Administration of PD-1 inhibitors for treating skin cancer

Abstract

To provide methods for treating or inhibiting the growth of a tumor in a patient with a skin cancer.SOLUTION: The present invention relates to a method comprising administering to a patient a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. In certain embodiments, the method comprises administering to a patient a therapeutically effective amount of a PD-1 inhibitor as adjuvant therapy after the patient has completed surgery and, optionally, radiation therapy for skin cancer, such as CSCC, and is at high risk of disease recurrence. In certain embodiments, the method comprises administering to a skin cancer patient a therapeutically effective amount of a PD-1 inhibitor as neoadjuvant therapy prior to planned surgery for skin cancer. In certain embodiments, the method comprises administering to a skin cancer patient a therapeutically effective amount of a PD-1 inhibitor as neoadjuvant therapy prior to planned surgery for skin cancer and, subsequently, administering a PD-1 inhibitor as adjuvant therapy after such surgery.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present disclosure relates to a method for treating or inhibiting tumor growth comprising selecting a patient with skin cancer and administering to the patient a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. [Background technology]

[0002] Skin cancer is the most common cancer in the United States (Non-Patent Document 1). An estimated 5.4 million cases of non-melanoma skin cancer, including basal cell carcinoma and squamous cell carcinoma, were diagnosed in the United States in 2012 (Non-Patent Document 2). Cutaneous squamous cell carcinoma (CSCC) is the second most common malignant lesion in the United States after basal cell carcinoma (BCC) (Non-Patent Document 3). Long-term sun exposure is the dominant risk factor for non-melanoma skin cancer.

[0003] CSCC is a malignant proliferation of epithelial keratinocytes with dermal invasion and is distinct from non-invasive precursor lesions such as actinic keratosis (Non-Patent Document 4). The global incidence varies widely, with Australia having the highest incidence and parts of Africa having the lowest incidence (Non-Patent Document 5). The exact incidence of CSCC is unknown, as it is not included in most cancer registries. However, the incidence of CSCC has increased in recent decades, according to estimates that do not include patients with only non-invasive precursor lesions (Non-Patent Document 5) (Non-Patent Document 6) (Non-Patent Document 7).

[0004] Risk factors for CSCC include UV exposure, advanced age, and immunosuppression (Non-Patent Document 8; Non-Patent Document 9). Although the majority of individuals diagnosed with CSCC or BCC have a very good prognosis, CSCC is more prone to aggressive recurrence than BCC. Furthermore, individuals diagnosed with CSCC, unlike those diagnosed with BCC, have a higher mortality rate compared to age-matched controls (Non-Patent Document 10).

[0005] Surgical resection is the centerpiece of the clinical management of CSCC or BCC. The primary goal is complete resection of the cancer, with an acceptable cosmetic outcome being the secondary goal. Factors associated with poor prognosis in CSCC include tumor size >2 cm, tumor depth >2 mm, perineural invasion, host immunosuppression, and recurrent disease. However, some patients who develop advanced CSCC, including both locally advanced and metastatic CSCC, are not candidates for surgery. Some such patients can be treated with postoperative radiation therapy or chemotherapy, but these may not be attractive options due to safety and tolerability concerns.

[0006] Widespread carcinogenesis, defined as multiple cancerous lesions at UV-exposed sites, is a feature of many CSCC patients. Moreover, recurrent CSCC increases the risk of subsequent recurrence. In a single-institution retrospective study of 212 patients, recurrent CSCC was twice as likely to recur again after excision surgery compared with primary CSCC (Non-Patent Document 11). Multiple surgeries over time can be disfiguring and can lead to postoperative fatigue, i.e., physical and emotional debilitation due to successive surgical procedures. Furthermore, CSCC at accessory sites of the head and neck, e.g., ears, temples, and lips, was associated with even worse clinical outcomes (Non-Patent Document 12; Non-Patent Document 11; Non-Patent Document 13).

[0007] The most common clinical subtype of BCC is nodular BCC. Less common clinical subtypes are superficial, morphoeic (fibrosing), and fibroepithelial. Although most patients are cured by surgery, a small proportion of patients experience recurrent disease or develop unresectable locally advanced or metastatic disease. Recognition of the oncogenic role of the G protein receptor Smoothened (SMO) in BCC has led to the development of a new class of cancer treatments, commonly referred to as hedgehog inhibition. This has led to the development of orally available inhibitors of SMO, vismodegib and sonidegib, termed hepatitis inhibitors (HHIs). In addition to the deleterious side effects of HHIs, it has been found that for patients who progress on HHIs (vismodegib), subsequent treatment with another HHI (sonidegib) does not result in tumor inhibition (Non-Patent Document 14).

[0008] Thus, there remains a need to provide safe and effective therapies for skin cancers, such as CSCC and BCC, especially skin cancers that have recurred despite prior surgery.

[0009] Furthermore, only a small proportion of CSCC patients are considered to have high-risk CSCC when assessed using several factors, including cancer staging using Non-Patent Document 15, immune status, lymphovascular invasion, extent of lymph node metastasis, presence of extracapsular spread, and treatment history. For high-risk cases, radiation therapy after surgery is recommended (Non-Patent Document 16) (Non-Patent Document 17). However, high-risk patients may relapse with locoregional recurrence or distant metastasis (Non-Patent Document 18). Thus, there is an unmet need to reduce the risk of CSCC recurrence, especially in high-risk patients. [Prior art documents] [Non-patent literature]

[0010] [Non-Patent Document 1] Guy et al., Am. J. Prev. Med. 48:183-87, 2015 [Non-Patent Document 2] Rogers et al., JAMA Dermatol., 151(10):1081-86, 2015 [Non-Patent Document 3] Karia et al., J. Am. Acad. Dermatol. 68:957-66, 2013 [Non-Patent Document 4] Fernandez et al., Immunol Allergy Clin North Am 37(2):315~27, 2017

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Non-licensed literature 9

Non-licensed literature 10

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Non-licensed Document 17

[0011] In one aspect, the disclosed technology relates to a method of treating or inhibiting tumor growth, comprising: (a) selecting a patient with skin cancer who has completed surgery and / or radiation therapy to treat the skin cancer; and (b) subsequently administering to the patient an adjuvant treatment comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. In one embodiment, the method of treating or inhibiting tumor growth comprises: (a) selecting a patient with skin cancer who has completed surgery and optionally radiation therapy to treat the skin cancer; and (b) subsequently administering to the patient an adjuvant treatment comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. In one embodiment, the skin cancer is cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma or melanoma. In another embodiment, the skin cancer is CSCC. In another embodiment, the patient is at high risk of recurrence of CSCC. In one embodiment, the patient has metastatic disease and has undergone resection surgery. In another embodiment, the patient has at least one of the following high-risk features: nodal disease with extracapsular extension and at least one nodule >20mm; in-transit metastasis (ITM); T4 lesion; perineural invasion (PNI); and recurrent CSCC+ with at least one of the following additional features: ≧N2b disease associated with a recurrent lesion; nominal ≧T3; and recurrent lesion ≧20mm in diameter.

[0012] In another embodiment, the therapeutically effective amount comprises 5 mg to 500 mg of the PD-1 inhibitor. In another embodiment, the therapeutically effective amount comprises 350 mg of the PD-1 inhibitor. In another embodiment, the PD-1 inhibitor is administered intravenously or intraperitoneally. In another embodiment, the PD-1 inhibitor is administered intravenously. In another embodiment, step (b) occurs 2 to 6 weeks after completion of surgery and / or radiation therapy. In one embodiment, one or more doses of the PD-1 inhibitor are administered as an adjuvant treatment, each dose being administered 2 to 12 weeks after the immediately preceding dose. In another embodiment, the PD-1 inhibitor is administered as an adjuvant treatment according to a dosing regimen disclosed herein. In another embodiment, administration of the PD-1 inhibitor leads to a reduced risk of subsequent skin cancer recurrence or eliminates the occurrence of subsequent skin cancer recurrence. In another embodiment, administration of the PD-1 inhibitor leads to at least about a 10% lower incidence of subsequent skin cancer recurrence compared to patients following completion of surgery and radiation therapy without the adjuvant skin cancer treatment. In another embodiment, the method further comprises administering a second therapeutic agent selected from a chemotherapeutic agent, a corticosteroid, an anti-inflammatory agent, and / or combinations thereof.

[0013] In another embodiment, the PD-1 inhibitor is selected from the group consisting of an anti-PD-1 antibody or antigen-binding fragment thereof, an anti-PD-L1 antibody or antigen-binding fragment thereof, and an anti-PD-L2 antibody or antigen-binding fragment thereof. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody or antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO: 1, and three CDRs (LCDR1, LCDR2 and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO: 2. In another embodiment, HCDR1 has the amino acid sequence of SEQ ID NO: 3; HCDR2 has the amino acid sequence of SEQ ID NO: 4; HCDR3 has the amino acid sequence of SEQ ID NO: 5; LCDR1 has the amino acid sequence of SEQ ID NO: 6; LCDR2 has the amino acid sequence of SEQ ID NO: 7; LCDR1 has the amino acid sequence of SEQ ID NO: 8; LCDR2 has the amino acid sequence of SEQ ID NO: 9; LCDR3 has the amino acid sequence of SEQ ID NO: 10; LCDR4 has the amino acid sequence of SEQ ID NO: 11; LCDR5 has the amino acid sequence of SEQ ID NO: 12; LCDR6 has the amino acid sequence of SEQ ID NO: 13; LCDR7 has the amino acid sequence of SEQ ID NO: 14; LCDR8 has the amino acid sequence of SEQ ID NO: 15; LCDR9 has the amino acid sequence of SEQ ID NO: 16; LCDR10 has the amino acid sequence of SEQ ID NO: 17; LCDR11 has the amino acid sequence of SEQ ID NO: 18; LCDR12 has the amino acid sequence of SEQ ID NO: 19; LCDR13 has the amino acid sequence of SEQ ID NO: 20; LCDR14 has and R3 has the amino acid sequence of SEQ ID NO: 8. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NOs: 1 / 2. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 9. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the light chain has the amino acid sequence of SEQ ID NO: 10. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO: 9 and the light chain has the amino acid sequence of SEQ ID NO: 10.

[0014] In another embodiment, the PD-1 inhibitor is cemiplimab or a biological equivalent thereof. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754091, PF-06801591, spartalizumab, camrelizumab, JNJ-63723283 and MCLA-134. In another embodiment, the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-072, KN035 and CK-301.

[0015] In another aspect, the disclosed technology relates to a pharmaceutical composition comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor for use in the adjuvant treatment of skin cancer after completion of surgery and, optionally, post-surgical radiation. In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody or antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 and three CDRs (LCDR1, LCDR2, and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2. In another embodiment, HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; LCDR3 has the amino acid sequence of SEQ ID NO:8. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NO:1 / 2. In another embodiment, the pharmaceutical composition comprises 5 mg to 500 mg of a PD-1 inhibitor.In another embodiment, the pharmaceutical composition comprises 350 mg of a PD-1 inhibitor.In another embodiment, the skin cancer is CSCC.

[0016] In another aspect, the disclosed technology relates to a method of treating or inhibiting tumor growth, comprising: (a) selecting a patient with skin cancer planned for surgical removal; and (b) administering to the patient a neoadjuvant treatment comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor prior to surgical removal. In one embodiment, the skin cancer is cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma or melanoma. In another embodiment, the skin cancer is CSCC. In another embodiment, the patient is at high risk of recurrence of CSCC. In another embodiment, the patient has at least one of the following high-risk features: nodal disease with extracapsular extension and at least one node of >20mm; in-transit metastasis (ITM); T4 lesion; perineural invasion (PNI); and at least one of the following additional features of recurrent CSCC+: disease with ≧N2b associated with recurrent lesion; nominal ≧T3; and recurrent lesion with a diameter of ≧20mm. In another embodiment, the patient has stage II to stage IV skin cancer and the patient is a candidate for surgery. In one embodiment, patients with resectable tumors are at increased risk of recurrence and / or risk of disfigurement or loss of function.

[0017] In another embodiment, the therapeutically effective amount comprises 5 mg to 500 mg of a PD-1 inhibitor administered as a neoadjuvant. In another embodiment, the therapeutically effective amount comprises 350 mg of a PD-1 inhibitor administered as a neoadjuvant. In one embodiment, the neoadjuvant treatment One or more doses of PD-1 inhibitor are administered, each dose being administered 2-12 weeks after the immediately preceding dose. In another embodiment, the method further comprises: (c) following the neoadjuvant treatment, surgically removing the skin cancer. In another embodiment, the method further comprises administering to the patient an adjuvant treatment comprising a therapeutically effective amount of a PD-1 inhibitor after step (c), the adjuvant PD-1 inhibitor being the same as or different from the neoadjuvant PD-1 inhibitor. In another embodiment, the adjuvant treatment comprises administering one or more doses of the PD-1 inhibitor, each dose comprising 5 mg to 500 mg of the PD-1 inhibitor. In another embodiment, each dose of the adjuvant treatment comprises 350 mg of the PD-1 inhibitor. In one embodiment, each dose of the adjuvant treatment is administered 2-12 weeks after the immediately preceding dose. In another embodiment, the PD-1 inhibitor is administered intravenously or intraperitoneally. In another embodiment, the PD-1 inhibitor is administered intravenously.

[0018] In another embodiment, administration of the PD-1 inhibitor leads to a reduced risk of subsequent skin cancer recurrence or eliminates the occurrence of subsequent skin cancer recurrence. In another embodiment, administration of the PD-1 inhibitor leads to at least about a 10% lower incidence of subsequent skin cancer recurrence compared to patients following completion of surgery and radiation therapy without the adjuvant skin cancer treatment. In another embodiment, the method further comprises administering a second therapeutic agent selected from a chemotherapeutic agent, a corticosteroid, an anti-inflammatory agent, and / or combinations thereof. In another embodiment, the PD-1 inhibitor is selected from the group consisting of an anti-PD-1 antibody or antigen-binding fragment thereof, an anti-PD-L1 antibody or antigen-binding fragment thereof, and an anti-PD-L2 antibody or antigen-binding fragment thereof.

[0019] In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody or antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three CDRs (LCDR1, LCDR2, and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2. In another embodiment, HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:8. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NO:1 / 2. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO:9. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the light chain has the amino acid sequence of SEQ ID NO:10. In another embodiment, the anti-PD-1 antibody comprises a heavy chain and a light chain, wherein the heavy chain has the amino acid sequence of SEQ ID NO:9 and the light chain has the amino acid sequence of SEQ ID NO:10.

[0020] In another embodiment, the PD-1 inhibitor is cemiplimab or a biological equivalent thereof. In another embodiment, the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754048, PF-06371548, spartalizumab, camrelizumab, JNJ-63313240, and MCLA-134. In another embodiment, the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-031, KN035, and CK-301.

[0021] In another aspect, the disclosed technology relates to a pharmaceutical composition comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor for use in neoadjuvant therapy prior to planned surgery to treat skin cancer. In one embodiment, the PD-1 inhibitor is a PD-1 inhibitor that binds to the three complementarity determining regions (CDRs) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 (HCDR1, HCDR2, HCDR3, HCDR4, HCDR5, HCDR6, HCDR7, HCDR8, HCDR9, HCDR10, HCDR11, HCDR12, HCDR13, HCDR14, HCDR15, HCDR16, HCDR17, HCDR18, HCDR19, HCDR110, HCDR111, HCDR192, HCDR112, HCDR13, HCDR14, HCDR15, HCDR16, HCDR17, HCDR18, HCDR192, HCDR193, HCDR194, HCDR195, HCDR196, HCDR197, HCDR198, HCDR199, HCDR199, HCDR111, HCDR120, HCDR121, HCDR122, HCDR123, HCDR124, HCDR125, HCDR130, HCDR131, HCDR132, HCDR133, HCDR134, HCDR140, HCDR141, HCDR142, HCDR143, HCDR144, HCDR145, HCDR146, HCDR147, HCDR148, HCDR149, HCDR151, HCDR152, HCDR153, , HCDR2 and HCDR3) and three CDRs (LCDR1, LCDR2 and LCDR3) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2. In another embodiment, HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:8. In another embodiment, the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NO:1 / 2. In another embodiment, the pharmaceutical composition comprises 5 mg to 500 mg of the PD-1 inhibitor. In another embodiment, the pharmaceutical composition comprises 350 mg of the PD-1 inhibitor. In another embodiment, the skin cancer is CSCC.

[0022] As used herein, a "PD-1 inhibitor" can refer to at least one of a neoadjuvant PD-1 inhibitor and an adjuvant PD-1 inhibitor. [Brief description of the drawings]

[0023] [Figure 1] FIG. 1 outlines the study described in Example 1. [Diagram 2] FIG. 1 outlines the study described in Example 2. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] It is to be understood that this disclosure is not limited to the particular methods and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, and that the scope of the disclosure will be limited only by the appended claims.

[0025] Unless otherwise specified, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which the disclosed invention belongs.As used herein, the term "about" means that when used in conjunction with a specific recited value, the value may vary from the recited value by 1% or less.For example, as used herein, the expression "about 100" includes 99 and 101 and all values ​​therebetween (e.g., 99.1, 99.2, 99.3, 99.4, etc.).

[0026] Although any methods and materials similar or equivalent to those described herein can be used in the practice of this disclosure, the preferred methods and materials are now described.

[0027] Methods of Treating or Inhibiting Tumor Growth The present disclosure includes methods for treating or inhibiting the growth of skin cancer, comprising selecting a patient with skin cancer and administering to the patient a therapeutically effective amount of a PD-1 inhibitor (e.g., an antibody or antigen-binding fragment thereof that specifically binds to PD-1, PD-L1 and / or PD-L2, or any other "PD-1 inhibitor" described herein). In this disclosure, reference to anti-PD-1 antibodies specifically is provided to illustrate representative PD-1 inhibitors and does not limit the scope of the disclosure. In one embodiment, the PD-1 inhibitor is administered prior to treating the patient with surgery and / or radiation therapy. In certain embodiments, the method comprises administering the PD-1 inhibitor to a subject in need thereof as an adjuvant treatment following completion of surgery and optionally radiation therapy, e.g., post-surgical radiation therapy, to treat the skin cancer. In certain embodiments, the methods include administering a PD-1 inhibitor to a subject in need thereof as a neoadjuvant treatment prior to planned surgery to treat the skin cancer, and the methods can optionally further include subsequently administering a PD-1 inhibitor to the patient as an adjuvant therapy after surgery to treat the skin cancer.

[0028] In certain embodiments, the disclosure includes a method for treating or inhibiting the growth of skin cancer comprising: (a) selecting a patient with skin cancer, the patient having at least one of the following high-risk features: nodal disease with extracapsular spread and at least one nodule >20mm; in-transit metastasis (ITM); T4 lesion; perineural invasion (PNI); and recurrent CSCC+ with at least one of the following additional features: ≧N2b disease associated with a recurrent lesion; nominal ≧T3; and recurrent lesion ≧20mm in diameter; and (b) administering a therapeutically effective amount of a PD-1 inhibitor as an adjuvant or neoadjuvant treatment to a patient in need thereof.

[0029] As used herein, the terms "treating," "treat," and the like refer to alleviating or reducing the severity of at least one symptom or sign to temporarily or permanently eliminate the cause of a condition, to slow or inhibit tumor growth, to reduce tumor cell burden or tumor mass, to promote tumor regression, to cause tumor shrinkage, necrosis, and / or disappearance, to prevent tumor recurrence, to prevent or inhibit metastasis, to inhibit metastatic tumor growth, to eliminate the need for surgery, and / or to prolong the survival of a subject. In many embodiments, the terms "tumor," "lesion," "tumor lesion," "cancer," and "malignant lesion" are used interchangeably to refer to one or more growths.

[0030] In some embodiments, the skin cancer is cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma or melanoma. In some embodiments, the skin cancer is squamous cell carcinoma of the head and neck. In some embodiments, the skin cancer is advanced CSCC. In some embodiments, the skin cancer is metastatic, resectable, unresectable, recurrent, or locally advanced. In some embodiments, the skin cancer is CSCC, including but not limited to metastatic CSCC, locally advanced CSCC, resectable CSCC, unresectable CSCC, or recurrent CSCC. In one embodiment, the skin cancer is resectable recurrent CSCC.

[0031] As used herein, the term "recurrent" refers to frequent or repeated diagnoses of skin cancer (e.g., CSCC) in a patient, or frequent or repeated occurrence of individual tumor lesions, e.g., new tumor lesions, which may indicate a recurrence of a primary tumor lesion and / or a previous tumor lesion.

[0032] As used herein, the term "recurrence" is defined as the appearance of one or more new skin cancer (e.g., CSCC) lesions, local, regional, or distant. In many cases, new lesions on the skin are new primary tumors due to widespread carcinogenesis from long-term UV-mediated skin damage (Christensen, F1000Res, 7, 2018). With respect to CSCC, local or regional (locoregional) recurrence is defined by any of the following sites of disease recurrence: (a) for HN CSCC, recurrence in nodal or soft tissue above the clavicle; (b) for non-HN CSCC, recurrence within the first draining nodal basin of the resected tumor (or associated soft tissue within the first draining nodal basin); (c) in-transit metastases, defined as cutaneous or subcutaneous metastases >2 cm from the primary lesion but not beyond the regional nodal basin. Distant recurrence is defined by any of the following sites of disease recurrence: (a) subclavian lymph node recurrence for HN CSCC; (d) recurrence beyond the first draining lymph node basin of the resected tumor bed for non-HN CSCC. Recurrence in two lymph node basins is considered distant recurrence even if they are close together (i.e., two mediastinal lymph node basins, two pelvic lymph node basins); (e) recurrence in non-lymph node tissues (including but not limited to lung, liver, bone, brain); (f) epidermotropic metastasis, defined as distant disease of the dermis without epithelial involvement.

[0033] As used herein, the phrase "subject in need thereof" or "patient in need thereof" refers to a human or non-human mammal that exhibits one or more symptoms or signs of skin cancer and / or has been diagnosed with skin cancer, including solid tumors, and is in need of treatment for skin cancer. In many embodiments, the terms "subject" and "patient" are used interchangeably. The phrase includes subjects with primary, established, or recurrent tumor lesions. In certain embodiments, the phrase includes human subjects with solid tumors and / or in need of treatment for solid tumors. The phrase also includes subjects with primary or metastatic tumors (advanced malignancies). In certain embodiments, the phrase includes patients with solid tumors that are resistant or refractory to or inadequately controlled by previous therapy (e.g., surgery or treatment with anticancer agents such as carboplatin or docetaxel). In certain embodiments, the phrase includes patients with tumor lesions that have been treated (e.g., surgically removed) with one or more lines of previous therapy, but have subsequently recurred. In certain embodiments, the term includes subjects with skin cancer tumor lesions who are not candidates for curative surgery or curative radiation, or for whom conventional anti-cancer therapy is not advisable, e.g., due to toxic side effects. In other embodiments, the term includes subjects with skin cancer tumor lesions planned for surgical removal. In other embodiments, the term includes subjects at high risk of recurrence due to a prior history of recurrence after surgery.

[0034] In certain embodiments, the method of the present disclosure is used in subjects with solid tumors. As used herein, the term "solid tumor" refers to an abnormal mass of tissue that does not usually contain cysts or liquid areas. A solid tumor can be benign (not cancer) or malignant (cancer). In the present disclosure, the term "solid tumor" refers to a malignant solid tumor. The term includes different types of solid tumors that are named after the cell type that forms them, namely, sarcoma, carcinoma, and lymphoma. In certain embodiments, the term "solid tumor" includes more than one tumor lesion located away from each other in a subject in need of treatment, for example, 2 or more, 5 or more, 10 or more, 15 or more, 20 or more, 25 or more lesions. In certain embodiments, more than one lesion is located distal to each other.

[0035] In certain embodiments, the disclosed methods include administering a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) in combination with an anti-tumor therapy. As used herein, the phrase "in combination with" means that the PD-1 inhibitor is administered before, after, or simultaneously with the anti-tumor therapy. The anti-tumor therapy includes, but is not limited to, conventional anti-tumor therapy, such as chemotherapy, radiation, surgery, or those described elsewhere herein. In one embodiment, the anti-tumor therapy includes surgery. In one embodiment, the PD-1 inhibitor is administered as a neoadjuvant treatment prior to surgery and / or radiation therapy. In another embodiment, the PD-1 inhibitor (e.g., an anti-PD-1 antibody) is administered as an adjuvant treatment after surgery, after radiation therapy, or after surgery and radiation therapy after surgery. In certain embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody) is administered after surgery and optionally radiation therapy after surgery.

[0036] In certain embodiments, the method of the disclosure includes: (a) selecting a patient with skin cancer, the skin cancer being selected from CSCC, BCC, Merkel cell carcinoma, or melanoma, and the patient having one of the following high-risk features: nodal disease with extracapsular spread and at least one nodule >20 mm; in-transit metastasis (ITM); T4 lesion; perineural invasion (PNI); and recurrent CSCC+ at least one of the following additional features: disease ≧N2b associated with recurrent lesion; nominal ≧T3; and recurrent lesion ≧20 mm in diameter); and (b) administering a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to the patient in need thereof. In some embodiments, the patient has an advanced solid tumor, e.g., CSCC. In certain embodiments, the advanced solid tumor is indolent or aggressive. In one embodiment, the patient has had previous surgery and / or surgery to treat the skin cancer. In some embodiments, the disclosed methods include selecting high risk CSCC patients for recurrence of CSCC based on patients exhibiting one of the following high risk features: nodal disease with extracapsular extension and at least one node >20mm; in-transit metastasis (ITM); T4 lesion; perineural invasion (PNI); and recurrent CSCC+ at least one of the following additional features: ≧N2b disease associated with the recurrent lesion; nominal ≧T3; and recurrent lesion ≧20mm in diameter (optionally the patient has undergone resection surgery).

[0037] In certain embodiments, the subject is unresponsive to a previous therapy or surgery or has relapsed (e.g., experienced a recurrent disease) after a previous therapy or surgery. In certain embodiments, the PD-1 inhibitor (e.g., an anti-PD-1 antibody) is administered intravenously or intraperitoneally to the subject. In some embodiments, the subject has a CSCC with high-risk tumor configuration, e.g., high-risk nodal disease, T4 tumor, perineural invasion, in-transit metastasis, or history of recurrence plus at least one other risk factor.

[0038] As used herein, the expression "high risk" for CSCC recurrence or disease recurrence in CSCC patients refers to at least one of the following factors: (a) extracapsular spread (ECE) and nodal disease with at least one node >20 mm in surgical pathology report (ECE is defined as spread through the lymph node capsule into the surrounding connective tissue with or without associated stromal reaction including, for example, invasion of the skin and infiltration / fixation to adjacent structures in clinical examination); (b) in-transit metastasis (ITM), defined as cutaneous or subcutaneous metastasis >2 cm from the primary lesion but not beyond the regional lymph node basin (Leitenberger et al., J. Cancer Res. 2014; 13: 1111-1122, 2014). Am Acad Dermatol, 75(5):1022-31, 2016; (c) T4 lesions, including HN lesions (AJCC, 2017) and non-HN lesions (UICC, 2015); (d) perineural invasion (PNI), defined as clinical and / or radiological involvement of a specified nerve (UICC, Manual of Clinical Oncology, 9th ed., 2015) (e) recurrent CSCC, defined as CSCC arising within a previously resected area of ​​tumor, with at least one of the following additional configurations (AJCC, 2017): (i) ≥N2b disease associated with the recurrent lesion, (ii) nominal ≥T3 (recurrent lesion ≥4 cm in diameter or minimal bone erosion or >6 mm deep invasion as measured from the granular layer of normal adjacent epithelium), or (iii) recurrent lesion with poorly differentiated histology and ≥20 mm in diameter (recurrent tumor must be documented to be within a previously resected area of ​​CSCC by a final radial measurement of the longest radius of the abnormality measured from the estimated center of the original surgical wound).

[0039] The methods of the disclosure, according to certain embodiments, include administering to a subject a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) in combination with an additional therapeutic agent, therapeutic regimen, or procedure that can be administered to enhance anti-tumor efficacy, to reduce toxic effects of one or more therapies, and / or to reduce the dosage of one or more therapies. In various embodiments, the additional therapeutic agent, therapeutic regimen, or therapeutic procedure can include one or more of the following: chemotherapy, cyclophosphamide, surgery, a cancer vaccine, a programmed death-ligand 1 (PD-L1) inhibitor (e.g., an anti-PD-L1 antibody), a lymphocyte-activation gene 3 (LAG3) inhibitor (e.g., an anti-LAG3 antibody), a cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) inhibitor (e.g., ipilimumab), a glucocorticoid-inducible tumor necrosis factor receptor (GITR) agonist (e.g., an anti-GITR antibody), a T-cell immunoglobulin and mucin-containing-3 (TIM3) inhibitor, a B-and T-lymphocyte attenuator (BTLA) inhibitor, a T-cell immunoreceptor with Ig and ITIM domains (T- cell immunoreceptor with Ig and ITIM domains (TIGIT) inhibitors, CA28 activators, 4-1BB agonists, CD38 inhibitors, CD47 inhibitors, indoleamine-2,3-dioxygenase (IDO) inhibitors, vascular endothelial growth factor (VEGF) antagonists, angiopoietin-2 (Ang2) inhibitors, anti-CD3 antibodies, transforming growth factor beta (TGFβ) inhibitors, epidermal growth factor receptor (EGFR) inhibitors, tumor-specific antigens [e.g., CA9, CA125, melanoma-associated antigen 3 (MAGE3), carcinoembryonic antigen (CEA), vimentin, tumor-specific antigens (TGF-β), ... Antibodies against 2-PK, prostate specific antigen (PSA), mucin-1, MART-1 and CA19-9, anti-CD3 / anti-CD20 bispecific antibodies, vaccines (e.g., bacillus Calmette-Guerin), granulocyte-macrophage colony stimulating factor, cytotoxins, chemotherapeutic agents, IL-6R inhibitors, IL-4R inhibitors, IL-10 inhibitors, cytokines such as IL-2, IL-7, IL-21 and IL-15, anti-inflammatory agents such as corticosteroids, nonsteroidal anti-inflammatory drugs (NSAIDs), and nutritional supplements such as antioxidants. In certain embodiments, PD-1 inhibitors (e.g., anti-PD-1 antibodies) can be administered in combination with therapies including chemotherapeutic agents and / or surgery.

[0040] In certain embodiments, administration of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as an adjuvant treatment following completion of surgery and optionally radiation therapy, e.g., post-surgical radiation therapy, leads to the complete disappearance of all evidence of tumor cells (a "complete response"), leads to at least a 30% or greater reduction in tumor cells or tumor size (a "partial response"), or leads to the complete or partial disappearance of tumor cells / lesions, including new measurable lesions. Tumor reduction can be measured by any method known in the art, e.g., by x-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0041] In certain embodiments, administering a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as an adjuvant treatment following completion of surgery and optionally radiation therapy, e.g., post-surgery radiation therapy, leads to an increase in the subject's overall survival (OS) or progression-free survival (PFS) compared to subjects receiving a "standard of care" (SOC) therapy (e.g., chemotherapy, surgery or radiation). In certain embodiments, PFS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects receiving any one or more SOC therapies. In certain embodiments, OS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects administered any one or more SOC therapies.

[0042] In certain embodiments, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as an adjuvant treatment following completion of surgery and optionally post-surgical radiation therapy, where the subject is at high risk of recurrence, leads to a reduced risk of subsequent recurrence of the skin cancer or eliminates the occurrence of recurrence of the skin cancer. In certain embodiments, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a CSCC patient following completion of surgery and / or radiation therapy to treat the skin cancer leads to a reduced risk of subsequent recurrence of the CSCC or eliminates the occurrence of recurrence of the CSCC. In certain embodiments of the disclosed methods, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a CSCC patient following completion of surgery and radiation therapy to treat the skin cancer leads to an adjuvant treatment for the skin cancer. and eliminating the occurrence of recurrence of CSCC for at least 6 months, or for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years or more following administration of the adjuvant PD-1 inhibitor.

[0043] In certain embodiments, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a skin cancer patient following completion of surgery and radiation therapy to treat the skin cancer leads to at least about a 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, or about 90% lower incidence of subsequent recurrence of CSCC compared to patients treated with surgery and radiation therapy without an adjuvant skin cancer treatment.

[0044] In certain embodiments, administering a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as a neoadjuvant therapy prior to planned surgery to treat the skin cancer, and optionally subsequently administering a PD-1 inhibitor to the patient as an adjuvant therapy after surgery to treat the skin cancer, leads to the complete disappearance of all evidence of tumor cells (a "complete response"), leads to at least a 30% or greater reduction in tumor cells or tumor size (a "partial response"), or leads to the complete or partial disappearance of tumor cells / lesions, including new measurable lesions. Tumor reduction can be measured by any method known in the art, for example, by x-ray, positron emission tomography (PET), computed tomography (CT), magnetic resonance imaging (MRI), cytology, histology, or molecular genetic analysis.

[0045] In certain embodiments, administering a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as a neoadjuvant therapy prior to planned surgery to treat the skin cancer, and optionally subsequently administering the PD-1 inhibitor to the patient as an adjuvant therapy after surgery to treat the skin cancer, results in an increase in the subject's overall survival (OS) or progression-free survival (PFS) compared to subjects receiving a "standard of care" (SOC) therapy (e.g., chemotherapy, surgery, or radiation). In certain embodiments, PFS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects receiving any one or more SOC therapies. In certain embodiments, OS is increased by at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months, at least 8 months, at least 9 months, at least 10 months, at least 11 months, at least 1 year, at least 2 years, or at least 3 years compared to subjects administered any one or more SOC therapies.

[0046] In certain embodiments, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject with skin cancer as a neoadjuvant treatment prior to planned surgery leads to a reduced risk of subsequent recurrence of skin cancer or eliminates the occurrence of recurrence of skin cancer. In certain embodiments, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a CSCC patient prior to surgical removal of a skin cancer lesion to treat skin cancer leads to a reduced risk of subsequent recurrence of CSCC or eliminates the occurrence of recurrence of CSCC. In certain embodiments of the disclosed methods, administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a CSCC patient prior to planned surgery to treat skin cancer as a neoadjuvant treatment eliminates the occurrence of recurrence of CSCC for at least 6 months, or for at least 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 years or more following surgery.

[0047] In certain embodiments, neoadjuvant therapy is administered prior to planned surgery to treat skin cancer. Administration of a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a skin cancer patient as an adjuvant treatment leads to at least about a 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80% or about 90% lower incidence of subsequent recurrence of CSCC compared to patients not treated with surgery without a neoadjuvant skin cancer treatment.

[0048] PD-1 inhibitors The methods disclosed herein include administering a therapeutically effective amount of a PD-1 inhibitor. As used herein, "PD-1 inhibitor" refers to any molecule capable of inhibiting, blocking, abrogating or preventing the activity or expression of PD-1. In some embodiments, the PD-1 inhibitor may be an antibody, a small molecule compound, a nucleic acid, a polypeptide, or a functional fragment or variant thereof. Non-limiting examples of suitable PD-1 inhibitor antibodies include anti-PD-1 antibodies and their antigen-binding fragments, anti-PD-L1 antibodies and their antigen-binding fragments, and anti-PD-L2 antibodies and their antigen-binding fragments. Other non-limiting examples of suitable PD-1 inhibitors include RNAi molecules, such as anti-PD-1 RNAi molecules, anti-PD-L1 RNAi and anti-PD-L2 RNAi, antisense molecules, such as anti-PD-1 antisense RNA, anti-PD-L1 antisense RNA and anti-PD-L2 antisense RNA, and dominant negative proteins, such as dominant negative PD-1 proteins, dominant negative PD-L1 proteins and dominant negative PD-L2 proteins. Some examples of the aforementioned PD-1 inhibitors are described, for example, in US9308236, US10011656, and US20170290808, the portions thereof identifying PD-1 inhibitors are incorporated herein by reference.

[0049] As used herein, the term "antibody" refers to immunoglobulin molecules and multimers thereof (e.g., IgM) that contain four polypeptide chains, i.e., two heavy (H) chains and two light (L) chains, interconnected by disulfide bonds. In a typical antibody, each heavy chain contains a heavy chain variable region (HCVR or V H The heavy chain constant region includes three domains: H 1. C H 2 and C H 3. Each light chain contains a light chain variable region (LCVR or V L The light chain constant region comprises one domain (C L 1) is included.H and V L The regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). H and V L is composed of three CDRs and four FRs, arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In different embodiments of the present invention, the FRs of the anti-IL-4R antibody (or antigen-binding portion thereof) may be identical to human germline or may be naturally or artificially modified. An amino acid consensus sequence may be defined based on a side-by-side analysis of two or more CDRs. As used herein, the term "antibody" also includes antigen-binding fragments of an intact antibody molecule.

[0050] As used herein, the terms "antigen-binding portion" of an antibody, "antigen-binding fragment" of an antibody, and the like, include any naturally occurring, enzymatically derived, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Antigen-binding fragments of antibodies can be generated, for example, from intact antibody molecules using any suitable standard technique, for example, proteolytic digestion or recombinant genetic engineering techniques, including the manipulation and expression of DNA encoding the variable and optionally constant domains of an antibody. Such DNA is known and / or readily available, for example, from commercial sources, DNA libraries (including, for example, phage antibody libraries), or can be synthesized. DNA can be sequenced and engineered chemically or by using molecular biology techniques to, for example, modify one or more variable and / or constant domains. Alternatively, one can place the constant domains in an appropriate configuration, or introduce codons, create cysteine ​​residues, modify, add or delete amino acids, and the like.

[0051] Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab')2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of amino acid residues that mimic the hypervariable regions of an antibody (e.g., isolated complementarity determining regions (CDRs), e.g., CDR3 peptides), or constrained FR3-CDR3-FR4 peptides. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression "antigen-binding fragments" as used herein.

[0052] Antigen-binding fragments of antibodies typically contain at least one variable domain. The variable domain may be of any size or amino acid composition and generally contains at least one CDR adjacent to or in frame with one or more framework sequences. L V bound to the domain H For antigen-binding fragments containing domains, V H and V L The domains may be positioned relative to each other in any suitable configuration. For example, the variable region may be a dimer, with the V H -V H , V H -V L or V L -V L Alternatively, the antigen-binding fragment of the antibody may comprise a monomeric V H or V L It may include a domain.

[0053] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found within an antigen-binding fragment of an antibody of the present disclosure include: (i) a V H -C H 1;(ii)V H -C H 2;(iii)V H -C H 3;(iv)V H -C H 1-C H 2;(v)V H -C H 1-C H 2-C H 3;(vi)V H -C H 2-C H 3;(vii)V H -C L ;(viii)V L -C H 1;(ix)V L -C H 2;(x)V L -C H 3;(xi)V L -C H 1-C H 2;(xii)V L -C H 1-C H 2-C H 3;(xiii)V L -C H 2-C H 3; and (xiv) V L -C LIn any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly linked to each other or may be linked by a complete or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids that provide a flexible or semi-flexible bond between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments of antibodies of the present disclosure may be linked to each other and / or to one or more monomeric V H Or V L It may comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above in non-covalent association (eg, via disulfide bonds) of the domains.

[0054] The antibody used in the methods disclosed herein may be a human antibody. As used herein, the term "human antibody" refers to an antibody having variable and constant regions derived from human germline immunoglobulin sequences. Nevertheless, the human antibody of the present disclosure may contain amino acid residues (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation), e.g., in the CDRs, particularly CDR3, that are not encoded by human germline immunoglobulin sequences. However, as used herein, the term "human antibody" refers to an antibody derived from the germline of another mammalian species, e.g., a mouse. It is not intended to include antibodies in which the CDR sequences of the present invention have been grafted onto human framework sequences.

[0055] The antibodies used in the methods disclosed herein may be recombinant human antibodies. As used herein, the term "recombinant human antibody" refers to any human antibody that is prepared, expressed, produced, or isolated by recombinant means, e.g., antibodies expressed using a recombinant expression vector transfected into a host cell (described further below), antibodies isolated from a recombinant combinatorial human antibody library (described further below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes [see, e.g., Taylor et al. (1992) Nucl. Acids. Res. 20:6287-6295], or any other means involving splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis) to thereby modify the V and constant regions of the recombinant antibodies. H and V L The amino acid sequence of the region is H and V L These are sequences that are derived from and related to a sequence, but may not naturally occur within the human antibody germline repertoire in vivo.

[0056] Anti-PD-1 antibodies and antigen-binding fragments thereof In some embodiments, the PD-1 inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to PD-1. The term "specifically binds" and the like means that the antibody or antigen-binding fragment thereof forms a relatively stable complex with the antigen under physiological conditions. Methods for determining whether an antibody specifically binds to an antigen are well known in the art and include, for example, equilibrium dialysis, surface plasmon resonance, and the like. For example, as used in this disclosure, an antibody that "specifically binds" to PD-1 has a K of less than about 500 nM, less than about 300 nM, less than about 200 nM, less than about 100 nM, less than about 90 nM, less than about 80 nM, less than about 70 nM, less than about 60 nM, less than about 50 nM, less than about 40 nM, less than about 30 nM, less than about 20 nM, less than about 10 nM, less than about 5 nM, less than about 4 nM, less than about 3 nM, less than about 2 nM, less than about 1 nM, or less than about 0.5 nM, as measured by a surface plasmon resonance assay. D An isolated antibody that specifically binds human PD-1 may, however, have cross-reactivity to other antigens, such as PD-1 molecules from other (non-human) species.

[0057] According to certain exemplary embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR) and / or a complementarity determining region (CDR) comprising the amino acid sequence of any of the anti-PD-1 antibodies described in U.S. Patent No. 9,987,500, the entire contents of which are incorporated herein by reference. In certain exemplary embodiments, the anti-PD-1 antibody or antigen-binding fragment thereof that can be used in the present disclosure comprises a heavy chain complementarity determining region (HCDR) of the heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 and a light chain complementarity determining region (LCDR) of the light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:2. According to certain embodiments, the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), where HCDR1 comprises the amino acid sequence of SEQ ID NO:3; HCDR2 comprises the amino acid sequence of SEQ ID NO:4; HCDR3 comprises the amino acid sequence of SEQ ID NO:5; LCDR1 comprises the amino acid sequence of SEQ ID NO:6; LCDR2 comprises the amino acid sequence of SEQ ID NO:7; and LCDR3 comprises the amino acid sequence of SEQ ID NO:8. In yet other embodiments, the anti-PD-1 antibody, or antigen-binding fragment thereof, comprises a HCVR comprising SEQ ID NO:1 and a LCVR comprising SEQ ID NO:2. In certain embodiments, the methods of the disclosure involve the use of an anti-PD-1 antibody. and the antibody comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 9. In some embodiments, the anti-PD-1 antibody comprises a light chain comprising the amino acid sequence of SEQ ID NO: 10. An exemplary antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO: 9 and a light chain comprising the amino acid sequence of SEQ ID NO: 10 is the fully human anti-PD-1 antibody known as cemiplimab (REGN2810, also known as LIBTAYO®).

[0058] According to certain exemplary embodiments, the methods of the present disclosure include the use of REGN2810 or a bioequivalent thereof. As used herein, the term "bioequivalent" refers to an anti-PD-1 antibody or PD-1 binding protein or fragment thereof that is a pharmaceutical equivalent or pharmaceutical substitute that exhibits no significant difference in rate and / or extent of absorption from a reference antibody (e.g., REGN2810) when administered in single or multiple doses at the same molar dose under similar experimental conditions. In the present disclosure, the term "bioequivalent" includes an antigen binding protein that binds to PD-1 and has no clinically meaningful difference from REGN2810 in terms of safety, purity and / or potency.

[0059] According to certain embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR having 90%, 95%, 98% or 99% sequence identity to SEQ ID NO:1.

[0060] According to certain embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises an LCVR having 90%, 95%, 98% or 99% sequence identity to SEQ ID NO:2.

[0061] According to certain embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a HCVR comprising the amino acid sequence of SEQ ID NO: 1, with no more than five amino acid substitutions. According to certain embodiments of the present disclosure, the anti-human PD-1 or antigen-binding fragment thereof comprises a LCVR comprising the amino acid sequence of SEQ ID NO: 2, with no more than two amino acid substitutions.

[0062] Sequence identity can be measured by methods known in the art (eg, GAP, BESTFIT and BLAST).

[0063] The disclosure also includes the use of anti-PD-1 antibodies in methods for treating skin cancer, where the anti-PD-1 antibodies comprise variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein having one or more conservative amino acid substitutions. For example, the disclosure includes the use of anti-PD-1 antibodies having HCVR, LCVR and / or CDR amino acid sequences with, e.g., 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0064] Other anti-PD-1 antibodies that can be used in the methods of the present disclosure include, for example, nivolumab, pembrolizumab, MEDI0608, pidilizumab, BI 754091, spartalizumab (also known as PDR001), camrelizumab (also known as SHR-1210), JNJ-63723283, MCLA-134, or any of the anti-PD-1 antibodies described in U.S. Patent Nos. 6,808,710, 7,488,802, 8,008,449, 8,168,757, 8,354,509, 8,609,089, 8,686,119, 8,779,105, 8,900,587, and 9,987,500, and in patent publications WO2006 / 121168, WO2009 / 114335. All portions of the foregoing publications identifying anti-PD-1 antibodies are incorporated herein by reference.

[0065] The anti-PD-1 antibodies used in the methods of the present disclosure may have pH-dependent binding properties. For example, an anti-PD-1 antibody for use in the methods of the present disclosure may exhibit reduced binding to PD-1 at acidic pH compared to neutral pH. Alternatively, an anti-PD-1 antibody of the present invention may exhibit enhanced binding to its antigen at acidic pH compared to neutral pH. The phrase "acidic pH" includes pH values ​​less than about 6.2, e.g., about 6.0, 5.95, 5.9, 5.85, 5.8, 5.75, 5.7, 5.65, 5.6, 5.55, 5.5, 5.45, 5.4, 5.35, 5.3, 5.25, 5.2, 5.15, 5.1, 5.05, 5.0 or less. As used herein, the phrase "neutral pH" refers to a pH of about 7.0 to about 7.4. The expression "neutral pH" includes pH values ​​of about 7.0, 7.05, 7.1, 7.15, 7.2, 7.25, 7.3, 7.35 and 7.4.

[0066] In certain cases, "reduced binding to PD-1 at acidic pH compared to neutral pH" refers to the K D K value vs. K for antibody binding to PD-1 at neutral pH D For example, an antibody or antigen-binding fragment thereof may be expressed in terms of a ratio of its acidic / neutral K value (or its inverse) of about 3.0 or greater. D When a ratio is presented, it can be considered in the present disclosure to indicate "reduced binding to PD-1 at acidic pH compared to neutral pH." In certain exemplary embodiments, the acidic / neutral K for an antibody or antigen-binding fragment of the present disclosure is D The ratio can be about 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 10.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 14.5, 15.0, 20.0, 25.0, 30.0, 40.0, 50.0, 60.0, 70.0, 100.0 or more.

[0067] Antibodies with pH-dependent binding properties can be obtained, for example, by screening a population of antibodies for reduced (or enhanced) binding to a particular antigen at acidic pH compared to neutral pH. Furthermore, modification of the antigen-binding domain at the amino acid level can result in an antibody with pH-dependent properties. For example, by replacing one or more amino acids in the antigen-binding domain (e.g., in the CDR) with histidine residues, an antibody can be obtained that has reduced antigen binding at acidic pH compared to neutral pH. As used herein, the expression "acidic pH" refers to a pH of 6.0 or less.

[0068] Anti-PD-L1 antibodies and antigen-binding fragments thereof In some embodiments, the PD-1 inhibitor used in the methods disclosed herein is an antibody or antigen-binding fragment thereof that specifically binds to PD-L1. For example, as used in this disclosure, an antibody that "specifically binds" to PD-L1 is an antibody that specifically binds to PD-L1, such as an antibody that binds to PD-L1 in an amount of about 1×10 -8 K below M D (For example, K D A "high affinity" anti-PD-L1 antibody includes an antibody that binds to PD-L1 or a portion thereof with a affinity of at least 10 as measured by surface plasmon resonance, e.g., BIACORE™, or solution affinity ELISA. -8 M, preferably 10 -9 M, more preferably 10 -10 M, and even more preferably 10 -11 M, and even more preferably 10 -12 K of M D " refers to a mAb that has binding affinity for PD-L1, expressed as " ". However, an isolated antibody that specifically binds human PD-L1 may have cross-reactivity to other antigens, such as PD-L1 molecules from other (non-human) species.

[0069] According to certain exemplary embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof comprises a heavy chain variable region (HCVR), a light chain variable region (LCVR) and / or a complementarity determining region (CDR) comprising the amino acid sequence of any of the anti-PD-L1 antibodies described in U.S. Patent No. 9,938,345, the contents of which are incorporated herein by reference in their entirety. In certain exemplary embodiments, the anti-PD-L1 antibodies that can be used in the present disclosure or an antigen-binding fragment thereof comprising a heavy chain complementarity determining region (HCDR) of the heavy chain variable region (HCVR) and a light chain complementarity determining region (LCDR) of the light chain variable region (LCVR), wherein the HCVR and LCVR comprise the amino acid sequence of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345. According to certain embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof comprises three HCDRs (HCDR1, HCDR2, and HCDR3) and three LCDRs (LCDR1, LCDR2, and LCDR3), wherein HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 comprise the amino acid sequence of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345. In yet other embodiments, the anti-PD-L1 antibody or antigen-binding fragment thereof comprises a HCVR and a LCVR that comprise the amino acid sequence of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345.

[0070] According to certain embodiments of the disclosure, the anti-human PD-L1 or antigen-binding fragment thereof comprises an LCVR that has 90%, 95%, 98% or 99% sequence identity to the LCVR amino acid sequence of the anti-PD-L1 antibody designated H2M8314N in U.S. Patent No. 9,938,345.

[0071] According to certain embodiments of the present disclosure, the anti-human PD-L1 or antigen-binding fragment thereof comprises a HCVR that includes the amino acid sequence of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345, with five or fewer amino acid substitutions. According to certain embodiments of the present disclosure, the anti-human PD-L1 or antigen-binding fragment thereof comprises a LCVR that includes the amino acid sequence of the anti-PD-L1 antibody designated as H1H8314N in U.S. Patent No. 9,938,345, with two or fewer amino acid substitutions.

[0072] Sequence identity can be measured by methods known in the art (eg, GAP, BESTFIT and BLAST).

[0073] The disclosure also includes the use of anti-PD-L1 antibodies in methods for treating skin cancer, where the anti-PD-L1 antibodies comprise variants of any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein having one or more conservative amino acid substitutions. For example, the disclosure includes the use of anti-PD-L1 antibodies having HCVR, LCVR and / or CDR amino acid sequences with, for example, 10 or less, 8 or less, 6 or less, 4 or less, etc., conservative amino acid substitutions relative to any of the HCVR, LCVR and / or CDR amino acid sequences disclosed herein.

[0074] Other anti-PD-L1 antibodies that can be used in the methods of the disclosure include, for example, MDX-1105, atezolizumab (TECENTRIQ™), durvalumab (IMFINZI™), avelumab (BAVENCIO™), LY3300054, FAZ053, STI-1014, CX-072, KN035 (Zhang et al., Cell Discovery, 3, 170004 (March 2017)), CK-301 (Gorelik et al., American Association for Cancer Research, 2014). Anti-PD-L1 antibodies include those known in the art (see, for example, the American Association of Clinical Chemists and Neurosurgeons (AACR), 2016-04-04 Abstract 4606), or any of the other anti-PD-L1 antibodies described in patent publications US7943743, US8217149, US9402899, US9624298, US9938345, WO2007 / 005874, WO2010 / 077634, WO2013 / 181452, WO2013 / 181634, WO2016 / 149201, WO2017 / 034916, or EP3177649. All portions of the foregoing publications identifying anti-PD-L1 antibodies are incorporated herein by reference.

[0075] Pharmaceutical Compositions and Administration The PD-1 inhibitors disclosed herein can be included in pharmaceutical compositions that can be formulated with suitable carriers, excipients, buffers, and other agents that provide suitable migration, delivery, tolerability, etc. Numerous suitable formulations can be found in the formulary known to every pharmacist: Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA. These formulations include, for example, powders, pastes, ointments, jellies, waxes, oils, lipids, lipid (cationic or anionic)-containing vesicles (e.g., LIPOFECTIN™), DNA conjugates, anhydrous absorption pastes, oil-in-water and water-in-oil emulsions, emulsions carbowax (polyethylene glycols of various molecular weights), semi-solid gels, and carbowax-containing semi-solid mixtures. See also Powell et al., "Compendium of excipients for parenteral formulations," PDA, J Pharm Sci Technol 52:238-311 (1998).

[0076] Various delivery systems are known and can be used to administer the pharmaceutical compositions of the present invention, such as encapsulation in liposomes, microparticles, microcapsules, recombinant cells capable of expressing mutant viruses, and receptor-mediated endocytosis. See, e.g., Wu et al., J. Biol. Chem. 262:4429-32 (1987).

[0077] The pharmaceutical compositions comprising the PD-1 inhibitors disclosed herein are suitable for intravenous or intraperitoneal administration.

[0078] The injectable formulation of the pharmaceutical composition can be prepared by known methods. For example, the injectable formulation can be prepared by dissolving, suspending, or emulsifying the above-mentioned antibody or its salt in a sterile aqueous or oily medium that is conventionally used for injection. Examples of aqueous media for injection include isotonic solutions containing physiological saline, glucose, and other auxiliary agents, which can be used in combination with suitable solubilizers, such as alcohol (e.g., ethanol), polyalcohols (e.g., propylene glycol, polyethylene glycol), nonionic surfactants [e.g., polysorbate 80, HCO-50 (polyoxyethylene (50 moles) adduct of hydrogenated castor oil)], etc. Examples of oily media include sesame oil, soybean oil, etc., which can be used in combination with solubilizers, such as benzyl benzoate, benzyl alcohol, etc. The injectable formulation thus prepared is preferably filled into a suitable injection ampoule. In some embodiments, the injectable formulation may be in the form of an injectable solution that includes a concentrate of the PD-1 inhibitor and one or more solvents (eg, distilled water, saline, etc.).

[0079] In certain embodiments, the disclosure provides a pharmaceutical composition or formulation comprising a therapeutic amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) and a pharma- ceutical acceptable carrier. In certain embodiments, the disclosure provides a PD-1 inhibitor (e.g., an anti-PD-1 antibody) formulated in a pharmaceutical composition suitable for administration by intravenous injection.

[0080] Exemplary pharmaceutical compositions comprising anti-PD-1 antibodies that can be used in the present disclosure are disclosed, for example, in US2019 / 0040137.

[0081] Dosing regimen In certain embodiments, the methods disclosed herein involve administering a therapeutically effective amount of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) to a subject, e.g., as part of a particular therapeutic dosing regimen. For example, suitable therapeutic dosing regimens include administering the compound in multiple doses to a tumor in a subject in need thereof, for as long as a therapeutic response is achieved, such as about once a day, once every 2 days, once every 3 days, once every 4 days, once every 5 days, once every 6 days, once a week, once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 5 weeks, once every 6 weeks, once every 8 weeks, once every 12 weeks, once a month, once every 2 months, once every 3 months, once every 4 months, twice a day, twice every 2 days, twice every 3 days, twice every 4 days, twice every 5 days, twice every 6 days, twice a week, twice every 2 weeks, twice every 3 weeks, twice every 4 weeks, twice every 5 weeks, twice every 6 weeks ... This can include administering one or more doses of a PD-1 inhibitor to a subject twice every week, twice every 8 weeks, twice every 12 weeks, twice monthly, twice every 2 months, twice every 3 months, twice every 4 months, 3 times daily, 3 times every 2 days, 3 times every 3 days, 3 times every 4 days, 3 times every 5 days, 3 times every 6 days, 3 times weekly, 3 times every 2 weeks, 3 times every 3 weeks, 3 times every 4 weeks, 3 times every 5 weeks, 3 times every 6 weeks, 3 times every 8 weeks, 3 times every 12 weeks, 3 times monthly, 3 times every 2 months, 3 times every 3 months, 3 times every 4 months, or less frequently, or as needed. In one embodiment, one or more doses of an anti-PD-1 antibody are administered once a week.

[0082] In certain embodiments, one or more doses are administered in at least one treatment cycle. The method according to this aspect includes administering to a subject in need thereof at least one treatment cycle comprising administration of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more doses of a PD-1 inhibitor (e.g., an anti-PD-1 antibody). In one embodiment, the treatment cycle includes 3 doses of a PD-1 inhibitor. In one embodiment, the treatment cycle includes 12 doses of a PD-1 inhibitor. In one embodiment, the treatment cycle includes 24 doses of a PD-1 inhibitor. In one embodiment, the treatment cycle includes 3 doses of a PD-1 inhibitor, each dose being administered 2 weeks after the immediately preceding dose. In one embodiment, the treatment cycle includes 10 doses of a PD-1 inhibitor, each dose being administered 1 week after the immediately preceding dose. In one embodiment, the treatment cycle includes 12 doses of a PD-1 inhibitor, each dose being administered 1 week after the immediately preceding dose.

[0083] In one embodiment, all doses administered in a treatment cycle contain the same amount of PD-1 inhibitor. In another embodiment, a treatment cycle includes administration of at least two doses containing different amounts of PD-1 inhibitor. In one embodiment, the first dose of a treatment cycle includes a larger amount of PD-1 inhibitor than subsequent doses of the treatment cycle. In one embodiment, the first dose of a treatment cycle includes a smaller amount of PD-1 inhibitor than subsequent doses of the treatment cycle.

[0084] In one embodiment, all doses of a treatment cycle are administered using the same route of administration, hi another embodiment, the doses of a treatment cycle are administered using different routes of administration, including two or more routes of administration.

[0085] In one embodiment, the treatment cycle is repeated, hi some embodiments, the treatment cycle is repeated 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 or more times.

[0086] In certain embodiments, a dose of the PD-1 inhibitor is administered to a subject in a single session or patient visit.

[0087] As used herein, the terms "first", "second", "third", etc. refer to the temporal order of administration. Thus, the "first dose" is the dose administered at the beginning of a treatment regimen (also referred to as the "baseline dose"); the "second dose" is the dose administered after the first dose; and the "third dose" is the dose administered after the second dose. The first, second and third doses may all contain the same amount of PD-1 inhibitor (anti-PD-1 antibody). However, in certain embodiments, the amount contained in the first, second and / or third dose may be 0.01 to 0.05%. , which may vary from one another (e.g., adjusted up or down as appropriate) during the course of treatment. In certain embodiments, one or more (e.g., 1, 2, 3, 4, or 5) doses are administered at the beginning of a treatment regimen as a "loading dose," followed by subsequent doses administered on a less frequent basis (e.g., "maintenance doses"). For example, an anti-PD-1 antibody can be administered to a cancer patient at a loading dose of about 1 mg / kg to about 3 mg / kg of the patient's body weight, followed by one or more maintenance doses of about 0.1 mg / kg to about 20 mg / kg.

[0088] In an exemplary embodiment of the disclosure, each second and / or third dose is administered ½ to 4 or more weeks (e.g., ½, 1, 1 and ½, 2, 2 and ½, 3, 3 and ½, 4 or more weeks) after the immediately preceding dose. As used herein, the phrase "immediately preceding dose" refers to a dose of an anti-PD-1 antibody in a multiple dose sequence that is administered to a subject prior to administration of the next dose in the sequence, without any intervening doses.

[0089] Similarly, a "first treatment cycle" is a treatment cycle administered at the beginning of a treatment regimen; a "second treatment cycle" is a treatment cycle administered after the first treatment cycle; and a "third treatment cycle" is a treatment cycle administered after the second treatment cycle. In the present disclosure, treatment cycles may be the same as or different from one another.

[0090] Dosage In certain embodiments, each dose of the PD-1 inhibitor comprises 0.1, 1, 0.3, 3, 4, 5, 6, 7, 8, 9, or 10 mg / kg of patient weight. In certain embodiments, each dose comprises 5-500 mg of the PD-1 inhibitor, e.g., 5, 10, 15, 20, 25, 40, 45, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450 mg or more of the PD-1 inhibitor. In one embodiment, the PD-1 inhibitor is REGN2810 (cemiplimab).

[0091] The amount of PD-1 inhibitor administered to a subject (e.g., intravenously or intraperitoneally) according to the methods disclosed herein is generally a therapeutically effective amount. As used herein, the term "therapeutically effective amount" refers to an amount of PD-1 inhibitor that results in one or more of the following: (a) reducing the severity or duration of symptoms or signs of skin cancer, e.g., tumor lesions; (b) inhibiting tumor growth or increasing tumor necrosis, tumor shrinkage, and / or tumor disappearance; (c) slowing tumor growth and development; (d) inhibiting tumor metastasis; (e) preventing recurrence of tumor growth; and / or (f) increasing the survival of a subject with cancer.

[0092] In the case of a PD-1 inhibitor (e.g., an anti-PD-1 antibody), a therapeutically effective amount can be about 5 mg to about 500 mg, about 10 mg to about 450 mg, about 50 mg to about 400 mg, about 75 mg to about 350 mg, or about 100 mg to about 300 mg of the antibody. For example, in various embodiments, the amount of PD-1 inhibitor is about 5 mg, about 10 mg, about 15 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 110 mg, about 120 mg, about 130 mg, about 140 mg, about 150 mg, about 160 mg, about 170 mg, about 180 mg, about 190 mg, about 200 mg, about 210 mg, about 220 mg, about 230 mg, about 240 mg, about 250 mg, about 260 mg, about 270 mg, about 280 mg, about 290 mg, about 300 mg, about 310 mg, about 320 mg, about 330 mg, about 340 mg, about 350 mg, about 360 mg, about 370 mg, about 380 mg, about 390 mg, about 400 mg, about 410 mg, about 420 mg, about 430 mg, about 440 mg, about 450 mg, about 460 mg, about 470 mg, about 480 mg, about 490 mg, about 500 mg, about 510 mg, about 520 mg, about 530 mg, about 540 mg, about 550 mg, about 560 mg, about 570 mg, about 580 mg, about 590 mg, or about 600 mg.

[0093] In one embodiment, according to certain methods disclosed herein, a therapeutically effective amount of 350 mg of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) can be administered intravenously as an adjuvant treatment following surgery and optionally post-surgical radiation therapy. In another embodiment, according to certain methods disclosed herein, a therapeutically effective amount of 350 mg of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) can be administered intravenously as a neoadjuvant treatment prior to a planned surgery to treat skin cancer.

[0094] The amount of PD-1 inhibitor included in an individual dose can be expressed in milligrams of antibody per kilogram of subject body weight (i.e., mg / kg). In certain embodiments, the PD-1 inhibitor used in the methods disclosed herein can be administered to a subject at a dose of about 0.0001 to about 100 mg / kg of the subject's body weight. In certain embodiments, the anti-PD-1 antibody can be administered at a dose of about 0.1 mg / kg to about 20 mg / kg of the patient's body weight. In certain embodiments, the methods of the disclosure include administration of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) at a dose of about 1 mg / kg, 3 mg / kg, 5 mg / kg, or 10 mg / kg of the patient's body weight.

[0095] In certain embodiments, the individual dose of a PD-1 inhibitor (e.g., an anti-PD-1 antibody) administered to a patient (e.g., intravenously or intraperitoneally) may be less than a therapeutically effective amount, i.e., a sub-therapeutic dose. For example, if a therapeutically effective amount of a PD-1 inhibitor comprises 3 mg / kg, a sub-therapeutic dose comprises an amount less than 3 mg / kg, e.g., 2 mg / kg, 1.5 mg / kg, 1 mg / kg, 0.5 mg / kg, or 0.3 mg / kg. As defined herein, a "sub-therapeutic dose" refers to an amount of a PD-1 inhibitor that does not cause a therapeutic effect by itself. However, in certain embodiments, multiple sub-therapeutic doses of a PD-1 inhibitor are administered to collectively achieve a therapeutic effect in a subject.

[0096] In certain embodiments, each dose contains 0.1-10 mg / kg (e.g., 0.3 mg / kg, 1 mg / kg, 3 mg / kg, or 10 mg / kg) of the subject's body weight. In certain other embodiments, each dose contains 5-600 mg of a PD-1 inhibitor (e.g., an anti-PD-1 antibody), e.g., 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 40 mg, 45 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, or 500 mg of a PD-1 inhibitor. EXAMPLES

[0097] The disclosed technology will now be described with the following examples. The use of these and other examples anywhere in this specification is merely illustrative and in no way limits the scope and meaning of the invention or any exemplified form. Likewise, the invention is not limited to any particular preferred embodiment described herein. Indeed, modifications and variations of the invention may become apparent to those skilled in the art upon reading this specification and may be made without departing from its spirit and scope. Thus, the present invention should only be limited by the terms of the claims, along with the full scope of equivalents to which the claims are entitled. Furthermore, efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric pressure. EXAMPLES

[0098] A clinical trial comparing anti-PD-1 antibodies versus placebo as adjuvant treatment for patients with CSCC after surgery and radiotherapy This is a randomized, placebo-controlled, double-blind, multicenter, phase 3 study comparing anti-PD-1 antibody versus placebo as adjuvant treatment for patients with CSCC, a configuration associated with a high risk of recurrent disease, who have completed surgery and postoperative radiotherapy (RT). The study population included patients with CSCC with high-risk configurations on surgical pathology who had completed surgery and postoperative RT (see Figure 1).

[0099] An exemplary anti-PD-1 antibody used in this study is REGN2810 (also known as cemiplimab, or H4H7798N as disclosed in US9987500), which is a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; a HCVR / LCVR amino acid sequence pair comprising SEQ ID NO:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0100] Research purpose The primary objective of the study is to compare disease-free survival (DFS) of patients with high-risk CSCC treated with adjuvant REGN2810 versus those treated with placebo after surgery and RT.

[0101] Secondary objectives of the study include: (1) to compare the overall survival (OS) of high-risk CSCC patients treated with adjuvant REGN2810 versus placebo after surgery and RT; (2) to compare the effect of adjuvant REGN2810 with that of placebo on patients' freedom from locoregional recurrence (FFLRR) after surgery and RT; (3) to compare the effect of adjuvant REGN2810 with that of placebo on patients' freedom from distant recurrence (FFDR) after surgery and RT; (4) to compare the effect of adjuvant REGN2810 with that of placebo on the cumulative incidence of second primary CSCC tumors (SPTs) after surgery and RT; and (5) to evaluate the safety of adjuvant REGN2810 versus placebo in high-risk CSCC patients after surgery and RT.

[0102] Research duration Part 1 of the study (the blinded treatment period) will last up to 48 weeks. Part 2 of the study (the open-label REGN2810 treatment period) will last up to 96 weeks.

[0103] Study population Approximately 412 patients will be randomized into two treatment arms of approximately 206 patients each. The target patient population will consist of adult high-risk CSCC patients undergoing surgical resection followed by RT. Complete macroscopic resection of high-risk CSCC of head and neck (HN) and non-HN sites will be given followed by post-operative radiation therapy (PORT) prior to enrollment and randomization for the study according to the following inclusion criteria:

[0104] Inclusion Criteria: Patients must meet the following criteria to be eligible for inclusion in the study: (1) at least 18 years of age (Japan only, at least 21 years of age); (2) patients with resection of pathologically confirmed CSCC (primary CSCC lesion only, or primary CSCC with lymph node metastasis, or CSCC nodal metastasis with a known previously treated primary CSCC lesion in the draining lymph node echelon) with gross gross resection of all disease; (3) high-risk CSCC defined by at least one of the following: (a) extracapsular extension (ECE) and at least 20 mm in the surgical pathology report; (b) nodal disease with at least one nodule (ECE is defined as extension through the lymph node capsule into the surrounding connective tissue with or without associated stromal reaction. Clear evidence of macroscopic ECE (defined as invasion of the skin, infiltration of the musculature / fixation to adjacent structures on clinical examination) is a high enough threshold to classify these as ECE positive (AJCC, 2017)); (b) in-transit metastases (ITM), defined as cutaneous or subcutaneous metastases >2 cm from the primary lesion but not beyond the regional lymph node basin (Leitenberger et al., J Am Acad Dermatol, 75(5):1022-31, 2016); (c) HN lesions (AJCC, 2 (d) T4 lesions, including T1 lesions (UICC, Manual of Clinical Neuropathy, 2017) and non-HN lesions (UICC, 2015); (e) perineural invasion (PNI), defined as clinical and / or radiological involvement of a specified nerve (UICC, Manual of Clinical Neuropathy, 2017); (e) recurrent CSCC, defined as CSCC arising within a previously resected area of ​​tumor, with at least one of the following additional constructs (AJCC, 2017): (i) ≥N2b disease associated with the recurrent lesion, (ii) nominal ≥T3 (recurrent lesion ≥4 cm in diameter or minimal bone erosion or >6 mm deep invasion as measured from the granular layer of normal adjacent epithelium), or (iii) recurrent lesion with poorly differentiated histology and ≥20 mm in diameter (the recurrent tumor must be documented to be within a previously resected area of ​​CSCC by a radial measurement of the longest radius of the final abnormality measured from the estimated center of the original surgical wound); (4) completion of postoperative RT with curative intent within 2–6 weeks of randomization. Patients must have received a minimum biologically equivalent dose of 2 Gy per fraction (EQD2) to sites of previous gross disease, with 60 Gy for head and neck primary sites and 50 Gy for non-head and neck primary sites; (5) Eastern Cooperative Oncology Group performance status (ECOG PS) ≤ 1; (6) adequate liver function: a. total bilirubin ≤ 1.5 × upper limit of normal (ULN), b. transaminases (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]) ≤ 3 × ULN, c. alkaline phosphatase (ALP) ≤ 2.5 × ULN; (7) adequate renal function: serum creatinine ≤ 1.5 × ULN or estimated creatinine clearance (CrCl) > 30 mL / min by the method of Cockcroft and Galt; (8) adequate bone marrow function: a. hemoglobin ≥ 9.0 g / dL, b. absolute neutrophil count (ANC) ≥ 1.0 × 10 9 / L, c. Platelet count ≥75×10 9(9) must be willing and able to provide informed consent signed by the study patient or legally acceptable representative, if specified by health authorities and institutional guidelines; (10) all toxicities from radiation therapy, except for dysgeusia, fatigue, xerostomia, trismus, alopecia, fibrosis, or edema in the irradiated area, must have resolved to Grade 1 or less; (11) must be willing and able to comply with clinic visits and study-related procedures; and (12) must be able to understand and complete study-related questionnaires.

[0105] Exclusion Criteria: Patients who met any of the following criteria would be excluded from the study: (1) squamous cell carcinoma (SCC) arising in a noncutaneous site (e.g., dry red lips [vermillion], oral cavity, oropharynx, paranasal sinuses, larynx, hypopharynx, nasopharynx, salivary glands, nasal mucosa, anogenital area, or SCC nodal metastasis of unknown primary site); (2) tumors with only negligible risk of metastasis or death, e.g., adequately treated BCC of the skin, carcinoma in situ of the cervix, or ductal carcinoma in situ of the breast, or low-risk early stage prostate adenocarcinoma (T1-T2) in which the management plan is active surveillance. aProstate adenocarcinoma with biochemical recurrence with a reported PSA doubling time >12 months and a management plan of active surveillance (D'Amico et al., JAMA, 294(4):440-47, 2005) (Pham et al., JAMA, 294(4):440-47, 2005) (Urol, 196(2):392-98, 2016); (3) patients with hematologic malignancies (e.g., chronic lymphocytic leukemia [CLL]); (4) patients with a history of distant metastatic CSCC (visceral or distant nodal) unless the disease-free interval was at least 3 years (regional nodal involvement of disease in draining lymph node basins that were resected and irradiated prior to enrollment would not be excluded by Exclusion Criterion 2); (5) ongoing or recent (within 5 years from date of randomization) evidence of significant autoimmune disease that has required treatment with systemic immunosuppressive therapy, which may indicate a risk of immune-related adverse events (irAEs). The following were not excluded: vitiligo that required hormone replacement only, resolved childhood asthma, type 1 diabetes, residual hypothyroidism, or psoriasis not requiring systemic treatment; (6) participation in a study of an investigational drug or device within 4 weeks or 5 half-lives (whichever is longer) from the date of randomization. However, patients who have undergone or are enrolled in a study involving treatment with an investigational immunoPET agent will not be excluded; (7) have received a live vaccine within 28 days of the randomization date; (8) have received previous systemic anti-cancer immunotherapy for CSCC. Examples of immune modulating agents include, but are not limited to, blockers of CTLA-4, 4-1BB (CD137) or OX-40, therapeutic vaccines, anti-PD-1 / PD-L1 or PI3Kδ inhibitors; 9) immunosuppressive corticosteroid administration (>10 mg prednisone daily or equivalent) within 4 weeks prior to the first dose of REGN2810 / placebo (Note: Patients who require a short course of steroids (e.g., prophylaxis for imaging evaluation due to contrast sensitivity) will not be excluded. People taking steroids for physiological replacement (i.e., adrenal insufficiency) will not be excluded); (10) have received treatment with an approved anti-cancer systemic therapy within 4 weeks of the randomization date or have any changes in the laboratory tests listed in inclusion criteria 6, 7 and 8. Have not yet recovered from any acute toxicity (i.e., ≤ grade 1 or baseline) except for (Note: patients receiving bisphosphonates or denosumab are not excluded); (11) prior allogeneic or autologous stem cell transplant; (12) patients who have permanently discontinued anti-cancer immunomodulatory therapy due to drug-related toxicity; (13) encephalitis, meningitis, or uncontrolled seizures in the year prior to screening / enrollment; (14) patients with myocardial infarction within 6 months prior to the randomization date; (15) any infection requiring hospitalization and / or intravenous antibiotic therapy within 2 weeks of the randomization date; (16) active tuberculosis; (17) uncontrolled infection with human immunodeficiency virus (HIV), hepatitis B virus, or hepatitis C virus (HBV or HCV) infection; or a diagnosis of immunodeficiency. (Notes: · Patients with known HIV infection who have controlled their infection (undetectable viral load (HIV RNA PCR) and CD4 count >350, either spontaneously or on a stable antiviral regimen) will be admitted. For patients with controlled HIV infection, monitoring will occur according to local standards. · Patients will be tested for HBV and HCV at screening.·Patients with HBV (hepatitis B surface antigen positive; HepBsAg+) who have controlled infection (serum HBV DNA PCR below the limit of detection and receiving antiviral therapy for HBV) are allowed. Patients with controlled infection must undergo regular monitoring of HBV DNA. Patients must continue antiviral therapy for at least 6 months past the last dose of investigational study drug. ·Patients who are HCV antibody positive (HCV Ab+) and have controlled infection (HCV RNA undetectable by PCR, either spontaneously or in response to a successful prior course of anti-HCV therapy) are allowed;(18) History of immune-related interstitial pneumonia within the past 5 years;(19) History of interstitial lung disease (e.g., idiopathic pulmonary fibrosis, organizing pneumonia) or active noninfectious interstitial pneumonia requiring immunosuppressive doses of glucocorticoids to aid management. History of radiation pneumonitis in the irradiated field is permitted, but only for pneumonitis that has resolved ≥ 6 months prior to the randomization date; (20) history of documented allergic or acute hypersensitivity reactions due to antibody treatment; (21) known hypersensitivity or allergy to any of the excipients in the REGN2810 drug product; (22) patients with a history of solid organ transplantation (patients with a previous corneal transplant are not excluded); (23) any medical comorbidity, physical examination findings or metabolic dysfunction, or laboratory test abnormality that, in the opinion of the investigator, makes the patient unsuitable for participation in the clinical trial because of a high safety risk and / or the potential to affect interpretation of the results of the study; (24) known psychiatric or substance abuse disorder that would prevent participation with the requirements of the study; (25) any member of the study team or his / her immediate family at the clinical site; (26) women with a positive serum β-human chorionic gonadotropin (HCG) pregnancy test at the Screening / Baseline visit. If positive, pregnancy must be ruled out by patient ultrasound in order for the patient to be included; (27) breastfeeding women; (28) women of childbearing potential (WOCBP) or sexually active men who are reluctant to use highly effective contraception prior to the first dose of study therapy, throughout the study, and for at least 180 days after the last dose.Highly effective contraceptive measures include: a. Screening. Stable use of combined (estrogen and progestogen-containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with suppression of ovulation that has initiated at least two previous menstrual cycles; b. Intrauterine device (IUD); Intrauterine hormone-releasing system (IUS); c. Bilateral tubal ligation; d. Vasectomized partner; and / or e. Sexual abstinence. WOCBP is defined as a woman who has had one episode of menstruation and has not yet reached menopause or has been surgically sterilized, as follows: Menopause is defined as at least 12 consecutive months without any episodes of menstruation (not hormonally induced). Postmenopausal women must be amenorrhea for at least 12 months to be considered for childbearing potential. Pregnancy testing and contraception are not required for women with documented hysterectomy, bilateral oophorectomy, or tubal ligation. Sexual abstinence is considered highly effective only if defined as abstinence from heterosexual intercourse for the entire period of risk associated with the study treatment. Periodic abstinence (calendar, symptomatic temperature, postovulatory), withdrawal (abortive intercourse), spermicide only, and lactational amenorrhea method (LAM) are not acceptable methods of contraception. Female and male condoms should not be used together.

[0106] Study variables The primary endpoint of this study is DFS, defined as the time from randomization to first documented disease recurrence (local, regional and / or distant) or death from any cause. For patients without tumor recurrence or death, DFS will be censored at the date of last disease assessment. The estimated time frame for DFS assessment is up to approximately 54 months.

[0107] Secondary endpoints of this include:· Overall survival, defined as the time from randomization to the date of death. Patients who have not died will be censored at their last known date as alive. The estimated time frame for OS assessment is up to approximately 78 months;· Freedom from locoregional recurrence, defined as the time from randomization to the date of first locoregional recurrence (LRR). Patients who died without a preceding LRR will be censored at the date of death. For patients without an LRR or death, it will be censored at the last disease date of FFLRR assessment. The estimated time frame for FFLRR assessment is up to approximately 54 months;· Freedom from distant recurrence, defined as the time from randomization to the date of first distant recurrence (DR). Patients who died without a preceding DR will be censored at the date of death. For patients without a DR or death, FFDR will be censored at the date of last disease assessment. The estimated time frame for FFDR assessment is up to approximately 54 months;· Cumulative accrual of SPT for each patient from randomization to the occurrence of the first primary endpoint event or the end of the study. The estimated time frame for evaluation of cumulative accrual of SPT is up to approximately 54 months; safety as measured by incidence and severity of treatment-emergent adverse events (TEAEs), deaths, and laboratory abnormalities. The estimated time frame for safety evaluation is up to approximately 78 months.

[0108] Pharmacokinetic (PK) variables are REGN2810 concentrations at each time point. Samples for this study will be collected using a sparse sampling schedule (e.g., only one blood sample for drug concentration measurement will be collected at any one clinic visit).

[0109] Study design This study is a randomized, double-blind, placebo-controlled Phase 3 trial evaluating REGN2810 as an adjuvant treatment for patients with CSCC who have completed surgery and post-operative RT, configurations associated with high risk of recurrent disease. The study population will include patients who have completed surgery and post-operative RT for CSCC and who have at least one factor that places them at high risk for recurrence of CSCC. Patients will receive REGN2810 every 3 weeks (Q3W) for up to 48 weeks. Patients will be randomized 1:1 to N2810 350 mg versus placebo. The primary endpoint is disease-free survival (DFS). For patients who experience documented disease recurrence during the study, there is the option of subsequent REGN2810 therapy after the first recurrence. Figure 1 provides a flow diagram of the general overview of the study.

[0110] REGN2810 will be supplied as a liquid in a sterile, single-use vial. Each vial will contain REGN2810 at a concentration of 50 mg / mL. Placebo will be prepared using the same formulation used for REGN2810, without the addition of active. Placebo will be supplied as a liquid in a sterile, single-use vial. REGN2810 350 mg or placebo will be administered every 3 weeks in the outpatient setting as a 30-minute (± 10 minutes) IV infusion.

[0111] The study includes two parts. Part 1 (blinded) includes a screening period up to 28 days prior to randomization, a treatment period up to 48 weeks, and a follow-up period. During the treatment period (up to 48 weeks), patients undergo imaging assessment for tumor recurrence at the end of each 12-week cycle for the planned treatment period of approximately 1 year (48 weeks). Patients undergo post-treatment follow-up until disease recurrence or end of the study. Part 1 of the study supports the primary endpoint. Part 2 (unblinded) includes optional REGN2810 treatment for patients in the placebo arm who experience disease recurrence, and optional subsequent treatment with REGN2810 for patients in the REGN2810 arm who experience disease recurrence ≧3 months after completing 48 weeks of planned REGN2810 treatment.

[0112] Study Part 1 (blinded): Patients may begin screening once surgery and post-operative RT for CSCC is completed. Patients who meet the eligibility criteria and have high-risk configurations on surgical pathology of the resected tumor sample will be randomized 1:1 to receive REGN2810 350mg or placebo intravenously (IV). The first dose of REGN2810 or placebo will be administered within 5 days of randomization, which will occur between 2 and 6 weeks after completion of RT. REGN2810 or placebo will be administered IV every 3 weeks (Q3W) for up to 48 weeks or until unacceptable toxicity, disease recurrence, death, or withdrawal of consent. Patients will be evaluated in clinic prior to each REGN2810 or placebo treatment. Cycle length will be 12 weeks (Q3W schedule, 4 treatments per cycle). The follow-up period begins after patients discontinue treatment, either due to completion of the planned 48-week treatment period, or premature discontinuation of treatment for any other reason.

[0113] DFS is the primary endpoint of Part 1. For patients with high-risk CSCC, patterns of failure include locoregional recurrence, distant recurrence, locoregional and distant recurrence, or death (Porceddu et al., J Clin Oncol, 36(13):1275-83, 2018). DFS was chosen as the primary endpoint of the study because it encompasses all of these patterns of failure.

[0114] To evaluate the efficacy of adjuvant REGN2810 in patients at high risk of disease recurrence, REGN2810 will be compared to placebo in a 1:1 randomization. Since the current standard of care after RT is surveillance, placebo is a suitable control arm to compare with REGN2810 to allow for evaluation of the efficacy of REGN2810 as an adjuvant treatment after surgery and RT.

[0115] Study Part 2 (unblinded): Optional REGN2810 treatment for patients in the placebo arm who experience disease recurrence and optional subsequent treatment with REGN2810 for patients in the REGN2810 arm who experience disease recurrence ≥ 3 months after completing 48 weeks of planned REGN2810 treatment. Patients may be treated for up to 96 weeks.

[0116] Patients assigned to the placebo arm in Part 1 will be eligible to receive open-label subsequent REGN2810 therapy in Part 2 of the study if they meet the following criteria for demonstrating disease recurrence: (i) have documentation of disease recurrence; (ii) provide separate consent for subsequent REGN2810 therapy; (iii) have not discontinued placebo treatment due to unacceptable toxicity (if a patient discontinued study treatment due to unacceptable toxicity and was then found to be on placebo due to open labeling, such a patient would not have the opportunity to receive REGN2810 in Part 2 because such a patient would have met the criteria for permanent study treatment discontinuation); and (iv) continue to meet the study eligibility criteria (except for selected eligibility criteria) prior to the repeat screening procedure.

[0117] Patients assigned to the REGN2810 arm in Part 1 will also be eligible to receive open-label subsequent REGN2810 therapy in Part 2 of the study if they meet the following criteria for demonstrating disease recurrence: (i) documentation of disease recurrence at least 3 months (90 days ± 3 days) after completion of 48 weeks of planned REGN2810 treatment (even if one or more doses of the planned REGN2810 treatment are missed during the 48-week treatment period); (ii) provide separate consent for subsequent REGN2810 treatment in Part 2; (iii) prior REGN2810 was not discontinued due to unacceptable toxicity; and (iv) repeat screening procedures continue to meet the study eligibility criteria (except for selected eligibility criteria).

[0118] Patients eligible for subsequent REGN2810 therapy in Part 2 of the study may receive REGN2810 350 mg Q3W for up to 96 weeks (Part 2) or until disease progression, unacceptable toxicity, withdrawal of consent, death, or loss to follow-up.

[0119] The severity of AEs (including laboratory findings classified as AEs) will be graded using the NCI-CTCAE staging system (NCI-CTCAE v5). Adverse events not listed in NCI-CTCAE v5 will be graded according to the scale described above in Table 1.

[0120] Concomitant medication applications and procedures Prohibited Medication and Procedures: While participating in this study, patients may not receive any of the following from the time of informed consent through the end of the follow-up period, unless otherwise specified below: (a) standard or investigational agents for the treatment of tumors, other than REGN2810 and placebo; (b) agents that block the PD-1 / PD-L1 pathway (other than patients assigned to receive REGN2810 in this study); (c) radiation therapy; and (d) live vaccines for at least 3 months after the last dose of study drug.

[0121] Allowed medications and procedures: The following medications and procedures are allowed under the following conditions: (a) any medications required to treat AEs and / or irAEs, including systemic corticosteroids, (b) systemic corticosteroids for physiological replacement (even if >10 mg / day prednisone equivalent), (c) short-course corticosteroids for prevention or treatment of non-autoimmune conditions, (d) bisphosphonates and denosumab, (e) systemic corticosteroids in physiological replacement doses even if >10 mg / day prednisone equivalent, (f) oral contraceptives, hormone replacement therapy or other maintenance therapy may be continued, (g) acetaminophen in doses ≦2 g / day, (h) surgical removal of non-index lesions if clinically indicated, and (i) other medications and procedures may be allowed on an individual basis. As this is an adjuvant study, no surgery is planned. However, if surgery for any urgent medical problem is clinically indicated for an individual patient, this will be allowed.

[0122] Procedures and Evaluation Screening / baseline procedures included: coagulation tests, serum β-HCG assay, brain imaging (computed tomography [CT] or magnetic resonance imaging [MRI]), and screening for hepatitis B virus (HBV), hepatitis C virus (HCV), and human immunodeficiency virus (HIV).

[0123] Efficacy procedures include: radiological imaging evaluation for tumor recurrence. Computed tomography imaging of the chest, abdomen and pelvis is required at each imaging evaluation. Head and neck imaging is obtained for patients with resected HN lesions. Tumor biopsy is performed when pathologic (histologic or cytologic) evidence of recurrent disease or SPT can be obtained. Recurrent lesions are evaluated for patterns of failure, with recurrence defined as ≥ 1 lesion that can be categorized as local, regional or distant recurrence.

[0124] Safety procedures will include: a comparison of the safety and tolerability of REGN2810 versus placebo, as assessed by taking adverse events (AEs), physical examination (complete or limited), weight, 12-lead electrocardiogram (ECG), vital sign assessments, and clinical laboratory tests including hematology, blood chemistry, and urinalysis.

[0125] PK / Drug Concentrations: PK samples will be collected for assessment of serum concentrations of REGN2810.

[0126] result Administration of REGN2810 as an adjuvant treatment for CSCC patients who have completed surgery and postoperative RT and are at high risk of recurrence is expected to lead to a reduced risk of subsequent disease recurrence or to eliminate the occurrence of subsequent disease recurrence. Adjuvant REGN2810 treatment administered to high-risk CSCC patients after surgery and RT is also expected to improve disease control. EXAMPLES

[0127] Clinical trial of anti-PD-1 antibody administered as neoadjuvant treatment for stage II to IV (M0) cutaneous squamous cell carcinoma (CSCC) This is a phase 2, single-arm, open-label, multicenter study of patients with stage II to IV (M0) CSCC who are candidates for surgery but are at increased risk of recurrence and / or disfigurement or functional deficit.

[0128] An exemplary anti-PD-1 antibody used in this study is REGN2810 (also known as cemiplimab or H4H7798N as disclosed in US9987500), which is a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; a HCVR / LCVR amino acid sequence pair comprising SEQ ID NOs:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0129] Research purpose The primary objective of the study is to evaluate the efficacy of neoadjuvant treatment with REGN2810, as measured by pathologic complete response (pCR) rate per independent central pathology review.

[0130] Secondary objectives of the study included: (1) (a) major pathologic response (mPR) rate per independent central pathology review, (b) pCR and mPR rates per local pathology review, and (c) pre-surgery follow-up according to local assessment using RECIST 1.1. (1) to evaluate the efficacy of neoadjuvant REGN2810 on measures of disease response, including objective response rate (ORR); (2) to evaluate the efficacy of neoadjuvant REGN2810 on event-free survival (EFS), disease-free survival (DFS), and overall survival (OS); (3) to evaluate the safety profile of neoadjuvant REGN2810; (4) to evaluate changes in surgical plan (resection and reconstructive procedures) from the screening period to definitive surgery, according to both the investigator's review and a separate surgical expert review; and (5) to evaluate changes in post-operative management plan (radiation, chemoradiation, or observation), according to both the investigator's review and a separate surgical expert review, from the screening period to post-operative pathological review.

[0131] The exploratory objectives of this study include: (1) to explore baseline tumor markers for association with treatment response, peripheral and tumor measures related to the mechanism of action of REGN2810 and discovery of other potential predictive markers of efficacy or safety; (2) to describe patterns of failure (locoregional vs. distant) in patients who experience disease recurrence; (3) to evaluate the cost impact of changes in surgical plan during the screening period compared to the actual surgical procedure performed; (4) to evaluate the cost impact of changes in post-operative management plan during the screening period compared to actual post-operative management; (5) to evaluate the immunogenicity of REGN2810; and (6) to evaluate health-related quality of life in patients with CSCC receiving neoadjuvant REGN2810.

[0132] Research duration The first part of the study (pre-surgery treatment period) will last up to 12 weeks. The second part of the study (optional post-surgery treatment period) will last up to 48 weeks. The follow-up period will be up to 3 years. Patients will be followed until disease recurrence or end of the study (whichever occurs first).

[0133] Study population Approximately 76 patients will be enrolled. The target population will consist of adult patients with stage III to IV (M0) CSCC of the head / neck, extremities or trunk, with patients with stage II CSCC selected for surgery being recommended in routine practice.

[0134] Inclusion Criteria: Patients must meet the following criteria to be eligible for inclusion in the study: (1) at least 18 years of age; (2) stage II to IV (M0) CSCC for which surgery would be routinely recommended. For stage II patients, lesions must be ≥ 3 cm in longest diameter (Note: staging is defined according to the AJCC 8th edition for HN tumors (Amin MB, American Joint Committee on C, American Cancer S. AJCC cancer staging manual. 8th edition Springer International Publishing; 2017) and according to the UICC 9th edition for non-HN tumors (O'Sullivan B, Union for International Cancer C. UICC manual of clinical oncology. 9th edition John Wiley & Sons, Ltd; 2015). (3) at least one lesion measurable by RECIST 1.1; (4) Eastern Cooperative Oncology Group (ECOG) performance status of 0 or 1; (5) (a) hemoglobin > 9.0 g / dL; (b) absolute neutrophil count (ANC) > 1.5 × 10 9 / L;(c) Platelet count>75×10 9 / L; (d) adequate organ and bone marrow function, documented by serum creatinine <1.5 upper limit of normal (ULN) or estimated creatinine clearance (CrCl) >30 mL / min; (e) adequate liver function (total bilirubin <1.5 × upper limit of normal (ULN); both aspartate aminotransferase (AST) and alanine aminotransferase (ALT) <3 × ULN; alkaline phosphatase (ALP) <2.5 × U LN) (Note: For patients with Gilbert's syndrome, total bilirubin ≤ 3 x ULN. Gilbert's syndrome must be appropriately documented in the medical history); (6) willing and able to comply with clinic visits and study-related procedures; (7) willing and able to provide informed consent signed by the study patient or legally acceptable representative; and (8) able to understand and complete study-related questionnaires.

[0135] Exclusion Criteria: Patients who meet any of the following criteria will be excluded from the study: (1) solid malignancies within 5 years of the planned enrollment date, or hematologic malignancies (chronic lymphocytic leukemia [CLL] at any time) will be included (Note: Patients with nonmelanoma skin cancer undergoing potentially curative therapy, or cervical intraepithelial neoplasia, or prostate intraepithelial neoplasia with undetectable prostate-specific antigen, or any other tumor being treated will not be excluded if the patient is considered in complete remission for at least 2 years prior to enrollment and will not require further therapy for the duration of the study); (2) distant metastatic disease (M1), visceral and / or distant nodal; (3) previous radiation therapy for CSCC; (4) patients with a condition requiring corticosteroid therapy (>10 mg prednisone / day or equivalent) within 14 days of the first dose of study drug (Note: physiological replacement doses are permitted even if they are >10 mg prednisone / day or equivalent, as long as this is not administered for immunosuppressive purposes. Inhaled or topical steroids are not permitted if these are autologous. (5) Patients with active, known, or suspected autoimmune disease that has required systemic therapy within 5 years of the planned enrollment date (Note: Patients with vitiligo, type I diabetes mellitus, and endocrine disease (including hypothyroidism due to autoimmune thyroiditis) requiring only hormone replacement, resolved childhood asthma, or psoriasis not requiring systemic treatment will be permitted); (6) History of interstitial lung disease (e.g., idiopathic pulmonary fibrosis, organizing pneumonia) or active non-infectious interstitial pneumonia requiring immunosuppressive doses of glucocorticoids to aid in management; (7) Uncontrolled infection with human immunodeficiency virus (HIV), hepatitis B virus, or hepatitis C virus (HBV or HCV) infection; or diagnosis of immunodeficiency (Note: (a) Patients will be tested for HIV, HBV, and HCV at screening; (b) Controlled infection (undetectable viral load [HIV measured via polymerase chain reaction] either spontaneously or on a stable antiviral regimen) (c) Patients with known HIV infection who have a serum hepatitis B virus DNA measured via polymerase chain reaction below the limit of detection and who are receiving antiviral therapy for hepatitis B (HBsAg+) will be admitted. Patients with controlled infection must undergo regular monitoring for HBV DNA. Patients must remain on antiviral therapy for at least 6 months past the last dose of investigational study drug; (d) Patients who are hepatitis C virus antibody positive (HCV Ab+) and have controlled their infection (HCV undetectable by polymerase chain reaction, either spontaneously or in response to a previous successful course of anti-HCV therapy). (RNA) patients are permitted;(8) active tuberculosis;(9) myocardial infarction within 6 months of enrollment;(10) any medical comorbidity, physical examination findings or metabolic dysfunction, or laboratory abnormalities that, in the opinion of the investigator, make the patient unfit for participation in the clinical trial because of a high safety risk and / or the potential to affect interpretation of the results of the study;(11) documented allergic or acute hypersensitivity reaction due to antibody treatment;(12) previous treatment with anti-cancer systemic therapy within the past 3 years prior to the planned enrollment date;(13) any previous treatment with an anti-PD1 / PD-L1 agent;(14) participation in an investigational drug or device study within 4 weeks of enrollment;(15) women with a positive serum chorionic gonadotropin HCG pregnancy test at the screening / baseline visit. Breastfeeding women are also excluded;(16) highly effective avoidance of study medication prior to the first dose, throughout the study, and for at least 6 months after the last dose. Women of childbearing potential (defined below) and sexually active men who are reluctant to undergo fertility treatment. Highly effective contraceptive measures included: (a) stable use of a combined (estrogen- and progestogen-containing) hormonal contraceptive method (oral, intravaginal, transdermal) or a progestogen-only hormonal contraceptive method (oral, injectable, implantable) associated with suppression of ovulation that has begun at least two menstrual cycles prior to screening; (b) an intrauterine device (IUD); an intrauterine hormone-releasing system (IUS); (c) bilateral tubal ligation; (d) a vasectomized partner (provided that the male vasectomized partner was the only sexual partner of the woman of childbearing potential (WOCBP) study participant and that the vasectomized partner was the only sexual partner of the WOCBP ... (17) having received a live vaccine within 28 days of enrollment; (18) previous allogeneic or autologous stem cell transplant; (19) recipients of a solid organ transplant (other than a corneal transplant); (20) a diagnosis of squamous cell carcinoma of unknown (or occult) primary source; (21) patients who have been institutionalized pursuant to an order issued by either a judicial or administrative authority; and (22) members of the study team at the clinical site or his / her immediate family, unless previously approved by the Sponsor.

[0136] A fertile woman is defined as a woman who is capable of becoming pregnant after menarche until postmenopause (defined below), unless permanently infertile. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. Postmenopausal status is defined as the absence of menstruation for 12 months without another medical cause. Elevated follicle-stimulating hormone (FSH) levels in the postmenopausal range can be used to confirm postmenopausal status in women who do not use hormonal sterilization methods or hormone replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of a postmenopausal status. The above definition follows guidance from the Clinical Trial Facilitation Group (CTFG). Pregnancy testing and contraception are not required for women with documented hysterectomy or tubal ligation.

[0137] Sexual abstinence is considered highly effective only if it is defined as abstinence from heterosexual intercourse for the entire period of risk associated with the study drug. The reliability of sexual abstinence needs to be evaluated in relation to the duration of the clinical trial and the patient's preferred and usual lifestyle. Periodic abstinence (calendar, symptomatic temperature, postovulatory), withdrawal (abortive intercourse), spermicide only, and lactational amenorrhea method (LAM) are not acceptable contraceptive methods. Female and male condoms should not be used together.

[0138] Study variables Patients will be evaluated at surgery (12 weeks) for endpoints measuring pCR, mPR and ORR (Part 1 of the study). Event-free survival (EFS) and OS will be assessed from the first dose of neoadjuvant REGN2810 to completion of follow-up. Disease-free survival (DFS) will be assessed from surgery to completion of follow-up. Event-free survival (EFS), DFS and OS assessments will continue until all enrolled patients have completed follow-up, with a total duration of approximately 4 years and 3 months.

[0139] The primary endpoints of the study included: pCR rate assessed by independent central pathology review;

[0140] Secondary endpoints of the study included: (1) mPR rate assessed by independent central pathology review; (2) pCR and mPR rates assessed by local pathology review; (3) ORR prior to surgery as assessed by investigator using RECIST 1.1; (4) event-free survival (EFS); (5) disease-free survival (DFS); (6) overall survival (OS); and (7) rates of adverse events (AEs), serious adverse events (SAEs), deaths, and laboratory abnormalities. (8) changes in surgical plan during the screening period compared to actual surgery after neoadjuvant REGN2810; and (9) changes in post-operative management plan during the screening period compared to actual post-operative management.

[0141] Exploratory variables in this study included: (1) patterns of failure in patients with local, regional, or distant disease recurrence, measured by descriptive statistics; (2) estimated change in costs due to changes in surgical plan during the screening period compared with actual surgical procedure performed after neoadjuvant REGN2810, (3) estimated change in costs due to changes in postoperative management plan during the screening period compared with actual postoperative management; (4) incidence of ADAs in response to REGN2810; and (5) health-related quality of life, assessed using the EORTC QLQ-C30.

[0142] Pharmacokinetic (PK) variables include serum concentrations of REGN2810 over time.

[0143] Immunogenicity variables included anti-drug antibody (ADA) status, dose titer, and time point / visit.

[0144] Safety variables included: (1) vital signs; (2) physical examination results; (3) electrocardiogram (ECG) results; (4) clinical laboratory results; (5) adverse events (AEs); and (6) immune-related adverse events (irAEs).

[0145] Pathologic complete response (pCR) is defined as the absence of viable cancer cells in the surgical pathology sample.

[0146] Major pathological response (mPR) is defined as ≦10% viable cancer cells in surgical pathology samples in patients who do not achieve pCR.

[0147] Objective response rate (ORR) will be assessed by the investigator using RECIST 1.1 (Eur J Cancer 2009;45(2):228-47).

[0148] Event-free survival (EFS) is defined as the time from the first dose of neoadjuvant REGN2810 to any of the following events: disease progression precluding surgery, inability to undergo complete resection (R0 or R1), disease recurrence (local, regional or distant) for patients who undergo complete resection (R0 or R1), or death from any cause.

[0149] Disease-free survival (DFS) is defined as the time from surgery to first recurrence (local, regional or distant) or death from any cause for patients who are disease-free at the completion of surgery (R0 or R1 resection).

[0150] Overall survival (OS) is measured as the time from the first dose of neoadjuvant REGN2810 to death from any cause.

[0151] Study design This is a single-arm, open-label, multicenter, phase 2 study in patients with stage II to IV (M0) CSCC who are candidates for surgery but are at increased risk of recurrence and / or disfigurement or functional deficit.

[0152] This study has two parts:

[0153] Part 1 (neoadjuvant): Screening period up to 28 days, treatment period up to 12 weeks, and post-treatment surgery up to 12 weeks. Part 1 of the study supports the primary endpoint.

[0154] Part 2 (adjuvant): Optional post-surgery REGN2810 treatment (or radiation therapy or observation only at the investigator's discretion) for up to 48 weeks.

[0155] After the second part of the study, patients will be followed for up to three years. Figure 2 provides a flow diagram of the general overview of the study.

[0156] REGN2810 is supplied as a sterile liquid in single-use glass vials. Each vial contains a concentration of 50 mg / mL REGN2810. REGN2810 350 mg is administered as an IV infusion over 30 minutes (± 10 minutes) every 3 weeks (Q3W) for up to 12 weeks (up to 4 doses) prior to Part 1 surgery and (optionally) up to 48 weeks (up to 16 doses) after Part 2 surgery.

[0157] Study Part 1: Patients with stage II to IV (M0) CSCC with planned surgery and who meet the eligibility criteria will receive REGN2810 350 mg intravenously (IV) every 3 weeks (Q3W) for up to 12 weeks (up to 4 doses) or until unacceptable toxicity, disease progression, or withdrawal of consent. Patients will be evaluated in clinic prior to each dose, and patients will undergo tumor response imaging assessment prior to receiving the third dose of REGN2810 (day 43 ± 3) and prior to surgery (day 85). The window for surgery is from day 75 to day 100. If patients meet the criteria for discontinuing REGN2810 during the 12-week neoadjuvant period, the treating physician may convert patients to surgery earlier. Following surgical tumor resection, the primary endpoint (pCR rate) will be assessed by an independent central pathology review committee.

[0158] Study Part 2: Patients will have the option to receive adjuvant REGN2810 treatment (350 mg IV Q3W) following surgery for up to 48 weeks (up to 16 doses) or until unacceptable toxicity, disease recurrence or withdrawal of consent. The first dose of adjuvant treatment will occur 3 weeks (± 3 days) after the end of treatment Part 1 (EOT1). At the investigator's discretion, patients can instead receive adjuvant radiotherapy (no concurrent or subsequent REGN2810 treatment will be permitted) or enter an observation-only period. During Part 2 of the study, patients will be evaluated every 15 weeks in the clinic. Patients receiving adjuvant REGN2810 will undergo a full evaluation as described in the schedule of events, whereas patients not receiving adjuvant REGN2810 will only undergo imaging evaluation according to a parallel schedule.

[0159] Follow-up: Follow-up will begin after patients have completed part 1 and part 2 of the study without disease progression (pre-surgery) or disease recurrence (post-surgery). Patients will be evaluated in the clinic for up to 3 additional years.

[0160] Concomitant medication applications and procedures Prohibited Medication and Procedures: While participating in this study (not including survival follow-up), unless otherwise specified below, patients may not receive any of the following medications from the time of informed consent through the end of the follow-up period: (1) standard or investigational agents for the treatment of tumors (other than REGN281); and (2) live vaccines for at least 3 months after the last dose of study drug.

[0161] Allowed Medication and Procedures: The following medications and procedures will be allowed under the following conditions: (1) AEs and / or irAEs, including systemic corticosteroids (2) any medications required to treat; (3) systemic corticosteroids for physiological replacement (even if >10 mg / day prednisone equivalent); (4) short-course corticosteroids for prevention or treatment of non-autoimmune conditions; (5) oral contraceptives, hormone replacement therapy, or other maintenance therapy may be continued; (6) acetaminophen at a dose ≤2 g / day; (7) surgical excision of premalignant or basal cell carcinoma (BCC) lesions; and (8) other medications and procedures may be allowed on an individual basis by the investigator and in consultation with the sponsor. Radiation therapy (with concurrent anticancer cytotoxic chemotherapy and / or epidermal growth factor receptor-directed therapy) will be allowed in the adjuvant portion of the study at the investigator's discretion. Such patients will be followed for disease recurrence but will not receive further REGN2810 in the study.

[0162] Procedures and Evaluation Screening / baseline procedures included: screening for Hepatitis B virus (HBV), Hepatitis C virus (HCV) and human immunodeficiency virus (HIV), height measurement, and serum β-human chorionic gonadotropin (HCG) testing (for women of childbearing potential).

[0163] Efficacy procedures included: (1) evaluation of surgically resected tumors for pathologic response; (2) radiological imaging evaluation (adjuvant portion of the study and follow-up) for tumor response and disease recurrence (adjuvant portion of the study); (3) images of externally visible lesions supplemented with digital medical photography; (4) tumor biopsy for histologic or cytologic evidence of disease recurrence or evidence of a second primary tumor (SPT); and (5) evaluation of the pattern of failure for recurrent lesions. Recurrence is defined as ≧1 lesion that can be categorized as local, regional, or distant recurrence.

[0164] Safety procedures included: taking of adverse events (AEs), physical examination (complete or limited), weight assessment, recording of a 12-lead electrocardiogram (ECG), vital signs assessment, and clinical testing including hematology, blood chemistry, and urinalysis.

[0165] Laboratory procedures include: (1) blood chemistry: sodium, potassium, chloride, carbon dioxide (bicarbonate), calcium, glucose (fasting or non-fasting), albumin, protein (serum), creatinine, blood urea nitrogen (BUN), aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase, total bilirubin); hematology: hemoglobin, white blood cells, platelet count, neutrophils, lymphocytes; (3) urinalysis: pH, specific gravity, ketones, glucose, blood, spot urine protein; and (4) other laboratory tests: (a) HBV, HCV, HIV tests; (b) pregnancy test: serum β-HCG or urine β-HCG; (c) thyroid stimulating hormone (TSH) with reflexes T3 and free T4; and (d) coagulation, by measurement of INR and aPTT.

[0166] result For the phase 1 study, it is expected that administration of REGN2810 as neoadjuvant treatment prior to surgery will result in significant pathologic complete response (pCR) rates in patients with CSCC.

[0167] For the phase 2 study, administration of REGN2810 as an adjuvant treatment for CSCC patients who have completed surgery and post-operative RT and are at high risk of recurrence is expected to lead to a reduced risk of subsequent disease recurrence or to eliminate the occurrence of subsequent disease recurrence. Adjuvant REGN2810 treatment given to high-risk CSCC patients after surgery and RT is also expected to improve disease control. EXAMPLES

[0168] A clinical trial of anti-PD-1 antibodies administered in patients with metastatic or unresectable advanced cutaneous squamous cell carcinoma (CSCC) This study is a phase 2, non-randomized, five-arm, pivotal trial in patients with advanced CSCC.

[0169] An exemplary anti-PD-1 antibody used in this study is REGN2810 (also known as cemiplimab or H4H7798N as disclosed in US9987500), which is a fully human monoclonal anti-PD-1 antibody comprising a heavy chain comprising the amino acid sequence of SEQ ID NO:9 and a light chain comprising the amino acid sequence of SEQ ID NO:10; a HCVR / LCVR amino acid sequence pair comprising SEQ ID NOs:1 / 2; and heavy and light chain CDR sequences comprising SEQ ID NOs:3-8.

[0170] research group Group 1 consists of patients with metastatic (nodal or distant) CSCC treated with REGN2810 (3 mg / kg / dose IV, once every 2 weeks).

[0171] Group 2 consists of patients with unresectable locally advanced CSCC treated with REGN2810 (3 mg / kg / dose IV, once every 2 weeks).

[0172] Group 3 consists of patients with metastatic (nodal or distant) CSCC treated with REGN2810 (350 mg / dose IV once every 3 weeks).

[0173] Arm 4 consisted of patients with advanced CSCC [metastatic (nodal or distant) or unresectable locally advanced] treated with REGN2810 (600 mg / dose, IV, once every 4 weeks).

[0174] Group 5 consisted of patients with advanced CSCC (metastatic or locally advanced) treated with REGN2810 (438 mg / dose, SC) 3 weeks followed by REGN2810 (350 mg / dose, IV, once every 3 weeks).

[0175] An overview of disease stages for the five study groups is provided in Table 3.

[0176] [Table 1]

[0177] Research purpose The primary objectives of the study for groups 1-4 include: to estimate the clinical benefit (overall response rate by central review) of REGN2810 monotherapy for patients with metastatic (nodal or distant) CSCC or unresectable CSCC. The objectives of the study for group 5 include: (1) to evaluate the subcutaneous bioavailability of REGN2810 by measuring the concentration of REGN2810 in serum after subcutaneous (SC) administration, and (2) to evaluate the safety, tolerability, and PK of a single dose of SC REGN2810.

[0178] Secondary objectives of the study include: (1) to estimate the objective response rate (ORR) according to investigator review; (2) to estimate duration of response, progression-free survival (PFS), and overall survival (OS) by central and investigator review; (3) to estimate the complete response (CR) rate by central review; (4) to evaluate the safety and tolerability of REGN2810; (5) to evaluate the pharmacokinetics (PK) of REGN2810; (6) to evaluate the immunogenicity of REGN2810; and (7) to evaluate the impact of REGN2810 on quality of life using the European Organisation for Research and Treatment of Cancer Quality of Life Questionnaire Core 30 (EORTC QLQ-C30, see Young et al., European Journal of Cancer, 1999, 35(13):1773-82).

[0179] Secondary objectives of the study for group 4 further included: using EORTC criteria: 18 F-fluorodeoxyglucose positron emission tomography ( 18 To assess ORR according to F-FDG-PET.

[0180] Further study objectives for groups 2 and 4 include: (1) to evaluate the pharmacodynamic effects of REGN2810 in patients with CSCC treated with REGN2810 in tumor biopsies obtained at baseline, during treatment, and at progression; (2) to evaluate the efficacy and safety of tumor-infiltrating lymphocytes (TILs) (CD8 + T cells, CD4 + (3) expression levels (mRNA and / or protein) of programmed death-ligand 1 (PD-L1), glucocorticoid-inducible TNFR family related gene (GITR) and lymphocyte activation gene-3 (LAG-3) and possibly other checkpoint modulators; (4) mutations in known oncogenes and potential tumor neoantigens; and (5) tumor mutational burden.

[0181] Research duration The duration of the study includes a screening period (up to 4 weeks) for all five groups. Groups 1 and 2 will each receive treatment up to 96 weeks. Group 3 will receive treatment up to 54 weeks. Group 4 will receive treatment up to 48 weeks. Group 5 will receive treatment up to 54 weeks. All groups will be eligible for retreatment for up to 2 years.

[0182] All groups will be followed up for approximately 1.5 years.

[0183] Study population The study includes eligible patients with metastatic (nodal and / or distant) CSCC (groups 1 and 3) and unresectable locally advanced CSCC (group 2). Group 3 for metastatic CSCC will be opened only after enrollment in group 1 is completed. Groups 4 and 5 will enroll patients with advanced CSCC, a term that includes both metastatic (nodal or distant) CSCC and locally advanced CSCC.

[0184] Inclusion Criteria: Patients must meet the following criteria to be eligible for inclusion in the study: (1) Histologically confirmed diagnosis of invasive CSCC; (a) Regarding tumor primary site: Patients whose primary site of squamous cell carcinoma was the dry red lip (vermilion) are not eligible. Patients with tumors arising in the cutaneous hairy (non-glabrous) lip that extend onto the dry red lip (vermilion) may be eligible after contacting and receiving permission from the medical monitor. Patients whose primary site of squamous cell carcinoma was in the anogenital area (penis, scrotum, and perianal area) are not eligible. Patients whose primary site was in the nose are not eligible for inclusion in the study. Eligible only if the physician can unequivocally demonstrate that the primary site is skin and not nasal mucosa extending outward to the skin; and (b) regarding tumor histology: Patients with mixed histology (e.g., sarcomatoid, adenosquamous) will generally not be eligible. Patients with predominantly invasive CSCC mixed histology (with only minimal components of mixed histology) may be eligible after contacting and receiving clearance from the medical monitor; (2) at least one lesion measurable by study criteria. If previously irradiated lesions are followed as target lesions, progression should be confirmed by biopsy after radiation therapy. Previously irradiated lesions may be followed as non-target lesions if there is at least one other measurable target lesion; (a) For patients with metastatic (nodal or distant) CSCC (all patients in groups 1 and 3 and patients in groups 4 and 5 with metastatic CSCC): There must be at least one baseline measurable lesion with a maximum diameter of ≥10 mm (1.5 cm for lymph nodes) according to Response Evaluation Criteria in Solid Tumors (RECIST) 1.1 criteria. For patients with metastatic disease that does not meet target lesion criteria according to RECIST 1.1 (e.g., bone-only lesions, perineural disease; see Eisenhauer et al., Eur J Cancer 2009;45:228-247). For externally visible CSCC target lesions, two-dimensional measurements by digital medical photography may be used (at baseline, both perpendicular diameters must be ≥10 mm). Patients will then be enrolled with a plan to measure externally visible target lesions by photography with two-dimensional measurements; metastatic lesions not measurable by RECIST 1.1 criteria will be followed with scans as non-target lesions; (b) For patients with unresectable locally advanced CSCC (all patients in group 2 and patients in groups 4 and 5 with locally advanced CSCC): If digital medical photography is to follow, there must be at least one measurable baseline lesion with both the longest diameter (LD) and perpendicular diameter ≥ 10 mm.Non-measurable disease for group 2 is defined as either unidimensionally measurable disease, tumors with poorly defined margins, or lesions with a maximum perpendicular diameter less than 10 mm. Patients without measurable disease at baseline were not eligible for the study; (3) Eastern Cooperative Oncology Group (ECOG) performance status ≤ 1 (ECOG PS 1 definition: physically strenuous activity is limited, but able to walk and perform work of a light or sedentary nature, e.g., light housework, clerical work. ECOG Patients with PS>1 are ineligible; (4) age ≥18 years; (5) liver function; (a) total bilirubin ≤1.5×upper limit of normal (ULN; if liver metastases ≤3×ULN). Patients with Gilbert's disease and total bilirubin up to 3×ULN may be eligible after contacting and receiving permission from the medical monitor; (b) transaminases ≤3×ULN (or ≤5.0×ULN if liver metastases); and (c) alkaline phosphatase (ALP) ≤2.5×ULN (or ≤5.0×ULN if liver or bone metastases). Patients with liver metastases and who are in group 1, group 3, group 4, or group 5 are eligible. For patients wishing to be enrolled in group 5: if transaminase levels (AST and / or ALT) are >3× but ≦5×ULN, then total bilirubin must be ≦1.5×ULN. If total bilirubin is >1.5× but ≦3×ULN, then both transaminases (AST and ALT) must be ≦3×ULN; (6) Renal function: serum creatinine ≦1.5×ULN or estimated creatinine clearance (CrCl) >30 mL / min; (7) Bone marrow function: (a) hemoglobin ≧9.0 g / dL; (b) absolute neutrophil count (ANC) ≧1.5×10. 9 / L; and (c) platelet count ≥ 75 × 10 9 / L;(8) Ability to provide signed informed consent;(9) Ability and willingness to comply with scheduled clinic visits, treatment plans, laboratory tests, and other study-related procedures;(10) Projected life expectancy >12 weeks;(11) Patients with locally advanced CSCC: All patients in Group 2 (and patients in Groups 4 and 5 with locally advanced CSCC): In the opinion of the Mohs dermatological, head and neck, or plastic surgeon, surgery should be considered contraindicated. A copy of the surgeon's consultation note from a clinical visit within 60 days of enrollment must be submitted. Acceptable contraindications in the surgeon's record include:(a) Two or more surgical procedures (b) CSCC that recurs in the same location after surgery and for which curative resection is deemed unlikely; (c) CSCC in a difficult anatomical location where surgery may result in severe disfigurement or functional impairment (e.g., removal of all or part of facial structures, e.g., nose, ears, or eyes; or the need for limb amputation); and (d) other conditions that are considered to contraindicate surgery should be discussed with the medical monitor before enrolling the patient; (12) Patients with locally advanced CSCC (all patients in group 2 and patients in groups 4 and 5 with locally advanced CSCC): Patients should be considered as not suitable for radiation therapy. In particular, patients must meet at least one of the following criteria: (a) Patients who have previously received radiation therapy for CSCC such that, according to the radiation oncologist, further radiation therapy would exceed the threshold of the acceptable cumulative dose. Copies of the radiation oncologist's consultation notes from a clinical visit within 60 days of enrollment must be submitted; (b) the radiation oncologist's judgment that such tumors are unlikely to respond to therapy. A copy of the radiation oncologist consultation notes from a clinical visit within 60 days of enrollment must be submitted; and (c) Investigator's individualized interest: Investigator's clinical notes indicating that a risk assessment was performed by a multidisciplinary team (consisting, at a minimum, of a radiation oncologist and a medical oncologist with expertise in cutaneous malignancies, either a radiation oncologist or a medical oncologist with expertise in cutaneous malignancies, or a cutaneous oncologist, or a head and neck surgeon) within 60 days prior to proposed study enrollment and that radiation therapy was deemed contraindicated. Acceptable contraindications to radiation therapy in the investigator's notes for patients who have not received any prior radiation include: (i) CSCC in an anatomically challenging location (e.g., a neck tumor where radiation therapy would result in the potential need for a percutaneous gastrostomy tube) where, in the opinion of the multidisciplinary team, radiation therapy would be associated with an unacceptable risk of toxicity in conjunction with the patient's overall medical condition.A copy of the investigator's consultation record demonstrating a multidisciplinary evaluation must be submitted; and (ii) other conditions considered to contraindicate radiation therapy should be discussed with the medical monitor before enrolling the patient; (13) all patients in either group must agree to provide archived or freshly obtained tumor material (either formalin-fixed, paraffin-embedded [FFPE] blocks or 10 unstained or stained slides) for central pathology review for confirmation of the diagnosis of CSCC. This material must be verified as received by a central laboratory prior to enrollment;(14) For Group 2 (patients with locally advanced CSCC) and Group 4 (patients with locally advanced CSCC and metastatic CSCC) only: Patients must consent to undergo biopsy of CSCC lesions at baseline, i.e., day 29 (± 3 business days) of Cycle 1, at the time of tumor progression, and at other time points that, in the opinion of the investigator, may be clinically indicated;(15) For all patients in Group 2, and for patients in Groups 4 and 5 with locally advanced CSCC only: Investigator's record stating that the natural history of the patient's progressive CSCC will be potentially life-threatening within 3 years with currently available management options outside of the clinical trial.

[0185] Exclusion Criteria: Patients who meet any of the following criteria will be excluded from the study: (1) Ongoing or recent (within 5 years) evidence of significant autoimmune disease that required treatment with systemic immunosuppressive therapy that may indicate risk of immune-related adverse events (irAEs). The following are not excluded: vitiligo that required only hormone replacement, resolved childhood asthma, type 1 diabetes, residual hypothyroidism, or psoriasis that did not require systemic treatment; (2) prior treatment with an agent that blocks the PD-1 / PD-L1 pathway; (3) prior treatment with another immune modulating agent that was (a) within less than 4 weeks (28 days) prior to the first dose of REGN2810, or (b) within 90 days prior to the first dose of REGN2810, associated with an immune-mediated adverse event that was ≥ Grade 1, or (c) associated with toxicity that led to discontinuation of the immune modulating agent. Examples of immune modulating agents include therapeutic anti-cancer vaccines, cytokine therapy (other than G-CSF and erythropoietin), or cytotoxic agents. Agents targeting cytotoxic T-lymphocyte antigen 4 (CTLA-4), 4-1BB (CD137), PI 3-K-δ, or OX-40; (4) untreated brain metastases that can be considered active. (Note: Patients with brain involvement of CSCC due to direct extension of invasive tumor rather than metastasis may be permitted to enroll after medical monitor discussion and authorization if they do not require more than 10 mg of prednisone daily). Patients with previously treated brain metastases may participate provided that their disease is stable (no evidence of progression for at least 6 weeks on imaging obtained during the screening period), there is no evidence of new or expanding brain metastases, and within 4 weeks of the first dose of REGN2810, the patient does not require any immunosuppressive doses of systemic corticosteroids for management of brain metastases; (5) immunosuppressive corticosteroid administration (>10 mg prednisone daily or equivalent) within 4 weeks prior to the first dose of REGN2810. Note: Patients requiring a short course of steroids (e.g., as prophylaxis for imaging due to hypersensitivity to contrast agents) are not excluded;(6) active infection requiring therapy, e.g., human immunodeficiency virus infection, or active infection with Hepatitis B virus (HBV) or Hepatitis C virus (HCV);(7) history of non-infectious interstitial pneumonia within the past 5 years. If the interstitial pneumonia was purely infectious in etiology, patients may be permitted to enroll in the protocol after discussion with the medical monitor;(8) hypercalcemia of grade ≥ 3 at the time of enrollment;(9) any systemic anti-cancer treatment (chemotherapy, targeted systemic therapy, photodynamic therapy), investigational or standard of care (patients receiving bisphosphonates or denosumab are not excluded) within 30 days of the first dose of REGN2810 or planned to be administered during the study period, radiation therapy within 14 days of the first dose of REGN2810 or planned to be administered during the study period.NOTE: For patients with multiple CSCCs at baseline that are not designated as target lesions by the investigator, treatment of these non-target CSCCs with surgery may be permitted but should be discussed with the medical monitor prior to any surgical procedure; (10) History of documented allergic or acute hypersensitivity reactions due to antibody treatment; (11) Patients with allergies or hypersensitivities to REGN2810 or to any of the excipients should be excluded. In particular, patients with allergies or hypersensitivity to doxycycline or tetracycline due to the presence of trace components of REGN2810 will be excluded; (12) breast-fed; (13) positive serum pregnancy test (false positive pregnancy tests will not be excluded if documented by serial measurements and negative ultrasound, and upon contact with and authorization from the medical monitor); (14) tumors with negligible risk of metastasis or death, e.g., adequately treated BCC of the skin, carcinoma in situ of the cervix, or ductal carcinoma in situ of the breast, or low-risk early stage prostate adenocarcinoma (T1-T2aN0M0 and Gleason score ≤ 6 and PSA ≤ 10 ng / mL) where the management plan is active surveillance, or prostate adenocarcinoma with only biochemical recurrence with a reported PSA doubling time > 12 months where the management plan is active surveillance (D'Amico 2005, Pham (13) concurrent malignancies other than CSCC and / or a history of malignancies other than CSCC within 3 years of the date of the first planned dose of REGN2810, except for those who have not completed the study (2016). Patients with hematological malignancies (e.g., chronic lymphocytic leukemia, CLL) will be excluded; (14) any acute or chronic psychiatric problem that, in the opinion of the investigator, makes the patient ineligible for participation; (15) ongoing sexual activity in men or women of childbearing potential with reluctance to practice highly effective contraception prior to the first dose, during the study, and for at least 6 months after the last dose of study drug.Highly effective contraceptive measures include: stable use of combined (estrogen- and progestogen-containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with suppression of ovulation that has begun at least two menstrual cycles prior to screening; intrauterine device; intrauterine hormone-releasing system; bilateral tubal ligation; vasectomized partner and sexual abstinence. Contraception is not required for men with a documented vasectomy. Postmenopausal women must be amenorrhea for at least 12 months to be considered for childbearing potential. Pregnancy testing and contraception are not required unless a hysterectomy has been performed. is not required for women with a documented tubal ligation;(17) patients with a history of solid organ transplant (patients with a previous corneal transplant may be permitted to enroll after discussion with and permission from the medical monitor);(18) previous treatment with a BRAF inhibitor;(19) any medical comorbidity, physical exam findings or metabolic dysfunction, or laboratory abnormalities that, in the investigator's opinion, make the patient unsuitable for participation in the clinical trial because of a high safety risk and / or the potential to affect interpretation of the study's results;(20) inability to undergo any contrast-enhanced radiation response assessment. Note regarding imaging options: A patient who cannot undergo CT with iodine contrast (e.g., because of contrast allergy) will not be excluded if his / her disease can be measured by MRI with gadolinium. A patient who cannot undergo MRI with gadolinium will not be excluded if his / her disease can be measured by CT scan with contrast. Note for Groups 2, 4, and 5 (locally advanced CSCC patients only): In selected cases, locally advanced CSCC patients (Groups 2, 4, or 5) who cannot undergo any contrast-enhanced radiographic imaging (neither CT with iodine contrast nor MRI with gadolinium) may be eligible after contacting and receiving permission from the medical monitor if their disease can be comprehensively assessed with digital medical photography.

[0186] Study variables The primary efficacy endpoints of the study include: ORR by central review during 12 treatment cycles (arms 1 and 2) or 6 treatment cycles (arms 3 and 4). Overall response rate will be assessed separately for patients with metastatic CSCC or patients with unresectable locally advanced CSCC.

[0187] For patients with metastatic disease (groups 1, 3, and 4), ORR will be determined using RECIST version 1.1 (Eisenhauer, EA, et al., Eur J Cancer 2009;45:228-247). In patients with all response assessments performed on radiological scans according to RECIST 1.1, the decision of an independent radiological response assessment committee will serve as the central response assessment. When not all metastatic disease is measurable by RECIST (as may occur, for example, in patients with bone-only metastases), clinical or composite response criteria may be used in patients with externally visible target disease.

[0188] For patients with unresectable locally advanced disease (groups 2 and 4), ORR will be determined using clinical response criteria for externally visible tumors requiring bidimensional measurements according to World Health Organization (WHO) criteria. ORR will be determined using composite response criteria for patients with both target lesions measurable by clinical response criteria and by RECIST 1.1. In patients achieving CR, tumor biopsy will be used in the final determination of complete vs PR.

[0189] Patients deemed not evaluable (NE) by RECIST version 1.1 (groups 1, 3, and 4) or unevaluable by clinical or composite response criteria (group 2) will be considered as not achieving a partial response (PR) / complete response (CR) for ORR.

[0190] Secondary endpoints of the study include: (1) ORR for Groups 1 to 5 as assessed by the investigator: (a) For patients in Groups 1, 3, 4 (if metastatic) and Group 5 (if metastatic), where all response assessments are performed on radiological scans according to RECIST 1.1, the term "composite response assessment" is not applicable. The investigator's response assessment for such patients will be the RECIST 1.1 assessment; (b) ORR for Groups 1 to 5 as assessed by the investigator; For patients in groups 2, 4 (unresectable locally advanced) and 5 (unresectable locally advanced) where all response assessments are performed on photographs according to the Clinical Response Criteria for Externally Visible Tumors, the term "composite response assessment" is not applicable. The investigator's response assessment for such patients will follow the Clinical Response Criteria for Externally Visible Tumors; (c) for patients where target lesion response assessments are performed on both scans (according to RECIST 1.1) and photographs (according to the Clinical Response Criteria for Externally Visible Tumors), the investigator's response assessment will follow the composite response criteria; (d) patients in group 4 will undergo PET-CT scans at baseline and at 6-month intervals. Investigator response assessment will be according to the EORTC criteria, Young et al., European Journal of Cancer, 1999, Vol. 35, No. 13, 1773-82; (2) duration of response; (3) PFS; (4) OS; (5) CR rate; (6) change in EORTC QLQ-C30 patient-reported outcome score; (7) AEs; (8) serum concentration of REGN2810; and (9) anti-REGN2810 antibodies.

[0191] Exploratory outcome measures for this study include: (1) fold change in mRNA expression of genes expressed in tumor tissue; (2) TIL (CD8 + T cells, CD4 +(3) percent changes in the numbers of T cells, regulatory T cells and other tissue-tolerant subtypes (e.g., B cells, bone marrow-derived cells, NK cells, etc.), and descriptive changes in the distribution of TILs with respect to tumor tissue and stroma; (4) percent changes in the expression levels (mRNA and / or protein) of PD-L1, GITR and LAG-3 and possibly other checkpoint modulators; (5) changes in the number and type of genetic mutations in known oncogenes and potential tumor neoantigens; and (6) changes in the tumor mutational burden.

[0192] Serum concentrations of REGN2810 will be assessed at multiple time points throughout the study treatment and follow-up period, and PK will be analyzed. Pharmacokinetic variables may include, but are not limited to, the following: (1) C eoi - Concentration at the end of infusion (IV); (2) C max - Peak concentration (SC); (3) C TROUGH -Pre-injection concentration; (4)t eoi - Time of end of injection; (5) t max - Time to peak concentration (SC); (6) AUC 3w - area under the plasma concentration-time curve after the first SC or IV dose; and (7) absolute bioavailability after F-SC administration.

[0193] The impact of immunogenicity of REGN2810 will be evaluated. Anti-drug antibody (ADA) variables include ADA status and titers: (1) treatment emergent - defined as any positive post-dose ADA assay response when baseline results are negative; (2) treatment boosted - defined as any post-dose ADA response that is at least 9-fold compared to baseline titer levels; (3) potency (potency category); (4) low (titer < 1,000); (5) intermediate (1,000 < titer < 10,000); high (titer > 10,000). The association between immunogenicity and PK of REGN2810 can be evaluated accordingly.

[0194] Study design Groups 1 and 2: After completion of a screening period of up to 28 days, each patient in Groups 1 and 2 will receive up to twelve 56-day (8-week) treatment cycles for up to 96 weeks total treatment. Each patient will receive REGN2810 3 mg / kg IV on days 1, 15±3, 29±3, and 43±3 during each treatment cycle. Tumor evaluations will be performed at the end of each treatment cycle. Extensive safety evaluations will be performed on day 1 of each cycle; routine safety evaluations will be performed at each REGN2810 dosing visit.

[0195] Arm 3: This cohort will enroll patients with metastatic CSCC. Arm 3 will begin enrollment only after enrollment in Arm 1 is complete. The dose regimen is 350 mg IV every 3 weeks for up to 54 weeks.

[0196] Arm 4: This cohort will enroll patients with advanced CSCC (metastatic [nodal or distant] or locally advanced). Arm 4 will begin enrollment only after enrollment in Arms 1 through 3 is complete. The dose regimen is 600 mg IV every 4 weeks for up to 48 weeks.

[0197] Group 5: This cohort will enroll patients with advanced CSCC. The regimen will be 438 mg SC, one dose, followed by 350 mg IV every 3 weeks for a total of 54 weeks of treatment. The first 3 patients in group 5 will receive doses divided into 3 days to monitor for injection site reactions (ISRs). If the first 3 patients are observed to be free of severe ISRs, the remaining cohorts may enroll without restrictions on enrollment dates (i.e., more than one patient may begin treatment on the same day). If one or more severe ISRs are observed, enrollment will be temporarily halted. In that situation, continued enrollment in group 5 may be permitted following review of all relevant data and agreement between the Medical Monitor and the designated Risk Management Lead in the Pharmacovigilance & Risk Management Department. Consultation may also be sought from the investigators involved in the care of these patients.

[0198] Patients will continue on REGN2810 until the planned duration of therapy is completed (96 weeks for Arms 1 and 2, 54 weeks for Arms 3 and 5, and 48 weeks for Arm 4) or until disease progression, unacceptable toxicity, or withdrawal of consent. (Patients in Arms 1 or 2 who experience a CR have the option to discontinue treatment after 48 weeks).

[0199] The dosing regimens for groups 1 to 5 are summarized in Tables 4 to 7.

[0200] Dose levels may be reduced according to Table 8.

[0201] Groups 1 to 5: Patients who do not experience progressive disease (PD) upon completion of the planned treatment period will enter a follow-up period of approximately 6 months. After completion of this follow-up period, patients will then be evaluated every 4 months and enter an extended follow-up period of approximately 1 year (Table 9).

[0202] [Table 2]

[0203] [Table 3]

[0204] [Table 4]

[0205] [Table 5]

[0206] [Table 6]

[0207] REGN2810 will be administered in an outpatient setting as an IV infusion of approximately 30 minutes (± 10 minutes). Longer infusion durations will be permitted if interruption is required. Group 1 and Group 2 patient dosing will depend on individual body weight. The dose of REGN2810 should be adjusted each cycle for a ≥ 10% body weight change. Dose adjustments for < 10% body weight change will be at the investigator's discretion.

[0208] REGN2810 is supplied as a liquid in sterile, single-use vials. Each vial of REGN2810 contains a concentration of 25 or 50 mg / mL (for IV infusion) or 175 mg / mL (for SC injection).

[0209] Procedures and Evaluation Screening procedures included: informed consent, genomics substudy informed consent (optional), inclusion / exclusion, medical / oncology history, demographics, complete physical exam and ECOG PS, 12-Lead ECG, vital signs and weight, brain MRI, viral serology, coagulation, urinalysis, laboratory tests (hematology, blood chemistry, serum HCG ≦72 hours predose, urinalysis, HBV, HCV, HIV), archived tissue for histological confirmation of CSCC, tumor biopsy for group 2, CT / MRI and / or digital photography, and treatment assignment.

[0210] Efficacy procedures include: imaging (computed tomography [CT] or magnetic resonance imaging [MRI]) and digital medical photography (for externally visible lesions) to measure tumor burden and to characterize the efficacy profile of the study treatment using response criteria. Further efficacy procedures include tumor measurements and tumor biopsies.

[0211] Safety procedures will include: assessment of vital signs, physical examination, electrocardiogram (ECG), immune safety assays, immunoglobulin levels (IgG, IgM, IgE), adverse event (AE) monitoring, and laboratory tests (including pregnancy testing for women of childbearing potential).

[0212] Immune safety assays include measurements of rheumatoid factor (RF), TSH, C-reactive protein (CRP) and antinuclear antibody (ANA) titers and patterns. If a 4-fold or greater increase from baseline in RF or ANA or abnormal levels of TSH or CRP are observed during the course of the study, the following tests may also be performed: anti-DNA antibodies, anti-Sjogren's syndrome antigen (SSA) antibodies (Ro), anti-Sjogren's syndrome B antigen (SSB) antibodies (La), anti-thyroglobulin antibodies, anti-LKM antibodies, antiphospholipid antibodies, anti-islet cell antibodies, anti-neutrophil cytoplasmic antibodies, C3, C4, CH50.

[0213] Laboratory procedures include: hematology, blood chemistry, pregnancy test (females only) and urinalysis.

[0214] PK / Drug Concentrations: PK samples will be collected for assessment of serum concentrations of REGN2810.

[0215] Anti-drug antibody measurements: Serum samples will be collected for assessment of immunogenicity to REGN2810.

[0216] Genomics substudy measures include: sequencing or single nucleotide polymorphism studies of candidate genes and surrounding genomic regions, whole exome sequencing, whole genome sequencing, and DNA copy number variation.

[0217] Concomitant medication applications and procedures Prohibited Medication and Procedures. While participating in this study, patients may not receive any standard or investigational agents for the treatment of the tumor other than REGN2810 as monotherapy. Once the patient has completed 24 weeks of study treatment, local palliative therapy (e.g., radiation) for local control of the tumor will be permitted after contact with the sponsor. Any other medications considered necessary for the patient's well-being and not expected to interfere with the evaluation of the study drug may be given at the discretion of the investigator.

[0218] Patients using systemic corticosteroids in immunosuppressive doses (>10 mg prednisone or equivalent per day) other than for the purpose of corticosteroid replacement will not be eligible for the study. Patients will not be receiving systemic corticosteroids, e.g., hydrocortisone, prednisone, prednisolone (Solumedrol®) or dexamethasone, at any time throughout the study, except in the case of a life-threatening emergency and / or to treat an irAE. It is recommended not to receive samethasone (Decadron®). Physiologic replacement doses of systemic corticosteroids are permitted, even if >10 mg / day prednisone equivalent. Short courses of corticosteroids for prophylaxis (e.g., contrast dye allergy) or for treatment of non-autoimmune conditions (e.g., delayed hypersensitivity reactions triggered by contact allergens) are permitted. Bisphosphonates and denosumab are not prohibited.

[0219] Surgery: For patients with locally advanced target lesions considered unresectable at baseline but subsequently deemed resectable during the course of the study due to tumor response to REGN2810, surgery with curative intent may be permitted but should be discussed with the medical monitor prior to any surgical procedure. (This statement does not apply to patients with urgent life-threatening situations requiring immediate surgery.) Patients with inoperable CSCC at baseline who become operable with clear margins will be considered to have experienced a PR.

[0220] If, during the course of the study, a patient develops a new cutaneous lesion suspected to be a nonmelanoma skin cancer other than CSCC (e.g., BCC), removal of the lesion and continuation of treatment under study may be permitted after discussion with the medical monitor.

[0221] Radiotherapy: Radiotherapy is not part of the study regimen. Patients for whom radiotherapy is planned will not be eligible. If, during the course of the study, a patient develops symptomatic disease for which palliative radiotherapy is deemed appropriate by the investigator, this will be considered PD and the patient will generally be removed from the study. Palliative radiotherapy may be permitted in certain circumstances for patients who have been on study for at least 24 weeks. Such cases should be discussed with the medical monitor prior to any radiotherapy if the investigator believes that resumption of REGN2810 after radiation is in the patient's best interest. Patients will be considered to have experienced disease progression if radiotherapy is initiated, but will be followed for OS.

[0222] WHO criteria for externally visible tumors requiring 2D measurements According to the WHO criteria, the criteria for clinical response of externally visible tumors requiring two-dimensional measurements are as follows:

[0223] Complete Response of Externally Visible Disease (vCR): All target and non-target lesions that are no longer visible, maintained for at least 4 weeks. Documentation of a vCR requires confirmation by biopsy of the site of the externally visible target lesion, with histologic confirmation of the absence of residual malignant disease, according to central pathology review (Appendix 5). In the absence of such histologic confirmation, the patient cannot be considered to have experienced a vCR, and the best response would be a partial response.

[0224] Partial response (vPR) of externally visible disease: A reduction of 50% or more (WHO criteria) in the sum of the products of the longest perpendicular dimensions of target lesions, sustained for at least 4 weeks.

[0225] Stable visible disease (vSD): Criteria for vCR, vPR, or progressive disease are not met.

[0226] Visible disease progression (vPD): An increase of ≥ 25% (WHO criteria) in the sum of the products of the longest perpendicular dimensions of target lesions. In rare cases, unsuspected progression of non-target lesions can be recognized as vPD.

[0227] result It is expected that administration of REGN2810 will lead to enhanced tumor regression and improved disease management.Furthermore, it is expected that administration of REGN2810 will lead to increased safety in patients with CSCC, with reduced or no incidence of adverse events and / or toxicity.For patients without metastatic CSCC, it is expected to reduce the likelihood of metastatic CSCC.

[0228] One hundred and ninety-three patients (pts) were enrolled (arm 1, n=59; arm 2, n=78; arm 3, n=56). 128 pts had not received prior anticancer systemic therapy and 65 pts had been previously treated. As of October 11, 2019 (data cutoff), the median duration of follow-up was 15.7 months (range: 0.6-36.1) among all pts; 18.5 months (range: 1.1-36.1) for arm 1, 15.5 months (range: 0.8-35.0) for arm 2, and 17.3 months (range: 0.6-26.3) for arm 3. ORR per INV was 54.4% (95% CI: 47.1-61.6) for all pts; for arm 1; 50.8% (95% CI: 37.5-64.1), for arm 2; 56.4% (95% CI: 44.7-67.6), and for arm 3; 55.4% (95% CI: 41.5-68.7). ORR per INV was 57.8% (95% CI: 48.8-66.5) among treatment-naive pts and 47.7% (95% CI: 35.1-60.5) among previously treated pts. Median duration of response (DOR) was not reached (observed DOR range: 1.8-34.2 months). In responding pts, estimated DOR at 24 months was 76.0% (95% CI: 64.1-84.4). Median OS was not reached. Estimated OS at 24 months was 73.3% (95% CI: 66.1-79.2). The most common treatment-emergent adverse events (TEAEs) of any grade were fatigue (34.7%), diarrhea (27.5%), and nausea (23.8%). The most common grade ≥3 TEAEs were hypertension (4.7%), anemia, and cellulitis (4.1% each).

[0229] For patients with advanced CSCC, cemiplimab achieved DOR and survival that was superior to that reported with other agents.

Claims

1. 1. A method of treating or inhibiting tumor growth comprising: (a) selecting patients with skin cancer who have completed surgery and / or radiation therapy to treat the skin cancer; and (b) subsequently administering to the patient an adjuvant treatment comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. The method comprising:

2. 10. The method of claim 1, wherein the patient has completed surgery and optionally post-operative radiation therapy.

3. 3. The method of claim 1 or 2, wherein the skin cancer is cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma or melanoma.

4. The method of any one of claims 1 to 3, wherein the skin cancer is CSCC.

5. The method of any one of claims 1 to 4, wherein the patient is at high risk of recurrence of CSCC or has suffered at least one incident of recurrence.

6. 6. The method of claim 5, wherein the patient has at least one of the following high-risk features: (a) nodular disease with extracapsular extension and at least one nodule >20 mm; (b) in-transit transfer (ITM); (c) T4 lesion; (d) perineural invasion (PNI); and (e) recurrent CSCC plus at least one of the following additional configurations: (i) ≥N2b disease associated with recurrent lesions; (ii) Nominal ≧T3; and (iii) Recurrent lesions ≥ 20 mm in diameter.

7. The method of any one of claims 1 to 6, wherein the therapeutically effective amount comprises 5 mg to 500 mg of the PD-1 inhibitor.

8. The method of any one of claims 1 to 6, wherein the therapeutically effective amount comprises 350 mg of the PD-1 inhibitor.

9. 9. The method of any one of claims 1 to 8, wherein the PD-1 inhibitor is administered in one or more doses, each dose being administered 2 to 12 weeks, preferably 3 weeks, after the immediately preceding dose.

10. 10. The method of claim 9, wherein each dose contains 5 mg to 500 mg, preferably 350 mg, of the PD-1 inhibitor.

11. The method of any one of claims 1 to 10, wherein the PD-1 inhibitor is administered intravenously.

12. The method according to any one of claims 1 to 11, wherein step (b) is performed 2 to 6 weeks after completion of radiotherapy.

13. 13. The method of any one of claims 1 to 12, wherein administration of a PD-1 inhibitor leads to a reduced risk of subsequent skin cancer recurrence or eliminates the occurrence of subsequent skin cancer recurrence.

14. 13. The method of any one of claims 1-12, wherein administration of the PD-1 inhibitor leads to at least about a 10% lower incidence of subsequent skin cancer recurrence as compared to patients following completion of surgery and radiation therapy without an adjuvant skin cancer treatment.

15. The method of any one of claims 1 to 14, further comprising administering an additional therapeutic agent selected from a chemotherapeutic agent, a corticosteroid, an anti-inflammatory agent and / or combinations thereof.

16. The method of any one of claims 1 to 15, wherein the PD-1 inhibitor is selected from the group consisting of an anti-PD-1 antibody or antigen-binding fragment thereof, an anti-PD-L1 antibody or antigen-binding fragment thereof, and an anti-PD-L2 antibody or antigen-binding fragment thereof.

17. The method of any one of claims 1 to 16, wherein the PD-1 inhibitor is an anti-PD-1 antibody or an antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three CDRs (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

2.

18. The method of claim 17, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:

8.

19. The method of claim 17 or 18, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NOs: 1 / 2.

20. The method of any one of claims 17 to 19, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and the heavy chain has the amino acid sequence of SEQ ID NO:

9.

21. The method of any one of claims 17 to 19, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and the light chain has the amino acid sequence of SEQ ID NO:

10.

22. 20. The method of any one of claims 17 to 19, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, the heavy chain having the amino acid sequence of SEQ ID NO:9 and the light chain having the amino acid sequence of SEQ ID NO:

10.

23. The method of any one of claims 1 to 16, wherein the PD-1 inhibitor is cemiplimab or a biological equivalent thereof.

24. The method of any one of claims 1 to 16, wherein the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754091, PF-06801591, spartalizumab, camrelizumab, JNJ-63723283, and MCLA-134.

25. The method of any one of claims 1 to 16, wherein the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-072, KN035, and CK-301.

26. A pharmaceutical composition comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor for use in the adjuvant treatment of skin cancer following completion of surgery and post-surgical radiation.

27. The pharmaceutical composition of claim 26, wherein the PD-1 inhibitor is an anti-PD-1 antibody or an antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 and three CDRs (LCDR1, LCDR2 and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

2.

28. The pharmaceutical composition of claim 27, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:

8.

29. 29. The pharmaceutical composition of claim 28, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NOs: 1 / 2.

30. 30. The pharmaceutical composition of any one of claims 26 to 29, comprising 5 mg to 500 mg of a PD-1 inhibitor.

31. The pharmaceutical composition of any one of claims 26 to 30, comprising 350 mg of a PD-1 inhibitor.

32. The pharmaceutical composition according to any one of claims 26 to 31, wherein the skin cancer is CSCC.

33. 1. A method of treating or inhibiting tumor growth comprising: (a) selecting patients with skin cancer for which surgical removal is planned; and (b) administering to the patient, prior to surgical removal, a neoadjuvant treatment comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor. The method comprising:

34. 34. The method of claim 33, wherein the skin cancer is cutaneous squamous cell carcinoma (CSCC), basal cell carcinoma (BCC), Merkel cell carcinoma or melanoma.

35. 35. The method of claim 33 or 34, wherein the skin cancer is CSCC.

36. The method of any one of claims 33 to 35, wherein the patient is at high risk of recurrence of CSCC.

37. 37. The method of claim 36, wherein the patient has at least one of the following high-risk configurations: (a) nodular disease with extracapsular extension and at least one nodule >20 mm; (b) in-transit transfer (ITM); (c) T4 lesion; (d) perineural invasion (PNI); and (e) recurrent CSCC plus at least one of the following additional configurations: (i) ≥N2b disease associated with recurrent lesions; (ii) Nominal ≧T3; and (iii) Recurrent lesions ≥ 20 mm in diameter.

38. A therapeutically effective dose is 5 mg to 500 mg of a PD-1 inhibitor administered as a neoadjuvant. The method of any one of claims 33 to 37, comprising:

39. 39. The method of any one of claims 33-38, wherein the therapeutically effective amount comprises 350 mg of the PD-1 inhibitor administered as a neoadjuvant.

40. 39. The method of any one of claims 33 to 38, wherein one or more doses of the PD-1 inhibitor are administered as a neoadjuvant treatment, each dose being administered 2 to 12 weeks, preferably 3 weeks, after the immediately preceding dose.

41. 41. The method of claim 40, wherein each dose comprises 5 mg to 500 mg, preferably 350 mg, of the PD-1 inhibitor.

42. 42. The method of any one of claims 33-41, further comprising (c) surgically removing the skin cancer following the neoadjuvant treatment.

43. 43. The method of claim 42, further comprising administering to the patient an adjuvant therapy comprising a therapeutically effective amount of a PD-1 inhibitor after step (c), wherein the adjuvant PD-1 inhibitor can be the same as or different from the neoadjuvant PD-1 inhibitor.

44. 44. The method of claim 43, wherein the adjuvant treatment comprises administering between 5 mg and 500 mg of the PD-1 inhibitor.

45. 45. The method of claim 43 or 44, wherein the adjuvant treatment comprises administering 350 mg of a PD-1 inhibitor.

46. The method of any one of claims 33 to 42, wherein the PD-1 inhibitor is administered intravenously.

47. 47. The method of any one of claims 33-46, wherein administration of the PD-1 inhibitor leads to a reduced risk of subsequent skin cancer recurrence or eliminates the occurrence of subsequent skin cancer recurrence.

48. 47. The method of any one of claims 33-46, wherein administration of the PD-1 inhibitor leads to at least about a 10% lower incidence of subsequent skin cancer recurrence as compared to patients following completion of surgery and radiation therapy without an adjuvant skin cancer treatment.

49. 47. The method of any one of claims 33 to 46, further comprising administering an additional therapeutic agent selected from a chemotherapeutic agent, a corticosteroid, an anti-inflammatory agent and / or combinations thereof.

50. 50. The method of any one of claims 33 to 49, wherein the PD-1 inhibitor is selected from the group consisting of an anti-PD-1 antibody or an antigen-binding fragment thereof, an anti-PD-L1 antibody or an antigen-binding fragment thereof, and an anti-PD-L2 antibody or an antigen-binding fragment thereof.

51. The method of any one of claims 33 to 50, wherein the PD-1 inhibitor is an anti-PD-1 antibody or an antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1, and three CDRs (LCDR1, LCDR2, and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

2.

52. HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:

6.

52. The method of claim 51 , wherein LCDR2 has the amino acid sequence of SEQ ID NO:7; LCDR3 has the amino acid sequence of SEQ ID NO:

8.

53. The method of claim 51 or 52, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NOs: 1 / 2.

54. The method of any one of claims 51 to 53, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and the heavy chain has the amino acid sequence of SEQ ID NO:

9.

55. The method of any one of claims 51 to 53, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, and the light chain has the amino acid sequence of SEQ ID NO:

10.

56. 54. The method of any one of claims 51 to 53, wherein the anti-PD-1 antibody comprises a heavy chain and a light chain, the heavy chain having the amino acid sequence of SEQ ID NO:9 and the light chain having the amino acid sequence of SEQ ID NO:

10.

57. 51. The method of any one of claims 33 to 50, wherein the PD-1 inhibitor is cemiplimab or a biological equivalent thereof.

58. The method of any one of claims 33 to 50, wherein the PD-1 inhibitor is an anti-PD-1 antibody selected from the group consisting of cemiplimab, nivolumab, pembrolizumab, pidilizumab, MEDI0608, BI 754048, PF-06371548, spartalizumab, camrelizumab, JNJ-63313240, and MCLA-134.

59. The method of any one of claims 33 to 50, wherein the PD-1 inhibitor is an anti-PD-L1 antibody selected from the group consisting of H1H8314N, avelumab, atezolizumab, durvalumab, MDX-1105, LY3300054, FAZ053, STI-1014, CX-031, KN035 and CK-301.

60. A pharmaceutical composition comprising a therapeutically effective amount of a programmed death 1 (PD-1) inhibitor for use in neoadjuvant therapy prior to planned surgery to treat skin cancer.

61. The pharmaceutical composition of claim 60, wherein the PD-1 inhibitor is an anti-PD-1 antibody or an antigen-binding fragment thereof comprising three complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) of a heavy chain variable region (HCVR) comprising the amino acid sequence of SEQ ID NO:1 and three CDRs (LCDR1, LCDR2 and LCDR3) of a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:

2.

62. The pharmaceutical composition of claim 61, wherein HCDR1 has the amino acid sequence of SEQ ID NO:3; HCDR2 has the amino acid sequence of SEQ ID NO:4; HCDR3 has the amino acid sequence of SEQ ID NO:5; LCDR1 has the amino acid sequence of SEQ ID NO:6; LCDR2 has the amino acid sequence of SEQ ID NO:7; and LCDR3 has the amino acid sequence of SEQ ID NO:

8.

63. The pharmaceutical composition of claim 62, wherein the anti-PD-1 antibody or antigen-binding fragment thereof comprises the HCVR / LCVR sequence pair of SEQ ID NOs: 1 / 2.

64. 64. The pharmaceutical composition of any one of claims 60 to 63, comprising 5 mg to 500 mg of a PD-1 inhibitor.

65. A pharmaceutical composition according to any one of claims 60 to 63, comprising 350 mg of a PD-1 inhibitor. thing.

66. The pharmaceutical composition of any one of claims 60 to 65, wherein the skin cancer is CSCC.

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