Compositions and uses of tumor activated antibodies targeting EGFR and effector cell antigens

Abstract

Disclosed herein are compositions and uses of tumor activated antibodies targeting EGFR and effector cell antigens. In particular, the present application provides isolated recombinant polypeptide complexes that comprise binding domains that selectively bind to an effector cell antigen and EGFR, in which one or more of the binding domains is selectively activated in the tumor microenvironment and the isolated polypeptide or polypeptide complex comprises a half-life extending molecule.

Description

COMPOSITIONS AND USES OF TUMOR ACTIVATED ANTIBODIES TARGETING EGFRAND EFFECTOR CELL ANTIGENSCROSS REFERENCE

[0001] The present application claims the benefit of U.S. Provisional Application No. 63 / 557,411 filed on February 23, 2024, and U.S. Provisional Application No. 63 / 686,588 filed on August 23, 2024, each of which is incorporated herein by reference in its entirety.SEQUENCE LISTING

[0002] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on February 18, 2025, is named 52426-770_601_SL.xml and is 30,078 bytes in size.SUMMARY

[0003] Disclosed herein, in one aspect, is a method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex, wherein the first dose is at least about 50 pg, and wherein the isolated recombinant polypeptide complex comprises a first chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1 and a second chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. In some embodiments, the first dose is about 50 pg to about 10 mg. In some embodiments, the first dose is at least about 100 pg. In some embodiments, the first dose is at least about 150 pg. In some embodiments, the first dose is at least about 200 pg. In some embodiments, the first dose is at least about 250 pg. In some embodiments, the first dose is at least about 300 pg. In some embodiments, the first dose is at least about 350 pg. In some embodiments, the first dose is at least about 400 pg. In some embodiments, the first dose is at least about 450 pg. In some embodiments, the first dose is at least about 500 pg. In some embodiments, the first dose is at least about 750 pg. In some embodiments, the first dose is at least about 1 mg. In some embodiments, the first dose is at least about 1.5 mg. In some embodiments, the first dose is at least about 2 mg. In some embodiments, the first dose is at least about 3 mg. In some embodiments, the first dose is at least about 4 mg. In some embodiments, the first dose is at least about 5 mg. In some embodiments, the first dose is at least about 6 mg. In some embodiments, the first dose is at least about 7 mg. In some embodiments, the first dose is at least about 8 mg. In some embodiments, the first dose is at least about 9 mg. In some embodiments, the first dose is at least about 10 mg. In some embodiments, the first dose is at least about 15 mg. In some embodiments, the first dose is at least about 20 mg. In some embodiments, the first dose is at least about 25 mg.

[0004] In some embodiments, the first dose is about 100 pg. In some embodiments, the first dose is about 150 pg. In some embodiments, the first dose is about 200 pg. In some embodiments, the first dose is about250 j g. In some embodiments, the first dose is about 300 pg. In some embodiments, the first dose is about350 pg. In some embodiments, the first dose is about 400 pg. In some embodiments, the first dose is about450 pg. In some embodiments, the first dose is about 500 pg. In some embodiments, the first dose is about750 pg. In some embodiments, the first dose is about 1 mg. In some embodiments, the first dose is about1.5 mg. In some embodiments, the first dose is about 2 mg. In some embodiments, the first dose is about 3 mg. In some embodiments, the first dose is about 4 mg. In some embodiments, the first dose is about 5 mg. In some embodiments, the first dose is about 6 mg. In some embodiments, the first dose is about 7 mg. In some embodiments, the first dose is about 8 mg. In some embodiments, the first dose is about 9 mg. In some embodiments, the first dose is about 10 mg. In some embodiments, the first dose is about 15 mg. In some embodiments, the first dose is about 20 mg. In some embodiments, the first dose is about 25 mg. In some embodiments, the first dose is about 30 mg.

[0005] In some embodiments, the method further comprises administering to the subject a target dose of the isolated recombinant polypeptide complex, wherein the target dose is higher than the first dose, and wherein the target dose is administered after the first dose. In some embodiments, the method further comprises administering to the subject a second dose of the isolated recombinant polypeptide complex, wherein the second dose is higher than the first dose and lower than the target dose, and wherein the second dose is administered between the first dose and the target dose. In some embodiments, the first dose is administered weekly. In some embodiments, the first dose is administered once every two weeks. In some embodiments, the first dose is administered once every three weeks. In some embodiments, the target dose is administered weekly. In some embodiments, the target dose is administered once every two weeks. In some embodiments, the target dose is administered once every three weeks. In some embodiments, the second dose is administered weekly. In some embodiments, the second dose is administered once every two weeks. In some embodiments, the second dose is administered once every three weeks. In some embodiments, the method comprises a treatment cycle that starts on day 1. In some embodiments, the first dose is administered on day 1 of the treatment cycle. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle. In some embodiments, the first dose, the second dose, and the target dose are administered on day 1, day 4, and day 8, respectively, of the treatment cycle. In some embodiments, the target dose is administered weekly after day 8 of the treatment cycle. In some embodiments, the target dose is administered once every two weeks after day 8 of the treatment cycle. In some embodiments, the target dose is administered once every three weeks after day 8 of the treatment cycle.

[0006] In some embodiments, the target dose is about 50 pg to about 10 mg. In some embodiments, the target dose is at least about 100 pg. In some embodiments, the target dose is at least about 250 pg. In some embodiments, the target dose is at least about 500 pg. In some embodiments, the target dose is at least about 750 pg. In some embodiments, the target dose is at least about 1 mg. In some embodiments, the target dose is at least about 1.25 mg. In some embodiments, the target dose is at least about 1.5 mg. In some embodiments, the target dose is at least about 2 mg. In some embodiments, the target dose is at least about 5 mg. In some embodiments, the target dose is at least about 7.5 mg. In some embodiments, the target doseis at least about 10 mg. In some embodiments, the target dose is at least about 15 mg. In some embodiments, the target dose is at least about 20 mg. In some embodiments, the target dose is at least about 25 mg. In some embodiments, the target dose is at least about 30 mg. In some embodiments, the target dose is at least about 50 mg. In some embodiments, the target dose is at least about 75 mg. In some embodiments, the target dose is at least about 100 mg. In some embodiments, the target dose is at least about 150 mg. In some embodiments, the target dose is at least about 200 mg. In some embodiments, the target dose is at least about 250 mg. In some embodiments, the target dose is at least about 300 mg.

[0007] In some embodiments, the target dose is about 100 pg. In some embodiments, the target dose is about 250 pg. In some embodiments, the target dose is about 500 pg. In some embodiments, the target dose is about 750 pg. In some embodiments, the target dose is about 1 mg. In some embodiments, the target dose is about 1.25 mg. In some embodiments, the target dose is about 1.5 mg. In some embodiments, the target dose is about 2 mg. In some embodiments, the target dose is about 5 mg. In some embodiments, the target dose is about 7.5 mg. In some embodiments, the target dose is about 10 mg. In some embodiments, the target dose is about 15 mg. In some embodiments, the target dose is about 20 mg. In some embodiments, the target dose is about 25 mg. In some embodiments, the target dose is about 30 mg. In some embodiments, the target dose is about 40 mg. In some embodiments, the target dose is about 50 mg. In some embodiments, the target dose is about 75 mg. In some embodiments, the target dose is about 100 mg. In some embodiments, the target dose is about 150 mg. In some embodiments, the target dose is about 200 mg. In some embodiments, the target dose is about 250 mg. In some embodiments, the target dose is about 300 mg.

[0008] In some embodiments, the first dose is at least about 500 pg and the target dose is at least about 1.25 mg. In some embodiments, the first dose is about 500 pg and the target dose is about 1.25 mg. In some embodiments, the first dose is about 500 pg and the target dose is about 5 mg. In some embodiments, the first dose is about 1.5 mg and the target dose is about 15 mg. In some embodiments, the first dose is about 5 mg and the target dose is about 50 mg. In some embodiments, the first dose is about 15 mg and the target dose is about 150 mg. In some embodiments, the first dose is about 30 mg and the target dose is about 300 mg. In some embodiments, the first dose, the second dose, or the target dose is administered at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, or more than 50 times. In some embodiments, the first dose is about 500 pg, the second dose is about 1.5 mg, and the target dose is about 5 mg. In some embodiments, the first dose is about 1.5 mg, the second dose is about 5 mg, and the target dose is about 15 mg. In some embodiments, the first dose is about 5 mg, the second dose is about 15 mg, and the target dose is about 50 mg. In some embodiments, the first dose is about 15 mg, the second dose is about 50 mg, and the target dose is about 150 mg. In some embodiments, the first dose is about 30 mg, the second dose is about 100 mg, and the target dose is about 300 mg. In some embodiments, the method further comprises at least 2 treatment cycles, at least 3 treatment cycles, at least 4 treatment cycles, at least 5 treatment cycles, at least 6 treatment cycles, at least 7 treatment cycles, at least 8 treatment cycles, at least9 treatment cycles, at least 10 treatment cycles, or more than 10 treatment cycles. In some embodiments, the cancer comprises a cell that expresses epidermal growth factor receptor (EGFR). In some embodiments, the cancer comprises a cell that overexpresses EGFR. In some embodiments, the cancer comprises colorectal cancer (CRC), squamous cell carcinoma of head and neck (SCCHN), non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), breast cancer, bladder cancer, ovarian cancer, liver cancer, pancreatic cancer, small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triplenegative breast cancer (TNBC), prostate cancer, or a combination thereof. In some embodiments, the cancer comprises advanced or metastatic NSCLC. In some embodiments, the cancer comprises advanced or metastatic SCCHN. In some embodiments, the cancer comprises advanced or metastatic CRC. In some embodiments, the cancer comprises advanced or metastatic RCC. In some embodiments, the cancer comprises advanced or metastatic SCLC. In some embodiments, the cancer comprises advanced or metastatic PDAC. In some embodiments, the cancer comprises advanced or metastatic TNBC. In some embodiments, the cancer comprises advanced or metastatic prostate cancer. In some embodiments, the cancer progresses after treatment with a prior cancer therapy. In some embodiments, the cancer is refractory, intolerant, non-responsive, or resistant to a prior cancer therapy. In some embodiments, the subject is at least 18 years old. In some embodiments, the subject is about 37 to about 81 years old. In some embodiments, the subject is about 65 years old. In some embodiments, the prior cancer therapy comprises 1 to 9 lines of cancer therapies. In some embodiments, the prior cancer therapy comprises about 4 lines of cancer therapies. In some embodiments, the prior cancer therapy comprises a treatment targeting programmed death ligand 1 (PD-L1), programmed cell death protein 1 (PD-1), or both. In some embodiments, the prior cancer therapy comprises pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, or a combination thereof. In some embodiments, the subject has adequate organ function. In some embodiments, the subject has resolved acute effects of any prior therapy to baseline severity of Common Terminology Criteria for Adverse Events (CTCAE) grade of no more than 1. In some embodiments, the subject has locally advanced or metastatic NSCLC, SCCHN, CRC, SCLC, PDAC, TNBC, prostate cancer, or RCC that is confirmed histologically or cytologically. In some embodiments, the subject has a tumor lesion measured by response evaluation criteria in solid tumors (RECIST) guidelines. In some embodiments, the subject is not treated with an anti-cancer therapy within 28 days before the administering. In some embodiments, the subject is not treated with an anti -cancer therapy that has no more than 5 elimination half-lives before the administering. In some embodiments, the subject is not treated with a bispecific T cell engager that targets EGFR before the administering. In some embodiments, the subject is not treated with a chimeric antigen receptor T (CAR-T) cell therapy that targets EGFR before the administering. In some embodiments, the subject is not treated with another cluster of differentiation 3 (CD3)-binding T cell engager before the administering. In some embodiments, the subject does not have a clinically significant cardiovascular disease. In some embodiments, the subject does not have an active and clinically significant infection. In some embodiments, the infection comprises abacterial infection, a viral infection, a fungal infection, a mycobacterial infection, or a combination thereof. In some embodiments, the subject is not treated with an oxygen therapy before the administering. In someembodiments, the administering comprises administering intravenously. In some embodiments, the subject exhibits a cytokine release syndrome (CRS) no more than grade 1. In some embodiments, the subject exhibits a cytokine release syndrome (CRS) no more than grade 2. In some embodiments, the subject exhibits a treatment related adverse event (TRAE) not related to a CRS. In some embodiments, the administering comprises administering to the subject a second treatment. In some embodiments, the administering results in a therapeutic effect in size in target lesion, hepatic metastasis, stable disease, partial response, complete response, size of renal mass, best RECIST response, overall response rate, duration of response, progression free survival, or a combination thereof, after the administering. In some embodiments, the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a 100% reduction in size of target lesion 12 weeks after the administering. In some embodiments, the administering comprises administering weekly the first dose of the isolated recombinant polypeptide complex in an amount of about 150 pg. In some embodiments, the subject exhibits a reduction or elimination of hepatic metastasis after the administering. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the subject exhibits no CRS or TRAE after the administering. In some embodiments, the subject has a mutation, and wherein the mutation comprises low microsatellite instability (MSI-L) / microsatellite stable (MSS), a high microsatellite instability (MSI-H), a MET gene mutation, a TP53 mutation, an epidermal growth factor receptor (EGFR) mutation, a Kirsten rat sarcoma virus (KRAS) mutation, an anaplastic lymphoma kinase (ALK) mutation, a Rearrangement of c-ros oncogene 1 (ROS1) mutation, an Adenomatous polyposis coli (APC) mutation, a B-type Raf protooncogene (BRAF) mutation, a pl 10a subunit of phosphatidylinositol-3 -kinase (PIK3CA) mutation, a cyclin-dependent kinase inhibitor 2A (CDKN2A) mutation, a Phosphatase and tensin homolog (PTEN) mutation, a NOTCH1 mutation, a breast cancer 1 early onset protein (BRAC1), a breast cancer 2 early onset protein (BRAC2), partner and localizer of BRCA2 (PALB2), or a combination thereof. In some embodiments, the subject is treated with four of the prior cancer therapy before the administering, wherein the four of the prior cancer therapy comprises a treatment targeting PD-L1 / PD-1, and wherein the cancer is refractory, non-responsive, or resistant to the treatment targeting PD-L1 / PD-1. In some embodiments, the subject has RCC of stage 4, and wherein the subject exhibits a reduction of about 12% in size of renal mass. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle. In some embodiments, the target dose is administered weekly after day 8. In some embodiments, the subject has MSI-L / MSS mutation and proficient mismatch repair (pMMR) mutation. In some embodiments, the subject is treated with three of the prior cancer therapy before the administering. In some embodiments, the subject has an extensive large renal mass. In some embodiments, the renal mass is about 11 cm. In some embodiments, the administering results in an elimination of a cancer related pain. In some embodiments, the administering results in an elimination of a cancer related pain after two doses of the isolated recombinant polypeptide complex. In some embodiments, the administering results in the reduction of about 12% in diameter of renal massmeasured by computed tomography (CT) scan on day 17 of the treatment cycle. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the administering comprises administering weekly the first dose in an amount of about 150 pg. In some embodiments, the subject exhibits a best RECIST response, a partial response (PR), or a complete response (CR). In some embodiments, the subject exhibits stable disease. In some embodiments, the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle. In some embodiments, the target dose is administered weekly after day 8.

[0009] In some embodiments, the method further comprises treating the subject with a therapy for arthralgia, anemia, CRS, dermatitis acneiform, nausea, rash maculopapular, back pain, diarrhea, dizziness, fatigue, headache, hyperglycemia, hypokalemia, hypophosphatemia, injection site irritation, lymphocyte count decrease, oedema peripheral, oral pain, pain in extremity, pyrexia, vomiting, or a combination thereof. In some embodiments, the subject receives 2 or 3 prior cancer therapies before the administering. In some embodiments, the subject is negative for HER2 or hormone receptor (HR). In some embodiments, the subject has never been diagnosed with gout before the administering. In some embodiments, the subject does not have arthritis or arthralgia before the administering.INCORPORATION BY REFERENCE

[0010] All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] The features of the disclosure are set forth with particularity in the appended claims. A better understanding of the features and advantages of the present invention will be obtained by reference to the following detailed description that sets forth illustrative embodiments, in which the principles of the disclosure are utilized, and the accompanying drawings (also “Figure” and “FIG.” herein), of which:

[0012] FIG. 1 illustrates a non-limiting example of design, structure and mechanism of action of a polypeptide complex 1 (PC-1) disclosed herein according to an embodiment of the present disclosure.

[0013] FIG. 2 illustrates a non -limiting example of an Outline of Planned Dose Escalation and Dose Expansion according to an embodiment of the present disclosure.

[0014] FIG. 3 illustrates a non-limiting example of an Algorithm for Continuation and Discontinuation of PC-1 Based on LVEF Assessments in Subjects with LVEF no less than 40% or nor more than 45% according to an embodiment of the present disclosure.

[0015] FIGs. 4A-4C illustrate non-limiting examples of example flat dose schemas according to an embodiment of the present disclosure.

[0016] FIGs. 5A-5D illustrate non-limiting examples of example step dose schemas according to anembodiment of the present disclosure.

[0017] FIG. 6 illustrates a non -limiting example of Time on treatment for all subjects according to an embodiment of the present disclosure.

[0018] FIG. 7 illustrates a non-limiting example of confirmed PR observed in a heavily pretreated subject with NSCLC according to an embodiment of the present disclosure.

[0019] FIG. 8 illustrates a non-limiting example of the treatment outcomes of patients according to an embodiment of the present disclosure.DETAILED DESCRIPTION OF THE INVENTION

[0020] Multispecific antibodies combine the benefits of different binding specificities derived from two or more antibodies into a single composition. Multispecific antibodies for redirecting T cells to cancers have shown promise in both pre-clinical and clinical studies. This approach relies on binding of one antigen interacting portion of the antibody to a tumor-associated antigen or marker, while a second antigen interacting portion can bind to an effector cell antigen on a T cell, such as CD3, which then triggers cytotoxic activity. One such tumor-associated antigen is epidermal growth factor receptor (EGFR). EGFR is a transmembrane protein that is a receptor for members of the epidermal growth factor family of extracellular protein ligands. EGFR is the most commonly overexpressed membrane protein in cancer. However, EGFR expression is not limited to tumors and is widely expressed throughout the body, resulting in systemic toxicities with EGFR-directed therapies.

[0021] T cell engagers (TCEs) therapeutics have several benefits including they are not cell therapies and thus can be offered as off-the-shelf therapies as opposed to chimeric antigen receptor T cell (CAR-T cell) therapies. While TCE therapeutics have displayed potent anti-tumor activity in hematological cancers, developing TCEs to treat solid tumors has faced challenges due to the limitations of prior TCE technologies, namely (i) overactivation of the immune system leading to cytokine release syndrome (CRS), (ii) on-target, healthy tissue toxicities and (iii) poor pharmacokinetics (PK) leading to short halflife. CRS arises from the systemic activation of T cells and can result in life-threatening elevations in inflammatory cytokines such as interleukin-6 (IL-6). Severe and acute CRS leading to dose limited toxicities and deaths have been observed upon the dosing of T cell engagers developed using other platforms to treat cancer patients in poor clinical studies. This toxicity restricts the maximum blood levels of T cell engagers that can be safely dosed. T cell engager effectiveness has also been limited because of on-target, healthy tissue toxicity. T cell engagers developed using a platform not designed for tumor-specification activation have resulted in clinical holds and dose-limiting toxicities resulting from target expression in healthy tissues. T cell engagers have also been limited by short half-lives. T cell engagers quickly reach sub-therapeutic levels after being administered as they are quickly eliminated from the body due to their short exposure half-lives. For this reason, T cell engagers such as blinatumomab are typically administered by a low-dose, continuous infusion pump over a period of weeks to overcome the challenge of a short half-life and to maintain therapeutic levels of drug in the body. A continuous dosing regimen represents a significant burden for patients.

[0022] To overcome these challenges associated with the effectiveness of T cell engagers, described herein are recombinant polypeptide complexes that comprise binding domains that selectively bind to an effector cell antigen and EGFR, in which one or more of the binding domains is selectively activated in the tumor microenvironment and the isolated polypeptide or polypeptide complex comprises a half-life extending molecule. Such modifications reduce CRS and on -target healthy tissue toxicity risk and improves stability in the bloodstream and serum half-life prior to activation. The recombinant polypeptide complexes described herein have activity at low levels of target expression, and can be easily manufactured and formulated.

[0023] In some embodiments, the recombinant polypeptide complexes described herein are used in a method of treating cancer. In some embodiments, the cancer has cells that express EGFR. In some embodiments, the recombinant polypeptide complexes described herein are used in a method of treating renal cell carcinoma, colorectal cancer (CRC), squamous cell carcinoma of the head and Neck (SCCHN), non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple -negative breast cancer (TNBC), prostate cancer, breast cancer, colon / rectum cancer, head and neck cancer, esophagogastric cancer, liver cancer, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, kidney cancer, or pancreatic cancer. In some embodiments, the polypeptides or polypeptide complexes described herein are used in a method of treating subjects who are resistant to EGFR inhibitor treatment. In some embodiments, the recombinant polypeptide complexes described herein are used in a method of treating subjects who harbor KRAS mutations. In some embodiments, the recombinant polypeptide complex described herein are used in a method of treating subjects who are resistant to EGFR inhibitor treatment and harbor KRAS mutations.Isolated Recombinant Polypeptide Complex Compositions

[0024] In some embodiments, the isolated recombinant polypeptide complex comprises a tumor-activated T-cell engager with EGFR and CD3 binding domains, an albumin binding domain to extend circulating half-life, a peptide mask that inhibits CD3 engagement on T-cells, and a tumor protease cleavable linker. Tumor specific proteolysis of the cleavable linker in the tumor microenvironment can separate the tandem mask and albumin-binding domain from the isolated recombinant polypeptide complex. The isolated recombinant polypeptide complex can comprise chain 1 and chain 2 as described herein. In some embodiments, chain 1 comprises an amino acid sequence having at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 1. In some embodiments, chain 2 comprises an amino acid sequence having at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 2.

[0025] Table 1A provides exemplary amino acid sequences of an isolated recombinant polypeptide complex. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to any one of the sequences in Table 1A.

[0026] In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 1.

[0027] In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence according to SEQ ID NO: 1.

[0028] In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 97% sequence identity to SEQID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2.

[0029] In some embodiments, the isolated recombinant polypeptide complex comprises an amino acid sequence according to SEQ ID NO: 2.

[0030] In some embodiments, the isolated recombinant polypeptide light chain (LC) comprises at least 85% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 90% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 95% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 99% sequence identity to SEQ ID NO:1. In some embodiments, the LC comprises the amino acid sequence according to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide heavy chain (HC) comprises at least 85% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 99% sequence identity to SEQ ID NO:2. In some embodiments, the HC comprises the amino acid sequence according to SEQ ID NO: 2. In some embodiments, the LC comprises at least 99% sequence identity to SEQ ID NO: 1, the HC comprises at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the LC comprises the amino acid sequence according to SEQ ID NO: 1, the HC comprises the amino acid sequence according to SEQ ID NO: 2.Table 1A. Exemplary Amino Acid Sequences of an Isolated Recombinant Polypeptide Complex (PC-1) that selectively binds to EGFR and effector cell antigens such as CD3 and its Metabolites and DerivativesPC-l=polypeptide complex 1, EGFR=Epidermal Growth Factor Receptor, HC=Heavy Chain, LC=Light Chain, CD3=Cluster of Differentiation 3, FAB=Fragment Antigen-Binding Region, scFv= Single Chain Variable Fragment, MMP9=Matrix Metallopeptidase 9, SP=Serine Protease, HisTag=Histidine Tag, NC=Non-Cleavable, TCE=T Cell Engager

[0031] In some embodiments, the CD3 binding domain of the isolated recombinant polypeptide complex comprises an antibody or antibody fragment. In some embodiments, the CD3 binding domain comprises an antibody or antibody fragment that is humanized. In some embodiments, the antibody or antibody fragment comprises a single chain variable fragment (scFv), a single domain antibody, or a Fab fragment. In some embodiments, the antibody or antibody fragment is the single change variable fragment (scFv). In some embodiments, the scFv comprises a scFv heavy chain polypeptide and a scFv light chain polypeptide. In some embodiments, the CD3 binding domain comprises a light chain and variable heavy chain each of which is capable of specifically binding to human CD3.

[0032] In some embodiments, the CD3 binding domain is a single chain variable fragment (scFv) that binds to CD3 comprises a scFv light chain variable domain and a scFv heavy chain variable domain. In some embodiments, the scFv heavy chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table IB or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the scFv light chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table IB or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0033] In some embodiments, the scFv heavy chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table IB or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and the scFv light chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table IB or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).Table IB. Anti-CD3 amino acid sequences (CDRs as determined by IMGT numbering system)

[0034] In some embodiments, the scFv light chain variable domain comprises complementarity determining regions (CDRs): LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC- CDR2, and the LC-CDR3 of the scFv light chain variable domain comprise LC-CDR1: SEQ ID NO 13, LC-CDR2: GTK, and LC-CDR3: SEQ ID NO: 15, and wherein the CDRs comprise from 0-2 amino acid modifications in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the scFv heavy chain variable domain comprises complementarity determining regions (CDRs): HC-CDR1, HC- CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the scFv heavy chain variable domain comprise: HC-CDR1: SEQ ID NO: 16, HC-CDR2: SEQ ID NO: 17, and HC- CDR3: SEQ ID NO: 18, and wherein the CDRs comprise from 0-2 amino acid modifications in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0035] In some embodiments, CD3 binding domain is a scFv, wherein the scFv comprises CDRs: LC- CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the scFv comprise LC-CDR1: SEQ ID NO: 13, LC-CDR2: GTK, and LC-CDR3: SEQ ID NO: 15; and the scFv comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the scFv comprise HC-CDR1: SEQ ID NO: 16, HC-CDR2: SEQ ID NO: 17, and HC- CDR3: SEQ ID NO: 18. In some embodiments, the CD3 binding domain is a scFv, and the scFv comprises an amino acid sequence according to SEQ ID NO: 19.

[0036] In some embodiments, the EGFR binding domain of the isolated recombinant polypeptide complex comprises an antibody or antibody fragment. In some embodiments, EGFR binding domain comprises an antibody or antibody fragment that is humanized. In some embodiments, the antibody or antibody fragment comprises a single chain variable fragment (scFv), a single domain antibody, a Fab, or a Fab’. In some embodiments the EGFR binding domain is the Fab or Fab’. In some embodiments, the Fab or Fab’ comprises (a) a Fab light chain polypeptide and (b) a Fab heavy chain polypeptide.

[0037] In some embodiments, the EGFR binding domain is a Fab or Fab’ that binds to EGFR comprises a Fab light chain polypeptide chain and a Fab heavy chain polypeptide. In some embodiment, the Fab light chain polypeptide comprises a Fab light chain variable domain. In some embodiments, the Fab heavy chain polypeptide comprises a Fab heavy chain variable domain. In some embodiments, the Fab heavy chain variable domain comprises at least one, two, or three complementarity determining regions(CDR)s disclosed in Table 1C or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity). In some embodiments, the Fab light chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table 1C or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).

[0038] In some embodiments, the Fab heavy chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table 1C or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity); and the Fab light chain variable domain comprises at least one, two, or three complementarity determining regions (CDR)s disclosed in Table 1C or a sequence substantially identical thereto (e.g., a sequence that has at least 90%, 95%, 96%, 97%, 98%, or 99% sequence identity).Table 1C. anti-EGFR amino acid sequences (CDRs as determined by IMGT numbering system)

[0039] In some embodiments, the EGFR binding domain is a Fab or Fab’. In some embodiments, the Fab comprises complementarity determining regions (CDR)s: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the Fab comprise LC-CDR1: SEQ ID NO: 20, LC- CDR2: YAS, and LC-CDR3: SEQ ID NO: 22, and wherein the CDRs comprise from 0-2 amino acid modifications in at least one of the LC-CDR1, LC-CDR2, or LC-CDR3. In some embodiments, the Fab comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the Fab comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, and HC- CDR3: SEQ ID NO: 25, wherein the CDRs comprise from 0-2 amino acid modifications in at least one of the HC-CDR1, HC-CDR2, or HC-CDR3.

[0040] In some embodiments, the EGFR binding domain is a Fab, wherein the Fab or Fab’ comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the Fab comprise LC-CDR1: SEQ ID NO: 20, LC-CDR2: YAS, and LC-CDR3: SEQ ID NO: 22; and wherein the Fab comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the Fab comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, and HC-CDR3: SEQ ID NO: 25.

[0041] In some embodiments, the Fab light chain polypeptide comprises the amino acid sequence according to SEQ ID NO: 26. In some embodiments, the Fab heavy chain polypeptide comprises the amino acid sequence according to SEQ ID NO: 27.

[0042] In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells comprises a sequence as disclosed in Table ID or a sequence substantially identical thereto (e.g., a sequence that has 0, 1, or 2 amino acid modifications).Table ID. Peptide Mask Sequences

[0043] In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells is bound to the CD3 binding domain through ionic interaction, electrostatic interactions, hydrophobic interactions, Pi- stacking interactions, and H-bonding interactions, or a combination thereof. In some embodiments, thepeptide mask that inhibits CD3 engagement on T-cells is bound to the CD3 binding domain at or near an antigen binding site. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells becomes unbound from the CD3 binding domain when the cleavable linker is cleaved by the tumor specific protease. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 70% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 80% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 85% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T- cells has less than 90% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 95% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 98% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells has less than 99% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells comprises a de novo amino acid sequence that shares less than 10% sequence identity to the CD3 antigen. In some embodiments, the peptide mask that inhibits CD3 engagement on T-cells comprises an amino acid sequence according to SEQ ID NO: 28.

[0044] In some embodiments, the isolated recombinant polypeptide complex comprises a tumor-activated T-cell engager with EGFR and CD3 binding domains, an albumin binding domain to extend circulating half-life, a peptide mask that inhibits CD3 engagement on T-cells, a peptide mask that inhibits EGFR engagement, a tumor protease cleavable linker that connects the CD3 binding domain to the peptide mask that inhibits CD3, and a tumor protease cleavable linker that connects the EGFR binding domain to the peptide mask that inhibits EGFR engagement.

[0045] In some embodiments, the peptide mask that inhibits EGFR engagement comprises a sequence as disclosed in Table ID or a sequence substantially identical thereto (e.g., a sequence that has 0, 1, or 2 amino acid modifications).

[0046] In some embodiments, the peptide mask that inhibits EGFR engagement is bound to the EGFR binding domain through ionic interaction, electrostatic interactions, hydrophobic interactions, Pi-stacking interactions, and H-bonding interactions, or a combination thereof. In some embodiments, the peptide mask that inhibits EGFR engagement on T-cells is bound to the EGFR binding domain at or near an antigen binding site. In some embodiments, the peptide mask that inhibits EGFR engagement becomes unbound from the EGFR binding domain when the cleavable linker is cleaved by the tumor specific protease. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 70% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 80% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 85% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 90% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 95% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFRengagement has less than 98% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement has less than 99% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement comprises a de novo amino acid sequence that shares less than 10% sequence identity to the EGFR antigen. In some embodiments, the peptide mask that inhibits EGFR engagement comprises an amino acid sequence according to SEQ ID NO: 29.Metabolic Products

[0047] Disclosed herein are metabolic products of an isolated recombinant polypeptide complex disclosed herein. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a protease to generate an enzymatic product of the isolated recombinant polypeptide complex after the administering. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a tumor specific protease to generate the enzymatic product of the isolated recombinant polypeptide complex after the administering. In some embodiments, the tumor specific protease comprises two or more proteases. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a first protease of the two or more proteases to generate a first metabolic product of the isolated recombinant polypeptide complex. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a second protease of the two or more proteases to generate a second metabolic product of the isolated recombinant polypeptide complex. In some embodiments, the first protease comprises a serine protease. In some embodiments, the second protease comprises a matrix metalloprotease. In some embodiments, the serine protease comprises human matriptase (MTSP1). In some embodiments, the matrix metalloprotease comprises human matrix metalloprotease 9 (MMP9).

[0048] In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the first metabolic product. In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the second metabolic product. In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the first metabolic product and the second metabolic product.

[0049] Disclosed herein is an isolated polypeptide that is an enzymatic product of an isolated recombinant polypeptide complex disclosed herein. Disclosed herein is an isolated polypeptide comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3 and wherein the isolated polypeptide is 221 amino acids in length. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%,93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQID NO: 3. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and wherein the isolated polypeptide is 221 amino acids in length.In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 3.

[0050] Disclosed herein is an isolated polypeptide comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4 and wherein the isolated polypeptide is229 amino acids in length. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%.84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 4. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%,77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and wherein the isolated polypeptide is 229 amino acids in length. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 4.

[0051] Disclosed herein is an isolated polypeptide comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 5 and wherein the isolated polypeptide is 484 amino acids in length. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%,77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5 and wherein the isolated polypeptide is 484 amino acids in length. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 5.

[0052] Disclosed herein is an isolated polypeptide comprising an amino acid sequence with at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 6 and wherein the isolated polypeptide is 492 amino acids in length. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the isolated polypeptide comprises an amino acid sequence with at least 70%, 71%, 72%, 73%, 74%, 75%, 76%,77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%,95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6 and wherein the isolated polypeptide is 492 amino acids in length. In some embodiments, the isolated polypeptide comprises the amino acid sequence of SEQ ID NO: 6.

[0053] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having at least 70%, 71%,72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 2 and 653 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 2 and 653 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 2.

[0054] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 5.

[0055] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having at least 70%, 71%,72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 3 and 221 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 3 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 6.

[0056] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 2. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 2 and 653 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 2 and 653 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 2.

[0057] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having at least 70%, 71%,72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 5.

[0058] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%,80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%,98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 4 and 229 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 4 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 6.

[0059] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 1 and a second amino acid sequence having at least 70%, 71%,72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%.83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least99% sequence identity to the amino acid sequence of SEQ ID NO: 1 and 256 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1 and 256 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 5 and 484 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 1 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 5.

[0060] Disclosed herein is an isolated polypeptide comprising a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 1 and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 1 and 256 amino acids in length and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 1 and 256 amino acids in length and a second amino acid sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 6 and 492 amino acids in length. In some embodiments, the isolated polypeptide comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 1 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 6.

[0061] Disclosed herein is a pharmaceutical composition comprising the isolated polypeptide disclosed herein, and a pharmaceutically acceptable excipient disclosed herein.Formulations of Isolated Recombinant Antibodies that Target EGFR and CD3

[0062] Disclosed herein includes formulation(s) comprising a population of antibodies or recombinantantibodies, such as comprising any one or a combination the antibodies or recombinant antibodies as described herein.

[0063] Disclosed herein is a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients, wherein the one or more pharmaceutically acceptable excipients comprise histidine, sucrose, polysorbate-20, sodium phosphate, citrate, acetate, sodium chloride, potassium chloride, magnesium chloride, and calcium chloride. Disclosed herein is a pharmaceutical composition, wherein the pharmaceutical composition comprises the isolated recombinant polypeptide complex (such as any described herein), sodium phosphate monobasic monohydrate, sodium phosphate dibasic, citrate, acetate, histidine, heptahydrate, sodium chloride, potassium chloride, histidine, citrate, acetate, sucrose, polysorbate-20, polysorbate 80, magnesium chloride hexahydrate and calcium chloride dihydrate. The pharmaceutical composition can have a pH less than or equal to 6.5, 6.0, 5.5, 5.0, or 4.5. The pharmaceutical composition can have a pH greater than or equal to 4.5, 5.0, 5.5, 6.0 or 6.5. The pharmaceutical composition can have a pH between 4.0 and 5.0, between 4.5 and 5.5, or between 5.0 and 6.0. The pharmaceutical composition can comprise greater than or equal to 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 1 ImM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, or 20 mM Histidine. The pharmaceutical composition can comprise less than or equal to 20 mM, 19 mM, 18 mM, 17 mM, 16 mM, 15 mM, 14 mM, 13 mM, 12 mM, 11 mM, 10 mM, 9 mM, 8 mM, 7 mM, 6 mM, 5 mM, 4 mM, 3 mM, 2 mM, or 1 mM Histidine. The pharmaceutical composition can comprise greater than or equal to 5%(w / v), 6% (w / v), 7% (w / v), 8% (w / v) , 9% (w / v) , or 10% (w / v) sucrose. The pharmaceutical composition can comprise less than or equal to 10% (w / v), 9% (w / v), 8% (w / v), 7% (w / v), 6% (w / v), or 5% (w / v) sucrose. The pharmaceutical composition can comprise less than or equal to 0.05% (w / v) polysorbate 20, 0.04% (w / v) polysorbate 20, 0.03% (w / v) polysorbate 20, 0.02% (w / v) polysorbate 20, or 0.01% polysorbate 20. The pharmaceutical composition can comprise greater than or equal to 0.01% (w / v) polysorbate 20, 0.02% (w / v) polysorbate 20, 0.03% (w / v) polysorbate 20, 0.04% (w / v) polysorbate 20, or 0.05% polysorbate 20. Disclosed herein is a pharmaceutical composition, wherein the pharmaceutical composition comprises the isolated recombinant polypeptide complex (such as any described herein), about 10 mM Histidine, about 8% (w / v) sucrose, about 0.01% (w / v) polysorbate 20, pH of about 5.3.

[0064] In some embodiments of the formulation, at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% (e.g., by mole or by mass) of the antibodies of the population is monomeric. In some embodiments of the formulation, less than or equal to 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% (e.g., by mole or by mass) of the antibodies of the population is aggregated.

[0065] Disclosed herein are a plurality of isolated recombinant antibodies that target EGFR and CD3 wherein the isolated recombinant antibodies comprise a light chain (LC) amino acid sequence that has at least 80% sequence identity to SEQ ID NO: 1, a heavy chain (HC) amino acid sequence that has at least 80 % sequence identity to SEQ ID NO: 2, wherein the plurality comprises greater than 90% monomer of the isolated recombinant antibodies.

[0066] In some embodiments, the plurality comprises greater than 91% monomer. In some embodiments,the plurality comprises greater than 92% monomer. In some embodiments, the plurality comprises greater than 93% monomer. In some embodiments, the plurality comprises greater than 94% monomer. In some embodiments, the plurality comprises greater than 95% monomer. In some embodiments, the plurality comprises greater than 96% monomer. In some embodiments, the plurality comprises greater than 97% monomer. In some embodiments, the plurality comprises greater than 98% monomer. In some embodiments, the plurality comprises greater than 99% monomer.

[0067] In some embodiments, the LC comprises at least 85% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 90% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 95% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the LC comprises the amino acid sequence according to SEQ ID NO: 1. In some embodiments, the HC comprises at least 85% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the HC comprises the amino acid sequence according to SEQ ID NO: 2.

[0068] In some embodiments, the LC comprises at least 99% sequence identity to SEQ ID NO: 1, the HC comprises at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the LC comprises the amino acid sequence according to SEQ ID NO: 1, the HC comprises the amino acid sequence according to SEQ ID NO: 2.

[0069] In some embodiments, the concentration of the isolated recombinant antibodies is greater than or equal to 1.0 mg / mL. In some embodiments, the concentration of the isolated recombinant antibodies is at least 2.0 mg / mL. In some embodiments, the concentration of the isolated recombinant antibodies is at least 5.0 mg / mL. In some embodiments, the concentration of the isolated recombinant antibodies is at least 10.0 mg / mL. In some embodiments, the concentration of the isolated recombinant antibodies is at least 15.0 mg / mL. In some embodiments, the concentration of the isolated recombinant antibodies is at least 20.0 mg / mL. In some embodiments, the plurality is in a buffered solution having a pH less than or equal to 5.5. In some embodiments, the buffered solution comprises one or more of acetate, phosphate, or histidine. In some embodiments, the buffered solution comprises histidine at a concentration greater than 5mM. In some embodiments, the buffered solution comprises sucrose. In some embodiments, the sucrose is at a concentration greater than or equal to 5% w / v. In some embodiments, the buffered solution comprises polysorbate 20. In some embodiments, the polysorbate 20 is at a concentration greater than or equal to 0.01% w / v. In some embodiments, the plurality comprises greater than 90% monomer at a concentration of greater than or equal to about 20.0 mg / mL in about 10 mM histidine, about 8 % (w / v) sucrose, about 0.01% (w / v) PS20 pH of about 5.3.

[0070] Disclosed herein is a pharmaceutical composition comprising an isolated recombinant polypeptide complex disclosed herein and a pharmaceutically acceptable excipient. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having at least 70%,71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 1 and a second amino acid sequence having at least 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and a second amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1 and a second amino acid sequence having at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises a second amino acid sequence having at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 1 and a second amino acid sequence having the amino acid sequence of SEQ ID NO: 2. In some embodiments, the isolated recombinant polypeptide complex comprises a first amino acid sequence having the amino acid sequence of SEQ ID NO: 1. In some embodiments, the isolated recombinant polypeptide complex comprises a second amino acid sequence having the amino acid sequence of SEQ ID NO: 2.

[0071] In some embodiments, the pharmaceutical composition comprises the isolated recombinant polypeptide complex at a concentration of about 2 mg / ml. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer, a stabilizing agent, a tonicity agent, a surfactant, or combinations thereof. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer. In some embodiments, the pharmaceutically acceptable excipient comprises a tonicity agent. In some embodiments, the pharmaceutically acceptable excipient comprises a surfactant. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer, a stabilizing agent, a tonicity agent, and a surfactant. In some embodiments, the buffer comprises an amino acid or a derivative thereof. In some embodiments, the amino acid or the derivative thereof comprises L-histidine, L-histidine monohydrochloride monohydrate, or combinations thereof. In some embodiments, the stabilizing agent comprises sugar. In some embodiments, the sugar comprises sucrose. In some embodiments, the tonicity agent comprises sugar. In some embodiments, the sugar comprises sucrose. In some embodiments, thesurfactant comprises a polysorbate. In some embodiments, the surfactant comprises polysorbate 20 (PS20).

[0072] In some embodiments, the pharmaceutical composition comprises about 1 millimolar (mM) to about 50 mM L-histidine in the form of L-histidine and / or L-histidine monohydrochloride monohydrate, e.g., about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, or about 50 mM, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 1 to about 50 mM L-histidine in the form of L-histidine and L-histidine monohydrochloride monohydrate, e.g., about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, or about 50 mM, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L- histidine and / or L-histidine monohydrochloride monohydrate. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L-histidine or L-histidine monohydrochloride monohydrate. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L-histidine and L-histidine monohydrochloride monohydrate.

[0073] In some embodiments, the pharmaceutical composition comprises about 1% weight / volume (w / v) to about 20% (w / v) sucrose, e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18 %, 19%, or about 20%, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 8% (w / v) sucrose.

[0074] In some embodiments, the pharmaceutical composition comprises about 0.001% (w / v) to about 0.1% (w / v) polysorbate 20 (PS20), e.g., about 0.001%, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.PC-1 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %,0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %,0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %,0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %0.051 %, 0.052 %, 0.053 %, 0.054 %, 0.055 %, 0.056 %, 0.057 %, 0.058 %, 0.059 %, 0.06 %, 0.061 %,0.062 %, 0.063 %, 0.064 %, 0.065 %, 0.066 %, 0.067 %, 0.068 %, 0.069 %, 0.07 %, 0.071 %, 0.072 %.0.073 %, 0.074 %, 0.075 %, 0.076 %, 0.077 %, 0.078 %, 0.079 %, 0.08 %, 0.081 %, 0.082 %, 0.083 %,0.084 %, 0.085 %, 0.086 %, 0.087 %, 0.088 %, 0.089 %, 0.09 %, 0.091 %, 0.092 %, 0.093 %, 0.094 %,0.095 %, 0.096 %, 0.097 %, 0.098 %, 0.099 %, or about 0.1 %, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 0.01% (w / v) polysorbate 20 (PS20).

[0075] In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in theform of L-histidine and L-histidine monohydrochloride monohydrate, about 8% (w / v) sucrose, and about 0.01% (w / v) polysorbate 20. In some embodiments, the pharmaceutical composition comprises a pH of about 5.3. In some embodiments, the pharmaceutical composition comprises an osmolality of about 276 mOsmol / kg.

[0076] Disclosed herein is a pharmaceutical composition comprising an isolated polypeptide disclosed herein, and a pharmaceutically acceptable excipient disclosed herein. In some embodiments, the isolated polypeptide is an enzymatic product of the isolated recombinant polypeptide complex disclosed herein.

[0077] In some embodiments, the isolated recombinant polypeptide complex is administered at least once weekly. In some embodiments, the isolated recombinant polypeptide complex is administered once weekly. In some embodiments, the isolated recombinant polypeptide complex is administered at least twice weekly. In some embodiments, the isolated recombinant polypeptide complex is administered for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks. In some embodiments, the isolated recombinant polypeptide complex is administered for at least 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. In some embodiments, the isolated recombinant polypeptide complex is administered for at least 1 year, 2 years, 3 years, 4 years, or 5 years. In some embodiments, the isolated recombinant polypeptide complex is administered for at most 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, or 10 weeks. In some embodiments, the isolated recombinant polypeptide complex is administered for at most 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, or 12 months. In some embodiments, the isolated recombinant polypeptide complex is administered for at most 1 year, 2 years, 3 years, 4 years, or 5 years.Methods of Treatment

[0078] Disclosed herein are methods of treating a subject having cancer, the method comprising: administering to the subject any of the antibodies that bind specifically to EGFR and CD3 as disclosed herein. In some embodiments, the cancer comprises cancer cells that express EGFR or CD3. In some embodiments, the cancer cells that express EGFR or CD3 are lysed. In some embodiments, the antibody induces antibody -dependent cellular phagocytosis (ADCP) of the cancer cells that express EGFR or CD3.

[0079] In some embodiments, the isolated recombinant polypeptide complex described herein is used in a method of treating cancer. In some embodiments, the isolated polypeptide disclosed herein is used in a method of treating cancer and the isolated polypeptide is an enzymatic product of the isolated recombinant polypeptide complex after the administering. In some embodiments, the cancer has cells that express EGFR. In some embodiments, the polypeptides or polypeptide complexes described herein are used in a method of treating renal cell carcinoma (RCC), colorectal cancer (CRC), squamous cell carcinoma of the head and neck (SCCHN), non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple -negative breast cancer (TNBC), prostate cancer, breast cancer, colon / rectum cancer, head and neck cancer, esophagogastric cancer, liver cancer, glioblastoma, cervical cancer, ovarian cancer, bladder cancer, kidney cancer, or pancreatic cancer. In some embodiments, the polypeptides or polypeptide complexes described herein are used in a method oftreating subjects who are resistant to EGFR inhibitor treatment. In some embodiments, the polypeptides or polypeptide complexes described herein are used in a method of treating subjects who harbor KRAS mutations. In some embodiments, the polypeptides or polypeptide complexes described herein are used in a method of treating subjects who are resistant to EGFR inhibitor treatment and harbor KRAS mutations.

[0080] Described herein, in some embodiments, is an isolated recombinant polypeptide complex administered as once weekly. In some embodiments, the isolated recombinant polypeptide complex is administered once weekly by intravenous, intramuscular, intralesional, topical, subcutaneous, infusion, or oral. In some embodiments, the isolated recombinant polypeptide complex is administered once weekly by bolus injection. In some embodiments, the isolated recombinant polypeptide complex is administered once weekly by continuous infusion. In some embodiments, the isolated recombinant polypeptide complex is administered to the subject once a week as a continuous infusion over a period of no more than 60 minutes. In some embodiments, the isolated recombinant polypeptide complex is administered to the subject once a week as a continuous intravenous infusion over a period of no more than 30 minutes. In some embodiments, the isolated recombinant polypeptide complex is administered to the subject once a week as a continuous intravenous infusion over a period of at least 10 minutes.

[0081] In some embodiments, the method further comprises administering to the subject an anti -cancer agent. In some embodiments, the anti -cancer agent is a chemotherapeutic agent or a biologic agent. In some embodiments, the administering is sufficient to reduce or eliminate the cancer as compared to a comparable method lacking the administering.

[0082] In some embodiments, are methods of treating cancer in a subject need in need thereof comprising administering to the subject an isolated recombinant polypeptide complex as described herein. In some embodiments, the cancer has cells that express EGFR.

[0083] For administration to a subject, the isolated recombinant polypeptide complex as disclosed herein, may be provided in a pharmaceutical composition together with one or more pharmaceutically acceptable carriers or excipients. The term "pharmaceutically acceptable carrier" includes, but is not limited to, any carrier that does not interfere with the effectiveness of the biological activity of the ingredients and that is not toxic to the patient to whom it is administered. Examples of suitable pharmaceutical carriers are well known in the art and include phosphate buffered saline solutions, water, emulsions, such as oil / water emulsions, various types of wetting agents, sterile solutions etc. Such carriers can be formulated by conventional methods and can be administered to the subject at a suitable dose. Preferably, the compositions are sterile. These compositions may also contain adjuvants such as preservative, emulsifying agents and dispersing agents. Prevention of the action of microorganisms may be ensured by the inclusion of various antibacterial and antifungal agents.

[0084] The pharmaceutical composition may be in any suitable form, (depending upon the desired method of administration). It may be provided in unit dosage form, may be provided in a sealed container and may be provided as part of a kit. Such a kit may include instructions for use. It may include a plurality of said unit dosage forms.

[0085] The pharmaceutical composition may be adapted for administration by any appropriate route, including a parenteral (e.g., subcutaneous, intramuscular, or intravenous) route. Such compositions may be prepared by any method known in the art of pharmacy, for example by mixing the active ingredient with the carrier(s) or excipient(s) under sterile conditions. In some embodiments, a pharmaceutical composition disclosed herein is administered intravenously to a subject in need thereof.

[0086] Dosages of the substances of the present disclosure can vary between wide limits, depending upon the disease or disorder to be treated, the age and condition of the individual to be treated, etc. and a physician will ultimately determine appropriate dosages to be used.

[0087] In some embodiments, disclosed herein is a method for treating cancer comprising administering to a subject in need thereof an effective amount of a pharmaceutical composition. In some embodiments, the pharmaceutical composition comprises a dose of the isolated recombinant polypeptide complex disclosed herein. In some embodiments, the pharmaceutical composition comprises a pharmaceutically acceptable excipient. In some embodiments, the cancer comprises a cancer cell expressing epidermal growth factor receptor (EGFR). In some embodiments, the cancer comprises a cancer cell overexpressing EGFR.

[0088] In some embodiments, the cancer comprises CRC, SCCHN, NSCLC, SCLC, renal cell carcinoma (RCC), TNBC, breast cancer, pancreatic cancer, pancreatic ductal adenocarcinoma (PDAC), ovarian cancer, prostate cancer, brain cancer, glioblastoma multiforme, or papillary carcinoma. In some embodiments, the cancer comprises CRC. In some embodiments, the cancer comprises SCCHN. In some embodiments, the cancer comprises NSCLC. In some embodiments, the cancer comprises SCLC. In some embodiments, the cancer comprises RCC. In some embodiments, the cancer comprises TNBC. In some embodiments, the cancer comprises breast cancer. In some embodiments, the cancer comprises pancreatic cancer. In some embodiments, the cancer comprises PDAC. In some embodiments, the cancer comprises ovarian cancer. In some embodiments, the cancer comprises prostate cancer. In some embodiments, the cancer comprises brain cancer. In some embodiments, the cancer comprises glioblastoma multiforme. In some embodiments, the cancer comprises papillary carcinoma. In some embodiments, the cancer is metastatic, refractory, or relapsed. In some embodiments, the cancer is metastatic. In some embodiments, the cancer is refractory. In some embodiments, the cancer is relapsed. In some embodiments, the cancer is advanced or non-advanced. In some embodiments, the cancer is advanced. In some embodiments, the cancer is non-advanced.

[0089] In some embodiments, disclosed herein is a method for treating cancer. In some embodiments, the method comprises administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex. In some embodiments, the first dose is at least about 50 pg. In some embodiments, the isolated recombinant polypeptide complex comprises a first chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1 and a second chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2.

[0090] In some embodiments, the first chain comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acidsequence having at least 71% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 72% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 73% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 74% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 75% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 76% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 77% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 78% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 79% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 81% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 82% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 83% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 84% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 86% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 87% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 88% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 89% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 1. In some embodiments, the first chain comprises the amino acid sequence of SEQ ID NO: 1.

[0091] In some embodiments, the second chain comprises an amino acid sequence having at least 70%sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 71% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 72% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 73% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 74% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 75% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 76% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 77% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 78% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 79% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 81% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 82% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 83% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 84% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 86% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 87% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 88% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 89% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 2. In some embodiments, the second chain comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, thesecond chain comprises the amino acid sequence of SEQ ID NO: 2.

[0092] In some embodiments, the first chain comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 70% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 71% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 71% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 72% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 72% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 73% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 73% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 74% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 74% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 75% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 75% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 76% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 76% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 77% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 77% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 78% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 78% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 79% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 79% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 81% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 81% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 82% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 82% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 83% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 83% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 84% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 84% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having atleast 85% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 86% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 86% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 87% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 87% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 88% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 88% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 89% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 89% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 91% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 92% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 93% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 94% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 95% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 96% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 97% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 98% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 1 and the second chain comprises an amino acid sequence having at least 99% sequence identity to SEQ ID NO: 2. In some embodiments, the first chain comprises the amino acid sequence of SEQ ID NO: 1 and the second chain comprises the amino acid sequence of SEQ ID NO: 2.

[0093] In some embodiments, the first dose is about 50 pg to about 30 mg, e.g., about 50 pg, 51 pg, 52 pg, 53 pg, 54 pg, 55 pg, 56 pg, 57 pg, 58 pg, 59 pg, 60 pg, 61 pg, 62 pg, 63 pg, 64 pg, 65 pg, 66 pg, 67 pg, 68 pg, 69 pg, 70 pg, 71 pg, 72 pg, 73 pg, 74 pg, 75 pg, 76 pg, 77 pg, 78 pg, 79 pg, 80 pg, 81 pg, 82j g, 83 pg, 84 pg, 85 g, 86 pg, 87 pg, 88 pg, 89 pg, 90 pg, 91 pg, 92 pg, 93 pg, 94 pg, 95 pg, 96 pg, 97 pg, 98 pg, 99 pg, 100 pg, 105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg,215 pg, 220 pg, 225 pg, 230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg, 275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg,340 pg, 345 pg, 350 pg, 355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg, 400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg,465 pg, 470 pg, 475 pg, 480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg, 525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg,590 pg, 595 pg, 600 pg, 605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg, 650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg,715 pg, 720 pg, 725 pg, 730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg, 775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg,840 pg, 845 pg, 850 pg, 855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg, 900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg,965 pg, 970 pg, 975 pg, 980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, or about 30 mg, or any dose therebetween. In some embodiments, the first dose is about 100 pg. In some embodiments, the first dose is about 150 pg. In some embodiments, the first dose is about 200 pg. In some embodiments, the first dose is about 250 pg. In some embodiments, the first dose is about 300 pg. In some embodiments, the first dose is about 350 pg. In some embodiments, the first dose is about 400 pg. In some embodiments, the first dose is about 450 pg. In some embodiments, the first dose is about 500 pg. In some embodiments, the first dose is about 750 pg. In some embodiments, the first dose is about 1 mg. In some embodiments, the first dose is about 1.5 mg. In some embodiments, the first dose is about 2 mg. In some embodiments, the first dose is about 3 mg. In some embodiments, the first dose is about 4 mg. In some embodiments, the first dose is about 5 mg. In some embodiments, the first dose is about 6 mg. In some embodiments, the first dose is about 7 mg. In some embodiments, the first dose is about 8 mg. In some embodiments, the first dose is about 9 mg. In some embodiments, the first dose is about 10 mg. In some embodiments, the first dose is about 15 mg. In some embodiments, the first dose is about 20 mg. In some embodiments, the first dose is about 25 mg. In some embodiments, the first dose is about 30 mg.

[0094] In some embodiments, the first dose is at least about 100 pg, 105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg, 215 pg, 220 pg, 225 pg, 230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg, 275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg, 340 pg, 345 pg, 350 pg, 355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg, 400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg, 465 pg, 470 pg, 475 pg, 480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg, 525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg, 590 pg, 595 pg, 600 pg, 605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg, 650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg, 715 pg, 720 pg, 725 pg, 730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg, 775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg, 840 pg, 845 pg, 850 pg, 855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg, 900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg, 965 pg, 970 pg, 975 pg, 980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1. 1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg,3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg,5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg,7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg,8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, or at least about 30 mg, or any dose therebetween. In some embodiments, the first dose is at least about 100 pg. In some embodiments, the first dose is at least about 150 pg. In some embodiments, the first dose is at least about 200 pg. In some embodiments, the first dose is at least about 250 pg. In some embodiments, the first dose is at least about 300 pg. In some embodiments, the first dose is at least about 350 pg. In some embodiments, the first dose is at least about 400 pg. In some embodiments, the first dose is at least about 450 pg. In some embodiments, the first dose is at least about 500 pg. In some embodiments, the first dose is at least about 750 pg. In some embodiments, the first dose is at least about 1 mg. In some embodiments, the first dose is at least about 1.5 mg. In some embodiments, the first dose is at least about 2 mg. In some embodiments, the first dose is at least about 3 mg. In some embodiments, the first dose is at least about 4 mg. In some embodiments, the first dose is at least about 5 mg. In some embodiments, the first dose is at least about 6 mg. In some embodiments, the first dose is at least about 7 mg. In some embodiments, the first dose is at least about 8 mg. In some embodiments, the first dose is at least about 9 mg. In some embodiments, the first dose is at least about 10 mg. In some embodiments, the first dose is at least about 15 mg. In some embodiments, the first dose is at least about 20 mg. In some embodiments, the first dose is at least about 25 mg. In someembodiments, the first dose is at least about 30 mg.

[0095] In some embodiments, the first dose is at least about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg,10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg,11.6 mg, 11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg,12.7 mg, 12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg,13.8 mg, 13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg,14.9 mg, 15 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg, 16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg,17.2 mg, 17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg,18.3 mg, 18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg,19.4 mg, 19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg,20.5 mg, 20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg,21.6 mg, 21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg,22.7 mg, 22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg,23.8 mg, 23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg,24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg, 26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg,27.2 mg, 27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg,28.3 mg, 28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg,29.4 mg, 29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, or at least about 100 mg, or any dose therebetween. In some embodiments, the first dose is at least about 10 mg. In some embodiments, the first dose is at least about 15 mg. In some embodiments, the first dose is at least about 20 mg. In some embodiments, the first dose is at least about 25 mg. In some embodiments, the first dose is at least about 30 mg. In some embodiments, the first dose is at least about 40 mg. In some embodiments, the first dose is at least about 50 mg. In some embodiments, the first dose is at least about 60 mg. In some embodiments, the first dose is at least about 70 mg. In some embodiments, the first dose is at least about 80 mg. In some embodiments, the first dose is at least about 90 mg. In some embodiments, the first dose is at least about 100 mg.

[0096] In some embodiments, the first dose is about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg, 10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg, 11.6 mg,11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg, 12.7 mg,12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg, 13.8 mg,13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg, 14.9 mg, 15mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg,16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg, 17.2 mg,17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg, 18.3 mg,18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg, 19.4 mg,19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg, 20.5 mg,20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg, 21.6 mg,21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg, 22.7 mg,22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg, 23.8 mg,23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg, 24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg,26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg, 27.2 mg,27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg, 28.3 mg,28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg, 29.4 mg,29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, or at least about 100 mg, or any dose therebetween. In some embodiments, the first dose is about 10 mg. In some embodiments, the first dose is about 15 mg. In some embodiments, the first dose is about 20 mg. In some embodiments, the first dose is about 25 mg. In some embodiments, the first dose is about 30 mg. In some embodiments, the first dose is about 40 mg. In some embodiments, the first dose is about 50 mg. In some embodiments, the first dose is about 60 mg. In some embodiments, the first dose is about 70 mg. In some embodiments, the first dose is about 80 mg. In some embodiments, the first dose is about 90 mg. In some embodiments, the first dose is about 100 mg.

[0097] In some embodiments, the first dose is at least about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg,117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg,129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg,141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg,153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg,165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg,177 mg, 178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg,189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg,201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg,213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg,225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg,237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg,249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg,261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg,273 mg, 274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg,285 mg, 286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg,297 mg, 298 mg, 299 mg, or at least about 300 mg, or any dose therebetween. In some embodiments, the first dose is at least about 100 mg. In some embodiments, the first dose is at least about 150 mg. In some embodiments, the first dose is at least about 200 mg. In some embodiments, the first dose is at least about250 mg. In some embodiments, the first dose is at least about 300 mg.

[0098] In some embodiments, the first dose is about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg,118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg,130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg,142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg,154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg,166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg,178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg,190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg,202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg,214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg,226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg,238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg,250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg,262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg, 273 mg,274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg, 285 mg,286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg, 297 mg,298 mg, 299 mg, or about 300 mg, or any dose therebetween. In some embodiments, the first dose is about 100 mg. In some embodiments, the first dose is about 150 mg. In some embodiments, the first dose is about 200 mg. In some embodiments, the first dose is about 250 mg. In some embodiments, the first dose is about 300 mg.

[0099] In some embodiments, the method further comprises administering to the subject a target dose of the isolated recombinant polypeptide complex, wherein the target dose is higher than the first dose, and wherein the target dose is administered after the first dose. In some embodiments, the method further comprises administering to the subject a second dose of the isolated recombinant polypeptide complex, wherein the second dose is higher than the first dose and lower than the target dose, and wherein the second dose is administered between the first dose and the target dose. In some embodiments, the first dose is administered weekly. In some embodiments, the first dose is administered once every two weeks. In some embodiments, the first dose is administered once every three weeks. In some embodiments, the target dose is administered weekly. In some embodiments, the target dose is administered once every twoweeks. In some embodiments, the target dose is administered once every three weeks. In some embodiments, the second dose is administered weekly.

[0100] In some embodiments, the second dose is administered once every two weeks. In some embodiments, the second dose is administered once every three weeks. In some embodiments, the method comprises a treatment cycle that starts on day 1. In some embodiments, the first dose is administered on day 1 of the treatment cycle. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle. In some embodiments, the first dose, the second dose, and the target dose are administered on day 1, day 4, and day 8, respectively, of the treatment cycle. In some embodiments, the target dose is administered weekly after day 8 of the treatment cycle. In some embodiments, the target dose is administered once every two weeks after day 8 of the treatment cycle. In some embodiments, the target dose is administered once every three weeks after day 8 of the treatment cycle.

[0101] In some embodiments, the target dose is at least about 50 pg, 51 pg, 52 pg, 53 pg, 54 pg, 55 pg,56 pg, 57 pg, 58 pg, 59 pg, 60 pg, 61 pg, 62 pg, 63 pg, 64 pg, 65 pg, 66 pg, 67 pg, 68 pg, 69 pg, 70 pg,71 pg, 72 pg, 73 pg, 74 pg, 75 pg, 76 pg, 77 pg, 78 pg, 79 pg, 80 pg, 81 pg, 82 pg, 83 pg, 84 pg, 85 pg,86 pg, 87 pg, 88 pg, 89 pg, 90 pg, 91 pg, 92 pg, 93 pg, 94 pg, 95 pg, 96 pg, 97 pg, 98 pg, 99 pg, 100 pg,105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg, 215 pg, 220 pg, 225 pg,230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg, 275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg, 340 pg, 345 pg, 350 pg,355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg, 400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg, 465 pg, 470 pg, 475 pg,480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg, 525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg, 590 pg, 595 pg, 600 pg,605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg, 650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg, 715 pg, 720 pg, 725 pg,730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg, 775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg, 840 pg, 845 pg, 850 pg,855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg, 900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg, 965 pg, 970 pg, 975 pg,980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg,4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg,5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg,8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg,9.7 mg, 9.8 mg, 9.9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20mg, 21 mg, 22 mg, 23 mg 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 35 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, or at least about 300 mg, or any dose therebetween. In some embodiments, the target dose is at least about 100 pg. In some embodiments, the target dose is at least about 250 pg. In some embodiments, the target dose is at least about 500 pg. In some embodiments, the target dose is at least about 750 pg. In some embodiments, the target dose is at least about 1 mg. In some embodiments, the target dose is at least about 1.25 mg. In some embodiments, the target dose is at least about 1.5 mg. In some embodiments, the target dose is at least about 2 mg. In some embodiments, the target dose is at least about 5 mg. In some embodiments, the target dose is at least about 7.5 mg. In some embodiments, the target dose is at least about 10 mg. In some embodiments, the target dose is at least about 15 mg. In some embodiments, the target dose is at least about 50 mg. In some embodiments, the target dose is at least about 100 mg. In some embodiments, the target dose is at least about 150 mg. In some embodiments, the target dose is at least about 300 mg.

[0102] In some embodiments, the target dose is about 50 pg to about 300 mg, e.g., about 50 pg, 51 pg,52 pg, 53 pg, 54 pg, 55 pg, 56 pg, 57 pg, 58 pg, 59 pg, 60 pg, 61 pg, 62 pg, 63 pg, 64 pg, 65 pg, 66 pg,67 g, 68 pg, 69 pg, 70 pg, 71 pg, 72 pg, 73 pg, 74 pg, 75 pg, 76 pg, 77 pg, 78 pg, 79 pg, 80 pg, 81 pg,82 pg, 83 pg, 84 pg, 85 pg, 86 pg, 87 pg, 88 pg, 89 pg, 90 pg, 91 pg, 92 pg, 93 pg, 94 pg, 95 pg, 96 pg,97 pg, 98 pg, 99 pg, 100 pg, 105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg, 215 pg, 220 pg, 225 pg, 230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg, 275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg, 340 pg, 345 pg, 350 pg, 355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg, 400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg, 465 pg, 470 pg, 475 pg, 480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg, 525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg, 590 pg, 595 pg, 600 pg, 605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg, 650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg, 715 pg, 720 pg, 725 pg, 730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg, 775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg, 840 pg, 845 pg, 850 pg, 855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg, 900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg, 965 pg, 970 pg, 975 pg, 980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, 10 mg, 11 mg, 12 mg, 13 mg, 14 mg, 15 mg, 16 mg, 17 mg, 18 mg, 19 mg, 20 mg, 21 mg, 22 mg, 23 mg 24 mg, 25 mg, 26 mg, 27 mg, 28 mg, 29 mg, 30 mg, 35 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, or about 300 mg, or any dose therebetween. In some embodiments, the target dose is about 100 gg. In some embodiments, the target dose is about 250 gg. In some embodiments, the target dose is about 500 gg. In some embodiments, the target dose is about 750 gg. In some embodiments, the target dose is about 1 mg. In some embodiments, the target dose is about 1.25 mg. In some embodiments, the target dose is about 1.5 mg. In some embodiments, the target dose is about 2 mg. In some embodiments, the target dose is about 5 mg. In some embodiments, the target dose is about 7.5 mg. In some embodiments, the target dose is about 10 mg. In some embodiments, the first dose is at least about 500 gg and the target dose is at least about 1.25 mg. In some embodiments, the first dose is about 500 gg and the target dose is about 1.25 mg. In some embodiments, the first dose is about 500 gg and the target dose is about 5 mg. In some embodiments, the first dose is about 1.5 mg and the target dose is about 15 mg. In some embodiments, the first dose is about 5 mg and the target dose is about 50 mg. In some embodiments, the first dose is about 15 mg and the target dose is about 150 mg. In some embodiments, the first dose is about 30 mg and the target dose is about 300 mg.

[0103] In some embodiments, the target dose is at least about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg, 10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg,11.6 mg, 11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg,12.7 mg, 12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg,13.8 mg, 13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg,14.9 mg, 15 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg, 16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg,17.2 mg, 17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg,18.3 mg, 18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg,19.4 mg, 19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg,20.5 mg, 20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg,21.6 mg, 21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg,22.7 mg, 22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg,23.8 mg, 23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg,24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg, 26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg,27.2 mg, 27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg,28.3 mg, 28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg,29.4 mg, 29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, or at least about 300 mg or any dose therebetween. In some embodiments, the target dose is at least about 10 mg. In some embodiments, the target dose is at least about 15 mg. In some embodiments, the target dose is at least about 20 mg. In some embodiments, the target dose is at least about 25 mg. In some embodiments, the target dose is at least about 30 mg. In some embodiments, the target dose is at least about 40 mg. In some embodiments, the target dose is at least about 50 mg. In some embodiments, the target dose is at least about 60 mg. In some embodiments, the target dose is at least about 70 mg. In some embodiments, the target dose is at least about 80 mg. In some embodiments, the target dose is at least about 90 mg. In some embodiments, the target dose is at least about 100 mg. In some embodiments, the target dose is at least about 150 mg. In some embodiments, the target dose is at least about 300 mg.

[0104] In some embodiments, the target dose is about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg, 10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg, 11.6 mg,11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg, 12.7 mg,12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg, 13.8 mg,13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg, 14.9 mg, 15 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg,16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg, 17.2 mg,17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg, 18.3 mg,18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg, 19.4 mg,19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg, 20.5 mg,20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg, 21.6 mg,21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg, 22.7 mg,22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg, 23.8 mg,23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg, 24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg,26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg, 27.2 mg,27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg, 28.3 mg,28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg, 29.4 mg,29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, 100 mg, , 110 mg, 120 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 260 mg, 270 mg, 280 mg, 290 mg, or at least about 300 mg, or any dose therebetween. In some embodiments, the target dose is about 10 mg. In some embodiments, the target dose is about 15 mg. In some embodiments, the target dose is about 20 mg. In some embodiments, the target dose is about 25 mg. In some embodiments, the target dose is about 30 mg. In some embodiments, the target dose is about 40 mg. In some embodiments, the target dose is about 50 mg. In some embodiments, the target dose is about 60 mg. In some embodiments, the target dose is about 70 mg. In some embodiments, the target dose is about 80 mg. In some embodiments, the target dose is about 90 mg. In some embodiments, the target dose is about 100 mg. In some embodiments, the target dose is about 150 mg. In some embodiments, the target dose is about 300 mg.

[0105] In some embodiments, the target dose is at least about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg, 178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg, 273 mg, 274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg, 285 mg, 286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg, 297 mg, 298 mg, 299 mg, or at least about 300 mg, or any dose therebetween. In some embodiments, the target dose is at least about 100 mg. In some embodiments, the target dose is at least about 150 mg. In some embodiments, the target dose is at least about 200 mg. In some embodiments, the target dose is at least about 250 mg. In some embodiments, the target dose is at least about 300 mg.

[0106] In some embodiments, the target dose is about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg, 178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg, 13 mg, 274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg, 285 mg, 286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg, 297 mg, 298 mg, 299 mg, or about 300 mg, or any dose therebetween. In some embodiments, the target dose is about 100 mg. In some embodiments, the target dose is about 150 mg. In some embodiments, the target dose is about 200 mg. In some embodiments, the target dose is about 250 mg. In some embodiments, the target dose is about 300 mg.

[0107] In some embodiments, the second dose is at least about 50 pg, 51 pg, 52 pg, 53 pg, 54 pg, 55 pg, 56 g, 57 pg, 58 pg, 59 pg, 60 pg, 61 pg, 62 pg, 63 pg, 64 pg, 65 pg, 66 pg, 67 pg, 68 pg, 69 pg, 70 pg, 71 pg, 72 pg, 73 pg, 74 pg, 75 pg, 76 pg, 77 pg, 78 pg, 79 pg, 80 pg, 81 pg, 82 pg, 83 pg, 84 pg, 85 pg, 86 pg, 87 pg, 88 pg, 89 pg, 90 pg, 91 pg, 92 pg, 93 pg, 94 pg, 95 pg, 96 pg, 97 pg, 98 pg, 99 pg, 100 pg, 105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg, 215 pg, 220 pg, 225 pg, 230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg, 275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg, 340 pg, 345 pg, 350 pg, 355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg, 400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg, 465 pg, 470 pg, 475 pg, 480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg, 525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg, 590 pg, 595 pg, 600 pg, 605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg, 650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg, 715 pg, 720 pg, 725 pg, 730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg, 775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg, 840 pg, 845 pg, 850 pg, 855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg, 900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg, 965 pg, 970 pg, 975 pg, 980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg,1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg,3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg,4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg,5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg,7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, or at least about 10 mg, or any dose therebetween. In some embodiments, the second dose is at least about 100 pg. In some embodiments, the second dose is at least about 250 pg. In some embodiments, the second dose is at least about 500 pg. In some embodiments, the second dose is at least about 750 pg. In some embodiments, the second dose is at least about 1 mg. In some embodiments, the second dose is at least about 1.25 mg. In some embodiments, the second dose is at least about 1.5 mg. In some embodiments, the second dose is at least about 2 mg. In some embodiments, the second dose is at least about 5 mg. In some embodiments, the second dose is at least about 7.5 mg. In some embodiments, the second dose is at least about 10 mg.

[0108] In some embodiments, the second dose is about 50 pg to about 10 mg, e.g., about 50 pg, 51 pg,52 pg, 53 pg, 54 pg, 55 pg, 56 pg, 57 pg, 58 pg, 59 pg, 60 pg, 61 pg, 62 pg, 63 pg, 64 pg, 65 pg, 66 pg,67 g, 68 pg, 69 pg, 70 pg, 71 pg, 72 pg, 73 pg, 74 pg, 75 pg, 76 pg, 77 pg, 78 pg, 79 pg, 80 pg, 81 pg,82 pg, 83 pg, 84 pg, 85 pg, 86 pg, 87 pg, 88 pg, 89 pg, 90 pg, 91 pg, 92 pg, 93 pg, 94 pg, 95 pg, 96 pg,97 pg, 98 pg, 99 pg, 100 pg, 105 pg, 110 pg, 115 pg, 120 pg, 125 pg, 130 pg, 135 pg, 140 pg, 145 pg, 150 pg, 155 pg, 160 pg, 165 pg, 170 pg, 175 pg, 180 pg, 185 pg, 190 pg, 195 pg, 200 pg, 205 pg, 210 pg, 215 pg, 220 pg, 225 pg, 230 pg, 235 pg, 240 pg, 245 pg, 250 pg, 255 pg, 260 pg, 265 pg, 270 pg,275 pg, 280 pg, 285 pg, 290 pg, 295 pg, 300 pg, 305 pg, 310 pg, 315 pg, 320 pg, 325 pg, 330 pg, 335 pg, 340 pg, 345 pg, 350 pg, 355 pg, 360 pg, 365 pg, 370 pg, 375 pg, 380 pg, 385 pg, 390 pg, 395 pg,400 pg, 405 pg, 410 pg, 415 pg, 420 pg, 425 pg, 430 pg, 435 pg, 440 pg, 445 pg, 450 pg, 455 pg, 460 pg, 465 pg, 470 pg, 475 pg, 480 pg, 485 pg, 490 pg, 495 pg, 500 pg, 505 pg, 510 pg, 515 pg, 520 pg,525 pg, 530 pg, 535 pg, 540 pg, 545 pg, 550 pg, 555 pg, 560 pg, 565 pg, 570 pg, 575 pg, 580 pg, 585 pg, 590 pg, 595 pg, 600 pg, 605 pg, 610 pg, 615 pg, 620 pg, 625 pg, 630 pg, 635 pg, 640 pg, 645 pg,650 pg, 655 pg, 660 pg, 665 pg, 670 pg, 675 pg, 680 pg, 685 pg, 690 pg, 695 pg, 700 pg, 705 pg, 710 pg, 715 pg, 720 pg, 725 pg, 730 pg, 735 pg, 740 pg, 745 pg, 750 pg, 755 pg, 760 pg, 765 pg, 770 pg,775 pg, 780 pg, 785 pg, 790 pg, 795 pg, 800 pg, 805 pg, 810 pg, 815 pg, 820 pg, 825 pg, 830 pg, 835 pg, 840 pg, 845 pg, 850 pg, 855 pg, 860 pg, 865 pg, 870 pg, 875 pg, 880 pg, 885 pg, 890 pg, 895 pg,900 pg, 905 pg, 910 pg, 915 pg, 920 pg, 925 pg, 930 pg, 935 pg, 940 pg, 945 pg, 950 pg, 955 pg, 960 pg, 965 pg, 970 pg, 975 pg, 980 pg, 985 pg, 990 pg, 995 pg, 1 mg, 1.1 mg, 1.2 mg, 1.3 mg, 1.4 mg, 1.5 mg, 1.6 mg, 1.7 mg, 1.8 mg, 1.9 mg, 2 mg, 2.1 mg, 2.2 mg, 2.3 mg, 2.4 mg, 2.5 mg, 2.6 mg, 2.7 mg, 2.8 mg, 2.9 mg, 3 mg, 3.1 mg, 3.2 mg, 3.3 mg, 3.4 mg, 3.5 mg, 3.6 mg, 3.7 mg, 3.8 mg, 3.9 mg, 4 mg, 4.1 mg, 4.2 mg, 4.3 mg, 4.4 mg, 4.5 mg, 4.6 mg, 4.7 mg, 4.8 mg, 4.9 mg, 5 mg, 5.1 mg, 5.2 mg, 5.3 mg, 5.4 mg, 5.5 mg, 5.6 mg, 5.7 mg, 5.8 mg, 5.9 mg, 6 mg, 6.1 mg, 6.2 mg, 6.3 mg, 6.4 mg, 6.5 mg, 6.6 mg, 6.7 mg, 6.8 mg, 6.9 mg, 7 mg, 7.1 mg, 7.2 mg, 7.3 mg, 7.4 mg, 7.5 mg, 7.6 mg, 7.7 mg, 7.8 mg, 7.9 mg, 8 mg, 8.1 mg, 8.2 mg, 8.3 mg, 8.4 mg, 8.5 mg, 8.6 mg, 8.7 mg, 8.8 mg, 8.9 mg, 9 mg, 9.1 mg, 9.2 mg, 9.3 mg, 9.4 mg, 9.5 mg, 9.6 mg, 9.7 mg, 9.8 mg, 9.9 mg, or about 10 mg, or any dose therebetween. In some embodiments, the second dose is about 100 pg. In some embodiments, the second dose is about 250 pg. In some embodiments, the second dose is about 500 pg. In some embodiments, the second dose is about750 |ig. In some embodiments, the second dose is about 1 mg. In some embodiments, the second dose is about 1.25 mg. In some embodiments, the second dose is about 1.5 mg. In some embodiments, the second dose is about 2 mg. In some embodiments, the second dose is about 5 mg. In some embodiments, the second dose is about 7.5 mg. In some embodiments, the second dose is about 10 mg. In some embodiments, the first dose is at least about 500 gg and the second dose is at least about 1.25 mg. In some embodiments, the first dose is about 500 gg and the second dose is about 1.25 mg.

[0109] In some embodiments, the second dose is at least about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg, 10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg,11.6 mg, 11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg,12.7 mg, 12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg,13.8 mg, 13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg,14.9 mg, 15 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg, 16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg,17.2 mg, 17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg,18.3 mg, 18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg,19.4 mg, 19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg,20.5 mg, 20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg,21.6 mg, 21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg,22.7 mg, 22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg,23.8 mg, 23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg,24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg, 26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg,27.2 mg, 27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg,28.3 mg, 28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg,29.4 mg, 29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, or at least about 100 mg, or any dose therebetween. In some embodiments, the second dose is at least about 10 mg. In some embodiments, the second dose is at least about 15 mg. In some embodiments, the second dose is at least about 20 mg. In some embodiments, the second dose is at least about 25 mg. In some embodiments, the second dose is at least about 30 mg. In some embodiments, the second dose is at least about 40 mg. In some embodiments, the second dose is at least about 50 mg. In some embodiments, the second dose is at least about 60 mg. In some embodiments, the second dose is at least about 70 mg. In some embodiments, the second dose is at least about 80 mg. In some embodiments, the second dose is at least about 90 mg. In some embodiments, the second dose is at least about 100 mg.

[0110] In some embodiments, the second dose is about 10 mg, 10.1 mg, 10.2 mg, 10.3 mg, 10.4 mg,10.5 mg, 10.6 mg, 10.7 mg, 10.8 mg, 10.9 mg, 11 mg, 11.1 mg, 11.2 mg, 11.3 mg, 11.4 mg, 11.5 mg,11.6 mg, 11.7 mg, 11.8 mg, 11.9 mg, 12 mg, 12.1 mg, 12.2 mg, 12.3 mg, 12.4 mg, 12.5 mg, 12.6 mg,12.7 mg, 12.8 mg, 12.9 mg, 13 mg, 13.1 mg, 13.2 mg, 13.3 mg, 13.4 mg, 13.5 mg, 13.6 mg, 13.7 mg,13.8 mg, 13.9 mg, 14 mg, 14.1 mg, 14.2 mg, 14.3 mg, 14.4 mg, 14.5 mg, 14.6 mg, 14.7 mg, 14.8 mg,14.9 mg, 15 mg, 15.1 mg, 15.2 mg, 15.3 mg, 15.4 mg, 15.5 mg, 15.6 mg, 15.7 mg, 15.8 mg, 15.9 mg, 16 mg, 16.1 mg, 16.2 mg, 16.3 mg, 16.4 mg, 16.5 mg, 16.6 mg, 16.7 mg, 16.8 mg, 16.9 mg, 17 mg, 17.1 mg,17.2 mg, 17.3 mg, 17.4 mg, 17.5 mg, 17.6 mg, 17.7 mg, 17.8 mg, 17.9 mg, 18 mg, 18.1 mg, 18.2 mg,18.3 mg, 18.4 mg, 18.5 mg, 18.6 mg, 18.7 mg, 18.8 mg, 18.9 mg, 19 mg, 19.1 mg, 19.2 mg, 19.3 mg,19.4 mg, 19.5 mg, 19.6 mg, 19.7 mg, 19.8 mg, 19.9 mg, 20 mg, 20.1 mg, 20.2 mg, 20.3 mg, 20.4 mg,20.5 mg, 20.6 mg, 20.7 mg, 20.8 mg, 20.9 mg, 21 mg, 21.1 mg, 21.2 mg, 21.3 mg, 21.4 mg, 21.5 mg,21.6 mg, 21.7 mg, 21.8 mg, 21.9 mg, 22 mg, 22.1 mg, 22.2 mg, 22.3 mg, 22.4 mg, 22.5 mg, 22.6 mg,22.7 mg, 22.8 mg, 22.9 mg, 23 mg, 23.1 mg, 23.2 mg, 23.3 mg, 23.4 mg, 23.5 mg, 23.6 mg, 23.7 mg,23.8 mg, 23.9 mg, 24 mg, 24.1 mg, 24.2 mg, 24.3 mg, 24.4 mg, 24.5 mg, 24.6 mg, 24.7 mg, 24.8 mg,24.9 mg, 25 mg, 25.1 mg, 25.2 mg, 25.3 mg, 25.4 mg, 25.5 mg, 25.6 mg, 25.7 mg, 25.8 mg, 25.9 mg, 26 mg, 26.1 mg, 26.2 mg, 26.3 mg, 26.4 mg, 26.5 mg, 26.6 mg, 26.7 mg, 26.8 mg, 26.9 mg, 27 mg, 27.1 mg,27.2 mg, 27.3 mg, 27.4 mg, 27.5 mg, 27.6 mg, 27.7 mg, 27.8 mg, 27.9 mg, 28 mg, 28.1 mg, 28.2 mg,28.3 mg, 28.4 mg, 28.5 mg, 28.6 mg, 28.7 mg, 28.8 mg, 28.9 mg, 29 mg, 29.1 mg, 29.2 mg, 29.3 mg,29.4 mg, 29.5 mg, 29.6 mg, 29.7 mg, 29.8 mg, 29.9 mg, 30 mg, 31 mg, 32 mg, 33 mg, 34 mg, 35 mg, 36 mg, 37 mg, 38 mg, 39 mg, 40 mg, 41 mg, 42 mg, 43 mg, 44 mg, 45 mg, 46 mg, 47 mg, 48 mg, 49 mg, 50 mg, 51 mg, 52 mg, 53 mg, 54 mg, 55 mg, 56 mg, 57 mg, 58 mg, 59 mg, 60 mg, 61 mg, 62 mg, 63 mg, 64 mg, 65 mg, 66 mg, 67 mg, 68 mg, 69 mg, 70 mg, 71 mg, 72 mg, 73 mg, 74 mg, 75 mg, 76 mg, 77 mg, 78 mg, 79 mg, 80 mg, 81 mg, 82 mg, 83 mg, 84 mg, 85 mg, 86 mg, 87 mg, 88 mg, 89 mg, 90 mg, 91 mg, 92 mg, 93 mg, 94 mg, 95 mg, 96 mg, 97 mg, 98 mg, 99 mg, or at least about 100 mg, or any dose therebetween. In some embodiments, the second dose is about 10 mg. In some embodiments, the second dose is about 15 mg. In some embodiments, the second dose is about 20 mg. In some embodiments, the second dose is about 25 mg. In some embodiments, the second dose is about 30 mg. In some embodiments, the second dose is about 40 mg. In some embodiments, the second dose is about 50 mg. In some embodiments, the second dose is about 60 mg. In some embodiments, the second dose is about 70 mg. In some embodiments, the second dose is about 80 mg. In some embodiments, the second dose is about 90 mg. In some embodiments, the second dose is about 100 mg.

[0111] In some embodiments, the second dose is at least about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176mg, 177 mg, 178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg, 273 mg, 274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg, 285 mg, 286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg, 297 mg, 298 mg, 299 mg, or at least about 300 mg, or any dose therebetween. In some embodiments, the second dose is at least about 100 mg. In some embodiments, the second dose is at least about 150 mg. In some embodiments, the second dose is at least about 200 mg. In some embodiments, the second dose is at least about 250 mg. In some embodiments, the second dose is at least about 300 mg.

[0112] In some embodiments, the second dose is about 100 mg, 101 mg, 102 mg, 103 mg, 104 mg, 105 mg, 106 mg, 107 mg, 108 mg, 109 mg, 110 mg, 111 mg, 112 mg, 113 mg, 114 mg, 115 mg, 116 mg, 117 mg, 118 mg, 119 mg, 120 mg, 121 mg, 122 mg, 123 mg, 124 mg, 125 mg, 126 mg, 127 mg, 128 mg, 129 mg, 130 mg, 131 mg, 132 mg, 133 mg, 134 mg, 135 mg, 136 mg, 137 mg, 138 mg, 139 mg, 140 mg, 141 mg, 142 mg, 143 mg, 144 mg, 145 mg, 146 mg, 147 mg, 148 mg, 149 mg, 150 mg, 151 mg, 152 mg, 153 mg, 154 mg, 155 mg, 156 mg, 157 mg, 158 mg, 159 mg, 160 mg, 161 mg, 162 mg, 163 mg, 164 mg, 165 mg, 166 mg, 167 mg, 168 mg, 169 mg, 170 mg, 171 mg, 172 mg, 173 mg, 174 mg, 175 mg, 176 mg, 177 mg, 178 mg, 179 mg, 180 mg, 181 mg, 182 mg, 183 mg, 184 mg, 185 mg, 186 mg, 187 mg, 188 mg, 189 mg, 190 mg, 191 mg, 192 mg, 193 mg, 194 mg, 195 mg, 196 mg, 197 mg, 198 mg, 199 mg, 200 mg, 201 mg, 202 mg, 203 mg, 204 mg, 205 mg, 206 mg, 207 mg, 208 mg, 209 mg, 210 mg, 211 mg, 212 mg, 213 mg, 214 mg, 215 mg, 216 mg, 217 mg, 218 mg, 219 mg, 220 mg, 221 mg, 222 mg, 223 mg, 224 mg, 225 mg, 226 mg, 227 mg, 228 mg, 229 mg, 230 mg, 231 mg, 232 mg, 233 mg, 234 mg, 235 mg, 236 mg, 237 mg, 238 mg, 239 mg, 240 mg, 241 mg, 242 mg, 243 mg, 244 mg, 245 mg, 246 mg, 247 mg, 248 mg, 249 mg, 250 mg, 251 mg, 252 mg, 253 mg, 254 mg, 255 mg, 256 mg, 257 mg, 258 mg, 259 mg, 260 mg, 261 mg, 262 mg, 263 mg, 264 mg, 265 mg, 266 mg, 267 mg, 268 mg, 269 mg, 270 mg, 271 mg, 272 mg, 273 mg, 274 mg, 275 mg, 276 mg, 277 mg, 278 mg, 279 mg, 280 mg, 281 mg, 282 mg, 283 mg, 284 mg, 285 mg, 286 mg, 287 mg, 288 mg, 289 mg, 290 mg, 291 mg, 292 mg, 293 mg, 294 mg, 295 mg, 296 mg, 297 mg, 298 mg, 299 mg, or about 300 mg, or any dose therebetween. In some embodiments, the second dose is about 100 mg. In some embodiments, the second dose is about 150 mg. In some embodiments, the second dose is about 200 mg. In some embodiments, the second dose is about 250 mg. In some embodiments, the second dose is about 300 mg. In some embodiments, the first dose is about 500 gg, the second dose is about 1.5 mg, and the target dose is about 5 mg. In some embodiments, the first dose is about 1.5 mg, the second dose is about 5 mg, and the target dose is about 15 mg. In some embodiments, the first dose is about 5 mg, the second dose is about 15 mg, and the target dose is about 50 mg. In someembodiments, the first dose is about 15 mg, the second dose is about 50 mg, and the target dose is about 150 mg. In some embodiments, the first dose is about 30 mg, the second dose is about 100 mg, and the target dose is about 300 mg.

[0113] In some embodiments, the first dose, the second dose, or the target dose is administered at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, or more than 50 times. In some embodiments, the method further comprises at least 2 treatment cycles, at least 3 treatment cycles, at least 4 treatment cycles, at least 5 treatment cycles, at least 6 treatment cycles, at least 7 treatment cycles, at least 8 treatment cycles, at least 9 treatment cycles, at least 10 treatment cycles, or more than 10 treatment cycles. In some embodiments, the cancer comprises a cell that expresses epidermal growth factor receptor (EGFR). In some embodiments, the cancer comprises a cell that overexpresses EGFR. In some embodiments, the cancer comprises colorectal cancer (CRC), squamous cell carcinoma of head and neck (SCCHN), non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), breast cancer, bladder cancer, ovarian cancer, liver cancer, pancreatic cancer, small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple -negative breast cancer (TNBC), prostate cancer, or a combination thereof. In some embodiments, the cancer comprises advanced or metastatic NSCLC. In some embodiments, the cancer comprises advanced or metastatic SCCHN. In some embodiments, the cancer comprises advanced or metastatic CRC. In some embodiments, the cancer comprises advanced or metastatic RCC. In some embodiments, the cancer comprises advanced or metastatic SCLC. In some embodiments, the cancer comprises advanced or metastatic PDAC. In some embodiments, the cancer comprises advanced or metastatic TNBC. In some embodiments, the cancer comprises advanced or metastatic prostate cancer.

[0114] In some embodiments, the cancer progresses after treatment with a prior cancer therapy. In some embodiments, the cancer is refractory, intolerant, non-responsive, or resistant to a prior cancer therapy. In some embodiments, the subject is at least 18 years old. In some embodiments, the subject is about 37 to about 81 years old. In some embodiments, the subject is about 65 years old. In some embodiments, the prior cancer therapy comprises 1 to 9 lines of cancer therapies. In some embodiments, the prior cancer therapy comprises about 4 lines of cancer therapies. In some embodiments, the prior cancer therapy comprises a treatment targeting programmed death ligand 1 (PD-L1), programmed cell death protein 1 (PD-1), or both. In some embodiments, the prior cancer therapy comprises pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, or a combination thereof. In some embodiments, the subject has adequate organ function. In some embodiments, the subject has resolved acute effects of any prior therapy to baseline severity of Common Terminology Criteria for Adverse Events (CTCAE) grade of no more than 1. In some embodiments, the subject has locally advanced or metastatic NSCLC, SCCHN, CRC, SCLC, PDAC, TNBC, prostate cancer, or RCC that is confirmed histologically or cytologically. In some embodiments, the subject has a tumor lesion measured by response evaluation criteria in solid tumors (RECIST) guidelines. In some embodiments, the subject is not treated with ananti -cancer therapy within about 28 days before the administering. In some embodiments, the subject is not treated with an anti -cancer therapy that has no more than 5 elimination half-lives before the administering. In some embodiments, the subject is not treated with a bispecific T cell engager that targets EGFR before the administering. In some embodiments, the subject is not treated with a chimeric antigen receptor T (CAR-T) cell therapy that targets EGFR before the administering. In some embodiments, the subject is not treated with another CD3 -binding T cell engager before the administering. In some embodiments, the subject does not have a clinically significant cardiovascular disease. In some embodiments, the subject does not have an active and clinically significant infection. In some embodiments, the infection comprises a bacterial infection, a viral infection, a fungal infection, a mycobacterial infection, or a combination thereof. In some embodiments, the subject is not treated with an oxygen therapy before the administering. In some embodiments, the administering comprises administering intravenously. In some embodiments, the subject exhibits a cytokine release syndrome (CRS) no more than grade 1. In some embodiments, the subject exhibits a cytokine release syndrome (CRS) no more than grade 2. In some embodiments, the subject exhibits a treatment related adverse event (TRAE) not related to a CRS. In some embodiments, the administering comprises administering to the subject a second treatment. In some embodiments, the administering results in a therapeutic effect in size in target lesion, hepatic metastasis, stable disease, partial response, complete response, size of renal mass, best RECIST response, overall response rate, duration of response, progression free survival, or a combination thereof, after the administering.

[0115] In some embodiments, the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a reduction of up to 100% in size of target lesion after the administering, e.g., about 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33%, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40 %, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49%, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56 %, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65%, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72 %, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81%, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88 %, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97%, 98 %, 99 %, or about 100 %, or any percentage therebetween. In some embodiments, the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a reduction of up to 100% in size of target lesion 12 weeks after the administering, e.g., about 1 %, 2 %, 3 %, 4 %, 5 %, 6 %, 7 %, 8 %, 9 %, 10 %, 11 %, 12 %, 13 %, 14 %, 15 %, 16 %, 17 %, 18 %, 19 %, 20 %, 21 %, 22 %, 23 %, 24 %, 25 %, 26 %, 27 %, 28 %, 29 %, 30 %, 31 %, 32 %, 33 %, 34 %, 35 %, 36 %, 37 %, 38 %, 39 %, 40%, 41 %, 42 %, 43 %, 44 %, 45 %, 46 %, 47 %, 48 %, 49 %, 50 %, 51 %, 52 %, 53 %, 54 %, 55 %, 56%, 57 %, 58 %, 59 %, 60 %, 61 %, 62 %, 63 %, 64 %, 65 %, 66 %, 67 %, 68 %, 69 %, 70 %, 71 %, 72%, 73 %, 74 %, 75 %, 76 %, 77 %, 78 %, 79 %, 80 %, 81 %, 82 %, 83 %, 84 %, 85 %, 86 %, 87 %, 88%, 89 %, 90 %, 91 %, 92 %, 93 %, 94 %, 95 %, 96 %, 97 %, 98 %, 99 %, or about 100 %, or any percentage therebetween. In some embodiments, the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a 100% reduction in size of target lesion 12 weeks afterthe administering.

[0116] In some embodiments, the administering comprises administering weekly the first dose of the isolated recombinant polypeptide complex in an amount of about 150 pg. In some embodiments, the subject exhibits a reduction or elimination of hepatic metastasis after the administering. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the subject exhibits no CRS or TRAE after the administering. In some embodiments, the subject has a mutation, and wherein the mutation comprises low microsatellite instability (MSI-L) / microsatellite stable (MSS), a high microsatellite instability (MSI-H), a MET gene mutation, a TP53 mutation, an EGFR mutation, a Kirsten rat sarcoma virus (KRAS) mutation, an anaplastic lymphoma kinase (ALK) mutation, a Rearrangement of c-ros oncogene 1 (ROS1) mutation, an Adenomatous polyposis coli (APC) mutation, a B-type Raf proto-oncogene (BRAF) mutation, a pl 10a subunit of phosphatidylinositol-3 -kinase (PIK3CA) mutation, a cyclin-dependent kinase inhibitor 2A (CDKN2A) mutation, a Phosphatase and tensin homolog (PTEN) mutation, a NOTCH1 mutation, a breast cancer 1 early onset protein (BRAC1), a breast cancer 2 early onset protein (BRAC2), partner and localizer of BRCA2 (PALB2), or a combination thereof.

[0117] In some embodiments, the subject is treated with four of the prior cancer therapy before the administering, wherein the four of the prior cancer therapy comprises a treatment targeting PD-L 1 / PD- 1 , and wherein the cancer is refractory, non-responsive, or resistant to the treatment targeting PD-L 1 / PD- 1. In some embodiments, the subject has RCC of stage 4, and wherein the subject exhibits a reduction of about 12% in size of renal mass. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle. In some embodiments, the target dose is administered weekly after day 8.

[0118] In some embodiments, the subject has MSI-L / MSS mutation and proficient mismatch repair (pMMR) mutation. In some embodiments, the subject is treated with three lines of the prior cancer therapy before the administering. In some embodiments, the subject has an extensive large renal mass. In some embodiments, the renal mass is about 11 cm. In some embodiments, the administering results in an elimination of a cancer related pain. In some embodiments, the administering results in an elimination of a cancer related back pain. In some embodiments, the administering results in a reduction of a cancer related pain. In some embodiments, the administering results in a reduction of a cancer related back pain. In some embodiments, the administering results in an elimination of a cancer related pain after two doses of the isolated recombinant polypeptide complex. In some embodiments, the administering results in an elimination of a cancer related back pain after two doses of the isolated recombinant polypeptide complex. In some embodiments, the administering results in a reduction of a cancer related pain after two doses of the isolated recombinant polypeptide complex. In some embodiments, the administering results in a reduction of a cancer related back pain after two doses of the isolated recombinant polypeptide complex .

[0119] In some embodiments, the administering results in the reduction of about 12% in diameter ofrenal mass measured by computed tomography (CT) scan on day 17 of the treatment cycle. In some embodiments, the subject exhibits a best RECIST response. In some embodiments, the administering comprises administering weekly the first dose in an amount of about 150 pg. In some embodiments, the subject exhibits a best RECIST response, a partial response (PR), or a complete response (CR).

[0120] In some embodiments, the subject exhibits stable disease. In some embodiments, the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg. In some embodiments, the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle . In some embodiments, the target dose is administered weekly after day 8.

[0121] In some embodiments, the method further comprises treating the subject with a therapy for arthralgia, anemia, CRS, dermatitis acneiform, nausea, rash maculopapular, back pain, diarrhea, dizziness, fatigue, headache, hyperglycemia, hypokalemia, hypophosphatemia, injection site irritation, lymphocyte count decrease, oedema peripheral, oral pain, pain in extremity, pyrexia, vomiting, or a combination thereof. In some embodiments, the subject receives 2 or 3 prior cancer therapies before the administering. In some embodiments, the subject is negative for HER2 or hormone receptor (HR). In some embodiments, the subject has never been diagnosed with gout before the administering. In some embodiments, the subject does not have arthritis or arthralgia before the administering.

[0122] In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least once every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least twice every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least three times every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most once every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most twice every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most three times every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered once every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered twice every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered three times every two weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least once every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least twice every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least three times every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most once every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most twice every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most three times every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered once every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex isadministered twice every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered three times every three weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least once every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least twice every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most once every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most twice every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered once every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered twice every four weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at least once every five or more weeks. In some embodiments, a dose of the isolated recombinant polypeptide complex is administered at most once every five or more weeks.

[0123] In some embodiments, the administering comprises administering a mixture comprising the pharmaceutical composition and dextrose, wherein the mixture comprises about 5% (w / v) dextrose. In some embodiments, the administering comprises administering a mixture comprising the pharmaceutical composition and dextrose, wherein the mixture comprises about 5% (v / v) dextrose.

[0124] In some embodiments, the pharmaceutical composition comprises the isolated recombinant polypeptide complex at a concentration of about 2 mg / ml. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer, a stabilizing agent, a tonicity agent, a surfactant, or combinations thereof. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer. In some embodiments, the pharmaceutically acceptable excipient comprises a tonicity agent. In some embodiments, the pharmaceutically acceptable excipient comprises a surfactant. In some embodiments, the pharmaceutically acceptable excipient comprises a buffer, a stabilizing agent, a tonicity agent, and a surfactant. In some embodiments, the buffer comprises an amino acid or a derivative thereof. In some embodiments, the amino acid or the derivative thereof comprises L-histidine, L-histidine monohydrochloride monohydrate, or combinations thereof. In some embodiments, the stabilizing agent comprises sugar. In some embodiments, the sugar comprises sucrose. In some embodiments, the tonicity agent comprises sugar. In some embodiments, the sugar comprises sucrose. In some embodiments, the surfactant comprises a polysorbate. In some embodiments, the surfactant comprises polysorbate 20 (PS20).

[0125] In some embodiments, the pharmaceutical composition comprises about 1 millimolar (mM) to about 50 mM L-histidine in the form of L-histidine and / or L-histidine monohydrochloride monohydrate, e.g., about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, or about 50 mM, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 1 to about 50 mM L-histidine in the form of L-histidine and L-histidinemonohydrochloride monohydrate, e.g., about 1 mM, 2 mM, 3 mM, 4 mM, 5 mM, 6 mM, 7 mM, 8 mM, 9 mM, 10 mM, 11 mM, 12 mM, 13 mM, 14 mM, 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, 30 mM, 31 mM, 32 mM, 33 mM, 34 mM, 35 mM, 36 mM, 37 mM, 38 mM, 39 mM, 40 mM, 41 mM, 42 mM, 43 mM, 44 mM, 45 mM, 46 mM, 47 mM, 48 mM, 49 mM, or about 50 mM, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L- histidine and / or L-histidine monohydrochloride monohydrate. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L-histidine or L-histidine monohydrochloride monohydrate. In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L-histidine and L-histidine monohydrochloride monohydrate.

[0126] In some embodiments, the pharmaceutical composition comprises about 1% weight / volume (w / v) to about 20% (w / v) sucrose, e.g., about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18 %, 19%, or about 20%, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 8% (w / v) sucrose.

[0127] In some embodiments, the pharmaceutical composition comprises about 0.001% (w / v) to about 0.1% (w / v) polysorbate 20 (PS20), e.g., about 0.001%, 0.002 %, 0.003 %, 0.004 %, 0.005 %, 0.006 %, 0.007 %, 0.PC-1 %, 0.009 %, 0.01 %, 0.011 %, 0.012 %, 0.013 %, 0.014 %, 0.015 %, 0.016 %, 0.017 %, 0.018 %, 0.019 %, 0.02 %, 0.021 %, 0.022 %, 0.023 %, 0.024 %, 0.025 %, 0.026 %, 0.027 %, 0.028 %,0.029 %, 0.03 %, 0.031 %, 0.032 %, 0.033 %, 0.034 %, 0.035 %, 0.036 %, 0.037 %, 0.038 %, 0.039 %,0.04 %, 0.041 %, 0.042 %, 0.043 %, 0.044 %, 0.045 %, 0.046 %, 0.047 %, 0.048 %, 0.049 %, 0.05 %,0.051 %, 0.052 %, 0.053 %, 0.054 %, 0.055 %, 0.056 %, 0.057 %, 0.058 %, 0.059 %, 0.06 %, 0.061 %,0.062 %, 0.063 %, 0.064 %, 0.065 %, 0.066 %, 0.067 %, 0.068 %, 0.069 %, 0.07 %, 0.071 %, 0.072 %,0.073 %, 0.074 %, 0.075 %, 0.076 %, 0.077 %, 0.078 %, 0.079 %, 0.08 %, 0.081 %, 0.082 %, 0.083 %,0.084 %, 0.085 %, 0.086 %, 0.087 %, 0.088 %, 0.089 %, 0.09 %, 0.091 %, 0.092 %, 0.093 %, 0.094 %,0.095 %, 0.096 %, 0.097 %, 0.098 %, 0.099 %, or about 0.1 %, or any concentration therebetween. In some embodiments, the pharmaceutical composition comprises about 0.01% (w / v) polysorbate 20 (PS20).

[0128] In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L-histidine and L-histidine monohydrochloride monohydrate. In some embodiments, the pharmaceutical composition comprises about 8% weight / volume (w / v) sucrose. In some embodiments, the pharmaceutical composition comprises about 0.01% (w / v) polysorbate 20 (PS20). In some embodiments, the pharmaceutical composition comprises about 10 mM L-histidine in the form of L- histidine and L-histidine monohydrochloride monohydrate, about 8% (w / v) sucrose, and about 0.01% (w / v) polysorbate 20. In some embodiments, the pharmaceutical composition comprises a pH of about 5.3. In some embodiments, the pharmaceutical composition comprises an osmolality of about 276 mOsmol / kg.

[0129] In some embodiments, the isolated recombinant polypeptide complex is cleaved by a protease to generate an enzymatic product of the isolated recombinant polypeptide complex after the administering.In some embodiments, the isolated recombinant polypeptide complex is cleaved by a tumor specific protease to generate the enzymatic product of the isolated recombinant polypeptide complex after the administering. In some embodiments, the tumor specific protease comprises two or more proteases. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a first protease of the two or more proteases to generate a first metabolic product of the isolated recombinant polypeptide complex. In some embodiments, the isolated recombinant polypeptide complex is cleaved by a second protease of the two or more proteases to generate a second metabolic product of the isolated recombinant polypeptide complex. In some embodiments, the first protease comprises a serine protease. In some embodiments, the second protease comprises a matrix metalloprotease. In some embodiments, the serine protease comprises human matriptase (MTSP1). In some embodiments, the matrix metalloprotease comprises human matrix metalloprotease 9 (MMP9).

[0130] In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the first metabolic product. In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the second metabolic product. In some embodiments, the enzymatic product of the isolated recombinant polypeptide comprises the first metabolic product and the second metabolic product.Certain Terminology

[0131] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the claimed subject matter belongs. It is to be understood that the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of any subject matter claimed. In this application, the use of the singular includes the plural unless specifically stated otherwise. It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. In this application, the use of “or” means “and / or” unless stated otherwise. Furthermore, use of the term “including” as well as other forms, such as “include,” “includes,” and “included,” is not limiting.

[0132] As used herein, ranges and amounts can be expressed as “about” a particular value or range. About also includes the exact amount. Hence “about 5 pL” means “about 5 pL” and also “5 pL.” Generally, the term “about” includes an amount that would be expected to be within experimental error.

[0133] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

[0134] ‘ ‘Antibodies” and “immunoglobulins” (IGs) are glycoproteins having the same structural characteristics. The terms are used synonymously. In some instances, the antigen specificity of the immunoglobulin is known.

[0135] The term “antibody” is used in the broadest sense, and covers fully assembled antibodies, antibody fragments that can bind antigen (e.g., Fab, F(ab’)2, Fv, single chain antibodies (scFv), diabodies, antibody chimeras, hybrid antibodies, bispecific antibodies, and the like), and recombinant peptides comprising the forgoing.

[0136] As used herein, the terms “individual(s),” “subject(s),” and “patient(s)” mean any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker).

[0137] As used herein, the term “percent (%) amino acid sequence identity” with respect to a sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as EMBOSS MATCHER, EMBOSS WATER, EMBOSS STRETCHER, EMBOSS NEEDLE, EMBOSS LALIGN, BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.

[0138] In situations where ALIGN-2 is employed for amino acid sequence comparisons, the % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B (which can alternatively be phrased as a given amino acid sequence A that has or comprises a certain % amino acid sequence identity to, with, or against a given amino acid sequence B) is calculated as follows: 100 times the fraction X / Y, where X is the number of amino acid residues scored as identical matches by the sequence alignment program ALIGN-2 in that program's alignment of A and B, and where Y is the total number of amino acid residues in B. It will be appreciated that where the length of amino acid sequence A is not equal to the length of amino acid sequence B, the % amino acid sequence identity of A to B will not equal the % amino acid sequence identity of B to A. Unless specifically stated otherwise, all % amino acid sequence identity values used herein are obtained as described in the immediately preceding paragraph using the ALIGN-2 computer program.

[0139] The terms “individual(s)”, “subject(s)” and “patient(s)” are used interchangeably herein and refer to any mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a non-human. None of the terms require or are limited to situations characterized by the supervision (e.g. constant or intermittent) of a health care worker (e.g. a doctor, a registered nurse, a nurse practitioner, a physician’s assistant, an orderly or a hospice worker).EMBODIMENTS

[0140] Embodiment 1 comprises a method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex, wherein the first dose is at least about 50 pg, and wherein the isolated recombinant polypeptide complex comprises a first chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1 and a second chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2.

[0141] Embodiment 2 comprises the method of embodiment 1, wherein the first dose is about 50 pg to about 30 mg.

[0142] Embodiment 3 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 100 pg.

[0143] Embodiment 4 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 150 pg.

[0144] Embodiment 5 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 200 pg.

[0145] Embodiment 6 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 250 pg.

[0146] Embodiment 7 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 300 pg.

[0147] Embodiment 8 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 350 pg.

[0148] Embodiment 9 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 400 pg.

[0149] Embodiment 10 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 450 pg.

[0150] Embodiment 11 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 500 pg.

[0151] Embodiment 12 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 750 pg.

[0152] Embodiment 13 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 1 mg.

[0153] Embodiment 14 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 1.5 mg.

[0154] Embodiment 15 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 2 mg.

[0155] Embodiment 16 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 3 mg.

[0156] Embodiment 17 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 4 mg.

[0157] Embodiment 18 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 5 mg.

[0158] Embodiment 19 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 6 mg.

[0159] Embodiment 20 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 7 mg.

[0160] Embodiment 21 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 8 mg.

[0161] Embodiment 22 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 9 mg.

[0162] Embodiment 23 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 10 mg.

[0163] Embodiment 24 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 15 mg.

[0164] Embodiment 25 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 20 mg.

[0165] Embodiment 26 comprises the method of embodiment 1 or 2, wherein the first dose is at least about 25 mg.

[0166] Embodiment 27 comprises the method of embodiment 1 or 2, wherein the first dose is about 100 fig-

[0167] Embodiment 28 comprises the method of embodiment 1 or 2, wherein the first dose is about 150 fig-

[0168] Embodiment 29 comprises the method of embodiment 1 or 2, wherein the first dose is about 200 fig-

[0169] Embodiment 30 comprises the method of embodiment 1 or 2, wherein the first dose is about 250 fig-

[0170] Embodiment 31 comprises the method of embodiment 1 or 2, wherein the first dose is about 300 fig-

[0171] Embodiment 32 comprises the method of embodiment 1 or 2, wherein the first dose is about 350 fig-

[0172] Embodiment 33 comprises the method of embodiment 1 or 2, wherein the first dose is about 400 fig-

[0173] Embodiment 34 comprises the method of embodiment 1 or 2, wherein the first dose is about 450 fig-

[0174] Embodiment 35 comprises the method of embodiment 1 or 2, wherein the first dose is about 500 fig-

[0175] Embodiment 36 comprises the method of embodiment 1 or 2, wherein the first dose is about 750 fig-

[0176] Embodiment 37 comprises the method of embodiment 1 or 2, wherein the first dose is about 1 mg.

[0177] Embodiment 38 comprises the method of embodiment 1 or 2, wherein the first dose is about 1.5 mg.

[0178] Embodiment 39 comprises the method of embodiment 1 or 2, wherein the first dose is about 2 mg.

[0179] Embodiment 40 comprises the method of embodiment 1 or 2, wherein the first dose is about 3 mg.

[0180] Embodiment 41 comprises the method of embodiment 1 or 2, wherein the first dose is about 4 mg.

[0181] Embodiment 42 comprises the method of embodiment 1 or 2, wherein the first dose is about 5 mg.

[0182] Embodiment 43 comprises the method of embodiment 1 or 2, wherein the first dose is about 6 mg.

[0183] Embodiment 44 comprises the method of embodiment 1 or 2, wherein the first dose is about 7 mg.

[0184] Embodiment 45 comprises the method of embodiment 1 or 2, wherein the first dose is about 8 mg.

[0185] Embodiment 46 comprises the method of embodiment 1 or 2, wherein the first dose is about 9 mg.

[0186] Embodiment 47 comprises the method of embodiment 1 or 2, wherein the first dose is about 10 mg.

[0187] Embodiment 48 comprises the method of embodiment 1 or 2, wherein the first dose is about 15 mg.

[0188] Embodiment 49 comprises the method of embodiment 1 or 2, wherein the first dose is about 20 mg.

[0189] Embodiment 50 comprises the method of embodiment 1 or 2, wherein the first dose is about 25 mg.

[0190] Embodiment 51 comprises the method of embodiment 1 or 2, wherein the first dose is about 30 mg.

[0191] Embodiment 52 comprises the method of any one of embodiments 1-51, further comprising administering to the subject a target dose of the isolated recombinant polypeptide complex, wherein the target dose is higher than the first dose, and wherein the target dose is administered after the first dose.

[0192] Embodiment 53 comprises the method of embodiment 52, further comprising administering to the subject a second dose of the isolated recombinant polypeptide complex, wherein the second dose is higher than the first dose and lower than the target dose, and wherein the second dose is administered between the first dose and the target dose.

[0193] Embodiment 54 comprises the method of any one of embodiments 1-53, wherein the first dose is administered weekly.

[0194] Embodiment 55 comprises the method of any one of embodiments 1-53, wherein the first dose is administered once every two weeks.

[0195] Embodiment 56 comprises the method of any one of embodiments 1-53, wherein the first dose is administered once every three weeks.

[0196] Embodiment 57 comprises the method of any one of embodiments 52-56, wherein the targetdose is administered weekly.

[0197] Embodiment 58 comprises the method of any one of embodiments 52-56, wherein the target dose is administered once every two weeks.

[0198] Embodiment 59 comprises the method of any one of embodiments 52-56, wherein the target dose is administered once every three weeks.

[0199] Embodiment 60 comprises the method of any one of embodiments 53-56, wherein the second dose is administered weekly.

[0200] Embodiment 61 comprises the method of any one of embodiments 52-56, wherein the second dose is administered once every two weeks.

[0201] Embodiment 62 comprises the method of any one of embodiments 52-56, wherein the second dose is administered once every three weeks.

[0202] Embodiment 63 comprises the method of any one of embodiments 1-62, wherein the method comprises a treatment cycle that starts on day 1.

[0203] Embodiment 64 comprises the method of embodiment 63, wherein the first dose is administered on day 1 of the treatment cycle.

[0204] Embodiment 65 comprises the method of any one of embodiments 52-64, wherein the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle.

[0205] Embodiment 66 comprises the method of any one of embodiments 53-65, wherein the first dose, the second dose, and the target dose are administered on day 1, day 4, and day 8, respectively, of the treatment cycle.

[0206] Embodiment 67 comprises the method of embodiment 65 or 66, wherein the target dose is administered weekly after day 8 of the treatment cycle.

[0207] Embodiment 68 comprises the method of embodiment 65 or 66, wherein the target dose is administered once every two weeks after day 8 of the treatment cycle.

[0208] Embodiment 69 comprises the method of embodiment 65 or 66, wherein the target dose is administered once every three weeks after day 8 of the treatment cycle.

[0209] Embodiment 70 comprises the method of any one of embodiments 52-69, wherein the target dose is about 50 pg to about 300 mg.

[0210] Embodiment 71 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 100 pg.

[0211] Embodiment 72 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 250 pg.

[0212] Embodiment 73 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 500 pg.

[0213] Embodiment 74 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 750 pg.

[0214] Embodiment 75 comprises the method of any one of embodiments 52-70, wherein the targetdose is at least about 1 mg.

[0215] Embodiment 76 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 1.25 mg.

[0216] Embodiment 77 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 1.5 mg.

[0217] Embodiment 78 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 2 mg.

[0218] Embodiment 79 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 5 mg.

[0219] Embodiment 80 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 7.5 mg.

[0220] Embodiment 81 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 10 mg.

[0221] Embodiment 82 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 15 mg.

[0222] Embodiment 83 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 20 mg.

[0223] Embodiment 84 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 25 mg.

[0224] Embodiment 85 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 30 mg.

[0225] Embodiment 86 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 50 mg.

[0226] Embodiment 87 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 75 mg.

[0227] Embodiment 88 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 100 mg.

[0228] Embodiment 89 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 150 mg.

[0229] Embodiment 90 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 200 mg.

[0230] Embodiment 91 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 250 mg.

[0231] Embodiment 92 comprises the method of any one of embodiments 52-70, wherein the target dose is at least about 300 mg.

[0232] Embodiment 93 comprises the method of any one of embodiments 52-70, wherein the target dose is about 100 pg.

[0233] Embodiment 94 comprises the method of any one of embodiments 52-70, wherein the targetdose is about 250 pg.

[0234] Embodiment 95 comprises the method of any one of embodiments 52-70, wherein the target dose is about 500 pg.

[0235] Embodiment 96 comprises the method of any one of embodiments 52-70, wherein the target dose is about 750 pg.

[0236] Embodiment 97 comprises the method of any one of embodiments 52-70, wherein the target dose is about 1 mg.

[0237] Embodiment 98 comprises the method of any one of embodiments 52-70, wherein the target dose is about L25 mg.

[0238] Embodiment 99 comprises the method of any one of embodiments 52-70, wherein the target dose is about L5 mg.

[0239] Embodiment 100 comprises the method of any one of embodiments 52-70, wherein the target dose is about 2 mg.

[0240] Embodiment 101 comprises the method of any one of embodiments 52-70, wherein the target dose is about 5 mg.

[0241] Embodiment 102 comprises the method of any one of embodiments 52-70, wherein the target dose is about 7.5 mg.

[0242] Embodiment 103 comprises the method of any one of embodiments 52-70, wherein the target dose is about 10 mg.

[0243] Embodiment 104 comprises the method of any one of embodiments 52-70, wherein the target dose is about 15 mg.

[0244] Embodiment 105 comprises the method of any one of embodiments 52-70, wherein the target dose is about 20 mg.

[0245] Embodiment 106 comprises the method of any one of embodiments 52-70, wherein the target dose is about 25 mg.

[0246] Embodiment 107 comprises the method of any one of embodiments 52-70, wherein the target dose is about 30 mg.

[0247] Embodiment 108 comprises the method of any one of embodiments 52-70, wherein the target dose is about 40 mg.

[0248] Embodiment 109 comprises the method of any one of embodiments 52-70, wherein the target dose is about 50 mg.

[0249] Embodiment 110 comprises the method of any one of embodiments 52-70, wherein the target dose is about 75 mg.

[0250] Embodiment 111 comprises the method of any one of embodiments 52-70, wherein the target dose is about 100 mg.

[0251] Embodiment 112 comprises the method of any one of embodiments 52-70, wherein the target dose is about 150 mg.

[0252] Embodiment 113 comprises the method of any one of embodiments 52-70, wherein the targetdose is about 200 mg.

[0253] Embodiment 114 comprises the method of any one of embodiments 52-70, wherein the target dose is about 250 mg.

[0254] Embodiment 115 comprises the method of any one of embodiments 52-70, wherein the target dose is about 300 mg.

[0255] Embodiment 116 comprises the method of any one of embodiments 52-70, wherein the first dose is at least about 500 pg and the target dose is at least about 1.25 mg.

[0256] Embodiment 117 comprises the method of any one of embodiments 52-70, wherein the first dose is about 500 pg and the target dose is about 1.25 mg.

[0257] Embodiment 118 comprises the method of any one of embodiments 52-70, wherein the first dose is about 500 pg and the target dose is about 5 mg.

[0258] Embodiment 119 comprises the method of any one of embodiments 52-70, wherein the first dose is about 1.5 mg and the target dose is about 15 mg.

[0259] Embodiment 120 comprises the method of any one of embodiments 52-70, wherein the first dose is about 5 mg and the target dose is about 50 mg.

[0260] Embodiment 121 comprises the method of any one of embodiments 52-70, wherein the first dose is about 15 mg and the target dose is about 150 mg.

[0261] Embodiment 122 comprises the method of any one of embodiments 52-70, wherein the first dose is about 30 mg and the target dose is about 300 mg.

[0262] Embodiment 123 comprises the method of any one of embodiments 52-122, wherein the first dose, the second dose, or the target dose is administered at least once, at least twice, at least 3 times, at least 4 times, at least 5 times, at least 6 times, at least 7 times, at least 8 times, at least 9 times, at least 10 times, at least 15 times, at least 20 times, at least 25 times, at least 30 times, at least 35 times, at least 40 times, at least 45 times, at least 50 times, or more than 50 times.

[0263] Embodiment 124 comprises the method of any one of embodiments 52-123, wherein the first dose is about 500 pg, the second dose is about 1.5 mg, and the target dose is about 5 mg.

[0264] Embodiment 125 comprises the method of any one of embodiments 52-123, wherein the first dose is about 1.5 mg, the second dose is about 5 mg, and the target dose is about 15 mg.

[0265] Embodiment 126 comprises the method of any one of embodiments 52-123, wherein the first dose is about 5 mg, the second dose is about 15 mg, and the target dose is about 50 mg.

[0266] Embodiment 127 comprises the method of any one of embodiments 52-123, wherein the first dose is about 15 mg, the second dose is about 50 mg, and the target dose is about 150 mg.

[0267] Embodiment 128 comprises the method of any one of embodiments 52-123, wherein the first dose is about 30 mg, the second dose is about 100 mg, and the target dose is about 300 mg.

[0268] Embodiment 129 comprises the method of any one of embodiments 1-128, further comprising at least 2 treatment cycles, at least 3 treatment cycles, at least 4 treatment cycles, at least 5 treatment cycles, at least 6 treatment cycles, at least 7 treatment cycles, at least 8 treatment cycles, at least 9 treatment cycles, at least 10 treatment cycles, or more than 10 treatment cycles.

[0269] Embodiment 130 comprises the method of any one of embodiments 1-129, wherein the cancer comprises a cell that expresses epidermal growth factor receptor (EGFR).

[0270] Embodiment 131 comprises the method of any one of embodiments 1-130, wherein the cancer comprises a cell that overexpresses EGFR.

[0271] Embodiment 132 comprises the method of any one of embodiments 1-131, wherein the cancer comprises colorectal cancer (CRC), squamous cell carcinoma of head and neck (SCCHN), non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), breast cancer, bladder cancer, ovarian cancer, liver cancer, pancreatic cancer, small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple-negative breast cancer (TNBC), prostate cancer, or a combination thereof.

[0272] Embodiment 133 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic NSCLC.

[0273] Embodiment 134 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic SCCHN.

[0274] Embodiment 135 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic CRC.

[0275] Embodiment 136 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic RCC.

[0276] Embodiment 137 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic SCLC.

[0277] Embodiment 138 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic PDAC.

[0278] Embodiment 139 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic TNBC.

[0279] Embodiment 140 comprises the method of any one of embodiments 1-132, wherein the cancer comprises advanced or metastatic prostate cancer.

[0280] Embodiment 141 comprises the method of any one of embodiments 1-139, wherein the cancer progresses after treatment with a prior cancer therapy.

[0281] Embodiment 142 comprises the method of any one of embodiments 1-141, wherein the cancer is refractory, intolerant, non-responsive, or resistant to a prior cancer therapy.

[0282] Embodiment 143 comprises the method of any one of embodiments 1-142, wherein the subject is at least 18 years old.

[0283] Embodiment 144 comprises the method of any one of embodiments 1-143, wherein the subject is about 37 to about 81 years old.

[0284] Embodiment 145 comprises the method of any one of embodiments 1-144, wherein the subject is about 65 years old.

[0285] Embodiment 146 comprises the method of any one of embodiments 141-145, wherein the prior cancer therapy comprises 1 to 9 lines of cancer therapies.

[0286] Embodiment 147 comprises the method of any one of embodiments 141-145, wherein the priorcancer therapy comprises about 4 lines of cancer therapies.

[0287] Embodiment 148 comprises the method of any one of embodiments 141-145, wherein the prior cancer therapy comprises a treatment targeting programmed death ligand 1 (PD-L1), programmed cell death protein 1 (PD-1), or both.

[0288] Embodiment 149 comprises the method of embodiment 148, wherein the prior cancer therapy comprises pembrolizumab, nivolumab, cemiplimab, atezolizumab, avelumab, durvalumab, or a combination thereof.

[0289] Embodiment 150 comprises the method of any one of embodiments 1-149, wherein the subject has adequate organ function.

[0290] Embodiment 151 comprises the method of any one of embodiments 1-149, wherein the subject has resolved acute effects of any prior therapy to baseline severity of Common Terminology Criteria for Adverse Events (CTCAE) grade of no more than 1.

[0291] Embodiment 152 comprises the method of any one of embodiments 1-150, wherein the subject has locally advanced or metastatic NSCLC, SCCHN, CRC, SCLC, PDAC, TNBC, prostate cancer, or RCC that is confirmed histologically or cytologically.

[0292] Embodiment 153 comprises the method of any one of embodiments 1-152, wherein the subject has a tumor lesion measured by response evaluation criteria in solid tumors (RECIST) guidelines.

[0293] Embodiment 154 comprises the method of any one of embodiments 1-153, wherein the subject is not treated with an anti -cancer therapy within 28 days before the administering.

[0294] Embodiment 155 comprises the method of any one of embodiments 1-154, wherein the subject is not treated with an anti -cancer therapy that has no more than 5 elimination half-lives before the administering.

[0295] Embodiment 156 comprises the method of any one of embodiments 1-155, wherein the subject is not treated with a bispecific T cell engager that targets EGFR before the administering.

[0296] Embodiment 157 comprises the method of any one of embodiments 1-156, wherein the subject is not treated with a chimeric antigen receptor T (CAR-T) cell therapy that targets EGFR before the administering.

[0297] Embodiment 158 comprises the method of any one of embodiments 1-157, wherein the subject is not treated with another CD3 -binding T cell engager before the administering.

[0298] Embodiment 159 comprises the method of any one of embodiments 1-158, wherein the subject does not have a clinically significant cardiovascular disease.

[0299] Embodiment 160 comprises the method of any one of embodiments 1-159, wherein the subject does not have an active and clinically significant infection.

[0300] Embodiment 161 comprises the method of embodiment 160, wherein the infection comprises a bacterial infection, a viral infection, a fungal infection, a mycobacterial infection, or a combination thereof.

[0301] Embodiment 162 comprises the method of any one of embodiments 1-161, wherein the subject is not treated with an oxygen therapy before the administering.

[0302] Embodiment 163 comprises the method of any one of embodiments 1-162, wherein the administering comprises administering intravenously.

[0303] Embodiment 164 comprises the method of any one of embodiments 1-163, wherein the subject exhibits a cytokine release syndrome (CRS) no more than grade 1.

[0304] Embodiment 165 comprises the method of any one of embodiments 1-164, wherein the subject exhibits a cytokine release syndrome (CRS) no more than grade 2.

[0305] Embodiment 166 comprises the method of any one of embodiments 1-165, wherein the subject exhibits a treatment related adverse event (TRAE) not related to a CRS.

[0306] Embodiment 167 comprises the method of any one of embodiments 1-166, wherein the administering comprises administering to the subject a second treatment.

[0307] Embodiment 168 comprises the method of any one of embodiments 1-167, wherein the administering results in a therapeutic effect in size in target lesion, hepatic metastasis, stable disease, partial response, complete response, size of renal mass, best RECIST response, overall response rate, duration of response, progression free survival, or a combination thereof, after the administering.

[0308] Embodiment 169 comprises the method of embodiment 168, wherein the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a 100% reduction in size of target lesion 12 weeks after the administering.

[0309] Embodiment 170 comprises the method of embodiment 169, wherein the administering comprises administering weekly the first dose of the isolated recombinant polypeptide complex in an amount of about 150 pg.

[0310] Embodiment 171 comprises the method of embodiment 169 or 170, wherein the subject exhibits a reduction or elimination of hepatic metastasis after the administering.

[0311] Embodiment 172 comprises the method of any one of embodiments 168-171, wherein the subject exhibits a best RECIST response.

[0312] Embodiment 173 comprises the method of any one of embodiments 168-172, wherein the subject exhibits no CRS or TRAE after the administering.

[0313] Embodiment 174 comprises the method of any one of embodiments 168-173, wherein the subject has a mutation, and wherein the mutation comprises low microsatellite instability (MSI- L) / microsatellite stable (MSS), a high microsatellite instability (MSI-H), a MET gene mutation, a TP53 mutation, an EGFR mutation, a Kirsten rat sarcoma virus (KRAS) mutation, an anaplastic lymphoma kinase (ALK) mutation, a Rearrangement of c-ros oncogene 1 (ROS1) mutation, an Adenomatous polyposis coli (APC) mutation, a B-type Raf proto-oncogene (BRAF) mutation, a pl 10a subunit of phosphatidylinositol-3 -kinase (PIK3CA) mutation, a cyclin -dependent kinase inhibitor 2A (CDKN2A) mutation, a Phosphatase and tensin homolog (PTEN) mutation, a NOTCH1 mutation, a breast cancer 1 early onset protein (BRAC1), a breast cancer 2 early onset protein (BRAC2), partner and localizer of BRCA2 (PALB2), or a combination thereof.

[0314] Embodiment 175 comprises the method of any one of embodiments 168-174, wherein the subject is treated with four of the prior cancer therapy before the administering, wherein the four of theprior cancer therapy comprises a treatment targeting PD-L1 / PD-1, and wherein the cancer is refractory, non-responsive, or resistant to the treatment targeting PD-L1 / PD-1.

[0315] Embodiment 176 comprises the method of any one of embodiments 1-175, wherein the subject has RCC of stage 4, and wherein the subject exhibits a reduction of about 12% in size of renal mass.

[0316] Embodiment 177 comprises the method of embodiment 176, wherein the subject exhibits a best RECIST response.

[0317] Embodiment 178 comprises the method of embodiment 176 or 177, wherein the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg.

[0318] Embodiment 179 comprises the method of embodiment 178, wherein the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle.

[0319] Embodiment 180 comprises the method of embodiment 179, wherein the target dose is administered weekly after day 8.

[0320] Embodiment 181 comprises the method of any one of embodiments 176-180, wherein the subject has MSI-L / MSS mutation and proficient mismatch repair (pMMR) mutation.

[0321] Embodiment 182 comprises the method of any one of embodiments 176-181, wherein the subject is treated with three of the prior cancer therapy before the administering.

[0322] Embodiment 183 comprises the method of any one of embodiments 176-181, wherein the subject has an extensive large renal mass.

[0323] Embodiment 184 comprises the method of embodiment 183, wherein the renal mass is about 11 cm.

[0324] Embodiment 185 comprises the method of any one of embodiments 176-184, wherein the administering results in an elimination of a cancer related pain.

[0325] Embodiment 186 comprises the method of embodiment 185, wherein the administering results in an elimination of a cancer related pain after two doses of the isolated recombinant polypeptide complex.

[0326] Embodiment 187 comprises the method of embodiment 186, wherein the administering results in the reduction of about 12% in diameter of renal mass measured by computed tomography (CT) scan on day 17 of the treatment cycle.

[0327] Embodiment 188 comprises the method of any one of embodiments 1-187, wherein the subject exhibits a best RECIST response.

[0328] Embodiment 189 comprises the method of embodiment 188, wherein the administering comprises administering weekly the first dose in an amount of about 150 pg.

[0329] Embodiment 190 comprises the method of any one of embodiments 1-189, wherein the subject exhibits a best RECIST response, a partial response (PR), or a complete response (CR).

[0330] Embodiment 191 comprises the method of any one of embodiments 1-190, wherein the subject exhibits stable disease.

[0331] Embodiment 192 comprises the method of embodiment 190 or 191, wherein the administeringcomprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg.

[0332] Embodiment 193 comprises the method of embodiment 192, wherein the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle.

[0333] Embodiment 194 comprises the method of embodiment 193, wherein the target dose is administered weekly after day 8.

[0334] Embodiment 195 comprises the method of any one of embodiments 1-194, wherein the method further comprises treating the subject with a therapy for arthralgia, anemia, CRS, dermatitis acneiform, nausea, rash maculopapular, back pain, diarrhea, dizziness, fatigue, headache, hyperglycemia, hypokalemia, hypophosphatemia, injection site irritation, lymphocyte count decrease, oedema peripheral, oral pain, pain in extremity, pyrexia, vomiting, or a combination thereof.

[0335] Embodiment 196 comprises the method of any one of embodiments 1-195, wherein the subject receives 2 or 3 prior cancer therapies before the administering.

[0336] Embodiment 197 comprises the method of any one of embodiments 1-196, wherein the subject is negative for HER2 or hormone receptor (HR).

[0337] Embodiment 198 comprises the method of any one of embodiments 1-196, wherein the subject has never been diagnosed with gout before the administering.

[0338] Embodiment 199 comprises the method of any one of embodiments 1-196, wherein the subject does not have arthritis or arthralgia before the administering.

[0339] Embodiment 200 comprises the method of any one of embodiments 1-199, wherein the isolated recombinant polypeptide complex comprises a CD3 binding domain.

[0340] Embodiment 201 comprises the method of embodiment 200, wherein the CD3 binding domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 16, HC- CDR2: SEQ ID NO: 17, and HC-CDR3: SEQ ID NO: 18; and the CD3 binding domain comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 13, LC-CDR2: GTK, LC-CDR3: SEQ ID NO: 15.

[0341] Embodiment 202 comprises the method of embodiment 200, wherein the CD3 binding domain comprises an antibody or antigen binding fragment thereof, wherein the CD3 binding domain is a single chain variable fragment (scFv) comprising an amino acid sequence according to SEQ ID NO: 19.

[0342] Embodiment 203 comprises the method of any one of embodiments 1-202, wherein the isolated recombinant polypeptide complex comprises a peptide mask that inhibits CD3 engagement on T-cells and a tumor protease cleavable linker that connects the CD3 binding domain, wherein the peptide mask comprises an amino acid sequence according to SEQ ID NO: 28.

[0343] Embodiment 204 comprises the method of any one of embodiments 1-199, wherein the isolated recombinant polypeptide complex comprises an EGFR binding domain.

[0344] Embodiment 205 comprises the method of embodiment 204, wherein the EGFR binding domaincomprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, HC-CDR3: SEQ ID NO 25; and the EGFR binding domain comprises CDRs LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 20, LC-CDR2: YAS, and LC-CDR3: SEQ ID NO: 22.

[0345] Embodiment 206 comprises the method of any one of embodiments 1-199, wherein the EGFR binding domain comprises an antibody or antibody fragment, wherein the EGFR binding domain is a Fab comprising a Fab light chain polypeptide comprising the amino acid sequence according to SEQ ID NO: 26 and a Fab heavy chain polypeptide comprising the amino acid sequence according to SEQ ID NO: 27.

[0346] Embodiment 207 comprises a method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex comprising a CD3 binding domain, wherein the first dose is at least about 50 pg, wherein (i) the CD3 binding domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC- CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 16, HC-CDR2: SEQ ID NO: 17, and HC-CDR3: SEQ ID NO: 18; and the CD3 binding domain comprises CDRs: LC-CDR1, LC- CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 13, LC-CDR2: GTK, LC-CDR3: SEQ ID NO: 15.

[0347] Embodiment 208 comprises the method of any one of embodiments 1-199 and 204-207, wherein the isolated recombinant polypeptide complex comprises a peptide mask that inhibits EGFR engagement and a tumor protease cleavable linker that connects the EGFR binding domain, wherein the peptide mask comprises an amino acid sequence according to SEQ ID NO: 29.

[0348] Embodiment 209 comprises a method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex comprising an EGFR binding domain, wherein the first dose is at least about 50 pg, wherein the EGFR binding domain comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, HC-CDR3: SEQ ID NO 25; and the EGFR binding domain comprises CDRs LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC- CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 20, LC-CDR2: YAS, and LC-CDR3: SEQ ID NO: 22.

[0349] Embodiment 210 comprises a method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex comprising an EGFR binding domain, wherein the first dose is at least about 50 pg, wherein the EGFR binding domain comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC- CDR3 comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, HC-CDR3: SEQ ID NO 25; and the EGFR binding domain comprises CDRs LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC- CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 20, LC-CDR2: YAS, and LC-CDR3: SEQ ID NO: 22.

[0350] Embodiment 211 comprises a method for treating cancer comprising administering to a subjectin need thereof a first dose of an isolated recombinant polypeptide complex comprising a CD3 binding domain and an EGFR binding domain, wherein the first dose is at least about 50 pg, wherein (i) the CD3 binding domain comprises complementarity determining regions (CDRs): HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 16, HC-CDR2: SEQ ID NO: 17, and HC-CDR3: SEQ ID NO: 18; and the CD3 binding domain comprises CDRs: LC-CDR1, LC-CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 13, LC-CDR2: GTK, LC-CDR3: SEQ ID NO: 15; and (ii) wherein the EGFR binding domain comprises CDRs: HC-CDR1, HC-CDR2, and HC-CDR3, wherein the HC-CDR1, the HC-CDR2, and the HC-CDR3 comprise: HC-CDR1: SEQ ID NO: 23, HC-CDR2: SEQ ID NO: 24, HC-CDR3: SEQ ID NO 25; and the EGFR binding domain comprises CDRs LC-CDR1, LC- CDR2, and LC-CDR3, wherein the LC-CDR1, the LC-CDR2, and the LC-CDR3 comprise: LC-CDR1: SEQ ID NO: 20, LC-CDR2: YAS, and LC-CDR3: SEQ ID NO: 22.EXAMPLESExample 1: Clinical Effects of Polypeptide Complex 1 (PC-1)

[0351] This Example illustrates the clinical effects of a polypeptide complex l(PC-l) disclosed herein.

[0352] SUMMARY

[0353] 1.1 PC-1

[0354] As illustrated in FIG. 1, PC-1 is a tumor activated T cell engager (TRACTr) comprising a humanized tri-specific protein that incorporates epidermal growth factor receptor (EGFR) and cluster of differentiation 3 (CD3) binding domains, an albumin-binding domain to extend circulating half-life, and two separate peptide masks. The peptide masks are fused to the molecule through tumor protease cleavable linkers. One peptide mask inhibits EGFR engagement on target cells, and the other peptide mask inhibits CD3 engagement on T cells.

[0355] 1.2 Pharmacology

[0356] Nonclinical pharmacology in vitro studies with PC-1 examined the following:1. PC-1 binding to EGFR, CD3, and albumin antigens from mouse, rat, cynomolgus monkey, and human2. PC-1 stability in serum of healthy human donors, and colorectal cancer (CRC), squamous cell carcinoma of head and neck (SCCHN), small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple -negative breast cancer (TNBC), and non-small cell lung cancer (NSCLC) patients, and cynomolgus monkeys3. The ability of PC-1 to induce T cell -mediated anti -tumor cytotoxic activity4. The ability of PC-1 to induce cytokine production

[0357] Results from the binding study concluded that PC-1 bound human and cynomolgus monkey EGFR, CD3, and albumin with low nanomolar affinity, while exhibiting minimal binding to mouse or rat antigens. Cleavage -dependent unmasking of PC-1 led to sub-nanomolar affinity to human andcynomolgus EGFR and CD3. Serum stability studies showed that PC-1 was more susceptible to cleavage in cynomolgus monkey serum compared to human serum or serum from CRC, SCCHN, and NSCLC patients. The cleavage rate of both masks was about 1% per day in healthy human serum and 2% in serum from CRC, SCCHN, and NSCLC patients, whereas cleavage of PC-1 in cynomolgus monkey serum was 11.5% for the EGFR mask and 6.6% for the CD3 mask.

[0358] Cellular cytotoxicity studies showed that PC-1 induced tumor cell killing is cleavage-and dosedependent. PC-1 exhibited much lower potency compared to cleaved forms (PC-1 serine protease [SP] - cleaved and PC-1 matrix metalloprotease [MMP] -cleaved) and non-masked, PC-l-T cell engager (TCE). Activity correlated with the density of EGFR expression on the surface of target tumor cells, with activity being lowest (-8,000 to 12,000x relative to SP cleaved PC-1) in the A549 cell line. Estimated decrease in tumor killing was ~5,500x and ~450x in HCT116 and CAL27 cell lines, respectively. The activity of PC- 1, ie, the ability to induce T cell -mediated tumor cell killing, depended on the expression level of EGFR on the surface of target tumor cells. None of the test articles induced cytotoxicity against a control EGFR-deficient A549 cell line. In in vitro cytokine release assays, cleaved (PC-l-SP and PC-l-MMP cleaved) and non-masked (PC- 1 -TCE) test articles exhibited potent, dose-dependent induction of interferon gamma (IFNy), tumor necrosis factor (TNF), and interleukin (IL)-6 release in the presence of EGFR positive cell lines HCT116 and A549, while dose -dependent production of IFNy and TNF, but not IL-6, was observed in the presence of the EGFR positive Cal27 cell line. Masked PC-1 showed a decreased ability to induce cytokine release in presence of all cell lines compared to PC- 1 -TCE.

[0359] A research version of PC-1 with a histidine tag (PC-l-Histag) demonstrated dose- and cleavagedependent anti -tumor activity in a mouse model of CRC. This preclinical proof of mechanism supports the requirement of protease activity in a human tumor to enable cleavage and anti -tumor activity of PC-1 in vivo. The nonclinical toxicity profde of PC-1 was investigated in non-human primates. There were no effects on the central nervous system (CNS) or respiratory systems based on clinical observation of the animals and detailed weekly examinations after administration of <0.6 mg / kg / dose of PC-1. There were no effects of PC-1 on qualitative electrocardiogram (ECG) parameters nor on QRS or heart rate-corrected QT intervals at PC-1 doses of <0.6 mg / kg / week. There was an increased incidence of animals with minimally increased heart rate and associated shortening of mean RR, PR, and QT intervals at > 0.2 mg / kg / dose at 1 to 3 hours post-end of infusion (EOI) on Days 1 and 22. These changes were not observed following a 4-week recovery period and were likely secondary to cytokine release and not considered to be a direct physiologic effect of PC-1.

[0360] 1.3 Pharmacokinetics

[0361] The pharmacokinetic (PK) properties of PC-1 were evaluated following single- and repeat-dose studies in cynomolgus monkeys. The PK characteristics of PC-1 were investigated as part of both nonclinical PK and toxicokinetic (TK) studies in cynomolgus monkeys. A total of 3 studies were performed: two single-dose, non-Good Laboratory Practice (GLP) PK studies in cynomolgus monkeys (one at doses of 0.1, 0.3, and 1 mg / kg and the other at 0.05, 0.2, and 0.6 mg / kg) and a repeat-dose GLP toxicology study in cynomolgus monkeys that examined weekly doses (4 weeks; total 5 doses) of PC-1 at0.05, 0.2, and 0.6 mg / kg / dose. When present, anti-drug antibodies (ADA) impacted the PK profile of PC- 1 in cynomolgus monkeys. In the first single-dose non-GLP PK study, following a single intravenous (IV) bolus administration of PC-1 to cynomolgus monkeys at doses of 0.1, 0.3, and 1 mg / kg, concentrations of PC-1 increased with increased dose. Mean Cmaxincreased in a more than doseproportional manner between doses of 0. 1 and 0.3 mg / kg and was dose-proportional between 0.3 and 1 mg / kg. Mean AUCo-2ieh was more than dose proportional between doses of 0. 1 and 0.3 mg / kg and was less than dose-proportional between doses of 0.3 and 1 mg / kg. Mean ti / 2 of PC-1 following a single dose ranged from 88.8 to 101 hours across all tested dose levels. The animal that was sacrificed after 24 hours had the highest observed Cmaxand AUCo-24h in this group. PC-l-TCE was not detectable in any samples. In the second single-dose non-GLP PK study, following a single 30-minute IV infusion of PC-1 to cynomolgus monkeys at doses of 0.05, 0.2, and 0.6 mg / kg, mean Cmaxvalues for PC-1 increased in an approximately dose-proportional manner. Mean AUCo-iesh increased in a less than dose-proportional manner from 0.05 to 0.2 mg / kg and increased in an approximately dose -proportional manner from 0.2 to 0.6 mg / kg. Mean PC-1 ti / 2 values were 81.3, 68.2, and 96.5 hours at 0.05, 0.2, and 0.6 mg / kg, respectively. PC-l-TCE was not detectable in any samples. Following five doses of weekly 30-minute IV infusions of PC-1 to cynomolgus monkeys at doses of 0.05, 0.2, and 0.6 mg / kg / dose, mean Cmax, mean AUCo-24hr, and mean AUCo-ieshr after Day 1 dose increased in a nearly dose-proportional manner. However, mean Cmax, mean AUCo-24hr, and / or mean AUCo-ieshr after Days 22 and 29 dose increased in a less than dose-proportional manner due to ADA presence in 3 out of 6 animals at 0.05 mg / kg dose, 6 out of 6 animals in 0.2 mg / kg dose, and 10 out of 10 animals at 0.6 mg / kg dose. Accumulation was not observed after multiple doses and could not be assessed fully with Groups 2, 3, and 4 due to the confirmed ADA presence. No marked accumulation was observed in ADA-negative animals at 0.05 mg / kg / dose (Group 2) and at 0.2 mg / kg / dose (Group 3). One of the ADA -positive animals in Group 2 (2503) and two of ADA -positive animals in Group 3 still attained substantial exposure after Days 22 and / or 29 doses with slightly lower exposure to Day 1 dose, as measured by Cmax, AUCo-24h, and AUCo- i68h. In addition, five of the ADA-positive animals in Group 4 also attained some exposure as measured by AUCo-24h, AUCo-iesh, and Cmaxafter Day 22 dose, however, with substantially lower exposure compared to Day 1 dose. PC-l-TCE was not detectable in the majority of samples, except one sample with measurable concentration that was close to the detection limit of the assay.

[0362] No pharmacokinetic drug interaction studies have been performed for PC-1. In general, molecules such as PC-1 are not metabolized by cytochrome P450 (CYP) enzymes or transported by P- glycoprotein (Pgp) or related adenosine triphosphate-binding cassette membrane transporters. Cytokines produced by activated lymphocytes may impact the levels of Pgp and the activity of CYP enzymes (Harvey and Morgan, 2014). The clinical relevance of PC-1 causing immune modulation and potential cytokine production that could impact Pgp and CYP is unknown, but a clinically relevant drug -drug interaction effect is considered highly unlikely. Thus, in accordance with guidelines and scientific evidence (FDA August 2020; Huang et al, 2010; Seitz and Zhou, 2007), no PK drug -drug interactions studies were conducted with PC-1.

[0363] 1.4 Toxicology

[0364] In the 4-week repeat-dose toxicity study in monkeys, transient clinical signs considered secondary to cytokine release were observed after PC-1 administration on Day 1 at 0.6 mg / kg / dose. PC- 1 -related clinical chemistry and hematology changes associated with an acute phase response were observed at >0.05 mg / kg / dose. Increases in IL-10 at >0.2 mg / kg / dose and IL-6 and IFNy at 0.6 mg / kg / dose were observed after PC-1 on Day 1. Most changes noted during the study were observed post the first dose and returned to the pre-dose levels before the next dose. Following a 4-week recovery period, any remaining minor changes were fully reversible. The no observed adverse effect level (NOAEL) was considered to be 0.6 mg / kg / dose, the highest dose tested. PC-l-induced concentration- or dose-dependent cytokine release (ie, IL-6, IFNy, and / or TNF) was observed in vitro in the presence of tumor cells and in vivo in normal monkeys with no tumors. In vivo, cytokine release was primarily observed after the first dose and correlated with clinical signs and clinical chemistry changes indicative of cytokine release syndrome.

[0365] 1.5 Phase 1 Clinical Study Design

[0366] The study is a first-in-human (FIH), Phase 1 / lb, open-label, multicenter dose escalation and dose expansion study to assess the safety, tolerability, PK, pharmacodynamic (PD), and preliminary antitumor activity of PC-1 in adult subjects with histologically confirmed advanced or metastatic CRC, NSCLC, renal cell carcinoma (RCC), and SCCHN. The study will be conducted in 3 parts: Dose Escalation (Part 1) with approximately 40 to 50 subjects, Cohort Backfill Expansion (Part 2) with up to approximately 40 subjects enrolled across 4 dose levels, and Dose Expansion (Part 3) with up to approximately 40 subjects enrolled at the recommended Phase 2 dose (RP2D). Dose Escalation (Part 1) will assess the safety, tolerability, PK, PD, and preliminary efficacy of PC-1 administered by IV infusion. Cohort Backfill Enrichment (Part 2) will allow for further characterization of safety and activity of dose levels. Dose Expansion (Part 3) will determine additional safety, tolerability, PK, PD, and preliminary clinical activity data with PC-1 at a dose and schedule to be determined by the Safety Review Committee after reviewing all available safety, PK, PD, and preliminary efficacy data. Depending on the data, randomization may be integrated for either two different RP2D doses or two different treatment intervals.

[0367] Cancer

[0368] The following is a non -limiting list of cancers that can be treated with PC-1.• SCLC: small cell lung cancer (SCLC)• PDAC: pancreatic ductal adenocarcinoma (PDAC)• TNBC: triple -negative breast cancer (TNBC)• CRC: colorectal cancer, adenocarcinoma of rectum, adenocarcinoma of colon.• SCCHN: squamous cell carcinoma of primary tumor location of oral cavity, oropharynx, hypopharynx, or larynx.• RCC: renal cell carcinoma with clear cell or papillary cell type (not chromophobe,• hereditary cancer syndrome or other types).• NSCLC: non-small cell lung cancer; squamous cell carcinoma, and adenocarcinoma.• Breast Cancer, Pancreatic Cancer, Ovarian Cancer, Prostate Cancer, Brain Cancer (glioblastoma multiforme)

[0369] 2.2 PC-1 Design, Structure, and Mechanism of Action

[0370] PC-1 is a TRACTr comprising a humanized tri-specific protein that incorporates EGFR and cluster of differentiation 3 (CD3)-binding domains, an albumin-binding domain to extend circulating half-life, and two separate peptide masks. The peptide masks are fused to the molecule through tumor protease cleavable linkers. One peptide mask inhibits EGFR engagement on target cells, and the other peptide mask inhibits CD3 engagement on T cells (FIG. 1). TRACTr target engagement requires proteolysis of its two cleavable linkers by proteases present in the tumor microenvironment (TME). Once the cleavage sequences undergo proteolysis, the EGFR mask and the tandem CD3 mask plus albuminbinding domain are released, which enables optimal EGFR and CD3 target engagement. This tumor- restricted binding and subsequent T cell activation by the EGFR x CD3 bispecific components of PC-1 promote T cell -mediated killing of EGFR-expressing cancer cells. In addition, loss of the albuminbinding domain ensures that any cleaved PC-1 that migrates out of the tumor will be cleared from the blood compartment rapidly to minimize its accumulation in healthy tissues that can contribute to longterm safety risks. PC-1 is being developed for the treatment of advanced or metastatic tumors known to overexpress EGFR, including metastatic CRC, NSCLC, SCCHN, and RCC in adults.

[0371] The conditional masking and half-life extension of TRACTrs are protease cleavage-dependent. Published work describes the upregulation of many proteases in tumors relative to healthy tissue, including MMPs and SPs. In addition, several protease-activated biologies and imaging agents have been clinically validated across a broad spectrum of tumor types. By design, TRACTr molecules are highly sensitive to tumor-selective proteases. Once the TRACTr reaches the TME, proteases cleave the specific substrates (one SP substrate and one MMP substrate) within the cleavable linker, releasing the CD3 mask and albumin-binding domain. The result of protease cleavage is the conversion of the TRACTr to its active form, a TCE.

[0372] Notably, the TCE form of PC-1 (PC- 1 -TCE) has a very short serum half-life, such that cleaved forms of PC-1 that escape the TME are predicted to be cleared from the body before they can generate significant off-tumor toxicity. Preclinical data indicate a TRACTr, via unmasking by proteases at the tumor site, can drive potent anti-tumor responses while producing 25-fold less systemic IL-6 (a key marker of cytokine release syndrome [CRS]) at a 1 Ox higher dose level relative to a non -masked TCE. We anticipate that the TRACTr approach will allow us to demonstrate differentiated safety and efficacy profiles in patients with metastatic and advanced NSCLC, SCCHN, and CRC. Accordingly, a FIH, Phase 1, multicenter, open-label study is planned to determine the safety, PK, RP2D, and preliminary antitumor activity of PC-1 administered as a single agent in adult subjects with metastatic or advanced NSCLC, SCCHN, CRC, and RCC

[0373] 2.3 Rationale for PC-1

[0374] Therefore, there is a significant unmet need to optimally leverage T cell mediated cytotoxicity intargeting tumor cells. Products that can selectively activate within the TME may have a significant advantage in developing a favorable risk-to-benefit profile. TRACTr-based approach is designed to offer a more focused way to activate T cells in the tumor, minimize systemic activation, enable higher dosing, and thereby increase anti-tumor efficacy. A TRACTr molecule (PC-1) was designed to improve the therapeutic profile of EGFR-targeted TCEs in patients with tumors known to overexpress EGFR, including metastatic CRC, NCSLC, SCCHN, and RCC. PC-1 consists of a core bispecific TCE that recognizes EGFR and CD3 on T cells that is modified by adding tumor protease cleavable linkers connected to peptide masks that specifically inhibit (1) the CD3 binding domain and (2) the EGFR binding domain of PC-1 (FIG. 1). The CD3 mask is designed to limit activity outside the TME by inhibiting CD3 binding in peripheral blood, therefore helping mitigate broad T cell activation that contributes to CRS. Similarly, the EGFR mask is designed to limit on -target, off-tumor EGFR binding and associated toxicity. In addition, PC-1 exhibits an extended half-life in plasma via incorporation of an albumin-binding domain, fused to the CD3 mask. The conditional masking and half-life extension of TRACTrs are protease cleavage-dependent. TRACTr molecules, by design, are highly sensitive to tumor- selective proteases. Once the TRACTr reaches the TME, two types of proteases can cleave the linkers (the linkers contain both an SP and an MMP substrate sequence), releasing the CD3 mask and albuminbinding domain as well as the EGFR mask. The result of protease cleavage is the conversion of the TRACTr to its active form, a TCE.

[0375] 3 PHYSICAL, CHEMICAL, AND PHARMACEUTICAL PROPERTIES AND

[0376] FORMULATION

[0377] 3.1 Drug Substance

[0378] PC-1 is a 97.1 kDa humanized tri-specific glycosylated protein comprised of:• Anti-EGFR antigen-binding fragment (Fab)• Anti-CD3 single-chain variable fragment (scFv)• Anti-albumin single domain antibody (SDA / sdAb)• A peptide mask that inhibits the anti-EGFR Fab from binding EGFR and is integrated into the molecule via a tumor protease cleavable amino acid linker• A second peptide mask that inhibits the anti-CD3 scFv from binding CD3 and is integrated into the molecule via a tumor protease cleavable amino acid linker

[0379] The light chain of the anti-CD3 scFv is fused to the N-terminal heavy chain of the anti-EGFR Fab via a short flexible linker. The EGFR inhibitory peptide mask is fused to the amino terminus of the anti-EGFR Fab light chain via a protease cleavable linker. Tandem albumin-binding sdAb and CD3 inhibitory peptide mask are fused to the amino terminus of the anti-CD3 scFv via a tumor protease cleavable linker. The albumin-binding SDA is connected to the amino terminus of the CD3 inhibitory peptide mask via a short flexible linker (FIG. 1).

[0380] The molecular formula for PC-1 is C4181H6437N1147O1339S26. The predicted average molecular weight of PC-1 without glycosylation is 95,027 Da. PC-1 TRACTr consists of 2 protein chains connected by a single intermolecular disulfide bond between the light chain (LC) and heavy chain (HC) of theTRACTr, and 9 intramolecular disulfide bonds. Each peptide mask contains a single internal disulfide bond.

[0381] 3.2 Drug Product

[0382] A PC-1 solution for injection (e.g., IV infusion (referred to as PC-1 drug product [DP])), will be provided for clinical investigational as a sterile aqueous solution formulated at a nominal concentration of 2 mg / mL. The solution is formulated in 10 mM histidine, 8% (w / v) sucrose, and 0.01% (w / v) polysorbate 20, and has an approximate pH of 5.3. The formulation of the PC-1 DP is outlined in Table 2. Abbreviations: cGMP = Current good manufacturing practice; BP = British Pharmacopoeia; Ch.P = Chinese Pharmacopoeia; JP = Japanese Pharmacopoeia; Ph Eur = European Pharmacopoeia; Q.S. = quantity sufficient; USP / NF = US Pharmacopeia / National Formulary.Table 2. Composition of PC-1 Drug Product per VialPharmacopoeia; JP = Japanese Pharmacopoeia; Ph Eur = European Pharmacopoeia; Q.S. = quantity sufficient; USP / NF = US Pharmacopeia / National Formulary.

[0383] The PC-1 DP comprises the drug substance filled at a target volume of 1.23 mL to enable an extractable volume of >1.0 mL in a single dose 2R, Type 1 borosilicate glass vial and sealed with a polypropylene nested cap that contains an embedded elastomeric stopper.

[0384] The physical and chemical properties of PC-1 are summarized in Table 3.Table 3. Physical and Chemical Properties of PC-1 Drug ProductCD3 = cluster of differentiation 3; DSC = differential scanning calorimetry; EGFR = epidermal growth factor receptor; ELISA = enzyme-linked immunosorbent assay; NTU = nephelometric turbidity unit; Tm = melting temperature.1Molecular weight measured by intact mass for the most abundant glycospecies (G2FS 1)2Relative potency of one batch compared to a second batch

[0385] 3.3 Storage and Handling

[0386] The PC-1 DP vials will be stored and shipped frozen (at -20 ± 5 °C). The vial contents will be diluted into an infusion solution that will be chosen based on the results of in-use compatibility studies.

[0387] 4.5 First-in-Human Dose Justification

[0388] The proposed starting dose in the FIH study is 50 pg, administered once weekly. The selection of the starting dose and regimen was based on a minimally anticipated biologic effect level (MABEL) approach integrating pharmacokinetic and pharmacodynamic data, including in vitro activity and in vivo safety data. The starting dose was calculated based on the expected Cmaxof PC-1 in humans translated from cynomolgus monkey PK studies and the most conservative PC-1 half-maximal effective concentration (EC50) derived from an in vitro cytotoxicity assay using an EGFR-expressing SCCHN tumor cell line co-cultured with human PBMCs. This PK-guided approach was used to compare the predicted human doses whose Cmaxmatched the PC-1 EC50 from in vitro cytotoxicity studies as well as the Cmaxfrom the cynomolgus monkey GLP toxicity study. Compared to the 2 rig / mL PC-1 Cmaxfrom the low dose (0.05 mg / kg) group in the GLP toxicity study where minimal to no cytokines were induced, the 142 ng / mL in vitro cytotoxicity PC-1 EC50 was more conservative. The proposed FIH dose includes an additional lOx safety factor to the 500 pg dose, which is based on the most conservative in vitro cytotoxicity assay, to further ensure safety. Using the additional lOx safety factor, the calculated FIH dose for PC-1 is 50 pg.

[0389] To ensure that PK, pharmacodynamic, and safety data are fully characterized, a starting dose of 50 pg once-weekly administration is selected, which is 10 times lower than MABEL ECso-based dose.

[0390] First-In-Human Study of PC-1

[0391] 5-EFFECTS IN HUMANS

[0392] 5. 1 Introduction

[0393] There is no clinical experience with PC-1. The current study will be the FIH Phase 1 clinical trial of PC-1. For complete patient eligibility criteria, please see the clinical study protocol. A brief summary of a planned clinical study is provided below.

[0394] 5.1.1 Study Design

[0395] Aa first-in-human (FIH), Phase 1 / lb, open-label, multicenter dose escalation and dose expansion study to assess the safety, tolerability, PK, PD, and preliminary anti -tumor activity of PC-1 in adult subjects with histologically confirmed advanced or metastatic CRC, NSCLC, SCLC, PDAC, TNBC, SCCHN and RCC will be conducted.

[0396] The study will be conducted in 3 parts: Dose Escalation (Part 1) with approximately 40 to 50 subjects, Cohort Backfill Expansion (Part 2) with up to approximately 40 subjects enrolled across 4 doselevels, and Dose Expansion (Part 3) with up to approximately 40 subjects enrolled at the RP2D.

[0397] Dose Escalation (Part 1) will assess the safety, tolerability, PK, PD, and preliminary efficacy of PC-1 administered by IV infusion.

[0398] Cohort Backfill Enrichment (Part 2) will allow for further characterization of safety and activity of dose levels. Dose Expansion (Part 3) will determine additional safety, tolerability, PK, PD, and preliminary clinical activity data with PC-1 at a dose and schedule to be determined by the Safety Review Committee after reviewing all available safety, PK, PD, and preliminary efficacy data. Depending on the data, randomization may be integrated for either 2 different PR2D doses or 2 different treatment intervals.

[0399] 6-SUMMARY OF DATA AND GUIDANCE

[0400] 6.1 Indication

[0401] PC-1 is in development for the treatment of advanced or metastatic solid tumors that are unresponsive to currently available therapies. PC-1 is currently not approved for any indication.

[0402] 6.2 Dosage and Administration

[0403] In the FIH study, the starting dose will be 50 pg, to be followed by dose escalation. PC-1 will be administered IV on Days 1, 8, and 15 of 21-day cycles. Different dosing intervals may be considered based on evolving PK, PD, safety, and efficacy data.

[0404] 6.3 Dosage Forms and Strengths

[0405] The PC-1 DP will be provided as a solution for injection for IV administration at a single strength presentation of 2 mg / mL.

[0406] 6.4 Contraindications

[0407] PC-1 should not be administered to patients who have had allergic or anaphylactic reactions to any component of PC-1. No other contraindications for PC-1 are currently known.

[0408] 6.5 Warnings and Precautions

[0409] There has been no prior clinical experience with PC-1. PC-1 is an experimental drug that should be administered only to patients within the context of a clinical study.

[0410] PC-1 is an antibody fragment based bispecific protein construct. Like other molecules in this class, it is highly specific for its targets. Although antibody therapeutics are well-tolerated, they are ‘foreign’ proteins, and some patients may experience infusion-related reactions or develop an immune response against them.

[0411] 6.6 Potential Adverse Reactions

[0412] The adverse reaction profile of PC-1 is unknown. However, safety measures should be considered based on nonclinical data and the mechanism of action.

[0413] Experience with other protein based biological therapies indicates that pyrexia, immunogenicity reactions (i.e., formation of ADAs), and / or hypersensitivity reactions may be observed. These reactions can be both serious and systemic (e.g., anaphylaxis) and may occur acutely or be delayed.

[0414] Because PC-1 is a T cell redirecting bispecific antibody, activation of T cell may induce CRS, neurotoxicity, and / or tumor lysis syndrome may occur.

[0415] Because PC-1 targets EGFR, adverse events reported with other EGFR targeting agents, such as cetuximab or panitumumab, may be observed. These risks include cardiac toxicity, pulmonary fibrosis or interstitial lung disease, dermatologic and soft tissue toxicities, photosensitivity, and ocular toxicity (ERBITUX® USPI; VECTIBIX® USPI).

[0416] 6.6.1 Infusion -Related Reactions

[0417] PC-1 is a recombinant protein based therapeutic, and administration of therapeutic proteins has been associated with infusion reactions with symptoms and signs including fever, rigors, rash, urticaria, dyspnea, hypotension, and / or nausea. To minimize the risk of infusion reactions, premedication with acetaminophen (or paracetamol) and an anti-histamine regimen should be administered during Cycle 1 and per standard institutional practice prior to administration of each dose of PC-1 afterwards as tolerated.

[0418] 6.6.2 Cytokine Release Syndrome

[0419] The identified risks of treatment with TCEs are primarily related to cytokine release and CRS. PC-1 is designed to reduce the risk of CRS by requiring protease cleavage for activation, focusing molecular activity to the TME where proteases are overexpressed, dysregulated, and activated. This approach has been shown to markedly reduce systemic cytokine exposure in preclinical models. However, CRS is a potential adverse reaction based on the mechanism of action of PC-1.

[0420] Symptoms associated with CRS vary greatly and may be difficult to distinguish from other conditions. The more common symptoms include fever, tachycardia, hypotension, hypoxia, fatigue, nausea, headache, dyspnea, rigors, myalgia / arthralgia, and anorexia. The severity of symptoms can be mild to life-threatening and thus, there should be a high suspicion for CRS if these symptoms occur. Grade 1 CRS according to the American Society for Transplantation and Cellular Therapy consensus grading scale does not require any intervention, but subjects should be monitored closely. Grade >2 CRS requires PC-1 dosing interruption and prompt symptomatic treatment per local standard institutional practice.

[0421] Preventive measures of cytokine release and CRS will include glucocorticoid premedication, IV pre-hydration, and holding of anti -hypertensive medication on day of the first infusion. Priming and step dosing will be initiated if CRS is seen during dose escalation.

[0422] 6.6.3 Other Potential Toxicities

[0423] 6.6.3. 1 Tumor Lysis Syndrome

[0424] Subjects with a high disease burden may be at risk for developing tumor lysis syndrome (TLS) with PC-1 treatment. Prophylactic treatment / measures are strongly recommended for subjects considered to be at risk for TLS, per institutional or clinical standards. Subjects should be closely monitored for laboratory evidence of TLS (hyperuricemia, hyperkalemia, hyperphosphatemia, and hypocalcemia).

[0425] In the case of evidence of TLS associated with PC-1, subjects will be admitted to the hospital, as clinically indicated. Standard management will include vigorous IV hydration, hypouricemic agents, and correction of acidosis, if present. Renal function, serum uric acid, calcium, phosphorus, and electrolytes should be closely monitored.

[0426] Subjects with Grade 3 to 4 TLS during Week 1 or Cycle 1 may also be hospitalized for >24 hours after the end of the administration of the subsequent dose, with considerations for dose reduction as described in the study protocol.

[0427] 6.6.3.2 Neurological Events Administration of solid tumor-targeted bispecifics has shown less potential for severe neurologic toxicity (compared with chimeric antigen receptor T cell therapy or anti- CD19 BiTE format antibodies such as blinatumomab) in early-stage clinical studies. Although PC-1 at active doses has not shown a propensity for induction of cytokine levels that may be associated with neurologic toxicity in in vitro human cell cultures or in vivo cynomolgus monkey studies, the risk of neurotoxicity is unknown and caution is warranted.

[0428] Neurological events associated with cytokine release may include tremor, mental status changes, confusion, speech difficulties, and potentially seizures. Monitor subjects for neurological events and exclude other causes for neurological symptoms. Neurological events should be graded according to the National Cancer Institute Common Terminology Criteria for Adverse Events (CTCAE) version 5.0. Mild disorientation or expressive aphasia (trouble word-finding) may be the earliest and most specific signs. Provide supportive care as needed for any neurological events. Workup may include head magnetic resonance imaging and electroencephalogram and may require corticosteroids, or anti-seizure medications, if severe. The medical monitor should be contacted if there is any potentially treatment- related neurological toxicity.

[0429] 6.6.3.3 Infections

[0430] As seen with other TCE-based immunotherapies, serious infections, including fatal bacterial, fungal, and new or reactivated viral infections, may occur during and / or following the completion of PC- 1 -based therapy. New or reactivated viral infections may include cytomegalovirus, herpes simplex virus, parvovirus B19, varicella zoster virus, West Nile virus, hepatitis B virus (HBV), and hepatitis C virus. PC-1 should be discontinued if serious infections develop, and appropriate anti -infective therapy instituted. PC-1 is not recommended for use in subjects with severe, active infections.

[0431] 6.6.3.4 Hepatitis B Reactivation

[0432] Hepatitis B virus reactivation can occur in patients treated with drugs classified as TCE. Cases have been reported in patients who are hepatitis B surface antigen (HBsAg) -negative but are hepatitis B core antibody (anti-HBc)-positive.

[0433] HBV reactivation is defined as an abrupt increase in HBV replication manifesting as a rapid increase in serum HBV DNA levels or detection of HBsAg in a person who was previously HBsAg - negative and anti-HBc-positive. Reactivation of HBV replication is often followed by hepatitis (i.e., increase in transaminase levels). In severe cases, increase in bilirubin levels, liver failure, and death can occur.

[0434] It is recommended to monitor subjects with evidence of prior HBV infection (anti-HBc-positive) with HBV DNA testing monthly and for clinical and laboratory signs of hepatitis during and for several months following PC-1 therapy.

[0435] 6.6.3.5 Immunization

[0436] The safety of immunization with live viral vaccines during or following PC-1 therapy has not been studied. Vaccination with live virus vaccines is not recommended for >2 weeks prior to the start of PC-1 treatment, during treatment, and until immune recovery following the last cycle of PC-1.

[0437] 6.6.4 Toxicities for Compounds Targeting EGFR

[0438] While not observed in preclinical studies with PC-1, cardiac, pulmonary, skin, and ocular toxicities have been observed with other compounds targeting EGFR. Subjects should be informed of these toxicities observed with other compounds targeting EGFR.

[0439] 6.6.4. 1 Cardiac Toxicity

[0440] Cardiac toxicity, including cardiac arrest, has been observed with other compounds targeting EGFR. Periodic echocardiograms and electrocardiograms will be conducted together with monitoring troponin I and brain natriuretic peptide as early signs of toxicity. In addition, electrolyte abnormalities should be carefully monitored.

[0441] 6.6.4.2 Pulmonary Toxicity

[0442] Events of interstitial lung disease or pulmonary fibrosis have been observed with other compounds targeting EGFR. Subjects should be advised to report any suggestive symptoms, such as dyspnea or cough.

[0443] 6.6.4.3 Skin Toxicity

[0444] Clinical manifestations of dermatologic and soft skin tissue toxicity, including but not limited to, acneiform dermatitis, pruritus, erythema, rash, skin exfoliation, paronychia, dry skin, and skin fissure, have been observed with other compounds targeting EGFR. Subjects should be monitored not only for infusion site reactions but also any skin lesions. Dermatological toxicity may be exacerbated by exposure to sunlight. Subjects should be advised to wear sunscreen and hats to limit sun exposure.

[0445] 6.6.4.4 Ocular Toxicity

[0446] Clinical manifestations of ocular toxicity including eye inflammation, lacrimation, light sensitivity, blurred vision, eye pain and / or red eye have been observed with other compounds targeting EGFR. Subjects should be informed about potential ocular toxicity and should be further examined at the first sign of possible ocular toxicity.

[0447] 6.7 Drug Interactions

[0448] The drug interaction profile of PC-1 is unknown, however, in general, antibodies such as PC-1 are not metabolized by CYP enzymes or transported by Pgp or related adenosine triphosphate-binding cassette membrane transporters. Cytokines produced by activated lymphocytes may impact the levels of Pgp and the activity of CYP enzymes. The clinical relevance of PC-1 immune modulation and potential cytokine production that could impact Pgp and CYP is unknown, but a clinically relevant drug-drug interaction effect is considered highly unlikely. Caution needs to be paid where subjects are receiving substrates of CYP enzymes with narrow therapeutic index window. Subject should be monitored closely and have doses for the concomitant treatments adjusted as necessary.

[0449] 6.8 Use in Specific Populations

[0450] 6.8.1 Fertility, Pregnancy, and Lactation

[0451] Nonclinical studies evaluating the effect of PC-1 on embryo-fetal development or reproductive parameters have not been conducted. Therefore, pregnant women and those breastfeeding will be excluded. Men and women who are biologically capable of having children must agree to commit to the use of highly effective methods of contraception as outlined in the study protocol.

[0452] 6.8.2 Geriatric Use

[0453] No studies in geriatric patients have been conducted. Elderly patients will be included in the initial clinical trials of PC-1. Elderly patients should, however, be carefully monitored.

[0454] 6.8.3 Renal Impairment

[0455] PK / functional alterations due to renal impairment are not anticipated.

[0456] 6.10 Clinical Studies

[0457] No prior clinical studies of PC-1 have been conducted.

[0458] 6.11 Reference Safety Information for Assessment of Expectedness of Serious Adverse Reactions

[0459] For regulatory reporting purposes, all adverse events (AEs) will be assessed as being unexpected at this stage of the development program. Therefore, AEs that are serious and possibly related to PC-1 will be reported to the health authorities per applicable regulations.EXAMPLE 2: Clinical Study Protocol of PC-1

[0460] This example illustrates an open label multicenter, phase 1 / lb study of PC-1 disclosed herein in subjects with advanced or metastatic solid tumor malignancies.

[0461] Protocol SynopsisTable 4: Study objective and endpointsADA = anti-drug antibody; AE = adverse event; ASTCT = American Society for Transplantation andCellular Therapy; AUClast = area under the concentration-time curve from time 0 to last timepoint prior to next dose; CL = clearance; Cmax= maximum concentration; CRS = cytokine release syndrome; ctDNA = circulating tumor DNA; DLT = dose-limiting toxicity; DOR = duration of response; EGFR = epidermal growth factor receptor; IHC = immunohistochemistry; IV = intravenous; MAD = maximally administered dose; MTD-R = maximum tolerated dose regimen; NCI CTCAE v5.0 = National Cancer Institute Common Terminology Criteria for Adverse Events Version 5.0; ORR = overall response rate; OS = overall survival; PFS = progression-free survival; PK = pharmacokinetics; RECIST = Response Evaluation Criteria in Solid Tumors; RP2D-R = recommended Phase 2 dose regimen; SAE = serious adverse event; ti / 2 = terminal half- life; Vd = volume of distribution

[0462] OVERALL STUDY DESIGN

[0463] This study is a first-in-human (FIH), Phase 1 / lb, open-label, multicenter dose escalation and dose expansion study to assess the safety, tolerability, pharmacokinetics (PK), pharmacodynamics, and preliminary anti-tumor activity of PC-1 in adult subjects with histologically confirmed metastatic or advanced non-small cell lung cancer (NSCLC), squamous cell carcinoma of the head and neck (SCCHN), colorectal carcinoma (CRC), small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple -negative breast cancer (TNBC), prostate cancer, or renal cell carcinoma (RCC). An outline of the study is shown in FIG. 2.

[0464] This study will be conducted in 3 parts: Dose Escalation (Part 1) with approximately 50 subjects, Backfill enrichment cohorts (Part 2) with up to approximately 60 subjects enrolled up to approximately 6 cohorts, and Dose Expansion (Part 3) with up to approximately 40 subjects at 2 or more preliminary recommended Phase 2 dose regimens (RP2D-R). In Part 1, transition from flat to step dosing regimens may be in part due to cytokine release syndrome or safety review committee recommendation. Indications for Part 1 include NSCLC, SCLC, SCCHN, PDAC, TNBC, CRC, and RCC, with CRC enrollment capped to keep less than 40%. An alternate regimen, for example every 2 weeks (Q2W) or every 3 weeks (Q3W) may be evaluated and intra-subject dose escalation may be allowed. An archival sample less than 24 months old or a baseline fresh biopsy may be obtained after clinical efficacy is observed. In Part 2, up to 6 backfill enrichment cohorts at candidate recommended phase 2 dose regimen (RP2D-R) no greater than maximum tolerated dose regimen (MTD-R) or maximum administered dose (MAD). Based on Part 1 data, alternate regimens, e.g., Q2W or Q3W, may be evaluated. Specific indications may be defined based on Part 1 data. Archival sample <24 months or baseline fresh biopsy may be obtained. In Part 3, preliminary RP2D-Rs are to be selected by Part 1 and Part 2 data. 2 or more regimens per indications may be selected with randomization. Populations in Part 3 may include second line SCCHN, second line SCLC, third line PDAC, second line TNBC, second line RCC, or third line CRC.

[0465] PC-1 will be administered intravenously (IV) weekly (QW) with step dosing schedule A and flat dosing schedule E. Alternate dose schedules for step dosing or flat dosing [e.g. step dosing schedules C and D (additional dose on Day 4 in Cycle 1), flat dosing schedule F and step schedule B (Q2W), flat dosing schedule G (Q3W), etc.] may also be evaluated (see FIGs. 4A-4C and FIGs. 5A-5D). Treatment with PC-1 will continue until the subject meets one of the criteria for treatment discontinuation in Section 6.1, such as progressive disease and unacceptable toxicities. Duration of treatment with study drug could be shortened if the study is terminated by the Sponsor (Section 6.3). Given the possibility of dose schemes involving different dose levels in the step dose schedules this protocol will refer to dose regimens instead of dose levels. The study will assess the safety and tolerability of different dose regimens of PC-1 which can be either flat dose or step dose. Moreover, the Maximum Tolerated Dose (MTD) will be referred to as the Maximum Tolerated Dose Regimen (MTD-R) and the Recommended Phase 2 Dose (RP2D) will be referred to as the Recommended Phase 2 Dose Regimen (RP2D-R). If theMTD-R is not defined during dose escalation, the highest dose administered in this study will be declared the Maximum Administered Dose (MAD).

[0466] Dose escalation of PC-1 in Part 1 is guided by a 3-step Bayesian Logistic Regression Model (BLRM) with escalation with overdose control (EWOC) that models cytokine release syndrome (CRS), dose limiting toxicities (DLTs) and non-CRS DLTs separately and calculates the joint probability of overall DLTs based on these two types of toxicity.

[0467] . The BLRM will be assessed for those subjects satisfying the requirements for inclusion in the DLT evaluable Analysis Set (DAS). After completion of a given dose regimen cohort, or at any time the BLRM is updated, the decision to dose-escalate and the actual regimen chosen will be guided by BLRM according to the escalation with overdose control (EWOC) principle and review of available clinical, PK, and laboratory data by the Safety Review Committee (SRC).

[0468] To minimize the number of subjects treated at potentially subtherapeutic dose regimens, the cohort size will be 1 to 3 subjects in the first cohort (dose 50 pg in a flat dose regimen) followed by cohorts of 3 to 6 subjects in the subsequent flat dose regimens. Cohorts of subjects will receive escalating doses regimens of PC-1 until the MTD-R / MAD and / or RP2D-R is reached or a suitable RP2D-R is identical. Each cohort will consist of newly enrolled subjects.

[0469] Step-up dose(s) may be introduced to allow subsequent higher level dose administration. Specific criteria to initiate step-up dosing are described in Section 3.4.2.2. If a step-up dose is implemented, the study will aim to establish the MTD of a dosing regimen that includes a step-up dose(s), and an independent MTD-R of a regimen that does not include a step, the SRC will review all available safety, PK, and data and will be guided by the BLRM to identify the MTD-R for both the flat and step dose regimens.

[0470] All subjects in Part 1 and Part 2 of this study will be required to be hospitalized as outlined in Section 5.2.

[0471] Safety assessments include AEs, vital signs, physical examinations, Eastern Cooperative Oncology Group (ECOG) scores, clinical laboratory tests, electrocardiograms (ECGs), and concomitant medications. Central labs for comprehensive assessment of cytokine and chemokine biomarker panels will also be collected.

[0472] DOSE-LIMITING TOXICITIES (DLT) AND DLT PERIOD

[0473] Subjects are classified as DLT evaluable if they experience a DLT or if they otherwise, in the absence of a DLT, receive at least 66% of intended dose of the study drug during Cycle 1 for a flat dose regimen, or receive all plarmed step doses and the target dose in Cycle 1 for a step dose regimen and have completed all scheduled safety assessments during the DLT observation period. Subjects who do not meet one of these criteria criterion may be replaced. For the purpose of dose regimen escalation, the DLT observation period is defined to be 21 days from the Cycle 1 Day 1 dose. For step dosing cohorts, an extension of the DLT observation period will be implemented to maintain 7-day observation after the first target dose level administration if more than 2-step doses are investigated. All AES will be collected from all subjects enrolled in the study. The severity of AES will be graded using the NCI CTCAE v5.0with the exception of CRS events, which will be graded per ASTCT criteria. Further details for DLTs are provided in Section 3.4.2.3.

[0474] INCLUSION CRITERIA

[0475] Each subject must meet the following criteria to be enrolled in this study:1. Males or females > 18 years of age who comprehend and are willing and able to provide written informed consent and can comply with scheduled visits, treatment schedule, laboratory tests, and other requirements for the study.2. ECOG performance status of 0 or 1.3. Adequate bone marrow function, including: a. Absolute neutrophil count (ANC) > 1500 / mm3or > 1.5 x 109 / L (ANC must be assessed at least 14 days from the last growth factor support) b. Platelet count > 75,000 / mm3or > 75 x 109 / L (platelet count must be assessed at least 7 days from the time of the prior transfusion) c. Hemoglobin > 9.0 g / dL (hemoglobin must be assessed at least 7 days from the time of the prior transfusion). Subjects requiring multiple transfusions in recent past (approximately 3 months) should be discussed with the Medical Monitor.4. Adequate renal function, including: a. Estimated creatinine clearance > 40 mL / min as calculated using the Cockcroft Gault equation, or method standard for the institution5. Adequate liver function, including: a. Total bilirubin < 1.5 x upper limit of normal (ULN) (except subjects with Gilbert syndrome, who must have total bilirubin < 3.0 mg / dL) b. Aspartate transaminase (AST) < 2.5 xULN (if subject has known liver metastases, then < 5 xULN) c. Alanine transaminase (ALT) < 2.5 xULN (if subject has known liver metastases, then <5 xULN) d. Serum albumin > 28 mg / mL6. Adequate pulmonary function as evidenced by baseline resting oxygen saturation > 92% on room air7. Adequate cardiac function as evidenced by cardiac ejection fraction > 50% by echocardiogram (ECHO).8. International normalized ratio (INR) or activated partial thromboplastin time (aPTT) < 1.5 xULN unless subject is receiving anticoagulation therapy as long as (prothrombin time) PT or aPTT is within the therapeutic range of intended use anticoagulants.9. Resolved acute effects of any prior therapy to baseline severity or CTCAE Grade < 1 except forAEs not constituting a safety risk by Investigator judgement. Subjects receiving ongoing replacement hormone therapy for endocrine immune-related adverse events without clinical symptoms will not be excluded.10. Male subjects able to father children and female subjects of childbearing potential must agree to use 2 highly effective methods of contraception (as defined in this protocol) during the treatment period and until 90 days after the last dose of study treatment.11. Have at least 1 measurable lesion per RECIST 1. 1 criteria.

[0476] Inclusion Criteria Specific to Part 112. Histologically or cytologically documented locally advanced or metastatic NSCLC, SCLC, SCCHN, CRC, PDAC, TNBC, prostate cancer, or RCC (for histology type, see Section 1.2.2. 1). Enrollment of subjects with CRC will be capped at approximately 40%.13. Progressed or was intolerant to all available therapies known to provide clinical benefit appropriate for the tumor type for which the subject was eligible and willing to receive. Subjects must not be amenable to local therapy with curative intent (surgery or radiation therapy with or without chemotherapy). Documentation of intolerance must be provided.14. A fresh tumor biopsy from a representative lesion is encouraged. If not, archival tumor biopsy (taken within 24 months prior to enrollment in this study) must be provided.

[0477] Inclusion Criteria Specific to Part 215. Histologically or cytologically documented locally advanced or metastatic NSCLC, SCLC, SCCHN, CRC, PDAC, TNBC, prostate cancer, or RCC (for histology type, see Section 1.2.2. 1).16. A fresh tumor biopsy from a representative lesion is encouraged. If not, an archival tumor biopsy (taken within 24 months prior to enrollment in this study) must be provided.Inclusion Criteria Specific to Part 3 (One to two of following indications to be selected)17. A fresh tumor biopsy from a representative lesion is encouraged. If not, an archival tumor biopsy (taken within 3 months prior to enrollment in this study) must be provided with confirmation of EGFR positivity by IHC in the biopsy sample.

[0478] Inclusion Criteria Specific to CRC18. Histologically or cytologically confirmed diagnosis of advanced / metastatic colorectal adenocarcinoma.19. Has been treated with fluoropyrimidine-, oxaliplatin- and irinotecan-based chemotherapy, an anti- vascular endothelial growth factor (VEGF) therapy in first line (IL) or second line (2L) settings.20. Has been treated with an anti -EGFR therapy in IL or 2L settings if colon cancer is left sided and RAS is wild-type.

[0479] Inclusion Criteria Specific to SCCHN21. Histological diagnosis of primary tumor location of oral cavity, oropharynx, hypopharynx, or larynx. Subject may not have a primary tumor site of nasopharynx (any histology). Results from testing of human papilloma virus (HPV) status (pl 6, ISH or PCR) must be available for subjects with oropharyngeal cancer.22. Subjects should be in the 2L or 2L+ setting and should have received platinum-based chemotherapy and anti-PD-Ll / PD-1 -therapy in combination or sequentially in any order for metastatic or advanced disease. Subject must be resistant to, or experience disease recurrence after these therapies.

[0480] Inclusion Criteria Specific to NSCLC23. Histologically or cytologically confirmed diagnosis of advanced / metastatic NSCLC (adenocarcinoma and squamous cell carcinoma subtypes).24. Subjects without an actionable driver mutation should be in the 2L or 2L+ setting and should have received platinum -based chemotherapy and anti-PD-Ll / PD-1 -therapy in combination or sequentially in any order for metastatic or advanced disease. Subject must be resistant to, or experience disease recurrence after these therapies.25. Subjects with an actionable mutation (e.g., EGFR, ALK, BRAF, ROS, NTRK, MET, RET, KRAS, or HER2) in the 2L or 3L setting may be enrolled. Such subjects should have received at least one treatment specific to the driver mutation for the disease and at least one line but no more than two lines of systemic chemotherapy for metastatic or advanced status.

[0481] Inclusion Criteria Specific to RCC26. Histologically or cytologically confirmed diagnosis of advanced / metastatic renal cell carcinoma (papillary or clear cell type).27. Subjects must have received at least 2 and no more than 3 prior lines of therapy for recurrent or metastatic disease, including both standard of care and investigational therapies. Subjects must have progressed / relapsed, be refractory, or intolerant to approved standard therapy.Inclusion Criteria Specific to PDAC28. Histologically or cytologically confirmed diagnosis of advanced / metastatic pancreatic ductal adenocarcinoma (PDAC).29. Subjects must have received at least 2 and no more than 3 prior lines of therapy for recurrent or metastatic disease, including both standard of care and investigational therapies.30. Subjects with an actionable mutation (e.g., BRCA1 / 2 or PALB2) setting may be enrolled. Such subjects should have received at least one target therapy specific to the driver mutation for the disease.

[0482] Inclusion Criteria Specific to TNBC31. Histologically or cytologically confirmed diagnosis of unresectable advanced / metastatic triple negative breast cancer (TNBC). Most recent biopsy must be HER2 and hormone receptor (HR) negative based on local testing. American Society of Clinical Oncology / College of American Pathologist (ASCO / CAP) guidelines should be utilized for assessing HER2 and HR status.32. Subjects must have received at least 2 and no more than 3 prior lines of therapy for recurrent or metastatic disease, including both standard of care and investigational therapies.Inclusion Criteria Specific to SCLC33. Histologically or cytologically confirmed diagnosis of advanced / metastatic SCLC.34. Subjects must have received at least 2 and no more than 3 prior lines of therapy for recurrent or metastatic disease, including both standard of care and investigational therapies.

[0483] EXCLUSION CRITERIA

[0484] Subjects are excluded from the study if any of the following criteria apply:History of prior malignancy other than the cancer under treatment in this study, except for adequately treated in situ cancer (excluding carcinoma in situ of the bladder), basal cell or squamous cell skin cancer or other cancers (e.g., breast, prostate) for which the subject has been disease-free for at least 3 years. Active brain or leptomeningeal metastasis. Subjects with known brain metastases are eligible if they have been treated and an MRI shows no evidence of progression for at least 8 weeks after treatment is completed and within 3 weeks prior to first dose of study drug. Subjects are not eligible if they require a high dose of systemic corticosteroids that could result in immunosuppression (> 10 mg / day prednisone equivalents) for at least 2 weeks prior to enrollment. Treatment with anti -neoplastic therapy (including but not limited to cytotoxic chemotherapy, major surgery, radiation, biologic agents, and investigational agents) within 28 days or < 5 elimination half-lives, whichever is earlier, before enrollment. Major surgery defined as any surgical procedure that involves general anesthesia and a significant incision (i.e., larger than what is required for the placement of a central venous access, percutaneous feeding tube, or biopsy) within 28 days of enrollment, or anticipated surgery during the study. Radiation therapy with treatment intent within 28 days prior to enrollment. Any palliative radiation therapy completed within the 7 days prior to enrollment is exclusionary. Females who are pregnant or breastfeeding. Prior treatment with EGFR-targeted bispecific T cell engager or chimeric antigen receptor T cell (CAR-T) therapy. Prior treatment with cluster of differentiation (CD)3 engaging bispecific antibodies. Prior solid organ transplant. Subjects with active, uncontrolled bacterial, fungal, or viral infection, including hepatitis B virus (HBV), hepatitis C virus (HCV), known human immunodeficiency virus (HIV), or acquired immune deficiency syndrome (AIDS)-related illness. In equivocal cases, subjects whose viral load is negative may be eligible. Subjects with elevated C-reactive protein (CRP) at screening should be discussed with the Medical Monitor and carefully evaluated to rule out active infection or autoimmune disease.HBV: Subjects who are hepatitis B surface antigen (HBsAg)-positive are eligible if they have received HBV antiviral therapy for at least 4 weeks and have undetectable HBV viral load prior to randomization.- Note: Subjects should remain on anti-viral therapy throughout study intervention and follow local guidelines for HBV anti-viral therapy post-completion of study intervention.HCV: Subjects with a history of HCV infection are eligible if their HCV viral load is undetectable at screening.Note: Subjects must have completed curative anti-viral therapy at least 4 weeks prior to randomization. Active bleeding disorder, including gastrointestinal bleeding, as evidenced by hematemesis, significant hemoptysis, or melena in the past 6 months. History of severe allergic or anaphylactic reactions to monoclonal antibodies (or recombinant antibody -related fusion proteins). Active autoimmune disease requiring systemic therapy (exceptions [s] : subjects with vitiligo, resolved childhood atopic dermatitis, hypothyroidism, or hyperthyroidism that is clinically euthyroid at screening are allowed). Subjects with a history of CTCAE Grade > 3 immune-related AEs that were considered related to prior immuno-oncology treatment. Uncontrolled hypertension, defined as systolic blood pressure > 160 mmHg or diastolic blood pressure > 100 mmHg which has been confirmed by 2 successive measurements despite optimal medical management. Baseline 12-lead ECG that demonstrates clinically relevant abnormalities that may affect subject safety or interpretation of study results (e.g., baseline QTc interval > 470 msec, complete LBBB, signs of an acute or indeterminate -age myocardial infarction, ST- interval changes suggestive of active myocardial ischemia, second- or third-degree atrioventricular block, or serious bradyarrhythmias or tachyarrhythmias.Note: If the baseline uncorrected QT interval is > 470 msec, this interval should be rate -corrected using the Fridericia method (QTcF), and the resulting QTcF should be used for decision making and reporting. Any of the following in the previous 6 months: myocardial infarction, symptomatic congestive heart failure (NYHA Class > II), unstable angina, coronary / peripheral artery bypass graft, or unstable cardiac arrhythmia requiring medication. Ongoing cardiac dysrhythmias of NCI CTCAE Grade > 2, atrial fibrillation of any grade (Grade > 2 in the case of asymptomatic lone atrial fibrillation). Subjects with cardiac rhythm device / pacemaker must be discussed in detail with the Medical Monitor to judge eligibility. Any of the following in the previous 6 months: cerebrovascular accident, transient ischemic attack, symptomatic pulmonary embolism, and / or other clinically significant episodes of thromboembolic disease. Venous thromboembolic events that are non -life threatening and stable on low molecular weight heparin or factor Xa inhibitors that can be reversed by andexanet-a are allowed or treated with 4-factor prothrombin complex concentrate (PCC) as direct oral anticoagulant reversal agent are allowed. Subjects with a history of interstitial lung disease, noninfectious pneumonitis that required steroid, radiation pneumonitis, active pulmonary tuberculosis, or evidence of active pneumonitis on screening chest CT scan. Subjects with radiation therapy to the lung that is > 30 Gy within 6months of the first dose of treatment are excluded. Subjects with active lung infections requiring treatment are also excluded.20. Subjects on supplemental oxygen.21 . Clinically significant and unmanageable ascites defined as requiring constant therapeutic paracentesis (limited medical treatment to control ascites is permitted, but all subjects who received paracentesis within 3 months of expected first dosing date will require review by the Medical Monitor).22. Subjects with a history of gout or active arthritis / arthralgia at screening must be excluded.23. Subjects with other inflammatory arthropathies at screening must be discussed with the Medical Monitor to judge eligibility.24. Any condition including medical, emotional, psychiatric, or logistical that, in the opinion of the Investigator, would preclude the subject from adhering to the protocol.

[0485] SAFETY REVIEW COMMITTEE (SRC)An SRC will oversee the conduct of the clinical study. The SRC will review cumulative safety data, as well as available PK, pharmacodynamic, and preliminary efficacy data, and make final decisions regarding dose regimen escalation and overall study conduct guided by BLRM to ensure subject safety while receiving study treatment. The SRC will meet to evaluate the safety of all cohorts through dose escalation and at regular intervals through dose expansion.

[0486] SAMPLE SIZE

[0487] The total number of subjects is estimated to be approximately 150.

[0488] Dose Escalation (Part 1) utilizes 3-step BLRM that combines two BLRMs. With this type of study design, the exact number of subjects needed to complete the dose escalation phase is unknown, as it depends on the number of subjects enrolled in each cohort and the number of cohorts required to reach MTD-R / RP2D-R. Approximately 50 subjects are expected to be treated in Part 1 for the Bayesian model to have reasonable operating characteristics relating to its MTD-R definition and / or RP2D-R recommendation.

[0489] Backfill enrichment cohorts (Part 2) will enroll up to approximately 60 subjects. Up to 6 tumor specific cohorts of no more than 12 subjects each will be enrolled to gain further characterization of the safety, tolerability, PK, pharmacodynamics, and preliminary efficacy of defined dose regimens of PC-1. These data will be used to better select the dose regimen for Dose Expansion (Part 3). The backfill enrichment cohorts can be used to further assess the safety of dose regimens with or without step doses.

[0490] Dose Expansion (Part 3) will enroll approximately 40 subjects to further evaluate the RP2D-R.

[0491] STATISTICAL METHODS

[0492] This is an open-label clinical study and, in general, descriptive statistics will be employed to analyze the data. More details will be provided in a Statistical Analysis Plan (SAP). No formal hypothesis testing is planned. The number of subjects in each part of the study is based on safety and tolerability. A sample size of up to approximately 40 subjects in Dose Expansion (Part 3) will furthercharacterize and provide an estimation precision (as the maximum half-width of the 95% confidence interval [CI]) of approximately ± 16% for any binary rate of outcome assessing safety, tolerability, exposure, immunogenicity, PD, or clinical activities of single-agent PC-1. Sample size of approximately 20 subjects in each of the two RP2D-Rs in Dose Expansion (Part 3) and similarly approximately 20 subjects in each of the two indications will provide an estimation precision (as the maximum half-width of the 95% confidence interval [CI]) of approximately ±22% for the binary rate of outcomes. These levels of precision are deemed sufficient in the comprehensive review of the study data to guide future clinical development.

[0493] STARTING DOSE

[0494] The FIH starting dose will be 50 pg PC-1 IV weekly during a 21 -day cycle.Table 5: List of Abbreviations

[0495] Clinical Overview

[0496] This is a first-in-human (FIH) study for PC-1; therefore, no clinical data are available.

[0497] 1.2.2. 1 Tumor Type Selection RationaleEGFR is the most commonly overexpressed transmembrane oncogenic protein found in cancer (Wells, 1999). EGFR is a tyrosine kinase receptor that induces cell proliferation and differentiation upon ligand binding, and dysregulated EGFR function is known to play a role in oncogenesis (Chen et al, 2016). The critical role of EGFR in cell signaling has made it a primary target in cancer therapy. Two modes of action to blockade of the EGFR axis have been applied to oncology drug development: tyrosine kinase inhibitors (TKIs) inhibit EGFR phosphorylation and its downstream cascade by blocking the adenosine triphosphate (ATP) pocket located in the intracellular catalytic domain of the receptor; and anti -EGFR monoclonal antibodies (mAbs) target the extracellular domain of the receptor and block ligand-induced EGFR dimerization and tyrosine kinase activation. Blockade of the receptor function has led to clinical benefit and regulatory approval in patients with NSCLC, SCCHN, and CRC, validating EGFR as a therapeutic target. However, in refractory / relapse populations, significant unmet medical need exists. Furthermore, despite a high expression level of EGFR in many other tumor types, such as breast, pancreatic, ovary, prostate, and brain (glioblastoma multiforme), results using EGFR blockade have been disappointing (Nabhan et al, 2009; Cortes and Roche, 2012; Tagawa et al, 2012; Qian et al, 2020; Peereboom et al, 2010). To identify cancer indications with a high-level of EGFR expression,

[0498] The Cancer Genome Atlas and Tempus database were queried for EGFR RNA expression. Mining of both databases indicated the following cancer types express high levels of EGFR;• SCCHN: squamous cell carcinoma with primary tumor location of oral cavity, oropharynx, hypopharynx, or larynx;• Renal cell carcinoma (RCC): renal cell carcinoma with clear cell (not chromophobe, Hereditary cancer syndrome or other types);• NSCLC: non-small cell lung cancer; squamous cell carcinoma and adenocarcinoma; and• CRC: adenocarcinoma of rectum, adenocarcinoma of colon.• SCLC: small cell lung cancer; neuroendocrine• PDAC: pancreatic ductal adenocarcinoma; acinar cell carcinoma; and• TNBC: Triple-negative breast cancer; invasive ductal, medullary, and metaplastic carcinoma.• Prostate cancerTreatment targeting specifically EGFR is approved for only 3 indications: CRC, SCCHN, and NSCLC. Despite the approvals of EGFR targeting treatments and other therapies, significant unmet medical need still exists in those indications.

[0499] STUDY DESIGN

[0500] 3.1 Overall Study DesignThis study is a FIH, Phase 1 / lb, open-label, multicenter, dose regimen escalation and dose regimen expansion study to assess the safety, tolerability, PK, pharmacodynamics, and the preliminary anti -tumoractivity of PC-1 in adult subjects with histologically confirmed metastatic or advanced NSCLC, SCCHN, CRC, SCLC, RCC, PDAC, TNBC, prostate cancer, or RCC. An outline of the study is shown in FIG. 2. Given the possibility of different dose schemes involving different dose levels in the step dose schedules this protocol will refer to dose regimens instead of dose levels. The study will assess the safety and tolerability of different dose regimens of PC-1 which can either be flat dose or step dose. Moreover, the Maximum Tolerated Dose (MTD) will be referred to as a Maximum Tolerated Dose Regimen (MTD-R) and the Recommended Phase 2 Dose (RP2D) will be referred to as Recommended Phase 2 Dose Regimen (RP2D-R).

[0501] This study will be conducted in 3 parts: Dose Escalation (Part 1) with approximately 50 subjects, Backfill enrichment cohorts (Part 2) with up to approximately 60 subjects enrolled in up to 6 cohorts and Dose Expansion (Part 3) with up to approximately 40 subjects enrolled at 2 more preliminary RP2D-R.

[0502] PC-1 will be administered IV weekly (QW) with step dosing schedule A and flat dosing schedule E. Alternate dose schedules for step dosing or flat dosing [e.g. step dosing schedule C and D (additional dose on Day 4 in Cycle 1), flat dosing schedule F and step schedule B [every 2 weeks (Q2W)], flat dosing schedule G [every 3 weeks (Q3W)], etc.] may also be evaluated. Examples of the flat dosing and step dosing schedules that may be evaluated are depicted in FIGs. 4A-4C and FIGs. 5A-5D respectively.

[0503] All subjects in Part 1 and Part 2 of this study will be required to be hospitalized (for at least 24 hours) following Cycle 1 Day 1 administration of PC-1 for observation. The requirement for hospitalization following subsequent administration of PC-1 will be determined based on the clinical course following the first administration of PC-1. Safety assessments include AEs, vital signs, physical examinations, Eastern Cooperative Oncology Group (ECOG) scores, clinical laboratory tests, ECGs, and concomitant medications. In addition to central labs for comprehensive assessment of cytokine and chemokine biomarker panels, post-infusion cytokine levels will also be measured to assess safety during dose escalation. Dose finding cohorts in combination with approved standard of care treatment may be evaluated after a protocol amendment in the future.

[0504] 3.1.1 Part 1 : Dose Escalation

[0505] Part 1 will investigate dose regimens of PC-1 as monotherapy in flat or sequentially escalating regimens with different dose levels to determine the RP2D-R and / or maximum tolerated dose regimen (MT-R) / maximum administered dose (MAD). The starting dose for Part 1 will be 50 pg in a flat dose regimen. Adult subjects with advanced or metastatic NSCLC, SCCHN, SCLC, CRC, PDAC, TNBC, prostate cancer, or RCC will be enrolled. Enrollment of subjects with CRC will be capped at approximately 40% in Part 1 and will be managed according to the cohort management plan. Dose escalation of PC-1 in Part 1 is guided by a 3-step Bayesian Logistic Regression Model (BLRM) with escalation with overdose control (EWOC) that models CRS DLTs and non-CRS-DLTs separately and calculates the joint probability of overall DLTs on these two types of toxicity. The BLRM will be assessed for those subjects satisfying the requirements for inclusion in the dose-limiting toxicity (DLT) evaluable Analysis Set (DAS) and will model CRS and non-CRS DLTs separately. After completion of agiven dose cohort, or at any time the BLRM is updated, the decision to dose -escalate and the actual dose and schedule chosen will be guided by the recommendation of the BLRM for the next admissible dose according to the EWOC principle and review of available clinical, PK, and laboratory data. The Safety Review Committee (SRC) will make decisions regarding dose escalation and overall study conduct guided by the BLRM. To minimize the number of subjects treated at potentially subtherapeutic dose levels, the cohort size will be 1 to 3 subjects in the first cohort (dose 50 pg in a flat dose regimen) followed by cohorts of 3 to 6 subjects in the subsequent dose levels. Cohorts of subjects will receive escalating doses of PC-1 until the MTD-R / MAD is reached or a suitable RP2D-R is identified. Each cohort will consist of newly enrolled subjects. Mandatory archival biopsy sample collection will be implemented in Part 1 to evaluate the correlation of baseline EGFR expression with efficacy and safety after clinical efficacy (e.g., at least one unconfirmed partial or complete response) is observed.

[0506] Dose regimens with step doses may be introduced to allow subsequent higher level dose administration. Criteria to initiate step dosing is described in Section 3.4.2.2. If step dosing is implemented, the study will aim to establish the MTD-R of a dosing regimen that includes step dose(s), and an independent. MTD-R of a regimen that does not include step dosing. Planned doses that may be investigated are shown in Table 18 and Table 19. Dosing of the first and second subjects in each cohort will be staggered for appropriate durations (e.g. 3 - 7 days) depending on the regimen employed (See Section 3.4.2). The initiation of treatment of all subsequent subjects will be staggered by at least 1 day. Investigation of alternative dosing schedules may also be initiated, e.g., step dosing schedule C and D (the addition of a step dose on Day 4 in Cycle 1), flat dosing schedule F (Q2W) and flat dosing scheduled G (Q3W), based on emerging PK, pharmacodynamics, biomarker and safety data. The totality of the data across indications will be considered to better inform the overall safety profile of PC-1 and decisions taken relative to dosing.

[0507] 3.1.2 Part 2: Backfill Enrichment Cohorts

[0508] Up to 6 enrichment cohorts will be evaluated, with up to 12 subjects each. Based on emerging PK, pharmacodynamic, safety and efficacy data, backfill enrichment cohort(s) may commence at dose levels where the safety has been confirmed in Part 1. As the subjects enrolled in Part 2 will receive a dose lower than the concurrent dose escalation cohort in Part 1, their potential DLT observations will not be used in the BLRM for the ongoing dose finding. However, the safety profile from the Part 2 subjects will be used to establish the RP2D. At no time will the PC-1 dose level studied in Part 2 exceed the highest dose level that qualifies as an MTD in Part 1.

[0509] Specific indication(s) may be selected based on Part 1 data. Archival biopsies will be collected during screening.

[0510] 3.1.3 Part 3: Dose Expansion

[0511] Approximately 40 subjects will be enrolled with the tumor type deemed to have the highest potential for benefit based on data from Part 1 and Part 2. Preliminary PK / pharmacodynamic modeling and dose-exposure-response analyses will be performed using available pre-clinical and clinical data after completion of Part 1 and Part 2 of the study. These datasets will be used to select 2 or more doseregimens for Part 3 for comparison to identify the RP2D-R (Section 3.7). To minimize bias, randomization may be integrated for each cohort. PK exposure, minimal dose expected to achieve efficacy, highest dose with acceptable safety profile and other aspects will be considered for dose regimen selection in Part 3. Subject selection with EGFR IHC, based on the Part 1 and Part 2 data, will be integrated. Expansion cohorts enrolling other tumor types expressing EGFR may be included after a protocol amendment.

[0512] 3.2 Scientific Rationale for Study Design

[0513] In vitro data demonstrated that killing of tumor by PC-1 is dependent on cleavage of masking molecules and the presence of T cells (Section 1.2.1.2). The intended mode of action design of PC-1 in vivo was successfully supported by GLP toxicology data showing significantly reduced cytokine production and longer half-life, compared to EGFR-TCE (Se...

Claims

CLAIMSWHAT IS CLAIMED IS:

1. A method for treating cancer comprising administering to a subject in need thereof a first dose of an isolated recombinant polypeptide complex, wherein the first dose is at least about 50 pg, and wherein the isolated recombinant polypeptide complex comprises a first chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 1 and a second chain with an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 2.

2. The method of claim 1, wherein the first dose is about 50 pg to about 30 mg.

3. The method of claim 1 or 2, wherein the first dose is at least about 100 pg.

4. The method of claim 1 or 2, wherein the first dose is at least about 150 pg.

5. The method of claim 1 or 2, wherein the first dose is at least about 200 pg.

6. The method of claim 1 or 2, wherein the first dose is at least about 250 pg.

7. The method of claim 1 or 2, wherein the first dose is at least about 300 pg.

8. The method of claim 1 or 2, wherein the first dose is at least about 350 pg.

9. The method of claim 1 or 2, wherein the first dose is at least about 400 pg.

10. The method of claim 1 or 2, wherein the first dose is at least about 450 pg.

11. The method of claim 1 or 2, wherein the first dose is at least about 500 pg.

12. The method of claim 1 or 2, wherein the first dose is at least about 750 pg.

13. The method of claim 1 or 2, wherein the first dose is at least about 1 mg.

14. The method of claim 1 or 2, wherein the first dose is at least about 1.5 mg.

15. The method of claim 1 or 2, wherein the first dose is at least about 2 mg.

16. The method of claim 1 or 2, wherein the first dose is at least about 3 mg.

17. The method of claim 1 or 2, wherein the first dose is at least about 4 mg.

18. The method of claim 1 or 2, wherein the first dose is at least about 5 mg.

19. The method of claim 1 or 2, wherein the first dose is at least about 6 mg.

20. The method of claim 1 or 2, wherein the first dose is at least about 7 mg.

21. The method of claim 1 or 2, wherein the first dose is at least about 8 mg.

22. The method of claim 1 or 2, wherein the first dose is at least about 9 mg.

23. The method of claim 1 or 2, wherein the first dose is at least about 10 mg.

24. The method of claim 1 or 2, wherein the first dose is at least about 15 mg.

25. The method of claim 1 or 2, wherein the first dose is at least about 20 mg.

26. The method of claim 1 or 2, wherein the first dose is at least about 25 mg.

27. The method of claim 1 or 2, wherein the first dose is about 150 pg.

28. The method of any one of claims 1-27, further comprising administering to the subject a target dose of the isolated recombinant polypeptide complex, wherein the target dose is higher than the first dose, and wherein the target dose is administered after the first dose.

29. The method of claim 28, further comprising administering to the subject a second dose of the isolated recombinant polypeptide complex, wherein the second dose is higher than the first dose and lower than the target dose, and wherein the second dose is administered between the first dose and the target dose.

30. The method of any one of claims 1-29, wherein the first dose is administered weekly.

31. The method of any one of claims 1 -29, wherein the first dose is administered once every two weeks.

32. The method of any one of claims 1-29, wherein the first dose is administered once every three weeks.

33. The method of any one of claims 28-32, wherein the target dose is administered weekly.

34. The method of any one of claims 28-32, wherein the target dose is administered once every two weeks.

35. The method of any one of claims 28-32, wherein the target dose is administered once every three weeks.

36. The method of any one of claims 29-32, wherein the second dose is administered weekly.

37. The method of any one of claims 28-32, wherein the second dose is administered once every two weeks.

38. The method of any one of claims 28-32, wherein the second dose is administered once every three weeks.

39. The method of any one of claims 1-38, wherein the method comprises a treatment cycle that starts on day 1.

40. The method of claim 39, wherein the first dose is administered on day 1 of the treatment cycle.

41. The method of any one of claims 28-40, wherein the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle.

42. The method of any one of claims 29-41, wherein the first dose, the second dose, and the target dose are administered on day 1, day 4, and day 8, respectively, of the treatment cycle.

43. The method of claim 41 or 42, wherein the target dose is administered weekly after day 8 of the treatment cycle.

44. The method of claim 41 or 42, wherein the target dose is administered once every two weeks after day 8 of the treatment cycle.

45. The method of claim 41 or 42, wherein the target dose is administered once every three weeks after day 8 of the treatment cycle.

46. The method of any one of claims 28-45, wherein the target dose is about 50 gg to about 300 mg.

47. The method of any one of claims 28-46, wherein the target dose is at least about 100 gg.

48. The method of any one of claims 28-46, wherein the target dose is at least about 250 gg.

49. The method of any one of claims 28-46, wherein the target dose is at least about 500 gg.

50. The method of any one of claims 28-46, wherein the target dose is at least about 750 gg.

51. The method of any one of claims 28-46, wherein the target dose is at least about 1 mg.

52. The method of any one of claims 28-46, wherein the target dose is at least about 1.25 mg.

53. The method of any one of claims 28-46, wherein the target dose is at least about1.5 mg.

54. The method of any one of claims 28-46, wherein the target dose is at least about 2 mg.

55. The method of any one of claims 28-46, wherein the target dose is at least about 5 mg.

56. The method of any one of claims 28-46, wherein the target dose is at least about7.5 mg.

57. The method of any one of claims 28-46, wherein the target dose is at least about 10 mg.

58. The method of any one of claims 28-46, wherein the target dose is at least about 15 mg.

59. The method of any one of claims 28-46, wherein the target dose is at least about60. The method of any one of claims 28-46, wherein the target dose is at least about 25 mg.

61. The method of any one of claims 28-46, wherein the target dose is at least about 30 mg.

62. The method of any one of claims 28-46, wherein the target dose is at least about 50 mg.

63. The method of any one of claims 28-46, wherein the target dose is at least about 75 mg.

64. The method of any one of claims 28-46, wherein the target dose is at least about 100 mg.

65. The method of any one of claims 28-46, wherein the target dose is at least about 150 mg.

66. The method of any one of claims 28-46, wherein the target dose is at least about 200 mg.

67. The method of any one of claims 28-46, wherein the target dose is at least about 250 mg.

68. The method of any one of claims 28-46, wherein the target dose is at least about 300 mg.

69. The method of any one of claims 28-46, wherein the target dose is about 1.25 mg.

70. The method of any one of claims 28-46, wherein the first dose is at least about 500 gg and the target dose is at least about 1.25 mg.

71. The method of any one of claims 28-46, wherein the first dose is about 500 gg and the target dose is about 1.25 mg.

72. The method of any one of claims 28-46, wherein the first dose is about 500 gg and the target dose is about 5 mg.

73. The method of any one of claims 28-46, wherein the first dose is about 1.5 mg and the target dose is about 15 mg.

74. The method of any one of claims 28-46, wherein the first dose is about 5 mg and the target dose is about 50 mg.

75. The method of any one of claims 28-46, wherein the first dose is about 15 mg and the target dose is about 150 mg.

76. The method of any one of claims 28-46, wherein the first dose is about 30 mg and the target dose is about 300 mg.

77. The method of any one of claims 28-76, wherein the first dose is about 500 gg, the second dose is about 1.5 mg, and the target dose is about 5 mg.

78. The method of any one of claims 28-76, wherein the first dose is about 1.5 mg, the second dose is about 5 mg, and the target dose is about 15 mg.

79. The method of any one of claims 28-76, wherein the first dose is about 5 mg, the second dose is about 15 mg, and the target dose is about 50 mg.

80. The method of any one of claims 28-76, wherein the first dose is about 15 mg, the second dose is about 50 mg, and the target dose is about 150 mg.

81. The method of any one of claims 28-76, wherein the first dose is about 30 mg, the second dose is about 100 mg, and the target dose is about 300 mg.

82. The method of any one of claims 1-81, wherein the cancer comprises a cell that expresses epidermal growth factor receptor (EGFR).

83. The method of any one of claims 1-82, wherein the cancer comprises a cell that overexpresses EGFR.

84. The method of any one of claims 1-83, wherein the cancer comprises colorectal cancer (CRC), squamous cell carcinoma of head and neck (SCCHN), non-small cell lung cancer (NSCLC), renal cell carcinoma (RCC), breast cancer, bladder cancer, ovarian cancer, liver cancer, pancreatic cancer, small cell lung cancer (SCLC), pancreatic ductal adenocarcinoma (PDAC), triple-negative breast cancer (TNBC), prostate cancer, or a combination thereof.

85. The method of any one of claims 1-84, wherein the cancer progresses after treatment with a prior cancer therapy.

86. The method of any one of claims 1-85, wherein the cancer is refractory, intolerant, non-responsive, or resistant to a prior cancer therapy.

87. The method of any one of claims 1-86, wherein the administering comprises administering intravenously.

88. The method of any one of claims 1-87, wherein the subject exhibits a cytokine release syndrome (CRS) no more than grade 1.

89. The method of any one of claims 1-88, wherein the subject exhibits a cytokine release syndrome (CRS) no more than grade 2.

90. The method of any one of claims 1-89, wherein the subject exhibits a treatment related adverse event (TRAE) not related to a CRS.

91. The method of any one of claims 1-90, wherein the administering comprises administering to the subject a second treatment.

92. The method of any one of claims 1-91, wherein the administering results in a therapeutic effect in size in target lesion, hepatic metastasis, stable disease, partial response, complete response, size of renal mass, best RECIST response, overall response rate, duration of response, progression free survival, or a combination thereof, after the administering.

93. The method of claim 92, wherein the subject has stage IIIB NSCLC, and wherein the subject exhibits a RECIST partial response and a 100% reduction in size of target lesion 12 weeks after the administering.

94. The method of claim 93, wherein the administering comprises administering weekly the first dose of the isolated recombinant polypeptide complex in an amount of about 150 pg.

95. The method of claim 93 or 94, wherein the subject exhibits a reduction or elimination of hepatic metastasis after the administering.

96. The method of any one of claims 92-95, wherein the subject exhibits a best RECIST response.

97. The method of any one of claims 92-96, wherein the subject exhibits no CRS or TRAE after the administering.

98. The method of any one of claims 1-97, wherein the subject has RCC of stage 4, and wherein the subject exhibits a reduction of about 12% in size of renal mass.

99. The method of claim 98, wherein the subject exhibits a best RECIST response.

100. The method of claim 97 or 98, wherein the administering comprises administering the first dose in an amount of about 500 pg and the target dose in an amount of about 1.25 mg.

101. The method of claim 100, wherein the first dose is administered on day 1 of the treatment cycle and the target dose is administered on day 8 of the treatment cycle.

102. The method of claim 101, wherein the target dose is administered weekly after day 8.

103. The method of any one of claims 1-102, wherein the subject exhibits a best RECIST response, a partial response (PR), or a complete response (CR).

104. The method of any one of claims 1-103, wherein the subject exhibits stable disease.

105. The method of any one of claims 1-104, wherein the target dose is administered weekly after day 8.

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