Combination Therapies Comprising Bispecific Antibodies Comprising NRP1-Binding Domains

Abstract

A method for enhancing the therapeutic effect of a therapeutic agent in a subject having a cancer that is resistant to the therapeutic agent includes administering to the subject (i) the therapeutic agent and (ii) a targeted protein degrader that synergistically enhances the therapeutic activity of the therapeutic agent by reversing or reducing the resistance of the cancer to the therapeutic agent; and thereby enhancing the therapeutic effect of the therapeutic agent in the subject.

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Application No. 63 / 423,456, filed November 7, 2022, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Targeted therapies are highly advanced and initially beneficial for patients with specific oncogenic mutations. However, despite initial clinical responses to therapeutic agents, long-term efficacy has not currently been achieved because acquired drug resistance to targeted therapies inhibits the effectiveness of these therapies. Effective treatments are needed. Summary of the Invention

[0003] overview The present disclosure relates to cancer treatment and the prevention or reduction of drug resistance in cancer. In particular, the present disclosure relates to combination therapy using a targeted protein degrader and one or more therapeutic agents. The present disclosure relates to a method for enhancing the therapeutic effect of a therapeutic agent in a subject with a cancer resistant to the therapeutic agent by administering the therapeutic agent in combination with a targeted protein degrader.

[0004] Accordingly, in certain aspects, the present disclosure relates to a method of enhancing the efficacy of a therapeutic agent in a subject having a cancer that is resistant or refractory to the therapeutic agent, comprising administering to the subject (i) the therapeutic agent; and (ii) a targeted protein degradation agent, wherein the efficacy of the therapeutic agent is enhanced compared to administration of the therapeutic agent alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to a target protein on a cancer cell of interest; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising:

[0005] In one embodiment, the target protein is a receptor tyrosine kinase (RTK). In another embodiment, the receptor tyrosine kinase is selected from epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), fibroblast growth factor receptor (FGFR), Met receptor tyrosine kinase (MET), and vascular endothelial growth factor receptor (VEGFR). In one embodiment, the receptor tyrosine kinase is EGFR. In one embodiment, the receptor tyrosine kinase is cMET. In one embodiment, the receptor tyrosine kinase is HER2. In one embodiment, the receptor tyrosine kinase is IGF1R.

[0006] In some embodiments, the target cells are cancer cells, e.g., cancer cells selected from the group consisting of lung cancer, breast cancer, colorectal cancer, head and neck cancer, esophagogastric cancer, liver cancer, glioblastoma, prostate cancer, cervical cancer, ovarian cancer, bladder cancer, renal cancer, and pancreatic cancer. In some embodiments, the cancer cells are non-small cell lung cancer (NSCLC) cells.

[0007] In certain embodiments, the target protein binding domain and the NRP1 binding domain are each independently selected from the group consisting of an IgG, a half antibody, a single domain antibody, a nanobody, a Fab, a monospecific Fab2, an Fc, an scFv, a minibody, an IgNAR, a V-NAR, an hcIgG, a VHH domain, a camelid antibody, and a peptibody.

[0008] In one embodiment, the NRP1-binding domain comprises: (i) an antibody heavy chain variable (VH) domain comprising CDR1, CDR2, and CDR3 regions (HCDR1, HCDR2, and HCDR3, respectively), wherein HCDR1 consists of the sequence set forth in SEQ ID NO: 79, HCDR2 consists of the sequence set forth in SEQ ID NO: 80, and HCDR3 consists of the sequence set forth in any of SEQ ID NOs: 81 to 84; and (ii) An antibody light chain variable (VL) domain comprising CDR1, CDR2 and CDR3 regions (LCDR1, LCDR2 and LCDR3, respectively), wherein LCDR1 consists of the sequence set forth in any one of SEQ ID NOs: 85 to 87, LCDR2 consists of the sequence set forth in SEQ ID NO: 88, and LCDR3 consists of the sequence set forth in SEQ ID NO: 89.

[0009] In one embodiment, (i) HCDR1 consists of the sequence set forth in SEQ ID NO: 79, HCDR2 consists of the sequence set forth in SEQ ID NO: 80, and HCDR3 consists of the sequence set forth in any of SEQ ID NOs: 84; and (ii) LCDR1 consists of the sequence shown in SEQ ID NO: 85, LCDR2 consists of the sequence shown in SEQ ID NO: 88, and LCDR3 consists of the sequence shown in SEQ ID NO: 89.

[0010] In some embodiments, the enhanced effect includes an increase in tumor growth suppression effect, hi some embodiments, the enhanced effect includes an increase in median survival time.

[0011] In some embodiments, the therapeutic agent targets the EGFR pathway, non-limiting examples of which are disclosed herein. In some embodiments, the therapeutic agent that targets the EGFR pathway is osimertinib.

[0012] In certain embodiments, the therapeutic agent targets the cMET pathway, non-limiting examples of which are disclosed herein. In certain embodiments, the therapeutic agent that targets the cMET pathway is crizotinib.

[0013] In some embodiments, the therapeutic agent targets a KRAS protein (e.g., a mutant KRAS protein), non-limiting examples of which are disclosed herein. In some embodiments, the therapeutic agent that targets a KRAS protein is sotorasib.

[0014] In other embodiments, the therapeutic agent targets a protein selected from the group consisting of, for example, HER2, IGF1R, ALK, Braf, VEGF, and PDGF.

[0015] In some embodiments, the method further comprises administering a second therapeutic agent to the subject, wherein the effect of the second therapeutic agent is enhanced by the protein-targeted degrader compared to administration of the second therapeutic agent alone. In some embodiments, the therapeutic agent targets EGFR and the second therapeutic agent targets cMET, for example, in some embodiments, the therapeutic agent is osimertinib and the second therapeutic agent is sotorasib.

[0016] In another aspect, the disclosure relates to a method of enhancing the effect of a receptor tyrosine kinase (RTK) inhibitor in a subject having a cancer that is resistant or refractory to an RTK inhibitor, the method comprising: administering to a subject (i) an RTK inhibitor; and (ii) a targeted protein degrader, thereby enhancing the effect of the RTK inhibitor compared to administering the RTK inhibitor alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to an RTK on a cancer cell of interest; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising:

[0017] In another aspect, the disclosure relates to a method of enhancing the effect of an epidermal growth factor receptor (EGFR) inhibitor in a subject having cancer that is resistant or refractory to an EGFR inhibitor, the method comprising: administering to a subject (i) an EGFR inhibitor; and (ii) a targeted protein degrader, thereby enhancing the effect of the EGFR inhibitor compared to administration of the EGFR inhibitor alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to EGFR on a cancer cell of interest; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising:

[0018] The present disclosure relates to methods for enhancing the therapeutic effect of a therapeutic agent in a subject having a cancer that is resistant to the therapeutic agent: (i) administering to the subject a targeted protein degrader and a therapeutic agent that synergistically enhance the therapeutic activity of the therapeutic agent by reversing or reducing the subject's resistance to the cancer therapeutic agent.

[0019] In some embodiments, the cancer is resistant to a chemotherapeutic agent.

[0020] The present disclosure provides a method for sensitizing a therapeutic drug-resistant cancer to a therapeutic drug, comprising administering to the subject a targeted protein degradation agent that degrades a target protein, thereby inducing tumor regression in the subject.

[0021] In some embodiments, the methods include a pharmaceutical composition comprising a bispecific binding molecule that degrades a target protein. In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the bispecific binding molecule and a pharmaceutically acceptable carrier or diluent.

[0022] In some embodiments, the pharmaceutical is provided as a kit, and the kit includes a package insert containing instructions for use of a pharmaceutical composition comprising a therapeutically effective amount of a bispecific binding molecule and a pharmaceutically acceptable carrier or diluent.

[0023] In some embodiments, the bispecific binding molecule comprises an EGFR degrading agent. In some embodiments, the bispecific binding molecule comprises a first polypeptide having the sequence of SEQ ID NO:11 and a second polypeptide having the sequence of SEQ ID NO:12.

[0024] In some embodiments, the cancer includes lung cancer, breast cancer, colorectal cancer, head and neck cancer, esophagogastric cancer, liver cancer, glioblastoma, prostate cancer, cervical cancer, ovarian cancer, bladder cancer, renal cancer, and pancreatic cancer. In some embodiments, the cancer is non-small cell lung cancer (NSCLC).

[0025] In some embodiments, the therapeutic agent comprises a receptor tyrosine kinase inhibitor, including epidermal growth factor receptor (EGFR) inhibitors, ERBB inhibitors, c-MET inhibitors, fibroblast growth factor receptor (FGFR) and platelet-derived growth factor receptor (PDGFR) inhibitors, CSF1R inhibitors, cKIT inhibitors, FTL3 inhibitors, vascular endothelial growth factor receptor (VEGFR) inhibitors, TGFIβ1R and TGFβ2R inhibitors, integrin inhibitors, and IGF1R and IR inhibitors.

[0026] In some embodiments, the EGFR inhibitor is lazertinib, osimertinib (AZD9291), WZ4002, cyasterone, erlotinib (OSI-774) HCl, efitinib (ZD1839), lapatinib (GW-572016) ditosylate, afatinib (BIBW2992), saracatinib (AZD0530), vandetanib ( ZD6474), neratinib (HKI-272), canertinib (CI-1033), lapatinib (GW-572016), AG-490 (tyrphostin B42), CP-724714, dacomitinib (PF-00299804), sapitinib (AZD8931), CUDC-101, AG-1478 (tyrphostin AG-1478), PD153035 HCl, pelitinib (EKB-569), AC480 (BMS-599626), AEE788 (NVP-AEE788), AP26113-analogue (ALK-IN-1), OSI-420, WZ3146, HER2-inhibitor-1, WZ8040, alitinib tosylate, rociletinib (CO-1686), genistein (NPI 031L), barlitinib, TQB33804 (EGFR-IN-7), icotinib (BPI-2009H), TAK-285, daphnetin, tyrphostin 9, AG-18, AG555, AZ5104, CL-387785 (EKI-785), tyrphostin AG-258, AG-556, tucatinib, erlotinib (OSI-774), gefitinib-based PROTAC 3, zolifertinib (AZD 3759), ErbB2, AV-412 free base, AST-1306, JND3229, BI-4020, ceriatinib (HMPL-309), BDTX-189, lifirafenib (BGB-283), pyrotinib (SHR-1258), O-demethyl-gefitinib, epertinib hydrochloride, SU5214, avitinib (AC0010), AG494, and poziotinib (HM781-36B). In certain embodiments, the EGFR inhibitor is N-(2-{2-dimethylaminoethyl-methylamino}-4-methoxy-5-{[4-(1-methylindol-3-yl)pyrimidin-2-yl]amino}phenyl)prop-2-enamide mesylate (osimertinib).

[0027] In some embodiments, the therapeutic agent comprises an ERBB inhibitor, in some embodiments, the ERBB inhibitor is selected from the group consisting of tucatinib, HER2-inhibitor-1, afatinib (BIBW2992), neratinib (HKI-272), CP-724714, mubritinib (TAK 165), AC480 (BMS-599626), AEE778, TAK-285, tyrphostin AG 879, tyrphostin AG-528, SU5204, poziotinib (HM781-36B), TAS0728, BDTX-189, pyrotinib, and epertinib hydrochloride.

[0028] In some embodiments, the therapeutic agent comprises a cMET inhibitor. In some embodiments, the cMET inhibitor is selected from the group consisting of crizotinib, cabozantinib, foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, tivantinib, JNJ-38877605, PF-04217903, amuvatinib (MP-470), MGCD-265 analogs, capmatinib, BMS-754807, BMS-794833, AMG-208, MK-2461, golvatinib, AMG-458, NVP-BVU972, AMG 337, merestinib, JNJ-38877618, crizotinib hydrochloride, ningetinib, AMG-1, UNC2025, pamfetinib, altiratinib, NPS-1304, and savolitinib.

[0029] In some embodiments, the therapeutic agent comprises a PDGFR inhibitor and an FGFR inhibitor. In some embodiments, the PDGFR inhibitor and the FGFR inhibitor are ponatinib (AP24534), infigratinib (BGJ398), nintedanib (BIBF 1120), pazopanib HCl (GW786034 HCl), pazopanib, AZD4547, tyrphostin AG 1296, SSR128129E, LY2874455, derazantinib (ARQ-087), SU5402, ODM-203, pemigatinib (INCB054828), lucitanib (E3810) hydrochloride, ferulic acid, masitinib mesylate, fisogatinib (BLU-554), PRN1371, ON123300, FIIN-3, lobritinib (FGF401), futibatinib (TAS-120), FIIN-2, zoligratinib (Debio-1347), nintedanib ethanesulfonate, BLU9931, surufatinib, and combinations thereof.

[0030] In some embodiments, the therapeutic agent comprises a CSF1R inhibitor, hi some embodiments, the CSF1R inhibitor is selected from the group consisting of CSF1R-IN-1, Ki20227, pazopanib, erzovantinib (TPX-0022), ARRY-382, PRN1371, ENMD-2076, PF-477736, surufatinib, and combinations thereof.

[0031] In some embodiments, the therapeutic agent comprises a cKIT inhibitor. In some embodiments, the cKIT inhibitor is selected from the group consisting of dasatinib (BMS-354825), sorafenib (BAY 43-9006) tosylate, imatinib (STI571) mesylate, sunitinib (SU11248) malate, ponatinib (AP24534), axitinib (AG 013736), imatinib (STI571), nintedanib (BIBF 1120), regorafenib (BAY 73-4506), pazopanib HCl (GW786034), and the like. HCl), linifanib (ABT-869), crenolanib (CP-868596), masitinib (AB1010), amuvatinib (MP-470), orantinib (SU6668), CP-673451, telatinib, PP121, pazopanib, tyrphostin AG 1296, tyrphostin 9, SU14813, regorafenib hydrochloride, tyrphostin AG1433, sunitinib (SU11248), ripretinib (DCC-2618), masitinib mesylate, AZD3229, ON123300, avapritinib (BLU-285), seraltinib (GB002), AZD2932, JNJ-10198409, nintedanib ethanesulfonate, flumatinib (HH-GV-678), regorafenib (BAY-734506) monohydrate, and combinations thereof.

[0032] In some embodiments, the therapeutic agent comprises a FLT3 inhibitor. In some embodiments, the FLT3 inhibitor is selected from the group consisting of pacritinib (SB1518), TCS 359, linifanib (ABT-869), zotiraclib, cediranib (AZD2171), dovitinib (TKI258) lactate, UNC2025 HCl, SU5614, FLT3-IN-2, FLT3-IN-4, FF-10101, merestinib (LY2801653), emabsertib (CA-4948), tandutinib (MLN518), R406 (free base), 5'-fluoroindirubin oxime, tozasertib, quizartinib (AC220), R406, AST-487 (NVP-AST487), sorafenib (BAY 43-9006) tosylate, BMS-794833, sorafenib (BAY 43-9006), 4SC-203, dovitinib (TKI-258), isoguanosine, TAK-659, ATH686, SGI-1776 free base, MK-2461, fostamatinib (R788) disodium salt, MRX-2843, brigatinib (AP26113), GW2580, revastinib (DCC-2036), BMS-754807, UNC2025, fedratinib (TG101348), FLT3-IN-3, G-749, crenolanib (CP-868596), BPR1K871, PHA-680632, entospletinib (GS-9973), HPK1-IN-2, SP600125, SK LB4771 (FLT3-IN-1), KW-2449, gilteritinib (ASP2215), CCT241736, PRT062607 (P505-15) HCl, ENMD-2076, FN-1501, ceritinib (LDK378), silmitasertib (CX-4945), AZD2932, fostamatinib (R788), tivozanib (AV-951), PF-477736, BPR1J-097, pexidartinib (PLX3397), Go6976, HM43239, OSI-930, TG101209, PLX5622, amuvatinib (MP-470), GNF-2, midostaurin (PKC412), AMG 925, and ENMD-2076 L-(+)-tartaric acid.

[0033] In some embodiments, the therapeutic agent comprises a VEGFR1, VEGFR2, VEGFR3, and VEGFR4 inhibitor. In some embodiments, the VEGFR1, VEGFR2, VEGFR3, or VEGFR4 inhibitor is sorafenib (BAY 43-9006) tosylate, sunitinib (SU11248) malate, lenalidomide (CC-5013), cabozantinib (BMS-907351), ponatinib (AP24534), axitinib (AG 013736), foretinib (GSK1363089), vandetanib (ZD6474), nintedanib (BIBF 1120), regorafenib (BAY 73-4506), pazopanib HCl (GW786034), or the like. HCl), cediranib (AZD2171), PD173074, dovatinib (TKI-258), linifanib (ABT-869), vatalanib (PTK787) 2HCl, RAF265 (CHIR-265), tivozanib (AV-951), motesanib diphosphate (AMG-706), lenvatinib (E7080), brivanib (BMS-540215), MGCD-265 analog, AEE788 (NVP-AEE788), ENMD-2076, OSI-930, CYC116, Ki8751, telatinib, PP121, pazopanib, KRN 633, SAR131675, BMS-794833, apatinib (YN968D1) mesylate, sorafenib (BAY 43-9006), cabozantinib malate, brivanib alaninate (BMS-582664), golvatinib (E7050), semaxanib (SU5416), ZM 306416, ZM 323881 HCl, ENMD-2076L-(+)-Tartaric acid, LY2874455, BAW2881 (NVP-BAW2881), WHI-P180, SU14813, ZD-4190, SU1498, SU5402, PDGFR inhibitor 1, Ki20227, dovatinib (TKI258) lactate, toceranib phosphate, cediranib maleate, apatinib, BFH772, lenvatinib (E7080) mesylate, SU5614, regorafenib hydrochloride, SU5204, SU5208, fruquintinib (HMPL-013), hVEGF-IN-1, ODM-203, erdafitinib (JNJ-42756493), tyrphostin AG1433, MAZ51, SKLB 610, sunitinib (SU11248), 4SC-203, sitravatinib (MGCD516), R1530, donafenib (sorafenib D3), emvodostat (PTC299), AG-13958, SKLB1002, motesanib (AMG-706), 4,4'-bis(4-aminophenoxy)biphenyl, lusitanib (E3810) hydrochloride, oglufanide, tiauranib, ningetinib, X-82 (boronib), pamfetinib (TAS-115), cassia seed extract, CS -2660 (JNJ-38158471), WAY-340935, (20R)-protopanaxadiol, Ganoderma lucidum kernel extract, altiratinib, vitamin E, SU5408, AZD2932, anolotinib (AL3818) dihydrochloride, nintedanib ethanesulfonate, chebulinic acid, SU5205, SU5214, regorafenib (BAY-734506) monohydrate, taxifolin (dihydroquercetin), surufatinib, XL092, and combinations thereof.

[0034] In some embodiments, the therapeutic agent comprises a TGFβ1R and a TGFβ2R inhibitor. In some embodiments, the TGFβ1R or TGFβ2R inhibitor is SD-208, GW788388, A-83-01, disitertide (P144), SRI-011381, TP0427736 HCl, LY2109761, ophiopogonin D, SB505124, SIS3 HCl, BIBF-0775, LY 3200882, LSKL, thrombospondin (TSP-1) inhibitor, galunisertib (LY2157299), ginsenoside Rh4, LDN-193189, A77-01, LDN-193189 2HCl, bactosertib (TEW-7197), halofuginone hydrobromide, halofuginone, sulfasalazine (NSC 667219), BMS-986260, XAV-939, LY364947, oxymatrine, pirfenidone (S-7701), hypaconitine, SB525334, ITD-1, gamabufotalin, TA-02, PD 169316, and combinations thereof.

[0035] In some embodiments, the therapeutic agent comprises an integrin inhibitor, in some embodiments, the integrin inhibitor is selected from the group consisting of cilengitide trifluoroacetate, RGD (Arg-Gly-Asp) peptide, A-205804, SB273005, cilengitide, RGD peptide (GRGDNP), OSU-T315, ILK-IN-3, cyclo(-RGDfK), A286982, cyclo(RGDyK), A286982, and combinations thereof.

[0036] In some embodiments, the therapeutic agent comprises an IGF1R inhibitor and an IR inhibitor. In some embodiments, the IGF1R inhibitor or the IR inhibitor is luminespib (NVP-AUY922), linsitinib (OSI-906), NVP-AEW541, GSK1904529A, NVP-ADW742, BMS-536924, ceritinib (LDK378), AG-1024, GSK1838705A, BMS-754807, PQ 401, ZD3463, nordihydroguaiaretic acid (NDGA), NT157, insulin (human), ceritinib dihydrochloride, Chuanshanlong Root (Dioscoreae Nipponicae) Rhizoma extract, MID-1, brigatinib (AP26113), picropodophyllin (PPP), MSDC-0160, insulin degludec, chrome picolinate, SBI-477, XL228, and combinations thereof.

[0037] In some embodiments, the therapeutic agent comprises a neuropilin-associated receptor inhibitor, which in some embodiments includes a receptor tyrosine kinase, a receptor serine / threonine kinase, a G protein-coupled receptor, an ion channel receptor, a CXCR, and an immune checkpoint modulator.

[0038] In some embodiments, the therapeutic agent comprises a PARP inhibitor. In some embodiments, the PARP inhibitor is selected from the group consisting of PJ34 HCl, AZD2461, olaparib (AZD2281), veliparib (ABT-888), XAV-939, rucaparib (AG-014699) phosphate, iniparib (BSI-201), talazoparib (BMN 673), AG-14361, 3-aminobenzamide, A-966492, niraparib (MK-4827), UPF 1069, ME0328, licochalcone D, DR2313, MN 64, 4',5,7-trimethoxyflavone, rucaparib, M2912, GeA-69, BYK204165, and BGP-15. 2HCl, atamparib (RBN-2397), venadaparib (IDX-1197), niraparib (MK-4827) tosylate, NU1025, rucaparib camsylate, berberine chloride (NSC 646666), pamiparib (BGB-290), fluzoparib (SHR-3162), G007-LK, NVP-TNKS656, berberine chloride hydrate, HI-TOPK-032, stenoparib (E7449), 4-hydroxyquinazoline, NMS-P118, WIKI4, RBN012759, AZD5305, AZD-9574, RK-287107, benzamide, JW55, picolinamide, and combinations thereof.

[0039] In some embodiments, the therapeutic agent comprises a Raf inhibitor. In some embodiments, the Raf inhibitor is selected from the group consisting of vemurafenib (PLX4032), B-Raf inhibitor 1 (compound 13) dihydrochloride, Raf inhibitor 1, Raf inhibitor 2, sorafenib (BAY 43-9006) tosylate, PLX-4720, dabrafenib (GSK2118436), regorafenib (BAY 73-4506), doramapimod (BIRB 796), GDC-0879, RAF265 (CHIR-265), AZ 628, NVP-BHG712, SB590885, ZM 336372, sorafenib (BAY 43-9006), GW5074, TAK-632, agerafenib (RXDX-105), encorafenib (LGX818), BAW2881 (NVP-BAW2881), PLX8394, TBAP-001, regorafenib hydrochloride, MCP110, naporafenib (LXH254), B-Raf IN 1, donafenib (sorafenib D3), CCT196969, RAF709, lifirafenib (BGB-283), L-779450, PLX7904, LY3009120, dabrafenib mesylate, RO5126766 (CH5126766), AZ304, belvarafenib (HM95573), regorafenib (BAY-734506) monohydrate, toborafenib (MLN2480), and combinations thereof.

[0040] In some embodiments, the therapeutic agent comprises an autophagy activator. In some embodiments, the autophagy activator is selected from the group consisting of enzalutamide (MDV3100), obatoclax mesylate (GX15-070), SRT1720 HCl, fulvestrant (ICI-182780), bicalutamide (ICI-176334), resveratrol (SRT501), colforsin, rosiglitazone (BRL-49653) maleate, rosiglitazone (BRL 49653), mifepristone (RU486), permorphine, clemastine (HS-592) fumarate, GW4064, chloroquine diphosphate, ivermectin (MK-933), loperamide HCl, melatonin (NSC), and the like. 113928), methylprednisolone (NSC-19987), clonidine HCl, flubendazole, fenofibrate (NSC-281319), montelukast sodium, LYN-1604, EN6, eprenetapopt (APR-246), 3BDO, MHY1485, methylprednisolone acetate, QX77, anisomycin, β-elemene, spermidine trihydrochloride, troglitazone (CS-045), corinoxin, obeticholic acid, SMER28, BC1618, monomethyl fumarate, PCNA-Il, CA77.1, spermidine, xylitol, isorhychophylline, MPP plus iodide, and combinations thereof.

[0041] In some embodiments, the therapeutic agent comprises an autophagy inhibitor. In some embodiments, the autophagy inhibitor is selected from the group consisting of MK-2206 2HCl, bortezomib (PS-341), olaparib (AZD2281), vemurafenib (PLX4032), vorinostat (SAHA), ABT-737, Y-27632 2HCl, dactolisib (BEZ235), sorafenib (BAY 43-9006) tosylate, dasatinib (BMS-354825), rapamycin (AY-22989), crizotinib (PF-02341066), erlotinib (OSI-774) HCl, everolimus (RAD001), gefitinib (ZD1839), veliparib (ABT-888), entinostat (MS-275), BI 2536, pictilisib (GDC-0941), raduviglusib (CHIR-99021) HCl, LY294002, trametinib (GSK1120212), ruxolitinib (INCB018424), panobinostat (LBH589), imatinib (STI571) mesylate, KU-55933 (ATM kinase inhibitor), alisertib (MLN8237), afatinib (BIBW2992), U0126-EtOH, idelalisib, tozasertib, AZD8055, saracatinib (AZD0530), paclitaxel (NSC 125973), SP600125, ponatinib (AP24534), tanespimycin (17-AAG), YM155 (sepantronium bromide), DAPT (GSI-IX), cisplatin (NSC 119875), imatinib (STI571), nilotinib (AMN-107), temsirolimus (CCI-779), PI-103, luminespib (NVP-AUY922), vandetanib (ZD6474), gemcitabine (LY-188011) HCl, mocetinostat (MGCD0103), regorafenib (BAY 73-4506), SRT1720 HCl, pazopanib HCl (GW786034)HCl), bosutinib (SKI-606), cediranib (AZD2171), belinstat (PXD101), GSK690693, SB202190 (FHPI), carfilzomib (PR-171), fulvestrant (ICI-182780), SB216763, SU11274, linifanib (ABT-869), pemetrexed (LY-231514) disodium, tolquinib (PP242), etoposide (VP-16), wortmannin (KY 12420), LY2109761, danusertib (PHA-739358), silmitasertib (CX-4945), venetoclax (ABT-199), flavopiridol (L86-8275), SGI-1776 freebase, lapatinib (GW-572016), vincristine (NSC-67574) sulfate, temozolomide (CCRG 81045), tacrolimus (FK506), metformin HCl, fasudil (HA-1077) HCl, oxaliplatin (NSC 266046), ravusertib (LY2603618), 3-methyladenine (3-MA), flavopiridol (L86-8275) HCl, (+)-JQ1, zoledronic acid (ZOL 446), momelotinib (CYT387), ixazomib citrate (MLN9708) analog, tamoxifen (ICI 46474) citrate, letrozole (CGS 20267), topotecan (NSC609699) HCl, Torin 1, omipalisib (GSK2126458), degracin (WP1130), 2-methoxyestradiol (2-MeOE2), azacitidine (5-azacitidine), BX-795, OSI-027, ixazomib (MLN2238), TWS119, apitolisib (GDC-0980), BI-D1870, resveratrol (SRT501), dexamethasone (MK-125), cytarabine (U-19920A), simvastatin (MK 733), Vistasertib (AZD2014), CCT128930, Idarubicin HCl, Gemcitabine (LY-188011), Verteporfin (CL 318952), PF-4708671, Torin 2, Streptozotocin (STZ), H 89 2HCl, Brefeldin A, Geldanamycin (NSC122750), MK-5108 (VX-689), colforsin, valproic acid (NSC 93819) sodium salt, pitavastatin (NK-104) calcium, dosomorphine (compound C) 2HCl, lovastatin (MK-803), rosiglitazone (BRL-49653) maleate, necrostatin-1, pazopanib, nocodazole (R17934), dapolinad, clofarabine, cabazitaxel (XRP6258), atorvastatin calcium, (R)-(-)-gossypol acetic acid, pifithrin-α (PFTa) HBr, SN-38, GSK343, BIX 01294, Natrin-3a, tigecycline (GAR-936), STF-62247, binimetinib (MEK162), itraconazole (R 51211), milciclib (PHA-848125), sorafenib (BAY 43-9006), 10058-F4, YM201636, C646, hydroxyurea (NSC-32065), bardoxolone methyl, sodium butyrate, PR-619, linagliptin (BI-1356), niclosamide (BAY2353), nitazoxanide (NSC 697855), UNC1999, heparin sodium, DynaSor, curcumin, celastrol (NSC 70931), GSK2606414, honokiol (NSC 293100), raduviglusib (CHIR-99021), GANT61, omeprazole, amiodarone (NSC 85442) HCl, dexamethasone sodium phosphate, aspirin (NSC 27223), AZD3463, GSK2656157, carbamazepine, bafilomycin Al (Baf-A1), azithromycin (CP-62993), nimodipine, IU1, sulfasalazine (NSC 667219), pifithrin-μ, PF-543 hydrochloride, tubastatin A, nitrendipine, DC661, KB-R7943 mesylate, tubastatin ATFA, PFK15, nordihydroguaiaretic acid (NDGA), nilotinib hydrochloride monohydrate, salirasib, neferine, emetine hydrochloride, SBI-0206965, apatinib, EAD1, lanatoside C, ruxolitinib phosphate, losmapimod (GW856553X), STO-609, IITZ-01, vacuolin-1, entrectinib (RXDX-101), sunitinib (SU11248), valproic acid (VPA), URMC-099, spautin-1, erlotinib (OSI-774), pemetrexed disodium hydrate, crizotinib hydrochloride, atorvastatin, berberine chloride (NSC 646666), quercetin (NSC 9221), VLX600, afatinib (BIBW2992) dimaleate, FL-411, pemetrexed, autofinib, cryptotanshinone, nortriptyline hydrochloride, dihydroartemisinin (DHA), KN-93 phosphate, telaglenastat (CB-839), pulvalanol A, brevilin A, chloroquine (NSC-187208), PFK158, sulfacetamide sodium salt hydrate, OTS964, sinomenine hydrochloride, MRT67307 HCl, Lys05, sulfacetamide sodium, resatorbid (TAK-242), DMH1, hydroxychloroquine sulfate (NSC 4375), MRT68921 HCl, PHY34, Vinorelbine Tartrate (KW-2307), CA-5f, VPS34-IN1, DMOG, ICCB-19 Hydrochloride, AS1842856, SAR405, PIK-III, ULK-101, Vinblastine (NSC-49842) Sulfate, SP2509, ROC-325, Codonopsis Radix Extract, ABTL-0812, ML-9 HCl, AZD1208, Lucanthone, Leonurine, VER155008, RA-190, LY3009120, 4-Phenylbutyric Acid (4-PBA), NSC 185058, E260, Daurisolin, Dorsomorphine (Compound C), Tamoxifen (ICI 46474), Deferoxamine Mesylate (Ba 33112), pepstatin A, trametinib DMSO solvate, and concanavalin A.

[0042] In some embodiments, the therapeutic agent comprises an mTOR inhibitor. In some embodiments, the mTOR inhibitor is selected from the group consisting of mTOR inhibitor-1, everolimus (RAD001), KU-0063794, dactolisib (BEZ235), rapamycin (AY-22989), AZD8055, temsirolimus (CCI-779), PI-103, NU7441 (KU-57788), tolquinib (PP242), ridaforolimus (deforolimus, MK-8669), sapanisertib (MLN0128), voxtalisib (XL 765) analog, Torin 1, omipalisib (GSK2126458), OSI-027, PF-04691502, apitolisib (GDC-0980), GSK1059615, gedatolisib (PKI-587), WYE-354, vistasetrib (AZD2014), Torin 2, WYE-125132 (WYE-132), BGT226 (NVP-BGT226) maleate, Palomid 529 (P529), PP121, WYE-687, nitazoxanide (NSC 697855), WAY-600, ETP-46464, GDC-0349, XL388, 4EGI-1, JR-AB2-011, rotonjic acid, lanatoside C, compound 401, astragaloside IV, ginkgolide K, CC-115, zotarolimus (ABT-578), paxalisib (GDC-0084), CZ415, SF2523, bimiralisib (PQR309), voxtalisib (XL765), chrysophanic acid, onatasertib (CC 223), 3-hydroxyanthranilic acid, samotricisib (LY3023414), MTI-31, ABTL-0812, PQR620, MHY-1685, GNE-477, GNE-493, and combinations thereof.

[0043] In some embodiments, the therapeutic agent comprises a PI3K activator and a PI3K inhibitor, hi some embodiments, the PI3K activator is selected from demethyl-coclaurine, cinobufagin, resibufogenin, 740 YP (PDGFR 740Y-P), erucic acid, amarogentin, YS-49, and combinations thereof. In some embodiments, the PI3K inhibitor is PI-103, XL147 analog, 3-methyladenine (3-MA), apitolisib (GDC-0980), TG100713, taselisib (GDC 0032), dactolisib (BEZ235), pictilisib (GDC-0941), LY294002, idelalisib, bupallisib (BKM120), NU7441 (KU-57788), TGX-221, IC-87114, wortmannin (KY 12420), ZSTK474, alpelisib (BYL719), AS-605240, PIK-75 HCl, rigosertib (ON-01910), A66, voxtalisib (XL765) analog, omipalisib (GSK2126458), PIK-90, AZD6482, PF-04691502, GSK1059615, duvelisib (IPI-145), gedatolisib (PKI-587), TG100-115, AS-252424, NU7026, BGT226 (NVP-BGT226) maleate, fimepinostat (CUDC-907), PIK-294, AS-604850, GSK2636771, copanilisib (BAY 80-6946), YM201636, CH5132799, CAY10505, PIK-293, PKI-402, VS-5584 (SB2343), KU-0060648, CZC24832, demethyl-coclaurine, oroxin B, homosalate, AMG319, cinobufagin, resibufogenin, hispidulin, GSK2292767, lanatoside C, zeaxanthin, disiteltide (P144), cafestol, paxalisib (GDC-0084), SKI-V, MTX-211, seletalisib (UCB-5857), Trichosanthis Pericarpium Extract, Chuanshanlong Root Extract, GDC-0326, 740 YP (PDGFR740Y-P), PIK-108, parsaclisib (INCB050465) hydrochloride, HS-173, SF2523, leniolisib (CDZ 173), lupenone, ceravelisib (TAK-117), eganelisib (IPI-549), quercetin (NSC 9221), bimiralisib (PQR309), VPS34 inhibitor 1 (compound 19), IHMT-PI3K6-372, voxtalisib (XL765), autofinib, GNE-317, (E)-Akt inhibitor-IV, notoginsenoside R1, tenalisib (RP6530), solasodine, gallein, pectolinarin, inavolisib (GDC-0077), SRX3207, α-linolenic acid, ambralisib (TGR-1202), acalisib (GS-9820), ME-401, 3-hydroxyanthranilic acid, nemiralisib, sa Selected from motricisib (LY3023414), VPS34-IN1, airantone, Tritergium wilfordii extract, IPI-3063, SAR405, PIK-III, IPI-3063, parsacrisib (INCB050465), quercetin dihydrate, pyralalisib (XL147), SPP-86, AZD8835, trigonelline, deguelin, selective PI3K6 inhibitor 1 (compound 7n), loureirin A, PF-4989216, AZD8186, GNE-477, GNE-493, and combinations thereof.

[0044] In some embodiments, the therapeutic agent comprises a proteasome inhibitor. In some embodiments, the proteasome inhibitor is selected from the group consisting of salinosporamide A (NPI-0052), bortezomib (PS-341), MG132, carfilzomib (PR-171), ixazomib citrate (MLN9708), ixazomib (MLN2238), ONX-0914 (PR-957), oprozomib (ONX 0912), delanzomib (CEP-18770), celastrol (NSC 70931), epoxomicin (BU-4061T), shikonin (CI 75535), VR23, isoginkgo ginseng, RA-190, and PI-1840.

[0045] In some embodiments, the therapeutic agent comprises a JNK inhibitor. In some embodiments, the JNK inhibitor is selected from the group consisting of JNK inhibitor VIII, JNK inhibitor IX, JNK-IN-8, BI-78D3, SP600125, dramapimod (BIRB 796), metformin HCl, DB07268, 3'-hydroxypterostilbene, KB-R7943 mesylate, JNK-IN-7, loureirin B, astragaloside IV, cucurbitacin Ilb, trans-zeatin, ezatiostat, IQ 3, berberine chloride (NSC 646666), SU3327, bentamapimod (AS602801), indirubin-3'-oxime, falcarindiol, tanzisertib (CC-930), NDMC101, malveloside A, C-401 hydrochloride, RPI-1, ginsenoside Re, IQ-1S, urolithin B, and combinations thereof.

[0046] In some embodiments, the therapeutic agent comprises an NF-κB inhibitor. In some embodiments, the NF-κB inhibitor is selected from the group consisting of phosphorylated IKB alpha (S32) rabbit recombinant monoclonal antibody, ophiopogonin D, cornuside, rubiadin 1-methyl ether, SM-7368, NF-κB-IN-1, chitosan oligosaccharide, chinanchia atrati extract, IAXO-102, adjudin, CBL0137, IQ 3, hyperforin, isolicritin apioside, jaceocidin, bortezomib (PS-341), urolithin B, sulfasalazine (NSCLC), and the like. 667219), Acetylcysteine ​​(N-acetylcysteine), Erdosteine, Curcumin, Andrographolide, Dihydroartemisinin (DHA), Indole-3-carbinol, Magnolol, (-)-Parthenolide, Evodiamine, Chondroitin Sulfate, TPCA-1, Sodium Salicylate, Methylthiouracil, Articaine HCl, L-Quebrachitol, UCB-9260, Zeaxanthin, Tectochrysin, Myrislignan, Gardenoside, Caulophilin (N-methylcytisine), Demethyleneberberine, 3-Hydroxyanthranilic Acid, (E / Z)-IT-603, Triptolide (PG490), Sarsasapogenin, Berbamine Dihydrochloride, Ammonium Pyrrolidinedithiocarbamate, Stachydrine Salt acid salt, sodium escinate, stachydrine, tyrosol, mangiferin, skatellarin, ginsenoside Re, dehydroevodiamine, (E)-cardamonin, muscone, guaiacol, curcumenol, schisantherin A, hederagenin, astragaloside IV, ginsenoside Rgl, ginsenoside Rbl, ginsenoside Rd, 4'-methoxyresveratrol, (+)-α-lipoic acid, sodium 4-aminosalicylate, IMM-H007, diethyl maleate, 4-hydroxychalcone, benfotiamine, QNZ (EVP4593), 4'-hydroxychalcone, neferine, vanillic acid, hyperoside, chelidonic acid, ethyl caffeate, sulforaphane, (R)-(-)-ibuprofen, SN50, and C25.-140, INH14, Licochalcone D, SC75741, JSH-23, Caffeic acid phenethyl ester, Rocaglamide, APX-3330, DTP3, Omaveloxolone (RTA-408), Bardoxolone methyl, Shikonin (CI 75535), TAK-243 (MLN7243), CBL0137 HCl, Withaferin A, NIK Selected from the group consisting of SMI1, allobresib (GS-5829), antatin, maslinic acid, marbelloside A, eleutheroside E, berbamine, engeletin, aristolochic acid A, ginsenoside Rb3, 8-O-acetylshangjiside methyl ester, dauricine, 2',5'-dihydroxyacetophenone, (+)-preruptorin A, homoplantaginin, madecassic acid, BTYNB, and combinations thereof.

[0047] In some embodiments, the therapeutic agent comprises an HSP90 inhibitor. In some embodiments, the HSP90 inhibitor is selected from the group consisting of luminespib (NVP-AUY922), tanespimycin (17-AAG), alvespimycin (17-DMAG) HCl, ganetespib (STA-9090), elesclomol (STA-4783), BIIB021, tamoxifen (ICI 46474) citrate, onarespib (AT13387), NVP-BEP800, geldanamycin (NSCLC), and tamoxifen (ICI 46474) citrate. 122750), SNX-2112 (PF-04928473), PF-04929113 (SNX-5422), KW-2478, XL888, pifithrin-μ, NMS-E973, zeravespib (PU-H71), teprenone, dimethyleneastron, apoptozole, KRIBB11, pseudolaric acid A, TRC051384, DTHIB, rocaglamide, KNK437, VER-49009, HA15, pimitespib (TAS-116), CH5138303, VER-50589, YUM70, triptolide (PG490), VER155008, JG98, tamoxifen (ICI 46474), NPX800, HSP990 (NVP-HSP990), and combinations thereof.

[0048] In some embodiments, the therapeutic agent comprises an E3 ligase inhibitor. In some embodiments, the E3 ligase inhibitor is selected from the group consisting of lenalidomide (CC-5013), pomalidomide (CC-4047), thalidomide (K17), NSC 207895, TAME, PRT4165, CC-885, dCBP-1, iberdomide (CC220), BC-1215, CC-90009, avadomide (CC-122), VL285, (S,R,S)-AHPC (MDK7526), ​​Smurfl-IN-A01, (S,R,S)-AHPC-PEG4-NH2 hydrochloride, SZL P1-41, VH298, thalidomide-OH, MuRF1-IN-1, Skp2 inhibitor Cl (SKPin C1), GMB-475, mezigdomide (CC-92480), Homo-PROTAC cereblon degrader 1, THAL-SNS-032, NSC232003, apsin, thalidomide-O-COOH (cereblon ligand 3), and combinations thereof.

[0049] In some embodiments, the therapeutic agent comprises a KRAS inhibitor. In some embodiments, the KRAS inhibitor is MRTX1133, sotorasib (AMG510), adagrasib (MRTX849), LC-2, deltalasin, BAY-293, ARS-1620, BI-2852, ARS-853 (ARS853), BI-3406, ASP2453, sotorasib (AMG510) racemate, Pan-RAS-IN-1, MRTX-1257, zoledronic acid (ZOL 446), lonafarnib (SCH66336), K-Ras(G12C) inhibitor 9, salirasib, alamandine, (Ras)-antineoplaston A10, K-Ras-IN-1, MCP110, 6H05, K-Ras(G12C) inhibitor 12, Kobe0065, K-Ras(G12C) inhibitor 6, BQU57, Kobe2602, NAV-2729, antineoplaston A10, fendiline hydrochloride, KRpep-2d, perillyl alcohol, RBC8, KY1220, CID-1067700, zoledronic acid monohydrate, and combinations thereof.

[0050] In some embodiments, the protein that the therapeutic agent targets and inhibits is the same protein that the targeted protein degrader binds to and degrades.

[0051] In certain embodiments, the targeted protein degradation agent and EGFR inhibitor are used in the manufacture of a medicament for administration in combination with an EGFR inhibitor to treat cancer in a subject.

[0052] In some embodiments, the therapeutic agent is an EGFR inhibitor and the target protein is an EGFR degrading agent. In some embodiments, the EGFR inhibitor is osimertinib. In some embodiments, the cancer is osimertinib-resistant NSCLC.

[0053] In some embodiments, the targeted protein degradation agent is administered intraperitoneally.

[0054] In some embodiments, the method comprises administering to the patient an amount of a targeted protein degrader that increases the therapeutic activity of the therapeutic agent by at least 2-fold. The method of claim 1 comprises administering to the patient an amount of a targeted degrader that increases the therapeutic activity of the therapeutic agent by at least 5-fold. The method of claim 1 comprises administering to the patient an amount of a targeted degrader that increases the therapeutic activity of the therapeutic agent by at least 10-fold. The method of claim 1 comprises administering to the patient an amount of a targeted degrader that increases the therapeutic activity of the therapeutic agent by at least 20-fold.

[0055] In some embodiments, the cancer is resistant to immunotherapeutic antibodies. In certain embodiments, the immunotherapeutic antibodies include anti-EGFR monoclonal antibodies. In certain embodiments, anti-EGFR monoclonal antibodies include amivantamab, cetuximab, depatuxizumab, depatuxizumab mafodotin, durigotuzumab, futuximab, GC1118, imagatuzumab, matuzumab, necitumumab, nimotuzumab, panitumumab, zalutumumab, and HumMR1.

[0056] In some embodiments, the subject is a human.

[0057] The present disclosure further provides various methods of administering pharmaceutical compositions comprising bispecific antibodies to a subject. In some embodiments, administration is intravenous. In some embodiments, administration is intraperitoneal, intrathecal, intracerebroventricular, or intraparenchymal.

[0058] The following figures are provided for illustrative purposes and are not intended to limit the scope of the present invention. [Brief explanation of the drawings]

[0059] [Figure 1] Figure 1 is a schematic diagram showing the development of resistance following targeted therapy. [Figure 2] FIG. 2 shows that existing EGFR inhibitors cannot achieve sustained efficacy due to the lack of EGFR degradation. [Figure 3] Figure 3 shows the enhanced antitumor effect of combined treatment with osimertinib and an EGFR-degrading agent in an osimertinib-resistant mouse xenograft model (H1975-OR). The mouse H1975-OR xenograft model was treated with the EGFR signal-blocking antibody panitumumab (10 mg / kg) or an EGFR-degrading agent (13.74 mg / kg) in combination with high-dose osimertinib (10 mg / kg). Tumor volume was quantified to evaluate the therapeutic effect of the EGFR-degrading agent. [Figure 4] FIG. 4 shows that combination therapy of a targeted protein and a targeted protein degrader prevents or minimizes acquired resistance to targeted protein inhibitors in targeted protein-resistant cancers. [Figure 5A] Figures 5A-5B are graphs showing the results of the indicated treatments in the H1975-NRP1 OE xenograft model. Figure 5A shows the mean tumor volume over time. Figure 5B shows the survival probability over time. [Figure 5B] Same as above. [Figure 6A]Figures 6A-6B are graphs showing the results of the indicated treatments in the H1975-HGF xenograft model, with Figure 6A showing the results for combination with osimertinib and Figure 6B showing the results for combination with osimertinib and / or crizotinib. [Figure 6B] Same as above. [Figure 7] 7 is a graph showing the results of the indicated treatments in a mutant KRAS xenograft model. The results show the change in mean tumor volume over time. DETAILED DESCRIPTION OF THE INVENTION

[0060] Detailed Description Provided herein are methods and compositions for treating cancer in a subject and preventing resistance to or enhancing the therapeutic effects of antiproliferative drugs. Using an osimertinib-resistant H1975 xenograft mouse model, the inventors found that treatment with an EGFP degrader (EGFPxNRP1 antibody) sensitizes osimertinib-resistant cancer to continued osimertinib treatment. The EGFP degrader (EGFRxNRP1 antibody) may be used alone or in combination with any EGFR inhibitor to prevent or minimize acquired resistance to cancer therapeutics. The inventors demonstrate that combined treatment with a targeted protein degrader and a targeted protein inhibitor is an effective therapeutic approach for treating cancers that have already acquired or will acquire resistance to the targeted protein inhibitor.

[0061] 1. Definition Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. Accordingly, the following terms are intended to have the following meanings:

[0062] As used herein, the singular forms "a," "an," and "the" include the plural forms as well, unless the context clearly indicates otherwise.

[0063] As used herein, the terms "comprise," "include," "having," "having," "obtaining," "containing," and variations thereof are intended to be open-ended transitional phrases, terms, or words that do not exclude the possibility of additional acts or structures.

[0064] As used herein, "administration" of a polypeptide of the present disclosure includes delivering the inventive polypeptide or composition disclosed herein, or a prodrug or other pharmaceutically acceptable derivative thereof, to a subject using any suitable formulation or route of administration (e.g., those described herein).

[0065] As used herein, the term "and / or," when used in a list of two or more items, means that any one of the listed items may be used alone, or that two or more items may be used in any combination.

[0066] As used herein, "cancer" refers to a neoplasm or tumor caused by the abnormal and uncontrolled proliferation of cells. Cancer may also be referred to as a cellular-proliferative disease. Cancer may include various tissue types, cell types, and stages of cancer progression (e.g., primary tumors or metastatic growths). Cancers include, for example, breast cancer, cholangiocarcinoma, colorectal cancer, endometriosis, esophageal cancer, gastric cancer, diffuse gastric cancer, pancreatic cancer, renal cell carcinoma, soft tissue tumors, testicular cancer, heart: sarcoma (angiosarcoma, fibrosarcoma, rhabdomyosarcoma, liposarcoma), myxoma, rhabdomyoma, fibroma, lipoma, teratoma; lung: bronchogenic carcinoma (squamous cell carcinoma, undifferentiated small cell carcinoma, undifferentiated large cell carcinoma, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, sarcoma, lymphoma, chondroitin hamartoma, mesothelioma, non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), and small cell lung cancer (SCLC). Cellular lung cancer (SCLC); gastrointestinal tract: esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (tubular adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP-secreting tumor), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma); genitourinary system: kidney (adenocarcinoma, Wilms' tumor) tumors [nephroblastoma], lymphoma, leukemia), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma), testis (seminoma, teratoma, germ cell carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma); liver: hepatoma (hepatocellular carcinoma), bile duct carcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, hemangioma; bone: osteosarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticuloendothelial cell alveolarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteochondroma, osteoid exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma and giant cell tumor; nervous system: skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meninges (meningiomas, meningeal sarcomas, gliomatosis), brain (astrocytoma, cerebellar blastoma, glioma, ependymoma, germinoma [pineal tumor], glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors), spinal neurofibroma, meningioma, glioma, sarcoma;Gynecological: Uterus (endometrial cancer), cervix (cervical cancer, precancerous lesions such as cervical dysplasia), ovaries (ovarian cancer, ovarian adenocarcinoma [serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma], granulosa-thecocytoma, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, acicular sarcoma [germ cell rhabdomyosarcoma]), eggs Blood system: blood (myeloid leukemia [acute and chronic], acute lymphocytic leukemia, chronic lymphocytic leukemia, myeloproliferative disorders, multiple myeloma, myelodysplastic syndrome), Hodgkin's disease, non-Hodgkin's lymphoma [malignant lymphoma], CML; skin: melanoma, malignant melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, nevi, dysplastic nevi, lipoma, hemangioma, dermatofibroma, keloid, psoriasis; and adrenal gland: may include neuroblastoma.

[0067] In some embodiments, the cancer comprises non-small cell lung cancer (NSCLC). In some embodiments, the cancer is resistant to treatment. In some embodiments, the cancer is not resistant to treatment.

[0068] "Carrier" refers to a vehicle used to formulate a composition and may be comprised of multiple excipients.

[0069] As used herein, the terms "administration in combination" or "co-administration" refer to the use of multiple therapies (e.g., one or more prophylactic and / or therapeutic agents) and may be used interchangeably. The use of these terms does not restrict the order in which therapies (e.g., prophylactic and / or therapeutic agents) are administered to a subject.

[0070] As used herein, the term "excipient" refers to a pharmacologically inactive natural or synthetic ingredient or substance that is formulated with (e.g., simultaneously with) or subsequently formulated with an active ingredient of the present invention. In some embodiments, an excipient can be any additive, adjuvant, binder, filler, carrier, coating, diluent, disintegrant, filler, glidant, preservative, vehicle, or combination thereof that can be administered with a recombinant polypeptide of the present invention or is useful in preparing a composition of the present invention. Excipients include materials known to those skilled in the art that are non-toxic and do not interact with other components of the composition. In some embodiments, an excipient may be formulated with a recombinant polypeptide for bulking purposes when preparing a composition (hence, they are often referred to as bulking agents, fillers, or diluents). In other embodiments, an excipient may be used to provide enhanced functionality, such as promoting absorption and / or solubility of the active ingredient in the final dosage form. In still other embodiments, an excipient may be used to impart stability or to prevent contamination (e.g., microbial contamination). In other embodiments, excipients may be used to impart physical characteristics to the composition (e.g., to provide a composition in the physical form of a dry granule or a dry, flowable powder). Reference to an excipient includes both one and more such excipients. Suitable pharmaceutical excipients are described in "Remington's Pharmaceutical Sciences" by E.W. Martin, the disclosure of which is incorporated herein by reference in its entirety.

[0071] An "inhibitor" refers to a compound that inhibits or reduces the activity of a polypeptide. An inhibitor may indirectly or directly bind to a polypeptide and inhibit the activity of the polypeptide, including binding or catalytic activity. For example, an inhibitor may prevent the expression of a polypeptide or inhibit the ability of a polypeptide to mediate binding to a ligand. An "allosteric inhibitor" refers to a compound that binds to a polypeptide at a secondary site distinct from the primary ligand binding site and inhibits or reduces the activity of the polypeptide. The term "inhibit" or "inhibiting" means that activity is reduced or prevented in the presence of an inhibitor compared to the absence of the inhibitor. "Inhibition" refers to the reduction or downregulation of a process or the removal of a stimulus for a process, thereby eliminating or minimizing the expression or activity of a biomolecule or polypeptide. Inhibition may be direct or indirect. Inhibition may be specific, in which case the inhibitor inhibits only one biomolecule or polypeptide and not others.

[0072] As used herein, an "EGFRxNRP1 bispecific antibody" or "anti-EGFRxNRP1 bispecific antibody" (for use as a targeted proteolytic agent) refers to a bispecific binding molecule having two distinct antigen-binding domains, one of which specifically binds to the EGFR1 antigen and the other specifically binds to NRP1. Similar nomenclature is used for other bispecific binding molecules that bind to targets other than EGFR; for example, "HER2xNRP1 bispecific antibody" and "cMETxNRP1 bispecific antibody" refer to bispecific binding molecules having an antigen-binding domain that specifically binds to HER2 or cMET, respectively, and another antigen-binding domain that specifically binds to NRP1.

[0073] As used herein, "receptor tyrosine kinase" or "receptor tyrosine kinase" or "RTK" refers to a protein that is a receptor (i.e., binds to a ligand) and phosphorylates tyrosine residues.

[0074] As used herein, "non-receptor tyrosine kinase" or "non-receptor tyrosine kinase" or "non-RTK" refers to a protein that is not an RTK, i.e., is not a receptor and / or does not phosphorylate tyrosine residues.

[0075] As used herein, "treatment," "treat," and "treating" are used interchangeably and refer to an approach to achieving therapeutic benefit. Therapeutic benefit is determined by tumor shrinkage, tumor maintenance, or progression-free survival compared to placebo. As used herein, "parenteral" includes, but is not limited to, subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques.

[0076] "Pharmaceutically acceptable salts" is meant to include salts of active bispecific antibodies prepared with relatively non-toxic acids or bases, depending on the particular substituents on the bispecific antibodies disclosed herein.

[0077] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acid residues. As used herein, a "polynucleotide" can be single-stranded or double-stranded, or can contain portions of both double-stranded and single-stranded sequences. Polynucleotides can be nucleic acids, natural or synthetic, DNA, genomic DNA, cDNA, RNA, or hybrids, and can contain combinations of deoxyribonucleotides and ribonucleotides, as well as combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine. Polynucleotides can be obtained by chemical synthesis or recombinant methods. The term "reduce," or other forms such as "reducing" or "reduction," generally refers to the reduction of an event or characteristic (e.g., one or more symptoms, or the binding of one protein to another). This typically refers to a relationship to some standard or predicted value, in other words, relative, but not necessarily to that standard or relative value.

[0078] The term "recombinant," when used with respect to, for example, a cell, nucleic acid, polynucleotide, protein, or vector, indicates that the cell, nucleic acid, polynucleotide, protein, or vector has been modified by the introduction of a heterologous nucleic acid or protein or the alteration of a naturally occurring polynucleotide or protein, or is derived from a cell so modified. Thus, for example, recombinant cells express genes that are not found in native (non-recombinant) cells, or express naturally occurring genes that are normally expressed aberrantly, at low levels, or not at all. For example, as used herein, a "recombinant DNA molecule" refers to a DNA molecule comprising DNA segments joined together by molecular biological techniques. A "recombinant protein" or "recombinant polypeptide" refers to a protein molecule expressed from a recombinant DNA molecule or recombinant polynucleotide.

[0079] "Specifically binds" generally means that a substance or polypeptide binds more readily to a target than to a random, unrelated target.

[0080] As used herein, the term "subject" may refer to any animal, such as a mammal (such as a laboratory animal, livestock, or pet), having cancer. In some embodiments, the animal is a primate, preferably a human. As used herein, the terms "subject" and "subject" are used interchangeably. The terms "subject" and "subject" refer to animals (e.g., birds such as chickens, quails, or turkeys, or mammals), particularly "mammals," including non-primates (e.g., cows, pigs, horses, sheep, rabbits, guinea pigs, rats, cats, dogs, and mice) and primates (e.g., monkeys, chimpanzees, and humans), and especially humans. In some embodiments, the subject is a non-human animal, such as a livestock animal (e.g., a horse, cow, pig, or sheep) or a pet (e.g., a dog, cat, guinea pig, or rabbit). In a preferred embodiment, the subject is a "human."

[0081] As used herein, the term "synergistic" refers to the effect of combining a polypeptide of the present invention with another therapeutic agent (e.g., a prophylactic or therapeutic agent) that is greater than the sum of the effects of each therapeutic agent.

[0082] As used interchangeably herein, the terms "therapeutically effective amount," "effective dosage," or "effective amount," unless otherwise defined, refer to an amount of a drug or agent effective for a period of time necessary to achieve a desired therapeutic outcome. An effective dosage can be determined by one skilled in the art and may vary depending on factors such as the individual's disease state, age, sex, and weight, as well as the ability of the agent to elicit a desired response in the individual. As used herein, the term may also refer to an amount effective to produce a desired in vivo effect in a subject. A therapeutically effective amount may be administered in one or more administrations (e.g., as a preventative treatment or therapeutically at any stage of disease progression, before or after symptom onset), application, or dosage, and is not intended to be limited to a particular composition, combination, or route of administration. Within the scope of the present disclosure, a drug may be administered at various times during the course of treatment of a subject. The timing and dosage of administration depend on several factors, such as the purpose of treatment (e.g., therapeutic or preventative), the condition of the subject, and can be readily determined by one skilled in the art. A therapeutically effective amount can also be said to be an amount in which any toxic or harmful effects of a substance are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective, at the dosage and for the period necessary, to achieve the desired prophylactic effect. Typically, prophylactic administration is used for subjects before or at an early stage of disease, so the prophylactically effective amount will be less than the therapeutically effective amount.

[0083] As used herein, the terms "treat," "treated," or "treating" refer to treatment to delay (alleviate) an undesirable physiological condition, disorder, or disease, or to obtain a beneficial or desired clinical outcome. For purposes of this invention, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, regression of the extent of the condition, disorder, or disease; stabilization (i.e., not worsening) of the condition, disorder, or disease; delay in the onset or progression of the condition, disorder, or disease; improvement and remission (whether partial or complete, detectable or undetectable) of the condition, disorder, or disease; or enhancement or amelioration of the condition, disorder, or disease. Treatment also includes prolonging survival compared to expected survival if not receiving treatment. The terms "treat," "treated," or "treating" also include prevention, suppression, arrest, amelioration, or complete elimination of disease. Prevention of disease may involve administering a composition of the invention to a subject before disease onset. Suppression of disease may involve administering a composition of the invention to a subject after disease induction but before clinical manifestation. Arresting or ameliorating the disease may involve administering a composition of the invention to a subject after clinical manifestation of the disease.

[0084] The term "variant" as used herein with respect to polynucleotides means (i) a portion or fragment of a referenced nucleotide sequence, (ii) the complement of a referenced nucleotide sequence or a portion thereof, (iii) a polynucleotide that is substantially identical to a referenced polynucleotide or its complement, or (iv) a polynucleotide that hybridizes under stringent conditions to a referenced polynucleotide, its complement, or a sequence substantially identical thereto.

[0085] As used herein, "percent identity" between two sequences (e.g., amino acid sequences or nucleotide sequences) refers to the percentage of positions (out of a maximum of 100%) that are identical when optimally aligned and compared (which may include insertions or deletions appropriate for optimal alignment). The percent identity between two sequences is a function of the number of identical positions shared between the sequences (i.e., % identity = number of identical positions / total number of positions × 100), taking into account the number of gaps and the length of each gap that need to be introduced for optimal alignment of the two sequences. Sequence comparison and determination of percent identity between two sequences can be performed using a mathematical algorithm, as described in the non-limiting examples below. Methods and algorithms for determining percent homology between two protein sequences are well established in the art.

[0086] For example, the percent identity between two amino acid sequences can be determined using the Needleman and Wunsch ((1970) J. Mol. Biol. (48):444-453) algorithm incorporated into the GAP program of the GCG software package (available at http: / / www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6. Furthermore, protein amino acid sequences can be used as "query sequences" to search public databases, e.g., to identify related sequences. Such searches can be performed using the XBLAST program (version 2.0) of Altschul et al. (1990) J. Mol. Biol. 215:403-10. BLAST protein searches can be performed with the XBLAST program, score = 50, word length = 3, to obtain amino acid sequences homologous to the protein molecules of the invention. To obtain gapped alignments for comparison purposes, Gapped BLAST, as described in Altschul et al. (1997) Nucleic Acids Res. 25(17):3389-3402, can be utilized. When utilizing BLAST and Gapped BLAST programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used.

[0087] All patent applications, patents, and publications referenced herein are incorporated by reference in their entirety to the same extent as if each individual publication, patent, or patent application was individually and specifically indicated to be incorporated by reference in its entirety, and all patent applications, patents, and publications cited herein are incorporated by reference in their entirety, except for definitions, subject matter disclaimers, or disclaimers, and except to the extent the incorporated content contradicts the express disclosure of this specification, which shall take precedence.

[0088] The present invention is not limited in scope by the specific embodiments described herein. Indeed, various modifications in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying drawings. Such modifications are intended to be within the scope of the appended claims. Furthermore, it should be understood that all values ​​are approximate and are provided for illustrative purposes.

[0089] 2. Targeted protein degraders and EGFR degraders Further provided herein is a targeted protein degradation agent. The targeted protein degradation agent comprises a target protein binding domain and a neuropilin binding domain. The target protein binding domain and the neuropilin binding domain of the targeted protein degradation agent specifically bind to the target protein and NRP1, and the internalized target protein is then degraded via lysosomes. The targeted protein degradation agent comprises a heavy chain polypeptide of any one of SEQ ID NOs: 2 to 11 and a light chain polypeptide of SEQ ID NO: 12. Also provided are SEQ ID NOs: 15 to 24 as nucleotide sequences encoding the corresponding heavy chain polypeptides of SEQ ID NOs: 2 to 11, and SEQ ID NO: 25 as a nucleotide sequence encoding the light chain polypeptide of SEQ ID NO: 12. All polypeptide, polynucleotide, and linker sequences disclosed in U.S. Provisional Application No. 63 / 325,312 are incorporated herein by reference.

[0090] For example, the EGFR degrading agent is a bispecific antibody (EGFRxNRP1 antibody) that specifically binds to EGFR and NRP1. The binding of the EGFR degrading agent to EGFR and NRP1 causes degradation of EGFR. The degradation of EGFR by the EGFR degrading agent occurs via the lysosome-dependent degradation pathway. The EGFR degrading agent comprises the polypeptide sequences set forth in SEQ ID NOs: 11 and 12. The polynucleotide sequences encoding the polypeptides of SEQ ID NOs: 11 and 12 are set forth in SEQ ID NOs: 15 and 25. See Table 1 and Table 2. [Table 1] [Table 2-1] [Table 2-2]

[0091] Additional exemplary bispecific antibody heavy chain polypeptide sequences for use in the proteolytic agents of the present disclosure are set forth in Table 3 and in SEQ ID NOs: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, and 39, and polynucleotide sequences encoding the bispecific antibody heavy chain polypeptide sequences are set forth in Table 3 and in SEQ ID NOs: 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, and 40, respectively. In some embodiments, the bispecific antibody further comprises a light chain polypeptide sequence comprising a variable light chain and a constant light chain paired with the heavy chain polypeptide, which is set forth in SEQ ID NO: 12. The polynucleotide sequence encoding the light chain polypeptide of SEQ ID NO: 12 is set forth in SEQ ID NO: 25. In some embodiments, the present disclosure provides polynucleotide sequences used to encode the GGGGS subunit, which are set forth in Table 3 and in SEQ ID NOs: 26-38.

[0092] In some embodiments, the proteolytic agent is a bispecific antibody comprising an anti-EGFR binding arm comprising (or consisting of) a VH amino acid sequence set forth in SEQ ID NO: 69 and a VL amino acid sequence set forth in SEQ ID NO: 70. In some embodiments, the bispecific antibody comprises an anti-EGFR binding arm comprising heavy chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 71, 72, and 73, respectively. In some embodiments, the bispecific antibody comprises an anti-EGFR binding arm comprising a heavy chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 73. In some embodiments, the bispecific antibody comprises an anti-EGFR binding arm comprising light chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 74, 75, and 76, respectively. In some embodiments, the bispecific antibody comprises an anti-EGFR binding arm comprising a light chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 76. In some embodiments, the anti-EGFR binding arm comprises (or consists of) one or more sequences that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.8% identical to any of the aforementioned VH, VL, HCDR, or LCDR sequences. These sequences are also shown in Table 3.

[0093] In some embodiments, the proteolytic agent is a bispecific antibody comprising an anti-NRP1 binding arm comprising (or consisting of) a VH amino acid sequence set forth in SEQ ID NO: 77 and a VL amino acid sequence set forth in SEQ ID NO: 78. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising heavy chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 79, 80, and 81, respectively. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising a heavy chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 81. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising light chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 82, 83, and 84, respectively. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising a light chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 84. In some embodiments, the anti-NRP1 binding arm comprises (or consists of) one or more sequences that are at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 99.8% identical to any of the aforementioned VH, VL, HCDR, or LCDR sequences. These sequences are also shown in Table 3.

[0094] The EGFR-degrading agent may reduce or prevent resistance of cancer to tyrosine kinase inhibitors. The anti-EGFR degrading agent may be, for example, an antibody, a polynucleotide, a small molecule, or a combination thereof. The EGFR degrading agent may cause degradation of the target protein, thereby reducing or preventing resistance of cancer to tyrosine kinase inhibitors. Acquired drug resistance to tyrosine kinase inhibitors may be reduced or prevented after administration of the EGFR-degrading agent. In some embodiments, tyrosine kinase inhibitor resistance may be reduced by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, or at least 20-fold. In some embodiments, tyrosine kinase inhibitor resistance is reduced by 2-fold to 10-fold. The activity of the tyrosine kinase inhibitor may be increased by at least 2-fold, at least 3-fold, at least 4-fold, at least 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, at least 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, at least 13-fold, at least 14-fold, at least 15-fold, at least 16-fold, at least 17-fold, at least 18-fold, at least 19-fold, or at least 20-fold. In some embodiments, the activity of the tyrosine kinase inhibitor is increased by at least 2-fold to at least 10-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a cancer patient resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least 2-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a cancer patient resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least 3-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a cancer patient resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least 4-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a cancer patient resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least 5-fold.In some embodiments, administering a predetermined amount of a targeted degradative agent to a patient with cancer resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least six-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a patient with cancer resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least seven-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a patient with cancer resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least eight-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a patient with cancer resistant to a therapeutic agent may increase the therapeutic activity of the therapeutic agent by at least nine-fold. In some embodiments, administering a predetermined amount of a targeted degradative agent to a patient may increase the therapeutic activity of the therapeutic agent by at least ten-fold.

[0095] a. Neuropilin There are two types of neuropilins: NRP1 and NRP-2. Neuropilins are transmembrane glycoproteins consisting of four domains: A (ala2), B (blb2), C (MAM), and a cytoplasmic domain. Neuropilins are known to regulate neurogenesis and angiogenesis by forming complexes with plexin receptors / class 3 semaphorin ligands and vascular endothelial growth factor (VEGF) receptors / VEGF ligands, respectively. Because neuropilins have very small cytoplasmic domains, they primarily function as coreceptors, relying on other cell surface receptors to transmit signals across the plasma membrane (Pellet et al., 2008; Schwarz, 2010).

[0096] Recent studies have demonstrated that neuropilins are multifunctional and can bind to a variety of transmembrane receptors. Neuropilins are closely associated with numerous signaling pathways, including those activated by epidermal growth factor (EGF), fibroblast growth factor (FGF), hepatocyte growth factor (HGF), insulin-like growth factor (IGF), platelet-derived growth factor (PDGF), and transforming growth factor beta (TGFI3) (Kofler, 2016; Roy et al., 2017). Neuropilins are generally present on the cell surface, but have also been reported to be present in mitochondria and the nucleus (Issitt et al., 2019; Mehta et al., 2018). Both neuropilin family members can also exist in soluble forms generated by alternative splicing or ectodomain shedding from the cell surface (Rossgnol et al., 2000; Werneberg et al., 2016).

[0097] Neuropilins bind to many receptors, including but not limited to receptor tyrosine kinases (e.g., EGFR), receptor serine / threonine kinases (e.g., TGFPR), integrins, GPCRs (G protein-coupled receptors), and ion channels.

[0098] The pleiotropic properties of NRP receptors implicate them in cellular processes such as axon guidance and angiogenesis, immune responses, and remyelination (Mecollari et al., 2014). Thus, dysregulation of NRP activity has been implicated in many pathological conditions, including many types of cancer and cardiovascular diseases (Niland et al., 2019; Kofler, 2016; Pellet-Many et al., 2019; Harma et al., 2020).

[0099] The NRP1-binding domain of the proteolytic agent used in the methods of the present disclosure includes an antibody or an NRP1-binding fragment thereof. For example, all or a portion of an NRP1 antibody known in the art or an anti-NRP1 antibody provided herein may be used. Non-limiting examples of anti-NRP1 monoclonal antibody (mAb) heavy chain polypeptide sequences are set forth in SEQ ID NOS: 41-47, which contain an N-terminal NRP1-binding domain. In some embodiments, the monoclonal antibody heavy chain polypeptide comprises an N-terminal NRP1-binding domain comprising a variable heavy chain (VH) and constant heavy chain 1 (CH1), and an Fc domain comprising constant heavy chain 2 (CH2) and constant heavy chain 3 (CH3). In some embodiments, the monoclonal antibody comprises a light chain polypeptide sequence set forth in any of SEQ ID NOS: 48-54. Thus, the heavy chains of SEQ ID NOS: 41-47 can be paired with the light chains of SEQ ID NOS: 48-54, respectively. Polynucleotide sequences encoding the heavy and light chain polypeptides are set forth in SEQ ID NOS: 55-61 and 62-68, respectively. These sequences are also shown in Table 3.

[0100] b. EGFR The epidermal growth factor receptor (EGFR) is one of the most frequently mutated oncogenes in solid tumors. Increased EGFR signaling promotes cell proliferation and survival in many cancer types. Mutations affecting EGFR expression or activity can lead to cancer. For example, mutations leading to EGFR overexpression are associated with the development of various cancers. All known EGFR inhibitors, as well as all known kinase inhibitors targeting oncogenic receptor tyrosine kinases and serine / threonine kinases, have ultimately led to drug resistance (Figure 1). Existing EGFR inhibitors fail to degrade EGFR, resulting in the continued presence of EGFR and failing to achieve a durable response (Figure 2).

[0101] c. Additional proteolytic agents cMETxNRP1 bispecific construct In certain embodiments, the proteolytic agent is a cMETxNRP1 bispecific construct. In some embodiments, the cMETxNRP1 bispecific antibody comprises an anti-NRP1 binding arm comprising heavy chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 79, 80, and 81, respectively. In certain embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising a heavy chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 81. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising light chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 82, 83, and 84, respectively. In certain embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising a light chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 84.

[0102] In some embodiments, the cMET-binding arm comprises the sequence of an anti-cMET monoclonal antibody (mAb) known and available in the art, such as the heavy and light chain CDRs (1-3) of the mAb or the VH / VL polypeptide sequence of the mAb. In some embodiments, the anti-cMET mAb is emibetuzumab (also known as LY2875358). In some embodiments, the anti-cMET mAb is onatuzumab. In some embodiments, the anti-cMET mAb is telisotuzumab.

[0103] HER2xNRP1 bispecific construct In certain embodiments, the proteolytic agent is a HER2xNRP1 bispecific antibody construct. In some embodiments, the HER2xNRP1 bispecific antibody comprises an anti-NRP1 binding arm comprising heavy chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 79, 80, and 81, respectively. In certain embodiments, the bispecific antibody comprises an anti-NRP1 binding arm having a heavy chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 81. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising light chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 82, 83, and 84, respectively. In certain embodiments, the bispecific antibody comprises an anti-NRP1 binding arm having a light chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 84.

[0104] In some embodiments, the HER2-binding arm comprises the sequence of an anti-HER2 monoclonal antibody (mAb) known and available in the art, such as the heavy and light chain CDRs (1-3) of the mAb or the VH / VL polypeptide sequence of the mAb. In some embodiments, the anti-HER2 mAb is trastuzumab. In some embodiments, the anti-HER2 mAb is pertuzumab.

[0105] IGF1RxNRP1 bispecific construct In certain embodiments, the proteolytic agent is an IGF1RxNRP1 bispecific antibody construct. In certain embodiments, the IGF1RxNRP1 bispecific antibody comprises an anti-NRP1 binding arm comprising heavy chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 79, 80, and 81, respectively. In another embodiment, the bispecific antibody comprises an anti-NRP1 binding arm comprising a heavy chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 81. In some embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising light chain CDR1, CDR2, and CDR3 regions comprising (or consisting of) the amino acid sequences set forth in SEQ ID NOs: 82, 83, and 84, respectively. In certain embodiments, the bispecific antibody comprises an anti-NRP1 binding arm comprising a light chain CDR3 region comprising (or consisting of) the amino acid sequence set forth in SEQ ID NO: 84.

[0106] In some embodiments, the IGF1R binding arm comprises the sequence of an anti-IGF1R monoclonal antibody (mAb) known and available in the art, such as the heavy and light chain CDR(1-3) of the mAb or the VH / VL polypeptide sequence of the mAb. In one embodiment, the anti-IGF1R mAb is ganitumab (also known as AMG 479). In another embodiment, the anti-IGF1R mAb is figitumumab. In one embodiment, the anti-IGF1R mAb is cixutumumab. In one embodiment, the anti-IGF1R mAb is dalotuzumab.

[0107] 3. Therapeutic drugs containing tyrosine kinase inhibitors tyrosine kinase Tyrosine kinases are enzymes that can transfer a gamma (terminal) phosphate group from ATP to a polypeptide within a cell. The phosphate group is added to the amino acid tyrosine on the polypeptide. Tyrosine kinases may phosphorylate one or more tyrosine residues on a polypeptide. Phosphorylation of tyrosines on a polypeptide can cause a change in the function of the polypeptide. Tyrosine kinases are a subgroup of a larger class of protein kinases that add phosphate groups to other amino acids (such as serine and threonine). Protein phosphorylation by kinases is one of the mechanisms involved in intracellular signal transduction and the regulation of cellular activities such as cell division.

[0108] B. tyrosine kinase inhibitor Tyrosine kinase inhibitors (TKIs) inhibit the activity or expression of tyrosine kinases. Tyrosine kinase inhibitors may include antibodies, polynucleotides such as interfering RNA, small molecules, or combinations thereof. In some embodiments, tyrosine kinase inhibitors include antibodies. In some embodiments, the antibodies are monoclonal antibodies. In some embodiments, tyrosine kinase inhibitors include small molecules. Tyrosine kinase inhibitors may be specific for a particular tyrosine kinase or a particular family or subfamily of tyrosine kinases. In some embodiments, tyrosine kinase inhibitors are specific for EGFR. Such inhibitors may be referred to as "EGFR inhibitors." Antibody-based EGFR inhibitors include, for example, cetuximab and panitumumab. Small molecule-based EGFR inhibitors include, for example, lapatinib (a combined EGFR and ERBB2 inhibitor), merletinib (osimertinib), gefitinib, icotinib, erlotinib, afatinib, and brigatinib. The EGFR inhibitor may bind to and inhibit the kinase domain of EGFR.Without kinase activity, EGFR may not be able to induce autoactivation or downstream activation and signal transduction.In some embodiments, the EGFR inhibitor comprises osimertinib.

[0109] All of the EGFR inhibitors listed below, but not limited to, target and inhibit specific tumorigenic EGFRs, ultimately resulting in drug resistance, which can be overcome by administration of any EGFR degrading agent, including the present invention (Compound 1 in Example 1 and Figure 3), alone or in combination with any EGFR inhibitor that causes drug resistance.

[0110] EGFR inhibitors include lazertinib, osimertinib (AZD9291), WZ4002, cyasterone, erlotinib (OSI-774) HCl, efitinib (ZD1839), lapatinib (GW-572016) ditosylate, afatinib (BIBW2992), saracatinib (AZD0530), and vandetanib (ZD6474). , neratinib (HKI-272), canetinib (CI-1033), lapatinib (GW-572016), AG-490 (tyrphostin B42), CP-724714, dacomitinib (PF-00299804), sapitinib (AZD8931), CUDC-101, AG-1478 (tyrphostin AG-1478), PD153035 HCl, pelitinib (EKB-569), AC480 (BMS-599626), AEE788 (NVP-AEE788), AP26113-analog (ALK-IN-1), OSI-420, WZ3146, HER2-inhibitor-1, WZ8040, alitinib tosylate, rociletinib (CO-1686), genistein (NPI 031L), barlitinib, TQB33804 (EGFR-IN-7), icotinib (BPI-2009H), TAK-285, daphnetin, tyrphostin, AG-18, AG-19 555, AZ5104, CL-387785 (EKI-785), tyrphostin AG-258, AG-556, tucatinib, erlotinib (OSI-774), gefitinib-based PROTAC 3, zolifertinib (AZD 3759), ErbB2, AV-412 free base, AST-1306, JND3229, BI-4020, ceriatinib (HMPL-309), BDTX-189, lifirafenib (BGB-283), pyrotinib (SHR-1258), O-demethyl-gefitinib, epertinib hydrochloride, SU5214, avitinib (AC0010), AG 494, and not limited to poziotinib (HM781-36B).

[0111] All of the ERBB2 (HER2), ERBB3 (HER3), and ERBB4 (HER4) inhibitors listed below, but not limited to, target and inhibit specific tumorigenic ERBBs, ultimately resulting in drug resistance, which can be overcome by any ERBB degrading agent and any of the present invention administered alone or in combination with any ERBB inhibitor that causes drug resistance.

[0112] ERBB inhibitors are tucatinib, HER2-inhibitor-1, afatinib (BIBW2992), neratinib (HKI-272), CP-724714, mubritinib (TAK 165), AC480 (BMS-599626), AEE778, TAK-285, tyrphostin AG 879, tyrphostin AG-528, SU5204, poziotinib (HM781-36B), TAS0728, BDTX-189, pyrotinib, and epertinib hydrochloride.

[0113] All of the cMET inhibitors listed below, but not limited to, target and inhibit specific oncogenic cMET, ultimately resulting in drug resistance, which is overcome by any cMET degrader and any cMET inhibitor of the present invention, administered alone or in combination with any drug resistance-causing cMET inhibitor.

[0114] cMET inhibitors include crizotinib, cabozantinib, foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, tivantinib, JNJ-38877605, PF-04217903, amuvatinib (MP-470), MGCD-265 analogs, capmatinib, BMS-754807, BMS-794833, AMG-208, MK-2461, golvatinib, AMG-458, NVP-BVU972, and AMG 337, merestinib, JNJ-38877618, crizotinib hydrochloride, ningetinib, AMG-1, UNC2025, pamfetinib, altiratinib, NPS-1304, and savolitinib.

[0115] All of the PDGFR and FGFR inhibitors listed below, but not limited to, target and inhibit specific tumorigenic PDGFRs and FGFRs, ultimately resulting in drug resistance, which can be overcome by the administration of any of the PDGFR-degrading agents and any of the FGFR-degrading agents of the present invention alone or in combination with any of the PDGFR inhibitors or any of the FGFR inhibitors that cause drug resistance.

[0116] PDGFR inhibitors and FGFR inhibitors include ponatinib (AP24534), infigratinib (BGJ398), nintedanib (BIBF 1120), pazopanib HCl (GW786034 HCl), pazopanib, AZD4547, and tyrphostin AG. 1296, SSR128129E, LY2874455, derazantinib (ARQ-087), SU5402, ODM-203, pemigatinib (INCB054828), lucitanib (E3810) hydrochloride, ferulic acid, masitinib mesylate, fisogatinib (BLU-554), PRN1371, ON123300, FIIN-3, lobritinib (FGF401), futibatinib (TAS-120), FIIN-2, zoligratinib (Debio-1347), nintedanib ethanesulfonate, BLU9931, and surufatinib.

[0117] All of the cKIT inhibitors listed below, but not limited to, target and inhibit specific oncogenic cKIT, ultimately resulting in drug resistance, which can be overcome by the administration of any cKIT degrading agent and the present invention alone or in combination with each cKIT inhibitor that causes drug resistance.

[0118] cKIT inhibitors include dasatinib (BMS-354825), sorafenib (BAY 43-9006) tosylate, imatinib (STI571) mesylate, sunitinib (SU11248) malate, ponatinib (AP24534), axitinib (AG 013736), imatinib (STI571), nintedanib (BIBF 1120), regorafenib (BAY 73-4506), pazopanib HCl (GW786034), and rituximab (RT-PCR). HCl), linifanib (ABT-869), crenolanib (CP-868596), masitinib (AB1010), amuvatinib (MP-470), orantinib (SU6668), CP-673451, telatinib, PP121, pazopanib, tyrphostin AG 1296, Tyrphostin 9, SU14813, Regorafenib Hydrochloride, Tyrphostin AG1433, Sunitinib (SU11248), Ripretinib (DCC-2618), Masitinib Mesylate, AZD3229, ON123300, Avapritinib (BLU-285), Seraltinib (GB002), AZD2932, JNJ-10198409, Nintedanib Ethanesulfonate, Flumatinib (HI-I-GV-678), and Regorafenib (BAY-734506) Monohydrate.

[0119] All of the FTL3 inhibitors listed below, but not limited to, target and inhibit specific tumorigenic FTL3, ultimately resulting in drug resistance, which can be overcome by any FTL3 degrading agent and the present invention, administered alone or in combination with an FTL3 inhibitor that causes drug resistance.FTL3 inhibitors include pacritinib (SB1518), TCS 359, linifanib (ABT-869), zotiraclib, cediranib (AZD2171), dovitinib (TKI258) lactate, UNC2025 HCl, SU5614, FLT3-IN-2, FLT3-IN-4, FF-10101, merestinib (LY2801653), emabsertib (CA-4948), tandutinib (MLN518), R406 (free base), 5'-fluoroindirubin oxime, tozasertib, quizartinib (AC220), R406, AST-487 (NVP-AST487), sorafenib (BAY 43-9006) tosylate, BMS-794833, sorafenib (BAY 43-9006), 4SC-203, dovitinib (TKI-258), isoguanosine, TAK-659, ATH686, SGI-1776 free base, MK-2461, fostamatinib (R788) disodium salt, MRX-2843, brigatinib (AP26113), GW2580, revastinib (DCC-2036), BMS-754807, UNC2025, fedratinib (TG101348), FLT3-IN-3, G-749, crenolanib (CP-868596), BPR1K871, PHA-680632, entospletinib (GS-9973), HPK1-IN-2, SP600125, SK LB4771 (FLT3-IN-1), KW-2449, gilteritinib (ASP2215), CCT241736, PRT062607 (P505-15) HCl, ENMD-2076, FN-1501, ceritinib (LDK378), silmitasertib (CX-4945), AZD2932, fostamatinib (R788), tivozanib (AV-951), PF-477736, BPR1J-097, pexidartinib (PLX3397), Go6976, HM43239, OSI-930, TG101209, PLX5622, amuvatinib (MP-470), GNF-2, midostaurin (PKC412), AMG 925, and ENMD-2076 L-(+)-tartaric acid.

[0120] All of the VEGFR1, VEGFR2, VEGFR3, and VEGFR4 inhibitors listed below, but not limited to, target and inhibit any one of the tumorigenic VEGFR1, VEGFR2, VEGFR3, and VEGFR4, ultimately resulting in drug resistance, which is overcome by combined treatment with any one of the VEGFR1, VEGFR2, VEGFR3, or VEGFR4 degrading agents of the present invention, either administered alone or in combination with a VEGFR1, VEGFR2, VEGFR3, or VEGFR4 inhibitor that causes drug resistance.

[0121] VEGFR1, VEGFR2, VEGFR3, and VEGFR4 inhibitors include sorafenib (BAY 43-9006) tosylate, sunitinib (SU11248) malate, lenalidomide (CC-5013), cabozantinib (BMS-907351), ponatinib (AP24534), axitinib (AG 013736), foretinib (GSK1363089), vandetanib (ZD6474), nintedanib (BIBF 1120), regorafenib (BAY 73-4506), pazopanib HCl (GW786034) HCl), cediranib (AZD2171), PD173074, dovatinib (TKI-258), linifanib (ABT-869), vatalanib (PTK787) 2HCl, RAF265 (CHIR-265), tivozanib (AV-951), motesanib diphosphate (AMG-706), lenvatinib (E7080), brivanib (BMS-540215), MGCD-265 analog, AEE788 (NVP-AEE788), ENMD-2076, OSI-930, CYC116, Ki8751, telatinib, PP121, pazopanib, KRN 633, SAR131675, BMS-794833, apatinib (YN968D1) mesylate, sorafenib (BAY 43-9006), cabozantinib malate, brivanib alaninate (BMS-582664), golvatinib (E7050), semaxanib (SU5416), ZM 306416, ZM 323881 HCl, ENMD-2076 L-(+)-Tartaric acid, LY2874455, BAW2881 (NVP-BAW2881), WHI-P180, SU14813, ZD-4190, SU1498, SU5402, PDGFR inhibitor 1, Ki20227, dovatinib (TKI258) lactate, toceranib phosphate, cediranib maleate, apatinib, BFH772, lenvatinib (E7080) mesylate, SU5614, regorafenib hydrochloride, SU5204, SU5208, fruquintinib (HMPL-013), hVEGF-IN-1, ODM-203, erdafitinib (JNJ-42756493), tyrphostin AG1433, MAZ51, SKLB610, sunitinib (SU11248), 4SC-203, sitravatinib (MGCD516), R1530, donafenib (sorafenib D3), emvodostat (PTC299), AG-13958, SKLB1002, motesanib (AMG-706), 4,4'-bis(4-aminophenoxy)biphenyl, lucitanib (E3810) hydrochloride, oglufanid, tiauranib, ningetinib, X-82 (boronib), pamfetinib (TAS-115), cassini These include Ganoderma lucidum seed extract, CS-2660 (JNJ-38158471), WAY-340935, (20R)-protopanaxadiol, Ganoderma lucidum kernel extract, altiratinib, vitamin E, SU5408, AZD2932, anolotinib (AL3818) dihydrochloride, nintedanib ethanesulfonate, chebulinic acid, SU5205, SU5214, regorafenib (BAY-734506) monohydrate, taxifolin (dihydroquercetin), surufatinib, and XL092.

[0122]

[0097] All of the TGFβ1R and TGFβ2R inhibitors listed below, but not limited to, target and inhibit specific tumorigenic TGFβ1R and TGFβ2R, ultimately resulting in drug resistance, which is overcome by combined treatment with any TGFβ1R and / or TGFβ2R degrading agent and any of the TGFβ1R and TGFβ2R inhibitors of the present invention, administered alone or in combination with the TGFβ1R and TGFβ2R inhibitors that cause drug resistance.

[0123] TGFβ1R and TGFβ2R inhibitors include SD-208, GW788388, A-83-01, disitertide (P144), SRI-011381, TP0427736 HCl, LY2109761, ophiopogonin D, SB505124, SIS3 HCl, BIBF-0775, LY 3200882, LSKL, thrombospondin (TSP-1) inhibitor, galunisertib (LY2157299), ginsenoside Rh4, LDN-193189, A77-01, LDN-193189 2HCl, bactosertib (TEW-7197), halofuginone hydrobromide, halofuginone, sulfasalazine (NSC 667219), BMS-986260, XAV-939, LY364947, oxymatrine, pirfenidone (S-7701), hypaconitine, SB525334, ITD-1, gamabufotalin, TA-02, and PD 169316.

[0124]

[0099] All of the integrin inhibitors listed below, but not limited to, target and inhibit specific tumorigenic integrins, ultimately resulting in drug resistance, which can be overcome by any integrin degrading agent and the present invention, administered alone or in combination with an integrin inhibitor that causes drug resistance.

[0125]

[0100] Integrin inhibitors are cilengitide trifluoroacetate, RGD (Arg-Gly-Asp) peptide, A-205804, SB273005, cilengitide, RGD peptide (GRGDNP), OSU-T315, ILK-IN-3, cyclo(-RGDfK), A286982, cyclo(RGDyK), and A286982.

[0126] All of the IGF1R and IR inhibitors listed below, but not limited to, target and inhibit specific tumorigenic IGF1R and IR, ultimately resulting in drug resistance, which is overcome by any IGF1R and IR degrading agent and the present invention, administered alone or in combination with an IGF1R or IR inhibitor that causes drug resistance.

[0127] IGF1R and IR inhibitors are luminespib (NVP-AUY922), linsitinib (OSI-906), NVP-AEW541, GSK1904529A, NVP-ADW742, BMS-536924, ceritinib (LDK378), AG-1024, GSK1838705A, BMS-754807, PQ 401, ZD3463, nordihydroguaiaretic acid (NDGA), NT157, insulin (human), ceritinib dihydrochloride, Chuanshanlong Root Extract, MID-1, brigatinib (AP26113), picropodophyllin (PPP), MSDC-0160, insulin degludec, chrome picolinate, SBI-477, and XL228.

[0128] 4. Additional target proteins Neuropilins are closely associated with many drug-resistant tumors. Neuropilins and their degradation are associated with receptor tyrosine kinases, receptor serine / threonine kinases, G protein-coupled receptors, ion channels, CXCRs, and immune checkpoint regulators for many biological functions. Therefore, co-degradation of neuropilins with other receptors is useful for treating neuropilin-related drug resistance.

[0129] All PARP inhibitors, including but not limited to those listed below, result in drug resistance, which is overcome by administration of any NRP-degrading compound alone or in combination with any PARP1 inhibitor that causes drug resistance.

[0130] PARP inhibitors include PJ34 HCl, AZD2461, olaparib (AZD2281), veliparib (ABT-888), XAV-939, rucaparib (AG-014699) phosphate, iniparib (BSI-201), talazoparib (BMN 673), AG-14361, 3-aminobenzamide, A-966492, niraparib (MK-4827), UPF 1069, ME0328, licochalcone D, DR2313, MN 64, 4',5,7-trimethoxyflavone, rucaparib, M2912, GeA-69, BYK204165, and BGP-15. 2HCl, atamparib (RBN-2397), venadaparib (IDX-1197), niraparib (MK-4827) tosylate, NU1025, rucaparib camsylate, berberine chloride (NSC 646666), pamiparib (BGB-290), fluzoparib (SHR-3162), G007-LK, NVP-TNKS656, berberine chloride hydrate, HI-TOPK-032, stenoparib (E7449), 4-hydroxyquinazoline, NMS-P118, WIKI4, RBN012759, AZD5305, AZD-9574, RK-287107, benzamide, JW55, and picolinamide.

[0131] All of the RAF inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any RAF inhibitor that causes drug resistance.

[0132] RAF inhibitors include vemurafenib (PLX4032), B-Raf inhibitor 1 (compound 13) dihydrochloride, Raf inhibitor 1, Raf inhibitor 2, sorafenib (BAY 43-9006) tosylate, PLX-4720, dabrafenib (GSK2118436), regorafenib (BAY 73-4506), doramapimod (BIRB 796), GDC-0879, RAF265 (CHIR-265), AZ 628, NVP-BHG712, SB590885, ZM 336372, sorafenib (BAY 43-9006), GW5074, TAK-632, agerafenib (RXDX-105), encorafenib (LGX818), BAW2881 (NVP-BAW2881), PLX8394, TBAP-001, regorafenib hydrochloride, MCP110, naporafenib (LXH254), B-Raf IN 1, donafenib (sorafenib D3), CCT196969, RAF709, lifirafenib (BGB-283), L-779450, PLX7904, LY3009120, dabrafenib mesylate, RO5126766 (CH5126766), AZ304, belbarafenib (HM95573), regorafenib (BAY-734506) monohydrate, and toborafenib (MLN2480).

[0133] All of the autophagy activators or inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any autophagy activator or inhibitor that causes drug resistance.

[0134] Autophagy activators include enzalutamide (MDV3100), obatoclax mesylate (GX15-070), SRT1720 HCl, fulvestrant (ICI-182780), bicalutamide (ICI-176334), resveratrol (SRT501), colforcin, rosiglitazone (BRL-49653) maleate, rosiglitazone (BRL 49653), mifepristone (RU486), permorphin, clemastine (HS-592) fumarate, GW4064, chloroquine diphosphate, ivermectin (MK-933), loperamide HCl, melatonin (NSC 113928), methylprednisolone (NSC-19987), clonidine HCl, flubendazole, fenofibrate (NSC-281319), montelukast sodium, LYN-1604, EN6, eprenetapopt (APR-246), 3BDO, MHY1485, methylprednisolone acetate, QX77, anisomycin, β-elemene, spermidine trihydrochloride, troglitazone (CS-045), corinoxin, obeticholic acid, SMER28, BC1618, monomethyl fumarate, PCNA-Il, CA77.1, spermidine, xylitol, isorhychophylline, and MPP + iodide.

[0135] Autophagy inhibitors include MK-2206 2HCl, bortezomib (PS-341), olaparib (AZD2281), vemurafenib (PLX4032), vorinostat (SAHA), ABT-737, Y-27632 2HCl, dactolisib (BEZ235), sorafenib (BAY 43-9006) tosylate, dasatinib (BMS-354825), rapamycin (AY-22989), crizotinib (PF-02341066), erlotinib (OSI-774) HCl, everolimus (RAD001), gefitinib (ZD1839), veliparib (ABT-888), entinostat (MS-275), and BI 2536, pictilisib (GDC-0941), raduviglusib (CHIR-99021) HCl, LY294002, trametinib (GSK1120212), ruxolitinib (INCB018424), panobinostat (LBH589), imatinib (STI571) mesylate, KU-55933 (ATM kinase inhibitor), alisertib (MLN8237), afatinib (BIBW2992), U0126-EtOH, idelalisib, tozasertib, AZD8055, saracatinib (AZD0530), paclitaxel (NSC 125973), SP600125, ponatinib (AP24534), tanespimycin (17-AAG), YM155 (sepantronium bromide), DAPT (GSI-IX), cisplatin (NSC 119875), imatinib (STI571), nilotinib (AMN-107), temsirolimus (CCI-779), PI-103, luminespib (NVP-AUY922), vandetanib (ZD6474), gemcitabine (LY-188011) HCl, mocetinostat (MGCD0103), regorafenib (BAY 73-4506), SRT1720 HCl, pazopanib HCl (GW786034)HCl), bosutinib (SKI-606), cediranib (AZD2171), belinstat (PXD101), GSK690693, SB202190 (FHPI), carfilzomib (PR-171), fulvestrant (ICI-182780), SB216763, SU11274, linifanib (ABT-869), pemetrexed (LY-231514) disodium, tolquinib (PP242), etoposide (VP-16), wortmannin (KY 12420), LY2109761, danusertib (PHA-739358), silmitasertib (CX-4945), venetoclax (ABT-199), flavopiridol (L86-8275), SGI-1776 freebase, lapatinib (GW-572016), vincristine (NSC-67574) sulfate, temozolomide (CCRG 81045), tacrolimus (FK506), metformin HCl, fasudil (HA-1077) HCl, oxaliplatin (NSC 266046), ravusertib (LY2603618), 3-methyladenine (3-MA), flavopiridol (L86-8275) HCl, (+)-JQ1, zoledronic acid (ZOL 446), momelotinib (CYT387), ixazomib citrate (MLN9708) analog, tamoxifen (ICI 46474) citrate, letrozole (CGS 20267), topotecan (NSC609699) HCl, Torin 1, omipalisib (GSK2126458), degracin (WP1130), 2-methoxyestradiol (2-MeOE2), azacitidine (5-azacitidine), BX-795, OSI-027, ixazomib (MLN2238), TWS119, apitolisib (GDC-0980), BI-D1870, resveratrol (SRT501), dexamethasone (MK-125), cytarabine (U-19920A), simvastatin (MK 733), Vistasertib (AZD2014), CCT128930, Idarubicin HCl, Gemcitabine (LY-188011), Verteporfin (CL 318952), PF-4708671, Torin 2, Streptozotocin (STZ), H 89 2HCl, Brefeldin A, Geldanamycin (NSC122750), MK-5108 (VX-689), colforsin, valproic acid (NSC 93819) sodium salt, pitavastatin (NK-104) calcium, dosomorphine (compound C) 2HCl, lovastatin (MK-803), rosiglitazone (BRL-49653) maleate, necrostatin-1, pazopanib, nocodazole (R17934), dapolinad, clofarabine, cabazitaxel (XRP6258), atorvastatin calcium, (R)-(-)-gossypol acetic acid, pifithrin-α (PFTa) HBr, SN-38, GSK343, BIX 01294, Natrin-3a, tigecycline (GAR-936), STF-62247, binimetinib (MEK162), itraconazole (R 51211), milciclib (PHA-848125), sorafenib (BAY 43-9006), 10058-F4, YM201636, C646, hydroxyurea (NSC-32065), bardoxolone methyl, sodium butyrate, PR-619, linagliptin (BI-1356), niclosamide (BAY2353), nitazoxanide (NSC 697855), UNC1999, heparin sodium, DynaSor, curcumin, celastrol (NSC 70931), GSK2606414, honokiol (NSC 293100), raduviglusib (CHIR-99021), GANT61, omeprazole, amiodarone (NSC 85442) HCl, dexamethasone sodium phosphate, aspirin (NSC 27223), AZD3463, GSK2656157, carbamazepine, bafilomycin Al (Baf-A1), azithromycin (CP-62993), nimodipine, IU1, sulfasalazine (NSC 667219), pifithrin-μ, PF-543 hydrochloride, tubastatin A, nitrendipine, DC661, KB-R7943 mesylate, tubastatin ATFA, PFK15, nordihydroguaiaretic acid (NDGA), nilotinib hydrochloride monohydrate, salirasib, neferine, emetine hydrochloride, SBI-0206965, apatinib, EAD1, lanatoside C, ruxolitinib phosphate, losmapimod (GW856553X), STO-609, IITZ-01, vacuolin-1, entrectinib (RXDX-101), sunitinib (SU11248), valproic acid (VPA), URMC-099, spautin-1, erlotinib (OSI-774), pemetrexed disodium hydrate, crizotinib hydrochloride, atorvastatin, berberine chloride (NSC 646666), quercetin (NSC 9221), VLX600, afatinib (BIBW2992) dimaleate, FL-411, pemetrexed, autofinib, cryptotanshinone, nortriptyline hydrochloride, dihydroartemisinin (DHA), KN-93 phosphate, telaglenastat (CB-839), pulvalanol A, brevilin A, chloroquine (NSC-187208), PFK158, sulfacetamide sodium salt hydrate, OTS964, sinomenine hydrochloride, MRT67307 HCl, Lys05, sulfacetamide sodium, resatorbid (TAK-242), DMH1, hydroxychloroquine sulfate (NSC 4375), MRT68921 HCl, PHY34, Vinorelbine Tartrate (KW-2307), CA-5f, VPS34-IN1, DMOG, ICCB-19 Hydrochloride, AS1842856, SAR405, PIK-III, ULK-101, Vinblastine (NSC-49842) Sulfate, SP2509, ROC-325, Codonopsis Radix Extract, ABTL-0812, ML-9 HCl, AZD1208, Lucanthone, Leonurine, VER155008, RA-190, LY3009120, 4-Phenylbutyric Acid (4-PBA), NSC 185058, E260, Daurisolin, Dorsomorphine (Compound C), Tamoxifen (ICI 46474), Deferoxamine Mesylate (Ba 33112), pepstatin A, trametinib DMSO solvate, and concanavalin A.

[0136] All of the mTOR inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any mTOR inhibitor that causes drug resistance.

[0137] mTOR inhibitors include mTOR inhibitor-1, everolimus (RAD001), KU-0063794, dactolisib (BEZ235), rapamycin (AY-22989), AZD8055, temsirolimus (CCI-779), PI-103, NU7441 (KU-57788), tolquinib (PP242), ridaforolimus (deforolimus, MK-8669), sapanisertib (MLN0128), voxtalisib (XL765), and analogs. G, Torin 1, omipalisib (GSK2126458), OSI-027, PF-04691502, apitolisib (GDC-0980), GSK1059615, gedatricisib (PKI-587), WYE-354, vistasetrib (AZD2014), Torin 2, WYE-125132 (WYE-132), BGT226 (NVP-BGT226) maleate, Palomid 529 (P529), PP121, WYE-687, nitazoxanide (NSC 697855), WAY-600, ETP-46464, GDC-0349, XL388, 4EGI-1, JR-AB2-011, rotonjic acid, lanatoside C, compound 401, astragaloside IV, ginkgolide K, CC-115, zotarolimus (ABT-578), paxalisib (GDC-0084), CZ415, SF2523, bimiralisib (PQR309), voxtalisib (XL765), chrysophanic acid, onatasertib (CC 223), 3-hydroxyanthranilic acid, samotricisib (LY3023414), MTI-31, ABTL-0812, PQR620, MHY-1685, GNE-477, and GNE-493.

[0138] All of the PI3K activators or PI3K inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any PI3K activator or PI3K inhibitor that causes drug resistance.

[0139] PI3K activators are demethyl-coclaurine, cinobufagin, resibufogenin, 740 YP (PDGFR 740Y-P), erucic acid, amarogentin, and YS-49.

[0140] PI3K inhibitors include PI-103, XL147 analogs, 3-methyladenine (3-MA), apitolisib (GDC-0980), TG100713, taselisib (GDC 0032), dactolisib (BEZ235), pictilisib (GDC-0941), LY294002, idelalisib, bupalisib (BKM120), NU7441 (KU-57788), TGX-221, IC-87114, wortmannin (KY 12420), ZSTK474, alpelisib (BYL719), AS-605240, and PIK-75. HCl, rigosertib (ON-01910), A66, voxtalisib (XL765) analog, omipalisib (GSK2126458), PIK-90, AZD6482, PF-04691502, GSK1059615, duvelisib (IPI-145), gedatolisib (PKI-587), TG100-115, AS-252424, NU7026, BGT226 (NVP-BGT226) maleate, fimepinostat (CUDC-907), PIK-294, AS-604850, GSK2636771, copanilisib (BAY 80-6946), YM201636, CH5132799, CAY10505, PIK-293, PKI-402, VS-5584 (SB2343), KU-0060648, CZC24832, demethyl-coclaurine, oroxin B, homosalate, AMG319, cinobufagin, resibufogenin, hispidulin, GSK2292767, lanatoside C, zeaxanthin, disiteltide (P144), cafestol, paxalisib (GDC-0084), SKI-V, MTX-211, seletalisib (UCB-5857), Trichosanthis Pericarpium Extract, Chuanshanlong Root Extract, GDC-0326, 740 YP (PDGFR 740Y-P), PIK-108, parsaclisib (INCB050465) hydrochloride, HS-173, SF2523, leniolisib (CDZ 173), lupenone, ceravelisib (TAK-117), eganelisib (IPI-549), quercetin (NSC9221), bimiralisib (PQR309), VPS34 inhibitor 1 (compound 19), IHMT-PI3K6-372, voxtalisib (XL765), autofinib, GNE-317, (E)-Akt inhibitor-IV, notoginsenoside R1, tenalisib (RP6530), solasodine, gallein, pectolinarin, inavolisib (GDC-0077), SRX3207, α-linolenic acid, ambralisib (TGR-1202), acalisib (GS-9820), ME-401, 3-hydroxyanthranilic acid, ne Miralisib, samotricisib (LY3023414), VPS34-IN1, airantone, Tritergium wilfordii extract, IPI-3063, SAR405, PIK-III, IPI-3063, parsaclisib (INCB050465), quercetin dihydrate, pyralalisib (XL147), SPP-86, AZD8835, trigonelline, deguelin, selective PI3K6 inhibitor 1 (compound 7n), loureirin A, PF-4989216, AZD8186, GNE-477, and GNE-493.

[0141] All of the proteasome inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any proteasome inhibitor that causes drug resistance.

[0142] Proteasome inhibitors are salinosporamide A (NPI-0052), bortezomib (PS-341), MG132, carfilzomib (PR-171), ixazomib citrate (MLN9708), ixazomib (MLN2238), ONX-0914 (PR-957), oprozomib (ONX 0912), delanzomib (CEP-18770), celastrol (NSC 70931), epoxomicin (BU-4061T), shikonin (CI 75535), VR23, isoginkgo, RA-190, and PI-1840.

[0143] All of the JNK inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any JNK inhibitor that causes drug resistance.

[0144] JNK inhibitors include JNK inhibitor VIII, JNK inhibitor IX, JNK-IN-8, BI-78D3, SP600125, dramapimod (BIRB 796), metformin HCl, DB07268, 3'-hydroxypterostilbene, KB-R7943 mesylate, JNK-IN-7, loureirin B, astragaloside IV, cucurbitacin Ilb, trans-zeatin, ezatiostat, IQ 3, and berberine chloride (NSC 646666), SU3327, bentamapimod (AS602801), indirubin-3'-oxime, falcarindiol, tanzisertib (CC-930), NDMC101, marbelloside A, C-401 hydrochloride, RPI-1, ginsenoside Re, IQ-1S, and urolithin B.

[0145] All of the NF-κB inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any neuropilin-degrading compound alone or in combination with any NF-κB inhibitor that causes drug resistance.

[0146] NF-κB inhibitors include phospho-IKB alpha (S32) rabbit recombinant mAb, ophiopogonin D, cornuside, rubiadin 1-methyl ether, SM-7368, NF-κB-IN-1, chitosan oligosaccharide, Cynanchi Atrati extract, IAXO-102, Adjudin, CBL0137, IQ 3, hyperforin, isolicritin apioside, jaceocidin, bortezomib (PS-341), urolithin B, sulfasalazine (NSC 667219), acetylcysteine ​​(N-acetylcysteine), erdosteine, curcumin, andrographolide, dihydroartemisinin (DHA), indole-3-carbinol, magnolol, (-)-parthenolide, evodiamine, chondroitin sulfate, TPCA-1, sodium salicylate, methylthiouracil, articaine HCl, L-quebrachitol, UCB-9260, zeaxanthin, tetracaine Ctocrisin, Myrislignan, Gardenoside, Caulophilin (N-Methylcytisine), Demethyleneberberine, 3-Hydroxyanthranilic Acid, (E / Z)-IT-603, Triptolide (PG490), Sarsasapogenin, Berbamine Dihydrochloride, Pyrrolidine Dithiocarbamate Ammonium, Stachydrine Hydrochloride, Sodium Escinate, Stachydrine, Tyrosol, Mangiferin, Skeleralin, Ginseng Ginsenoside Re, Dehydroevodiamine, (E)-Cardamonin, Muscone, Guaiacol, Curcumenol, Schisantherin A, Hederagenin, Astragaloside IV, Ginsenoside Rg1, Ginsenoside Rbl, Ginsenoside Rd, 4'-Methoxyresveratrol, (+)-α-Lipoic Acid, Sodium 4-Aminosalicylate, IMM-H007, Diethyl Maleate, 4-Hydroxychalcone, Benfotiamine, QNZ(E VP4593), 4'-hydroxychalcone, neferine, vanillic acid, hyperoside, chelidonic acid, ethyl caffeate, sulforaphane, (R)-(-)-ibuprofen, SN50, C25-140, INH14, licochalcone D, SC75741, JSH-23, caffeic acid phenethyl ester, rocaglamide, APX-3330, DTP3, omaveloxolone (RTA-408), bardoxolone methyl, shikonin (CI75535), TAK-243 (MLN7243), CBL0137 HCl, withaferin A, NIK SMI1, allobresib (GS-5829), antatin, maslinic acid, marbelloside A, eleutheroside E, berbamine, engeletin, aristolochic acid A, ginsenoside Rb3, 8-O-acetylshangjiside methyl ester, dauricine, 2',5'-dihydroxyacetophenone, (+)-praeruptorin A, homoplantaginin, madecassic acid, and BTYNB.

[0147] All of the HSP90 inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any of the neuropilin-degrading compounds alone or in combination with any of the HSP90 inhibitors that result in drug resistance.

[0148] HSP90 inhibitors include luminespib (NVP-AUY922), tanespimycin (17-AAG), alvespimycin (17-DMAG) HCl, ganetespib (STA-9090), elesclomol (STA-4783), BIIB021, tamoxifen (ICI 46474) citrate, onarespib (AT13387), NVP-BEP800, geldanamycin (NSC 122750), SNX-2112 (PF-04928473), PF-04929113 (SNX-5422), KW-2478, XL888, pifithrin-μ, NMS-E973, zeravespib (PU-H71), teprenone, dimethyleneastron, apoptozole, KRIBB11, pseudolaric acid A, TRC051384, DTHIB, rocaglamide, KNK437, VER-49009, HA15, pimitespib (TAS-116), CH5138303, VER-50589, YUM70, triptolide (PG490), VER155008, JG98, tamoxifen (ICI 46474), NPX800, and HSP990 (NVP-HSP990).

[0149] All of the E3 ligase inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any of the neuropilin-degrading compounds alone or in combination with any of the E3 ligase inhibitors that cause drug resistance.

[0150] E3 ligase inhibitors include lenalidomide (CC-5013), pomalidomide (CC-4047), thalidomide (K17), NSC 207895, TAME, PRT4165, CC-885, dCBP-1, iverdomide (CC220), BC-1215, CC-90009, avadomide (CC-122), VL285, (S,R,S)-AHPC (MDK7526), ​​Smurfl-IN-A01, (S,R,S)-AHPC-PEG4-NH2 hydrochloride, SZL P1-41, VH298, thalidomide-OH, MuRF1-IN-1, and Skp2 inhibitor Cl (SKP1). C1), GMB-475, mezigdomide (CC-92480), homo-PROTAC cereblon degrader 1, THAL-SNS-032, NSC232003, Apcin, and thalidomide-O-COOH (cereblon ligand 3).

[0151] All of the KRAS inhibitors listed below, but not limited to, result in drug resistance, which is overcome by administration of any of the neuropilin-degrading compounds alone or in combination with any of the KRAS ligase inhibitors that cause drug resistance.

[0152] KRAS inhibitors include MRTX1133, sotorasib (AMG510), adagrasib (MRTX849), LC-2, deltalasin, BAY-293, ARS-1620, BI-2852, ARS-853 (ARS853), BI-3406, ASP2453, sotorasib (AMG510) racemate, Pan-RAS-IN-1, MRTX-1257, and zoledronic acid (ZOL 446), lonafarnib (SCH66336), K-Ras(G12C) inhibitor 9, salirasib, alamandine, (Rac)-antineoplaston A10, K-Ras-IN-1, MCP110, 6H05, K-Ras(G12C) inhibitor 12, Kobe0065, K-Ras(G12C) inhibitor 6, BQU57, Kobe2602, NAV-2729, antineoplaston A10, fendiline hydrochloride, KRpep-2d, Pei - illy' alcohol, RBC8, KY1220, CID-1067700, and zoledronic acid monohydrate.

[0153] 5. Pharmaceutical Compositions The polypeptides described herein can be formulated as pharmaceutical compositions further comprising a pharmaceutically acceptable carrier, diluent, adjuvant, or vehicle. In certain embodiments, the present disclosure provides a pharmaceutical composition comprising the disclosed polypeptide and a pharmaceutically acceptable carrier, diluent, adjuvant, or vehicle. In certain embodiments, the present invention is a pharmaceutical composition comprising an effective amount of the disclosed bispecific antibody, or a pharmaceutically effective amount of the disclosed bispecific antibody.

[0154] Pharmaceutically acceptable carriers or excipients may contain inactive ingredients that do not excessively inhibit the biological activity of polypeptides. Pharmaceutically acceptable carriers should be biocompatible, for example, non-toxic, non-inflammatory, non-immunogenic, or have no other undesirable reactions or side effects on the subjects to which they are administered. Standard pharmaceutical formulation techniques can be used.

[0155] As used herein, pharmaceutically acceptable carriers, adjuvants, or vehicles include any solvents, diluents, or other liquid vehicles, dispersing or suspending agents, surfactants, isotonicity agents, thickening or emulsifying agents, preservatives, solid binders, lubricants, and the like, appropriate for the particular dosage form desired. Remington's Pharmaceutical Sciences, Sixteenth Edition, by E.W. Martin (Mack Publishing Co., Easton, Pa., 1980) discloses various carriers used in formulating pharmaceutically acceptable compositions and known techniques for their preparation. The use of conventional carrier media incompatible with the polypeptides described herein, e.g., those that produce any undesired biological effects or otherwise adversely interact with other components of the pharmaceutically acceptable composition, is contemplated within the scope of the present invention. As used herein, the phrase "side effects" encompasses undesired and adverse effects of treatment.

[0156] Materials that function as pharmaceutically acceptable carriers for antibodies have the effects of increasing conformational stability, reducing protein dynamics, inhibiting aggregation, and preventing protein adsorption to the liquid / air interface, and include cyclodextrin hydrogel, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins (e.g., human serum albumin), buffer substances (e.g., Tween 80, phosphate, glycine, sorbic acid, or potassium sorbate), partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes (e.g., protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts), colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, polyacrylate, wax, polyethylene-polyoxypropylene block polymer, methylcellulose, hydroxypropyl methylcellulose, wool fat, sugars (e.g., lactose, glucose, sucrose), and starch (corn). Other non-toxic and compatible lubricants, including, but not limited to, starch, sugar, corn starch, potato starch, etc.), cellulose and its derivatives (such as sodium carboxymethylcellulose, ethyl cellulose, cellulose acetate, etc.), powdered tragacanth, malt, gelatin, talc, excipients (such as cocoa butter and suppository wax), fats and oils (such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil), glycols (such as propylene glycol and polyethylene glycol), esters (such as ethyl oleate and ethyl laurate), agar, buffers (such as magnesium oxide and aluminum oxide), alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and phosphate buffer, as well as other non-toxic and compatible lubricants (such as sodium lauryl sulfate and magnesium stearate), coloring agents, release agents, coating agents, sweetening agents, flavoring agents, fragrances, preservatives, antioxidants, and the like, may also be present in the composition at the discretion of the formulator.

[0157] In some embodiments, the compositions of the present invention comprise pharmaceutically acceptable salts. When the polypeptides of the present invention contain relatively acidic functional groups, a base addition salt can be obtained by contacting the neutral form of such polypeptide with a sufficient amount of a desired base, either directly or in an inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium salts, potassium salts, calcium salts, ammonium salts, organic amino salts, magnesium salts, or similar salts. When the polypeptides of the present invention contain relatively basic functional groups, a base addition salt can be obtained by contacting the neutral form of such polypeptide with a sufficient amount of a desired acid, either directly or in an inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts derived from inorganic acids such as hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid, monohydrogencarbonic acid, phosphoric acid, monohydrogenphosphate, dihydrogenphosphate, sulfuric acid, monohydrogensulfuric acid, hydroiodic acid, or phosphorous acid, and salts derived from relatively non-toxic organic acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-tolylsulfonic acid, citric acid, tartaric acid, oxalic acid, methanesulfonic acid, etc. Also included are salts of amino acids such as arginine, and salts of organic acids such as glucuronic acid or galacturonic acid (see Berge et al., "Pharmaceutical Salts," Journal of Pharmaceutical Science, 1977, 66, 1-19). Certain polypeptides of the present disclosure contain both basic and acidic functional groups that allow the polypeptides to be converted into either base or acid addition salts.

[0158] Thus, the disclosed polypeptides may exist as salts with pharmaceutically acceptable acids. The present invention encompasses such salts. Non-limiting examples of such salts include hydrochloride, hydrobromide, phosphate, sulfate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, propionate, tartrate (e.g., (+)-tartrate, (-)-tartrate, or a mixture including the racemate), succinate, benzoate, and salts with amino acids such as glutamic acid, and quaternary ammonium salts (e.g., methyl iodide, ethyl iodide, etc.). These salts may be prepared by methods known to those skilled in the art.

[0159] The neutral form of the polypeptide is preferably regenerated by contacting the salt with a base or acid and isolating the parent polypeptide in the conventional manner. The parent form of the polypeptide may differ from the various salt forms in certain physical properties, such as solubility in polar solvents.

[0160] Certain polypeptides of the present invention can exist in unsolvated and solvated forms (including hydrated forms). Generally, solvated forms are equivalent to unsolvated forms and are within the scope of the present invention. Certain polypeptides of the present invention may also exist in multiple crystalline or amorphous forms. Generally, all physical forms are equivalent for the uses contemplated by the present invention and are within the scope of the present invention.

[0161] In some embodiments, the subcutaneous formulation may contain recombinant human PH2O hyaluronidase (rHuPH20) to facilitate diffusion of the antibody from the injection site.

[0162] 6. Combination Therapy In some embodiments, the compositions and methods described herein may be used in combination with other cancer treatments. These additional cancer treatments may be administered to a subject simultaneously with, before, or after the resistance suppressor or tyrosine kinase inhibitor. Additional cancer treatments may include, for example, chemotherapy, radiation therapy, and surgery.

[0163] Chemotherapy involves the killing of hyperproliferative cells using one or more chemotherapeutic agents. Chemotherapeutic agents include compounds or compositions administered to treat cancer or other hyperproliferative diseases. Chemotherapeutic agents may be classified by their mode of activity within cells, such as the stage of the cell cycle they affect. Alternatively, chemotherapeutic agents may be characterized based on their ability to directly crosslink DNA, intercalate DNA, or affect nucleic acid synthesis to induce chromosomal and mitotic abnormalities. Categories of chemotherapeutic agents include, for example, alkylating agents, antimetabolites, antitumor antibiotics, mitotic inhibitors, and nitrosoureas. Specific chemotherapeutic agents include, for example, cyclophosphamide, doxorubicin, daunorubicin, vinblastine, vincristine, bleomycin, etoposide, topotecan, irinotecan, taxotere, taxol, 5-fluorouracil, methotrexate, gemcitabine, cisplatin, carboplatin, and chlorambucil, as well as agonists of any of the above compounds.

[0164] Radiation therapy (also called radiotherapy) is a method of treating cancer and other diseases with ionizing radiation. Ionizing radiation provides cells with energy that damages or kills them by damaging their genetic material in the treated area, preventing them from continuing to grow. Radiation damages both cancer cells and normal cells, but normal cells are able to repair themselves and function normally. Radiation therapy may be used to treat localized solid tumors, such as cancers of the skin, tongue, larynx, brain, breast, or cervix. It may also be used to treat leukemia and lymphoma (cancers of the blood-forming cells and lymphatic system, respectively).

[0165] Immunotherapy may involve monoclonal antibodies that recognize cancer cells and target them for destruction by the immune system. Such antibodies include rituximab (targeting CD-20), trastuzumab (targeting HER-2), and cetuximab (targeting EGFR). In some embodiments, the anti-cancer antibody is selected from rituximab, trastuzumab, and cetuximab.

[0166] Surgery may involve total or partial removal of the tumor or cancerous tissue. Surgery may also include removal of surrounding tissue. Surgery may also include removal of any tissue that is believed to be at risk for cancer due to the spread or metastasis of cancer.

[0167] 7. Administration The compositions of the present invention may be administered to a subject in need of cancer treatment. The term "administering" or "administration" refers to the act of providing a composition of the present invention, such as a polypeptide or a pharmaceutically acceptable salt thereof, to a subject in need of cancer treatment.

[0168] As used herein, "intermittent administration" includes administering a drug for a period of time (which may be considered a "first administration period"), followed by a period during which the composition is not used or is administered at a lower maintenance dose (which may be considered a "drug holiday"), followed by re-administration of the composition for a period of time (which may be considered a "second administration period"). Generally, the amount of drug administered during the second administration period is the same as the amount administered during the first administration period, but may be increased or decreased depending on medical needs.

[0169] The resistance suppressor and / or tyrosine kinase inhibitor described in detail herein, or a pharmaceutical composition containing them, may be administered to a subject. The resistance suppressor and / or tyrosine kinase inhibitor described in detail herein may be formulated as a composition and administered at a dosage and by a method well known to those skilled in the medical field, taking into consideration factors such as the age, sex, weight, condition, and administration route of a specific subject.

[0170] The resistance suppressor and / or tyrosine kinase inhibitor can be administered prophylactically or therapeutically. In prophylactic administration, the resistance suppressor and / or tyrosine kinase inhibitor can be administered in an amount sufficient to induce a response. In therapeutic applications, the resistance suppressor and / or tyrosine kinase inhibitor is administered to a subject in need of treatment in an amount sufficient to exert a therapeutic effect. The resistance suppressor and / or tyrosine kinase inhibitor can be administered in a therapeutically effective amount.

[0171] For example, a therapeutically effective amount of a resistance suppressor and / or a tyrosine kinase inhibitor or a pharmaceutically acceptable salt thereof is about 1 mg / kg to about 1000 mg / kg, about 5 mg / kg to about 950 mg / kg, about 10 mg / kg to about 900 mg / kg, about 15 mg / kg to about 850 mg / kg, about 20 mg / kg to about 800 mg / kg, about 25 mg / kg to about 750 mg / kg, about 30 mg / kg to about 700 mg / kg, about 35 mg / kg to about 650 mg / kg, about 40 mg / kg to about 650 mg / kg, or about 750 mg / kg. The dose may be about 100 mg / kg to about 600 mg / kg, about 45 mg / kg to about 550 mg / kg, about 50 mg / kg to about 500 mg / kg, about 55 mg / kg to about 450 mg / kg, about 60 mg / kg to about 400 mg / kg, about 65 mg / kg to about 350 mg / kg, about 70 mg / kg to about 300 mg / kg, about 75 mg / kg to about 250 mg / kg, about 80 mg / kg to about 200 mg / kg, about 85 mg / kg to about 150 mg / kg, and about 90 mg / kg to about 100 mg / kg.

[0172] The tolerance suppressor and / or tyrosine kinase inhibitor can be administered by methods well known to those skilled in the art, such as those described in Donnelly et al. (Ann. Rev. Immunol. 1997, 15, 617-648), Feigner et al. (U.S. Patent No. 5,580,859, issued December 3, 1996), Feigner (U.S. Patent No. 5,703,055, issued December 30, 1997), and Carson et al. (U.S. Patent No. 5,679,647, issued October 21, 1997), the contents of which are incorporated herein by reference in their entirety. The tolerance suppressor and / or tyrosine kinase inhibitor can be conjugated to particles or beads and administered to an individual, for example, using a vaccine gun. Those skilled in the art will understand that the selection of a pharmaceutically acceptable carrier (including a physiologically acceptable compound) depends, for example, on the route of administration.

[0173] The tolerance suppressor and / or tyrosine kinase inhibitor can be delivered via various routes. Typical delivery routes include parenteral administration, such as intradermal, intramuscular, or subcutaneous administration. Other routes include oral, intranasal, vaginal, transdermal, intravenous, intraarterial, intratumoral, intraperitoneal, and epidermal routes. In some embodiments, the tolerance suppressor and / or tyrosine kinase inhibitor is administered to a subject intravenously, intraarterially, or intraperitoneally.

[0174] The resistance suppressor and / or tyrosine kinase inhibitor can be a liquid such as a suspension, syrup, or elixir. The resistance suppressor and / or tyrosine kinase inhibitor can be incorporated into liposomes, microspheres, or other polymer matrices (for example, by the method described in Feigner et al., U.S. Pat. No. 5,703,055; Gregoriadis, Liposome Technology, Vols. I-III (2nd ed., 1993), the entire contents of which are incorporated herein by reference). Liposomes can be composed of phospholipids or other lipids, and can be non-toxic, physiologically acceptable, and metabolizable carriers, making them relatively easy to prepare and administer.

[0175] Resistance suppressors and / or tyrosine kinase inhibitors may be used as vaccines. Vaccines can be administered via electroporation, such as by the method described in U.S. Patent No. 7,664,545, the contents of which are incorporated herein by reference. Electroporation can be performed by the methods and / or devices described in U.S. Patent Nos. 6,302,874, 5,676,646, 6,241,701, 6,233,482, 6,216,034, 6,208,893, 6,192,270, 6,181,964, 6,150,148, 6,120,493, 6,096,020, 6,068,650, and 5,702,359, the contents of which are incorporated herein by reference in their entirety. Electroporation can be performed using minimally invasive devices.

[0176] In some embodiments, the resistance inhibitor and / or tyrosine kinase inhibitor is administered in a controlled release formulation. For example, the resistance inhibitor and / or tyrosine kinase inhibitor may be released into the circulatory system. In some embodiments, the resistance inhibitor and / or tyrosine kinase inhibitor may be released for at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 6 days, at least about 7 days, at least about 1 week, at least about 1.5 weeks, at least about 2 weeks, at least about 2.5 weeks, at least about 3.5 weeks, at least about 4 weeks, or at least about 1 month.

[0177] Sterile injectable forms of the compositions described herein may be aqueous or oily suspensions. These suspensions may be formulated using suitable dispersing or wetting agents and suspending agents according to procedures known to those skilled in the art. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as 1,3-butanediol solutions. Suitable carriers and solvents that may be used include water, Ringer's solution, and isotonic saline solution. Additionally, sterile, fixed oils are conventionally used as solvents or suspending media. For this purpose, any bland, odorless, fixed oil may be used, including synthetic monoglycerides or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives are useful in the preparation of injectables, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially those polyoxyethylated. These oil solutions or suspensions may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other surfactants commonly used in the manufacture of pharmaceutically acceptable dosage forms, such as Tween, Span, and other emulsifiers or bioavailability enhancers, may also be used for formulation purposes.

[0178] In some embodiments, the formulation includes agents such as excipients, buffers, tonicity agents, preservatives, surfactants, and preferably zinc. The formulation may also include excipients or agents for polypeptide stabilization, such as buffers, reducing agents, bulk proteins, or carbohydrates. Bulk proteins useful in formulating at least one polypeptide composition include albumin, protamine, and the like. Typical carbohydrates useful in formulating at least one polypeptide include sucrose, mannitol, lactose, trehalose, glucose, and the like. The bispecific antibody formulation may also include a surfactant capable of reducing or preventing surface-induced aggregation of at least one polypeptide caused by micronization of the solution. Various conventional surfactants, such as polyoxyethylene fatty acid esters and alcohols, and polyoxyethylene sorbitol fatty acid esters, may be used. The amount generally ranges from about 0.001 to 4% by weight of the formulation. Particularly preferred surfactants for the purposes of the present invention include polyoxyethylene sorbitan monooleate, polysorbate 80, polysorbate 20, and the like. Additional components for formulation of polypeptides known to those skilled in the art, such as antibody proteins, may also be included in the formulation. [Example]

[0179] The examples herein are not intended to, and should not be used to, limit the invention, but are provided merely to illustrate the invention.

[0180] Example 1. Combination treatment of EGFR degraders with osimertinib enhanced antitumor activity in an osimertinib-resistant mouse xenograft model The bispecific EGFRxNRP1 antibody, an EGFR degrader, has been shown to degrade the tumorigenic receptor EGFR via lysosomes and reduce cell viability in vitro. Furthermore, EGFR degraders have been shown to significantly inhibit tumor growth in osimertinib-sensitive (H1975), osimertinib-resistant (H1975-OR), and osimertinib-refractory (H1975-HGF) xenograft mouse models, all of which harbor an EGFR double mutation (T970M / L858R) (unpublished data). In the osimertinib-resistant xenograft mice (H1975-OR) used herein, osimertinib at a dose of 3 mg / kg did not exhibit antitumor activity.

[0181] To investigate whether cotreatment with an EGFR-degrading agent and a therapeutic agent enhances the therapeutic effect in therapeutically resistant cancers, we treated an osimertinib-resistant mouse xenograft model (H1975-OR) with panitumumab (10 mg / kg, intraperitoneally, once weekly) plus osimertinib (10 mg / kg, intraperitoneally, once daily), an EGFR-degrading agent (13.74 mg / kg, intraperitoneally, once weekly) plus osimertinib (10 mg / kg), osimertinib alone, or IgG1 isotype (5 mg / kg, intraperitoneally, once weekly). Panitumumab is a fully humanized anti-EGFR monoclonal antibody that binds to EGFR and inhibits EGFR signaling. Osimertinib is an EGFR tyrosine kinase inhibitor.

[0182] As shown in Figure 3, no synergistic effect was observed in osimertinib-resistant tumors from mice treated with panitumumab and osimertinib. This result suggests that co-treatment of osimertinib and panitumumab inhibited EGFR signaling in osimertinib-resistant tumors, maintaining resistance to osimertinib. In contrast, osimertinib-resistant mice co-treated with an EGFR degrader and osimertinib showed greater tumor regression. This result suggests that EGFR degraders mediate lysosomal downregulation of EGFR. T790M / L858R degradation of EGFR and subsequentT790M / L858R This shows that removal of the target protein degrader resulted in a highly efficient synergistic effect against osimertinib-resistant tumors. Combination therapy with a therapeutic agent and a targeted protein degrader enhanced the therapeutic effect of the therapeutic agent in therapeutic-resistant mice. Overall, the combination therapy disclosed herein may provide a new approach to overcoming drug resistance and enhancing therapeutic efficacy in cancer treatment. A schematic diagram of combination therapy for enhancing the therapeutic effect of a therapeutic agent in patients with cancer resistant to the therapeutic agent is shown in Figure 4.

[0183] Example 2. Combination treatment of EGFR degraders with osimertinib enhanced antitumor activity in an NRP1-overexpressing mouse xenograft model In this example, a modified version of the H1975 xenograft model was used in which tumor cells were engineered to overexpress NRP1. In this xenograft model, called H1975-NRP1 OE, tumor cells express EGFR L858R / T790M , NRP1 +++ , EGFR + Regarding osimertinib resistance, treatment of the parental H1975 model with 3 mg / kg osimertinib resulted in potent and durable tumor regression, whereas treatment of the H1975-NRP1 OE model with the same dose of osimertinib resulted in significant tumor growth, suggesting that overexpression of NRP1 induced at least some degree of resistance to osimertinib treatment.

[0184] To investigate whether cotreatment with EGFR-degrading agents and osimertinib enhances the therapeutic effect of osimertinib in the H1975-NRP1 OE xenograft model, mice were treated with panitumumab (10 mg / kg, intraperitoneally, once weekly), EGFR-degrading agents (13.74 mg / kg, intraperitoneally, once weekly), osimertinib (3 mg / kg, intraperitoneally, once daily), EGFR-degrading agents plus osimertinib, or IgG1 isotype control (10 mg / kg, intraperitoneally, once weekly). The time course of tumor volume is shown in Figure 5A. The time course of survival probability is shown in Figure 5B.

[0185] The results demonstrated that administration of an EGFR-degrading agent alone resulted in significantly greater tumor growth inhibition than administration of panitumumab alone in this model. Furthermore, the combination of an EGFR-degrading agent with osimertinib demonstrated strong synergy, resulting in tumor regression. While not intending to be limited by mechanism of action, this suggests that elimination of EGFR mutants via degradation by the EGFR-degrading agent is necessary for optimal combined tumor growth inhibition. Administration of an EGFR-degrading agent alone also resulted in improved median survival compared with panitumumab or osimertinib alone, and the combination of an EGFR-degrading agent with osimertinib demonstrated strong synergy, resulting in significantly improved survival in mice.

[0186] Thus, in summary, similar to the results observed in Example 1, an EGFR inhibitor in combination with osimertinib was the most effective treatment in suppressing tumor growth and improving median survival in this NRP1-overexpressing xenograft model.

[0187] Example 3. Combination treatment of EGFR degrading agents with therapeutic agents enhanced antitumor activity in an HGF-overexpressing mouse xenograft model In this example, we used a modified H1975 xenograft model in which tumor cells were engineered to overexpress hepatocyte growth factor (HGF, the cMET ligand). The HGF / cMET pathway was demonstrated to be active in H1975 cells by inhibiting the pathway with either crizotinib, a small molecule inhibitor that inhibits cMET pathway signaling, or amivantamab, an EGFRxcMET bispecific antibody construct that inhibits cMET pathway signaling and induces cMET degradation (data not shown). The H1975-HGF cell line was prepared by recombinantly expressing HGF in the H1975 cell line using standard methods. Thus, the H1975-HGF xenograft model represents a dual activation model in which both the EGFR and cMET pathways are constitutively activated.

[0188] In the first series of experiments, to investigate whether cotreatment of EGFR-degrading agents with osimertinib enhances the therapeutic effect of osimertinib in the H1975-HGF xenograft model, mice were treated with panitumumab (5 mg / kg, i.p., twice weekly), EGFR-degrading agents (6.87 mg / kg, i.p., twice weekly), amivantamab (5.05 mg / kg, i.p., twice weekly), osimertinib (3 mg / kg, i.p., once daily), EGFR-degrading agents plus osimertinib, or an IgG1 isotype control (5 mg / kg, i.p., twice weekly). The results of tumor volume over time are shown in Figure 6A. Results demonstrated that the H1975-HGF model was resistant to both panitumumab and low-dose osimertinib monotherapy, whereas EGFR-degrading agent monotherapy and amivantamab monotherapy had similar tumor growth suppression effects. However, the combination of EGFR-degrading agent and osimertinib demonstrated the greatest tumor growth suppression effect over time.

[0189] In a second series of experiments, we investigated the effects of cotreatment with the EGFR degrader, osimertinib, and the cMET inhibitor crizotinib. Mice were treated with osimertinib (10 mg / kg, intraperitoneally, once daily), crizotinib (20 mg / kg, intraperitoneally, once daily), EGFR degrader (15 mg / kg, intraperitoneally, once weekly), EGFR degrader + osimertinib, EGFR degrader + crizotinib, EGFR degrader + osimertinib + crizotinib, or IgG1 isotype control (5 mg / kg, intraperitoneally, twice weekly). The time course of tumor volume is shown in Figure 6B. Results demonstrated that crizotinib monotherapy had limited efficacy in the H1975-HGF model, and even high-dose osimertinib monotherapy failed to suppress tumor growth over time. EGFR-degrading agents alone demonstrated tumor growth inhibition, and the combination of an EGFR-degrading agent with crizotinib or osimertinib demonstrated enhanced tumor growth inhibition compared with either agent alone. The triple combination of an EGFR-degrading agent, osimertinib, and crizotinib demonstrated the best efficacy over time.

[0190] Therefore, in summary, similar to the results observed in Examples 1 and 2, the combination of EGFR degraders with osimertinib, crizotinib, or both significantly improved tumor growth suppression in this HGF-overexpressing xenograft model in which the EGFR / cMET pathway was dually activated.

[0191] Example 4. Combination treatment with EGFR degraders and KRAS inhibitors enhanced antitumor activity in mutant KRAS mouse xenograft models In this example, we used a mutant KRAS mouse xenograft model to examine the effects of EGFR degrading agents, alone or in combination with KRAS inhibitors, on tumor growth in the mutant KRAS model H358. In the H358 model, tumor cells express KRAS G12C , NRP1 + , EGFR + and is responsive to sotorasib, an irreversible KRAS G12C selective inhibitor.

[0192] Mice were treated with sotorasib (5 mg / kg, intraperitoneally, once daily, five times per week), an EGFR-degrading agent (5 mg / kg, intraperitoneally, once per week), an EGFR-degrading agent plus sotorasib, or an IgG1 isotype control (3.66 mg / kg, intraperitoneally, once per week). The results of tumor volume over time are shown in Figure 7. The results demonstrated that while both sotorasib monotherapy and EGFR-degrading agent monotherapy exhibited similarly potent antitumor activity, the combination of an EGFR-degrading agent and sotorasib exhibited synergistic effects, resulting in the most significant tumor regression. These results further support the effects observed and reported in Examples 1-3 and demonstrate the effectiveness of combining an EGFR-degrading agent with a therapeutic agent that targets a tumor cell pathway to enhance the antitumor activity of both agents.

[0193] The above description of the specific embodiments fully reveals the general nature of the present invention, and those skilled in the art can easily modify and / or adapt the specific embodiments for various applications without undue experimentation and without departing from the general concept of the present disclosure. Therefore, such modifications and adaptations are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teachings and guidance provided herein. The terms or phrases used herein are for the purpose of description, not limitation, and it should be understood that the terms or phrases used herein should be interpreted by those skilled in the art in light of the teachings and guidance provided herein.

[0194] The breadth and scope of the present disclosure should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0195] All publications, patents, patent applications, and / or other documents cited herein are incorporated by reference in their entirety for all purposes to the same extent as if each were individually indicated to be incorporated by reference for all purposes.

[0196] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5] [Table 3-6] [Table 3-7] Table 3-8 Table 3-9 Table 3-10 Table 3-11 Table 3-12 Table 3-13 Table 3-14 Table 3-15 Table 3-16 Table 3-17 Table 3-18 Table 3-19 Table 3-20 Table 3-21 Table 3-22 Table 3-23

Claims

1. 1. A method of enhancing the effect of a therapeutic agent in a subject having a cancer that is resistant or refractory to the therapeutic agent, comprising: administering to a subject (i) a therapeutic agent and (ii) a targeted protein degrader, thereby enhancing the effect of the therapeutic agent compared to administering the therapeutic agent alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to a target protein on a cancer cell of a subject; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising method.

2. The method of claim 1, wherein the target protein is a receptor tyrosine kinase (RTK).

3. 3. The method of claim 2, wherein the receptor tyrosine kinase is selected from epidermal growth factor receptor (EGFR), platelet-derived growth factor receptor (PDGFR), fibroblast growth factor receptor (FGFR), Met receptor tyrosine kinase (MET), and vascular endothelial growth factor receptor (VEGFR).

4. The method of claim 2, wherein the receptor tyrosine kinase is EGFR.

5. 3. The method of claim 2, wherein the receptor tyrosine kinase is cMET.

6. 3. The method of claim 2, wherein the receptor tyrosine kinase is HER2.

7. The method of claim 2, wherein the receptor tyrosine kinase is IGF1R.

8. The method of claim 1 , wherein the target cells are cancer cells.

9. 9. The method of claim 8, wherein the cancer cells are selected from the group consisting of lung cancer, breast cancer, colorectal cancer, head and neck cancer, esophagogastric cancer, liver cancer, glioblastoma, prostate cancer, cervical cancer, ovarian cancer, bladder cancer, renal cancer, and pancreatic cancer.

10. 9. The method of claim 8, wherein the cancer cells are non-small cell lung cancer (NSCLC) cells.

11. 2. The method of claim 1, wherein the target protein binding domain and the NRP1 binding domain are each independently selected from the group consisting of an IgG, a half antibody, a single domain antibody, a nanobody, a Fab, a monospecific Fab2, an Fc, a scFv, a minibody, an IgNAR, a V-NAR, a hcIgG, a VHH domain, a camelid antibody, and a peptibody.

12. NRP1 binding domain (i) an antibody heavy chain variable (VH) domain comprising CDR1, CDR2 and CDR3 regions (HCDR1, HCDR2 and HCDR3, respectively), wherein HCDR1 consists of the sequence set forth in SEQ ID NO: 79, HCDR2 consists of the sequence set forth in SEQ ID NO: 80, and HCDR3 consists of the sequence set forth in any one of SEQ ID NOs: 81-84; and (ii) an antibody light chain variable (VL) domain comprising CDR1, CDR2 and CDR3 regions (LCDR1, LCDR2 and LCDR3, respectively), wherein LCDR1 consists of the sequence set forth in any one of SEQ ID NOs: 85-87, LCDR2 consists of the sequence set forth in SEQ ID NO: 88, and LCDR3 consists of the sequence set forth in SEQ ID NO: 89; The method of claim 1 , comprising:

13. (iii) HCDR1 consists of the sequence set forth in SEQ ID NO: 79, HCDR2 consists of the sequence set forth in SEQ ID NO: 80, and HCDR3 consists of the sequence set forth in any one of SEQ ID NOs: 84; and (iv) LCDR1 consists of the sequence shown in SEQ ID NO: 85, LCDR2 consists of the sequence shown in SEQ ID NO: 88, and LCDR3 consists of the sequence shown in SEQ ID NO: 89; The method of claim 12.

14. The method of claim 1 , wherein the enhanced effect comprises an increased tumor growth inhibitory effect.

15. The method of claim 1 , wherein the enhanced effect comprises an increase in median survival time.

16. 10. The method of claim 1, wherein the therapeutic agent targets the EGFR pathway.

17. Therapeutic drugs include lazertinib, osimertinib (AZD9291), WZ4002, cyasterone, erlotinib (OSI-774) HCl, efitinib (ZD1839), lapatinib (GW-572016) ditosylate, afatinib (BIBW2992), saracatinib (AZD0530), vandetanib (ZD6474), and nera. Tinib (HKI-272), canertinib (CI-1033), lapatinib (GW-572016), AG-490 (tyrphostin B42), CP-724714, dacomitinib (PF-00299804), sapitinib (AZD8931), CUDC-101, AG-1478 (tyrphostin AG-1478), PD153035 HCl, pelitinib (EKB-569), AC480 (BMS-599626), AEE788 (NVP-AEE788), AP26113-analogue (ALK-IN-1), OSI-420, WZ3146, HER2-inhibitor-1, WZ8040, alitinibutosylate, rociletinib (CO-1686), genistein (NPI 031L), barlitinib, TQB33804 (EGFR-IN-7), icotinib (BPI-2009H), TAK-285, daphnetin, tyrphostin 9, AG-18, AG555, AZ5104, CL-387785 (EKI-785), tyrphostin AG-258, AG-556, tucatinib, erlotinib (OSI-774), gefitinib-based PROTAC3, zolifertinib (AZD 3759), ErbB2, AV-412 free base, AST-1306, JND3229, BI-4020, ceriatinib (HMPL-309), BDTX-189, lifirafenib (BGB-283), pyrotinib (SHR-1258), O-demethyl-gefitinib, epertinib hydrochloride, SU5214, avitinib (AC0010), AG494, and poziotinib (HM781-36B).

18. 17. The method of claim 16, wherein the therapeutic agent is osimertinib.

19. 10. The method of claim 1, wherein the therapeutic agent targets the cMET pathway.

20. Therapeutic drugs include crizotinib, cabozantinib, foretinib, PHA-665752, SU11274, SGX-523, BMS-777607, tivantinib, JNJ-38877605, PF-04217903, amuvatinib (MP-470), MGCD-265 analogs, capmatinib, BMS-754807, BMS-794833, and A 20. The method of claim 19, wherein the medicament is selected from the group consisting of MG-208, MK-2461, golvatinib, AMG-458, NVP-BVU972, AMG337, merestinib, JNJ-38877618, crizotinib hydrochloride, ningetinib, AMG-1, UNC2025, pamfetinib, altiratinib, NPS-1304, and savolitinib.

21. 20. The method of claim 19, wherein the therapeutic agent is crizotinib.

22. 10. The method of claim 1, wherein the therapeutic agent targets the KRAS protein.

23. Therapeutic drugs include MRTX1133, sotorasib (AMG510), adagrasib (MRTX849), LC-2, deltalasin, BAY-293, ARS-1620, BI-2852, ARS-853 (ARS853), BI-3406, ASP2453, sotorasib (AMG510) racemate, Pan-RAS-IN-1, MRTX-1257, and zoledronic acid (ZOL 446), lonafarnib (SCH66336), K-Ras (G12C) inhibitor 9, salirasib, alamandine, (Rac) antineoplaston A10, K-Ras-IN-1, MCP110, 6H05, K-Ras (G12C) inhibitor 12, Kobe0065, K-Ras (G12C) inhibitor 6, BQU57, Kobe2602, NAV-2729, antineoplaston A10, fendiline hydrochloride, KRpep-2d, Pei － 23. The method of claim 22, wherein the anticoagulant is selected from the group consisting of illy' alcohol, RBC8, KY1220, CID-1067700, and zoledronic acid monohydrate.

24. 24. The method of claim 23, wherein the therapeutic agent is sotorasib.

25. 10. The method of claim 1, wherein the therapeutic agent targets a protein selected from the group consisting of HER2, IGF1R, ALK, Braf, VEGF, and PDGF.

26. 10. The method of claim 1, further comprising administering to the subject a second therapeutic agent, wherein the effect of the second therapeutic agent is enhanced by the targeted protein degradation agent compared to administration of the second therapeutic agent alone.

27. 27. The method of claim 26, wherein a therapeutic agent targets EGFR and a second therapeutic agent targets cMET.

28. 28. The method of claim 27, wherein the therapeutic agent is osimertinib and the second therapeutic agent is sotorasib.

29. 1. A method of enhancing the effect of a receptor tyrosine kinase (RTK) inhibitor in a subject having a cancer that is resistant or refractory to an RTK inhibitor, comprising: administering to a subject (i) an RTK inhibitor and (ii) a targeted protein degrader, thereby enhancing the effect of the RTK inhibitor compared to administering the RTK inhibitor alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to an RTK on a cancer cell of a subject; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising method.

30. 1. A method of enhancing the effect of an epidermal growth factor receptor (EGFR) inhibitor in a subject having a cancer that is resistant or refractory to an EGFR inhibitor, comprising: administering to a subject (i) an EGFR inhibitor and (ii) a targeted protein degradation agent, thereby enhancing the effect of the EGFR inhibitor compared to administering the EGFR inhibitor alone; wherein the targeted protein degradation agent is (a) a target protein binding domain that specifically binds to EGFR on cancer cells of a subject; and (b) a neuropilin-1 (NRP1)-binding domain, including an antibody or NRP1-binding fragment thereof, that binds to NRP1; and a bispecific binding molecule comprising method.

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