, TNF and/or any of its excipients
14. Has an active autoimmune disease that has required systemic treatment in past 2 years (i.e., with use of disease modifying agents, corticosteroids or immunosuppressive drugs).
Replacement therapy (e.g., thyroxine, insulin, or physiologic corticosteroid replacement therapy for adrenal or pituitary insufficiency) is not considered a form of systemic treatment and is allowed.
15. Has a history of (non-infectious) pneumonitis / interstitial lung disease that required steroids or has current pneumonitis / interstitial lung disease.
16. Has an active infection requiring systemic therapy.
17. Has a history or current evidence of any condition, therapy, or laboratory abnormality, or other circumstance that might confound the results of the study or interfere with the participant's participation for the full duration of the study, such that it is not in the best interest of the participant to participate, in the opinion of the treating investigator.
18. Has a known psychiatric or substance abuse disorder that would interfere with the participant's ability to cooperate with the requirements of the study.
19. Previous enrolment and randomization in this same study.
New York, New York, 10065, United States
[email protected] / No phone
Status: Recruiting