TW202002952A - ABBV-621 in combination with anti-cancer agents for the treatment of pancreatic cancer - Google Patents

Abstract

This invention pertains to a method for the treatment of pancreatic cancer in a subject comprising administering to the subject an effective amount of ABBV-621 in combination with anti-cancer agents.

Description

Combination of ABBV-621 and anti-cancer agent for the treatment of pancreatic cancer

The present invention relates to a synergistic method of using ABBV-621 in combination with an anticancer agent for the treatment of pancreatic cancer.

Cell death can be triggered by the activation of foreign and internal apoptosis signaling pathways. Tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL), a member of the TNF superfamily of interleukins, binds as a trimer to two closely related cell surface death receptors TRAIL-R1 ( DR4) and TRAIL-R2 (DR5) preferentially trigger the foreign cell apoptosis pathway. When these receptors undergo trimerization, they result in the formation of death-inducing signaling complexes (DISCs) to recruit and activate downstream apoptotic proteases that ultimately lead to apoptotic cell death. Since TRAIL was found to induce apoptosis, several TRAIL receptor agonists have been developed to treat cancer.

ABBV-621 is a novel second-generation TRAIL receptor agonist, which is currently being tested in Phase 1 clinical trials. It is an engineered fusion protein composed of IgG1-Fc, which is linked to a single chain trimer of TRAIL subunits, producing a total of six death receptor binding sites per molecule to maximize receptor clustering. After 24 hours of treatment, ABBV-621 showed potent single agent activity in acute myeloid lymphoma (AML) and diffuse large B-cell lymphoma (DLBCL) cell lines. This activity is rapid and mechanism-based, due to the observation of downstream apoptosis signaling events (apoptosis protease activation, mitochondrial depolarization, phospholipid amide serine acid exposure) as early as 1 hour after the addition of ABBV-621. And cell death can be completely blocked by the pan-apoptotic protease inhibitor z-VAD-FMK.

The activity of ABBV-621 has been established in a large number of cancer types in vitro and in vivo and it is potent as a single agent. Specifically, ABBV-621 has shown the antitumor activity of single agents in AML and DLBCL in an in vivo model. However, some unit types are resistant to ABBV-621 as a single agent. The present invention demonstrates the ability of ABBV-621 to work synergistically with a group of pancreatic cancer cell lines: six standard care agents (gemcitabine, 5-fluorouracil (5FU), Erlotinib, paclitaxel, docetaxel, and SN-38 (the active form of irinotecan) and several other anti-cancer agents that induce apoptosis (Navito Grams (navitoclax), venetoclax (venetoclax) and compound (I)).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 in combination with one or more anti-pancreatic cancer agents.

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and one or more anti-pancreatic cancer agents, the one or more anti-pancreatic cancer The agent is selected from gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, docetaxel and irinotecan.

The present invention relates to a method for treating pancreatic cancer, which comprises administering a synergistic combination of an anti-pancreatic cancer agent before, after or simultaneously with a therapeutically effective amount of TRAIL receptor agonist ABBV-621 The cancer agent is selected from gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, docetaxel and irinotecan.

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and one or more anti-cancer agents that induce apoptosis. The anti-cancer agent for apoptosis is selected from the group consisting of Navitoc, Venetoc and Compound (I).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine) and albumin-bound paclitaxel.

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (docetaxel).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (5-fluorouracil).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (erlotinib).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (paclitaxel).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (irinotecan).

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-cancer agent (Navitock) that induces apoptosis.

The invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-cancer agent (Venetoc) that induces apoptosis.

The present invention relates to a method for treating pancreatic cancer, which comprises a synergistic combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-cancer agent (compound (I)) that induces apoptosis.

The present invention relates to a method for treating pancreatic cancer in an individual in need, which comprises administering to the individual a synergistically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine) combination.

The present invention relates to a method for treating pancreatic cancer in an individual in need, which comprises administering to the individual a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine and white Protein-bound paclitaxel) combination.

The present invention relates to a method for treating pancreatic cancer in an individual in need, which comprises administering to the individual a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and a combination selected from the group consisting of paclitaxel and albumin-binding type A combination of anti-pancreatic cancer agents composed of paclitaxel.

The present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (SN-38) Of the combination.

The present invention relates to a method for treating pancreatic cancer in an individual in need, which comprises administering to the individual a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (docetaxel) Of the combination.

Cross-Reference to Related Applications This application claims the rights and interests of US Provisional Application No. 62/643,235 filed on March 15, 2018; and claims the US Provisional Application No. 62/815,549 filed on March 8, 2019 Interests, the contents of which are hereby incorporated by reference in full text.

The activity of ABBV-621 has been established in a large number of cancer types in vitro and in vivo and it is potent as a single agent. However, some unit types are resistant to ABBV-621 as a single agent. The ability of ABBV-621 in a group of pancreatic cancer cell lines to work synergistically with the following illustrates a method for treating pancreatic cancer: six standard care agents (gemcitabine, 5-fluorouracil ( 5FU), erlotinib, paclitaxel, docetaxel and SN-38 (the active form of irinotecan)) and several anti-cancer agents that induce apoptosis (navitoc, venetoc and compounds ( I)).

In one embodiment, the present invention relates to ABBV-621, a TNF-related apoptosis-inducing ligand (TRAIL) receptor agonist protein. A TRAIL receptor agonist protein is disclosed, which comprises a dimer of two polypeptides having an amino acid sequence as set forth in SEQ ID NO: 1, wherein the two polypeptides are formed by cysteine in each polypeptide The three interchain disulfide bonds between residues 513, 519 and 522 are covalently linked, and one or more of the asparagine residues at positions 168 and 337 of the polypeptide are N- glycosylated , And by post-translational modification by changing the N- terminal glutamic acid into pyrofuronic acid salt, as disclosed in US Patent Publication No. 2015/0337027, which is cited in full and out of all The purpose is incorporated into this article.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, comprising administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and one or more anti-pancreatic Dirty cancer agent.

In one embodiment, the present invention relates to a synergistic method for treating pancreatic cancer in an individual in need, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and one or more options From the following anti-pancreatic cancer agents: gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, docetaxel and irinotecan.

In one embodiment, the present invention relates to a synergistic method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of the TRAIL receptor agonist ABBV-621 before or simultaneously Or more anti-pancreatic cancer agents selected from the group consisting of gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, docetaxel and irinotecan.

In one embodiment, the present invention relates to a synergistic method for the treatment of pancreatic cancer in an individual in need, which comprises administering an TRAIL receptor agonist ABBV-621 to the individual before administering an effective amount Or multiple anti-pancreatic cancer agents selected from the group consisting of gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, docetaxel, and irinotecan, so that the anti-pancreatic cancer agent is administered before ABBV-621 With sufficient time to ensure that the individual can achieve synergy or optimal synergy. For this purpose, "sufficient time" may account for about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 , 21, 22, 23 or 24 hours ± about 15 to about 45 minutes.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Gemcitabine).

As known in the art, the CAS registration number for gemcitabine is 95058-81-4; the test formula is C ₉ H ₁₁ F ₂ N ₃ O ₄ ; the gram molecular weight is 263.20; and it is described in GB published on September 19, 1984 In Patent Publication No. GB2136425A, it is incorporated herein by reference in its entirety and for all purposes. The term "4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxetan-2-yl]pyrimidine The use of "-2(1 H )-one" (unless otherwise specified) covers solvates (including hydrates) and polycrystalline forms of gemcitabine or its salts. 4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl]pyrimidine-2 (1 H )-one pharmaceutical composition includes: all containing 4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl ) A pharmaceutically acceptable composition of oxolane-2-yl]pyrimidin-2(1 H )-one, and one or more diluents, vehicles and/or excipients. Contains 4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolane-2-yl]pyrimidine- An example of a pharmaceutical composition of 2(1 H )-one is GEMZAR ^® (Lilly). GEMZAR ^® contains 4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxolane as an active ingredient- 2-yl]pyrimidin-2(1 H )-one, which is also called gemcitabine. The GEMZAR ^® vial contains 200 mg or 1 g of gemcitabine HCl (expressed as a free base), which is formulated as a sterile lyophilized powder with mannitol (200 mg or 1 g, respectively) and sodium acetate (12.5 mg or 62.5 mg, respectively). Hydrochloric acid and/or sodium hydroxide may be added for pH adjustment.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Another combination of gemcitabine and albumin-bound paclitaxel).

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (5-fluorouracil).

As known in the art, the CAS registration number of 5-fluorouracil is 51-28-8; the test formula is C ₄ H ₃ FN ₂ O ₂ ; the gram molecular weight is 130.08; and it was described on August 6, 1957. U.S. Patent No. 2,802,005. The use of the term "5-fluorouracil" (unless otherwise specified) covers solvates (including hydrates) and 5-fluorouracil or its salts in polycrystalline form. The 5-fluorouracil pharmaceutical composition includes all pharmaceutically acceptable compositions containing 5-fluorouracil and one or more diluents, vehicles and/or excipients. 5-fluorouracil can be obtained in two pharmacy bulk vials (2.5 g/50 mL vial or 5 g/100 mL vial) in water for injection and the pH is adjusted to 8.6 to 9.4 with sodium hydroxide.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Erlotinib).

As known in this technology, the erlotinib CAS registration number is 183321-74-6; the test formula is C ₂₂ H ₂₃ N ₃ O ₄ ; the gram molecular weight is 393.44; and it is described as granted on December 29, 2009 No. USRE41065E1, which is incorporated herein by reference in its entirety and for all purposes. The term " N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine" (unless otherwise specified) covers solvates ( Including hydrates) and polycrystalline forms of N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine or its salts. The pharmaceutical composition of N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine includes: all containing N- (3-ethynylphenyl ) A pharmaceutically acceptable composition of 6,7-bis(2-methoxyethoxy)quinazolin-4-amine, and one or more diluents, vehicles, and/or excipients. An example of a pharmaceutical composition containing N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine is TARCEVA ^® (OSI Pharmaceuticals, LLC) . TARCEVA ^® lozenges for oral administration can be used in three dosage strengths and contain: erlotinib hydrochloride (27.3 mg, 109.3 mg and 163.9 mg), equivalent to 25 mg, 100 mg and 150 mg erlotinib Nigeria; and the following inactive ingredients: lactose monohydrate, hypromellose, hydroxypropyl cellulose, magnesium stearate, microcrystalline cellulose, sodium starch glycolate, sodium lauryl sulfate and titanium dioxide. The lozenges also contain trace amounts of coloring additives, including FD&C Yellow #6 (only 25 mg) for product identification.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Paclitaxel).

As known in the art, the paclitaxel CAS registration number is 33069-62-4; the test formula is C ₄₇ H ₅₁ NO ₁₄ ; and the gram molecular weight is 853.9. The term "(2a R ,4 S ,4a S ,6 R ,9 S ,11 S ,12 S ,12a R ,12b S )-6,12b-bis(ethyloxy)-9-{((2 R ,3 S )-3-benzylamino-2-hydroxy-3-phenylpropionyl]oxy}-4,11-dihydroxy-4a,8,13,13-tetramethyl-5 -Penoxy-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecane-1 H -7,11-methylenecyclodecane[3,4] The use of "benzo[1,2- b ]oxetane-12-ylbenzoate" (unless otherwise specified) covers solvates (including hydrates) and polymorphic forms (2a R , 4 S ,4a S ,6 R ,9 S ,11 S ,12 S ,12a R ,12b S )-6,12b-bis(ethyloxy)-9-{((2 R ,3 S )- 3-benzylamino-2-hydroxy-3-phenylpropionyl]oxy}-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-pentoxy-2a ,3,4,4a,5,6,9,10,11,12,12a,12b-dodecane-1 H -7,11-methylenecyclodecane[3,4]benzo[1,2 -b ] oxetane-12- yl benzoate or its salt. (2a R ,4 S ,4a S ,6 R ,9 S ,11 S ,12 S ,12a R ,12b S )-6,12b-bis(acetyloxy)-9-{((2 R , 3 S )-3-benzylamino-2-hydroxy-3-phenylpropionyl]oxy)-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-side Oxy-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecane-1 H -7,11-methylenecyclodec[3,4]benzo [1,2- b ]Oxetane-12- yl benzoate pharmaceutical composition includes: all containing (2a R , 4 S , 4a S , 6 R , 9 S , 11 S , 12 S , 12a R ,12b S )-6,12b-bis(ethyloxy)-9-{((2 R ,3 S )-3-benzylamino-2-hydroxy-3-phenylpropane Yl]oxy}-4,11-dihydroxy-4a,8,13,13-tetramethyl-5-pentoxy-2a,3,4,4a,5,6,9,10,11,12 ,12a,12b-dodecane-1 H -7,11-methylenecyclodecane[3,4]benzo[1,2- b ]oxetane-12-ylbenzoate, and A pharmaceutically acceptable composition of one or more diluents, vehicles and/or excipients. Pacific paclitaxel injection is a clear, colorless to slightly yellow viscous solution. It is supplied as a non-aqueous solution intended for dilution with a suitable parenteral fluid before intravenous infusion. Paclitaxel injection is available in 30 mg (5 mL), 100 mg (16.7 mL) and 300 mg (50 mL) multi-dose vials. Each milliliter of sterile pyrogen-free solution contains 6 mg paclitaxel, 527 mg pure Cremophor ^® EL* (polyoxyethylated castor oil) and 49.7% (v/v) absolute alcohol USP. Paclitaxel can also be used as an alternative formulation for ABRAXANE ^® (Celgene Corporation) for injectable suspensions (paclitaxel protein binding particles for injectable suspensions, albumin-bound paclitaxel) (albumin-bound type) Paclitaxel, the albumin-bound form, has an average particle size of about 130 nanometers. Paclitaxel is present in the particles in an amorphous, non-crystalline form. ABRAXANE ^{® is} supplied with 20 mL of 0.9% sodium chloride injection USP in the form of a white to yellow sterile lyophilized powder for reconstitution before intravenous infusion. Each disposable vial contains 100 mg paclitaxel (conjugated to human albumin) and approximately 900 mg human albumin (containing sodium caprylate and sodium acetyltryptamine). Each millilitre (mL) of reconstituted suspension contains 5 mg paclitaxel. ABRAXANE ^® does not contain solvents.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Docetaxel).

As known in the art, the therapeutic form of docetaxel is docetaxel injection concentrate (referred to herein as compound (IV)), and its CAS registration number is 114977-28-5; the test formula is C ₄₃ H ₅₃ NO ₁₄ • 3H 2 O; the gram molecular weight is 861.9; and is described in US Patent No. 4,814,470, published on March 21, 1989, which is incorporated herein by reference in its entirety and for all purposes. The term "(2R,3S)-N-carboxy-3-phenylisoserine N-third butyl ester 13-ester 5β-20-epoxy-1,2α,4,7β,10β,13α -The use of hexahydroxytaxel-11-ene-9-one 4-acetate 2-benzoate" (unless otherwise specified) covers solvates (including hydrates) and polyenes in polycrystalline form Paclitaxel or its salt. (2R,3S)-N-Carboxy-3-phenylisoserine trihydrate N-third butyl ester, 13-ester 5β-20-epoxy-1,2α,4,7β,10β,13α- The pharmaceutical composition of hexahydroxytaxel-11-en-9-one 4-acetate 2-benzoate includes: all containing (2R,3S)-N-carboxy-3-phenylisoseramine trihydrate N-tert-butyl ester 13-ester 5β-20-epoxy-1,2α,4,7β,10β,13α-hexahydroxytaxane-11-ene-9-one 4-acetate 2-benzene A pharmaceutically acceptable composition of formate, and one or more diluents, vehicles and/or excipients. Contains (2R,3S)-N-carboxy-3-phenylisoserine N-third butyl ester 13-ester 5β-20-epoxy-1,2α,4,7β,10β,13α- An example of a pharmaceutical composition of hexahydroxytaxel-11-en-9-one 4-acetate 2-benzoate is TAXOTERE ^® (RPR) injection concentrate. TAXOTERE ^® contains (2R,3S)-N-carboxy-3-phenylisoserine trihydrate N-third butyl ester 13-ester 5β-20-epoxy-1,2α,4 as the active ingredient, 7β,10β,13α-hexahydroxytaxel-11-en-9-one 4-acetate 2-benzoate, which is also called docetaxel for injection. TAXOTERE ^® is supplied in the form of a clear yellow to light brown-yellow viscous non-pyrogenic sterile solution. Each vial contains 20 mg (0.5 mL) or 80 mg (2 mL) docetaxel (anhydrous). Each milliliter contains 40 mg docetaxel (anhydrous) and 1040 mg polysorbate 80. TAXOTERE injection concentrate needs to be diluted with diluent before being added to the infusion bag. The diluent for TAXOTERE contains 13% ethanol/water for injection and is supplied in a vial.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (Irinotecan).

As known in the art, the irinotecan CAS registration number is 97682-44-5; the test formula is C ₃₃ H ₃₈ N ₄ O ₆ ; and the gram molecular weight is 586.68; and it is described in Japan published on January 31, 1985 Patent publication JP60019790, which is incorporated herein by reference in its entirety and for all purposes. The term "(4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-bi-sideoxy-3,4,12,14 -Tetrahydro-1 H -piperano[3',4':6,7]indolozino[1,2- b ]quinolin-9-yl ester" (unless otherwise specified) covers Solutes (including hydrates) and polycrystalline forms of (4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-di Pendantoxy-3,4,12,14-tetrahydro-1 H -piperano[3',4':6,7]indolozino[1,2- b ]quinolin-9-yl ester Or its salt. (4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-bi- pendantoxy-3,4,12,14-tetra Hydrogen- 1H -piperano[3',4':6,7]indazolino[1,2- b ]quinolin-9-yl ester pharmaceutical composition includes: all containing (4 S )- [1,4'-Dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-bi- pendantoxy-3,4,12,14-tetrahydro-1 H- Piperono[3',4':6,7]indolozino[1,2- b ]quinolin-9-yl ester, and one or more diluents, vehicles and/or excipients Academically acceptable composition. Contains (4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-bi-sideoxy-3,4,12,14- An example of a pharmaceutical composition of tetrahydro- 1H -piperano[3',4':6,7]indazolino[1,2- b ]quinolin-9-yl ester is CAMPTOSAR ^® (Pfizer Inc.). CAMPTOSAR ^® is provided in the form of a sterile, light yellow clear aqueous solution. Each milliliter of solution contains 20 mg of irinotecan hydrochloride (calculated as trihydrate salt), 45 mg of sorbitol NF and 0.9 mg of USP lactate. The pH of the solution is adjusted to 3.5 (range 3.0 to 3.8) by sodium hydroxide or hydrochloric acid. CAMPTOSAR ^{® is} intended to be diluted with 5% Dextrose Injection USP (D5W) or 0.9% Sodium Chloride Injection USP before intravenous infusion. The preferred diluent is 5% dextrose injection USP. Contains (4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-bi-sideoxy-3,4,12,14- Another example of a pharmaceutical composition of tetrahydro- 1H -piperano[3',4':6,7]indazolino[1,2- b ]quinolin-9-yl ester is ONIVYDE™ ( Merrimack Pharmaceuticals, Inc.). ONIVYDE is a sterile white to slightly yellow opaque isotonic lipid dispersion. Each 10 mL single-dose vial contains 43 mg of irinotecan free base at a concentration of 4.3 mg/mL. The liposome is a single-layer lipid bilayer vesicle with a diameter of about 110 nm, which encapsulates the aqueous solution space containing irinotecan (such as sucrose octasulfate) in gel or precipitated state. The vesicles are composed of 6.81 mg/mL 1,2-distearoyl-sn-glycerol-3-phosphate choline (DSPC), 2.22 mg/mL cholesterol, and 0.12 mg/mL methoxy-terminated polyethylene glycol (MW 2000)-Distearyl phosphatidylethanolamine (MPEG-2000-DSPE). Each milliliter also contains 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid (HEPES) in the form of 4.05 mg/mL buffer and isotonic reagent in the form of 8.42 mg/mL Of sodium chloride.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and inducing apoptosis Anticancer agent.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and selected from Navito Anti-cancer agent of gram, venetoc and compound (I) for inducing apoptosis.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and inducing apoptosis Anticancer agent (Navitac).

As known in the art, Navitok's CAS registration number is 923564-51-6; the test formula is C ₄₇ H ₅₅ ClF ₃ N ₅ O ₆ S ₃ ; and the gram molecular weight is 974.61; and it was described in June 2008 U.S. Patent No. 7,390,799, granted on the 24th, is incorporated herein by reference in its entirety and for all purposes. The term "4-{4-[(4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl] Piperazin-1-yl}- N -[4-{[(2 R )-4-(morpholin-4-yl)-1-(phenylthio)but-2-yl]amino}-3 -(Trifluoromethanesulfonyl)benzene-1-sulfonyl]benzamide” (unless otherwise specified) covers solvates (including hydrates) and polymorphic forms 4-{4- [(4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl} -N -[4-{[(2 R )-4-(morpholin-4-yl)-1-(phenylthio)but-2-yl]amino}-3-(trifluoromethanesulfonamide Group) benzene-1-sulfonyl] benzamide or salt thereof. 4-{4-[(4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazine -1-yl}- N -[4-{[(2 R )-4-(morpholin-4-yl)-1-(phenylthio)but-2-yl]amino}-3-( The pharmaceutical composition of trifluoromethanesulfonyl)benzene-1-sulfonyl]benzamide includes: all containing 4-{4-[(4'-chloro-4,4-dimethyl-3,4 ,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}- N -[4-{[(2 R )-4-(morpholine -4-yl)-1-(phenylthio)but-2-yl]amino]-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl]benzamide, and one or A pharmaceutically acceptable composition of various diluents, vehicles and/or excipients.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and inducing apoptosis Anticancer agent (Venetoc).

As is known in the art, Vinetoc's CAS registration number is 1257044-40-8; the test formula is C ₄₅ H ₅₀ ClN ₇ O ₇ S; and the gram molecular weight is 868.44; and the description was published on December 2, 2010 PCT Patent Application Publication WO 2010/138588, which is incorporated by reference in its entirety and for all purposes. The term "4-{4-[(4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl] Piperazin-1-yl} -N- (3-nitro-4-{[(oxetan-4-yl)methyl]amino}benzene-1-sulfonyl)-2-[( 1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxy]benzamide” (unless otherwise specified) covers solvates (including hydrates) and polymorphic forms 4 -{4-[(4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazine- 1-yl}- N -(3-nitro-4-{[(oxetan-4-yl)methyl]amino}benzene-1-sulfonyl)-2-[(1 H- Pyrrolo[2,3- b ]pyridin-5-yl)oxy]benzamide or its salt. 4-{4-[(4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazine -1-yl} -N- (3-nitro-4-{[(oxetan-4-yl)methyl]amino]benzene-1-sulfonyl)-2-[(1 H -Pyrrolo[2,3- b ]pyridin-5-yl)oxy]benzylamide pharmaceutical composition includes: all containing 4-{4-[(4'-chloro-5,5-dimethyl -3,4,5,6-tetrahydro[1,1'-biphenyl]-2-yl)methyl]piperazin-1-yl}- N -(3-nitro-4-{[( Oxetan-4-yl)methyl]amino]benzene-1-sulfonyl)-2-[(1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxy] A pharmaceutically acceptable composition of benzamide and one or more diluents, vehicles and/or excipients. The venetoc lozenges for oral administration are supplied in the form of light yellow or beige tablets, containing 10, 50, and 100 mg venetoc as active ingredients. Each lozenge also contains the following inactive ingredients: copovidone, colloidal silica, polysorbate 80, sodium stearyl fumarate and calcium hydrogen phosphate. In addition, 10 mg and 100 mg coated tablets include the following: yellow iron oxide, polyvinyl alcohol, polyethylene glycol, talc, and titanium dioxide. 50 mg coated tablets also include the following: yellow iron oxide, red iron oxide, black iron oxide, polyvinyl alcohol, talc, polyethylene glycol, and titanium dioxide.

In one embodiment, the present invention relates to a method for treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual an effective amount of TRAIL receptor agonist ABBV-621 and an anti-cancer cell that induces apoptosis Agent (compound (I)).

As known in the art, the CAS registration number of compound (I) is 1430845-59-2; the test formula is C ₄₃ H ₄₉ N ₇ O ₄ S; and the gram molecular weight is 759.96; and it was described on April 18, 2013 The published PCT Patent Application Publication WO 2013/055897 is incorporated herein by reference in its entirety and for all purposes. The term used "6-{8-[(1,3-benzothiazol-2-yl)aminecarboxamide]-3,4-dihydroisoquinoline-2(1 H )-yl}-3 -[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]adamantan-1-yl}methyl)-5-methyl-1 H -pyrazole "-4-yl]pyridine-2-carboxylic acid" (unless otherwise specified) covers solvates (including hydrates) and polymorphic forms of 6-{8-[(1,3-benzothiazol-2-yl )Aminecarboxamide]-3,4-dihydroisoquinoline-2(1 H )-yl}-3-[1-({3,5-dimethyl-7-[2-(methylamine Group) ethoxy]adamantan-1-yl}methyl)-5-methyl-1 H -pyrazol-4-yl]pyridine-2-carboxylic acid or a salt thereof. 6-{8-[(1,3-benzothiazol-2-yl)aminecarboxamide]-3,4-dihydroisoquinoline-2(1 H )-yl}-3-[1-( {3,5-Dimethyl-7-[2-(methylamino)ethoxy]adamantan-1-yl}methyl)-5-methyl- 1H -pyrazol-4-yl] The pharmaceutical composition of pyridine-2-carboxylic acid includes: all containing 6-{8-[(1,3-benzothiazol-2-yl)aminomethanyl]-3,4-dihydroisoquinoline-2( 1 H )-yl}-3-[1-({3,5-dimethyl-7-[2-(methylamino)ethoxy]adamantan-1-yl}methyl)-5- A pharmaceutically acceptable composition of methyl- 1H -pyrazol-4-yl]pyridine-2-carboxylic acid and one or more diluents, vehicles and/or excipients.

Unless otherwise defined herein, the scientific and technical terms used herein have the meanings generally understood by those who are familiar with the technology. If any potential divergence occurs, the definitions provided in this article take precedence over any dictionary or foreign definitions. Unless otherwise required by circumstances, singular terms shall include pluralities and plural terms shall include the singular. Unless otherwise stated, the use of "or" means "and/or". The use of the term "including" and other forms (such as "includes" and "included" is not limiting. Any ranges described herein should be understood to include the endpoints and all values between the endpoints.

Some of the headings used in this article are for organizational purposes only and should not be construed as limiting the subject matter described. All documents or parts of documents cited in this application (including but not limited to patents, patent applications, articles, books, and papers) are hereby expressly incorporated by reference in their entirety for any purpose. To the extent that documents incorporated by reference conflict with the disclosure contained in this specification, this specification will replace any opposing material.

In general, the nomenclature described in this document in conjunction with cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization is a well-known and commonly used nomenclature in the art law. The nomenclature used in conjunction with analytical chemistry, synthetic organic chemistry, and medicinal chemistry described herein is well known and commonly used in this technology, unless otherwise specified.

The selection terms are defined below to make the present invention easier to understand. definition

The term "Treat/treating/treatment" refers to a method of relieving or eliminating disease and/or its accompanying symptoms.

The term "individual" is defined herein to include animals, such as mammals, including but not limited to primates (eg, humans). In the preferred embodiment, the individual is a human.

The terms "patient" and "individual" are used interchangeably herein.

The term "biological activity" refers to any one or more biological characteristics of a molecule (naturally occurring as found in vivo or provided or achieved by recombinant methods). Biological properties include, but are not limited to, inhibition of tumor angiogenesis, inhibition of tumor initiation/cancer stem cell maintenance, and inhibition of tumor cell chemoresistance.

The term "effective amount" refers to an amount sufficient to induce a desired biological, pharmacological, or therapeutic result in an individual. A therapeutically effective amount means an amount of TRAIL receptor agonist protein and anticancer agent sufficient to treat or prevent pancreatic cancer at a reasonable benefit/risk ratio applicable to any medical treatment. However, it should be understood that the amount of the composition of the present invention will be determined by the attending physician within the scope of reasonable medical judgment.

"Specificity" refers to the ability of a binding protein to selectively bind to an antigen or receptor.

The term "efficacy" refers to the ability of the binding protein to achieve the desired effect and is a measure of its therapeutic efficacy. Methods known to those skilled in the art can be used to assess performance.

The term "biological function" refers to the specific in vitro or in vivo action of the binding protein. Binding proteins can target several classes of antigens and achieve the desired therapeutic outcome through multiple mechanisms of action. Binding proteins can target soluble proteins, cell surface antigens, and/or extracellular protein deposits. Binding proteins can promote, antagonize or neutralize the activity of their targets. The binding protein can help clear the target to which it binds, or can produce cytotoxicity when bound to the cell. Parts of two or more antibodies can be combined into a multivalent form to achieve more than one function in a single binding protein molecule. In vitro analysis and in vivo models for assessing biological functions are known to those skilled in the art.

A "stable" binding protein is one in which the binding protein substantially maintains its physical stability, chemical stability, and/or biological activity when stored. Multivalent binding proteins that are stable in vitro at various temperatures for an extended period of time are desirable. Methods for stabilizing binding proteins and assessing their stability at various temperatures are known to those skilled in the art.

The term "solubility" refers to the ability of a protein to remain dispersed in an aqueous solution. The solubility of a protein in an aqueous formulation depends on the proper distribution of hydrophobic and hydrophilic amino acid residues, and therefore the solubility can be related to the production of properly folded proteins. Those skilled in the art will be able to detect the increase or decrease in the solubility of the binding protein using conventional HPLC techniques and methods known to those skilled in the art.

"Control" refers to a composition that does not contain an analyte ("negative control") or does contain an analyte ("positive control"). The positive control may contain a known concentration of analyte. "Control", "positive control" and "calibrator" are used interchangeably herein to refer to a composition containing a known concentration of analyte. "Positive controls" can be used to establish analytical performance characteristics and are an indication of the integrity of reagents (eg, analytes).

The term "Fc region" defines the C-terminal region of an immunoglobulin heavy chain, which can be produced by papain digestion of intact antibodies. The Fc region may be a native sequence Fc region or a variant Fc region. The Fc region of an immunoglobulin generally contains two constant domains, namely the CH2 domain and the CH3 domain, and optionally the CH4 domain. Substitution of amino acid residues in the Fc portion to modify antibody effector functions is known in the art (for example, US Patent Nos. 5,648,260 and 5,624,821). The Fc region mediates several important effector functions, such as cytokine induction, antibody-dependent cell-mediated cytotoxicity (ADCC), phagocytosis, complement-dependent cytotoxicity (CDC), and the half-life of antibodies and antigen-antibody complexes/ clearance rate. In some cases, these effector functions are desirable in terms of therapeutic immunoglobulins, but in other cases, they may be unnecessary or even harmful depending on the treatment target.

The term "linking group" means an amino acid residue or a polypeptide comprising two or more amino acid residues joined by peptide bonds used to connect two polypeptides. Examples of such linking polypeptides are well known in the art (see, for example, Holliger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448; Poljak et al. (1994) Structure 2: 1121-1123 ).

"Navitoc" is 4-{4-[(4'-chloro-4,4-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2- Yl)methyl]piperazin-1-yl}- N -[4-{[(2 R )-4-(morpholin-4-yl)-1-(phenylthio)but-2-yl] Amino}-3-(trifluoromethanesulfonyl)benzene-1-sulfonyl]benzamide.

"Venetoc" is 4-{4-[(4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro[1,1'-biphenyl]-2- Yl)methyl]piperazin-1-yl}- N -(3-nitro-4-{[(oxetan-4-yl)methyl]amino}benzene-1-sulfonyl) -2-[(1 H -pyrrolo[2,3- b ]pyridin-5-yl)oxy]benzamide.

"Irinotecan" is (4 S )-[1,4'-dipiperidine]-1'-carboxylic acid 4,11-diethyl-4-hydroxy-3,14-dipentoxy-3,4 ,12,14-Tetrahydro- 1H -piperano[3',4':6,7] indozino[1,2- b ]quinolin-9-yl ester.

"SN-38" is (4 S )-4,11-diethyl-4,9-dihydroxy-1 H -piperano[3',4':6,7]indozino[1, 2- b ] Quinoline-3,14(4 H ,12 H )-dione. SN-38 is the active metabolite of irinotecan and is formed by the hydrolysis of irinotecan by decarboxylose esterase mainly in the liver. In vitro analysis requires the use of SN-38 to study the efficacy of irinotecan.

"Gemcitabine" is 4-amino-1-[(2 R ,4 R ,5 R )-3,3-difluoro-4-hydroxy-5-(hydroxymethyl)oxetan-2-yl ] Pyrimidine-2(1 H )-one.

"Erotinib" is N- (3-ethynylphenyl)-6,7-bis(2-methoxyethoxy)quinazolin-4-amine.

``Pacific paclitaxel'' is (2a R ,4 S ,4a S ,6 R ,9 S ,11 S ,12 S ,12a R ,12b S )-6,12b-bis(acetyloxy)-9-{ [(2 R ,3 S )-3-benzylamino-2-hydroxy-3-phenylpropionyl]oxy}-4,11-dihydroxy-4a,8,13,13-tetramethyl Yl-5-pentoxy-2a,3,4,4a,5,6,9,10,11,12,12a,12b-dodecane-1 H -7,11-methylenecyclodecane[3 , 4] Benzo[1,2- b ]oxetane-12-yl benzoate.

"Docetaxel" is (2 R ,3 S ) -N -carboxy-3-phenylisoserine trihydrate, N -third butyl ester, 13-ester and 5β-20-epoxy-1, 2α,4,7β,10β,13α-hexahydroxytaxel-11-en-9-one 4-acetate 2-benzoate.

"5-fluorouracil" is 5-fluoropyrimidine-2,4(1 H ,3 H )-dione.

化合物 (I) "Compound (I)" is 6-{8-[(1,3-benzothiazol-2-yl)aminecarboxamide]-3,4-dihydroisoquinolin-2(1 H )-yl} -3-[1-({3,5-Dimethyl-7-[2-(methylamino)ethoxy]adamantan-1-yl}methyl)-5-methyl-1 H- Pyrazol-4-yl]pyridine-2-carboxylic acid, an inhibitor of Bcl-xL.

Compound (I)

All references cited in this article (including publications, patent applications, and patents) are incorporated by reference to the same extent as if each reference individually and specifically indicated to be incorporated by reference and in full Explained in this article.

Unless otherwise indicated herein or clearly contradicted by context, the terms "a (an)" and "the" and similar indications are used in the context of describing the invention (especially above and below the scope of the patent application below) Objects should be interpreted as covering both singular and plural. Unless otherwise indicated, the terms "including", "having", "including" and "including" should be understood as open-ended terms (ie, meaning "including (but not limited to)"). Unless otherwise indicated herein, the recitation of numerical ranges herein is intended only to serve as a shorthand method of referring individually to individual values falling within the range, and each individual value is incorporated into this specification as if it were individually recited herein. Unless otherwise indicated herein or clearly contradicted by context, all methods described herein can be performed in any suitable order. Unless otherwise claimed, the use of any and all examples or illustrative language (eg, "such as") provided herein is merely intended to better clarify the invention and not to limit the scope of the invention. The language in this specification should not be understood as implying that any unclaimed elements are essential for practicing the invention.

序列表之簡要說明 Preferred embodiments of the invention are described herein, including the best mode known to the inventors for carrying out the invention. After reading the foregoing description, changes to their preferred embodiments will become apparent to those of ordinary skill in the art. The inventor expects those skilled in the art to adopt such changes as appropriate, and the inventor intends to implement the invention in other ways than those specifically described herein. Therefore, if permitted by applicable law, the present invention includes all modifications and equivalents of the subject matter described in the scope of the patent application accompanying this document. Furthermore, unless otherwise indicated herein or otherwise clearly contradicted by context, the invention encompasses any combination of the above elements in all possible variations thereof. Table 1 provides the amino acid sequence of the receptor agonist protein monomer for TRAIL in ABBV-621.

Brief description of the sequence table

The sequence listing named "ABV12441WO01_ST25" is incorporated herein by reference in its entirety. The sequence listing contains SEQ ID NO: 1, including the amino acid sequence disclosed herein. Therefore, the sequence listing has been submitted via EF in ASCII text format. The sequence table was first created on March 11, 2019 and is 7 KB in size. EXAMPLES Example 1 : Treatment of pancreatic cancer cell line materials and methods with ABBV-621 in combination with standard care agents and anti-cancer agents that induce apoptosis

Cell culture : All cell lines are derived from the American Type Culture Collection (ATCC) or German Microbial Culture Collection GmbH (DSMZ) and are obtained through the AbbVie Cancer Core Cell Line facility, which is repeated with short tandem (STR ) Atlas analysis (Table 2) to verify the identity of each cell line. All reagents used for cell culture are listed in Table 3. The frozen cells were thawed at 37°C and transferred to a 15 mL conical tube containing 10 mL of the specified medium (depending on the cell line). The cells were collected by centrifugation at 500×g for 3 minutes. Remove the medium; resuspend the cells in 5 mL medium and transfer to a Corning T75 flask containing 10 mL medium. Incubate the cells in a moisture-containing incubator at 37°C/5% CO ₂ until approximately 75% confluence. At this time, the medium is removed, and the cells are separated from the culture dish in 3 mL trypsin-EDTA and transferred To two T175 flasks containing 30 mL of appropriate medium. The cells were incubated in a moisture-containing incubator at 37°C/5% CO ₂ until about 75% confluence, separated from the culture plate using 5 mL trypsin-EDTA, and transferred to a 15 mL cone containing 7 mL medium tube. The cells were then counted on Beckman Coulter Vi-CELL™, and 3.6×10 ⁶ viable cells were diluted to 59 mL of medium in a 100 mL reservoir. Spread the cells at 25 µL/well in a 384-well tissue culture dish (1500 cells/well). The culture plate was centrifuged at 100×g in a floating bucket rotor for 1 minute to ensure that the cells were at the bottom, and then incubated in a moisture-containing incubator at 37°C/5% CO ₂ for 18 to 24 hours. abbreviation:

表3：用於細胞培養之試劑

DMEM: Dulbecco's Modified Eagle's Medium; FBS: Fetal Bovine Serum; IMDM: DMEM's Modified Eagle's; IMEM: Modified Minimum Essential Medium; MEM: Minimum Essential Medium: Pen Strep: Penicillin Streptomycin; RPMI : Culture medium of Losway Parker Memorial Institute; and Trypsin-EDTA: Trypsin-ethylenediaminetetraacetic acid. Table 2: Pancreatic cancer cell lines used for in vitro cytotoxicity analysis

Table 3: Reagents used for cell culture

Dose response : The maximum concentration of the compounds used and each of them is described in Table 4. Manufacture half-logarithms of ABBV-621 (10 dilutions and medium controls) and compounds (9 dilutions and medium controls) starting at a concentration seven times the maximum concentration indicated in Table 4 in cell line-specific medium Serial dilution. Add test compound at 5 µL/well. After compound addition, 5 µL/well of ABBV-621 was added. The final volume in each well is 35 µL and each compound is 5 µL. The culture plate was centrifuged at 100×g for 1 minute to ensure that the compound was in the medium, and then incubated at 37°C/5% CO ₂ . Table 4: Compounds used to treat pancreatic cancer cell lines and the highest doses of each

Viability determination : After approximately 24 hours of incubation, add 25 µL CellTiter Glo ^® reagent (Promega, Madison WI, catalog number G7571) to all wells. The plates were incubated at room temperature for 15-45 minutes, and analyzed on emitting Biotek ^® Synergy ™ Neo2 disc reader.

Calculation : Synergy was determined based on the Bliss independence model (Borisy et al., Proc Natl Acad Sci USA 2003; 100: 7977-7982). Using GraphPad Prism ^® , a curve fitting algorithm based on logarithm (inhibitor) and normalized response was used, and the IC ₅₀ value was determined using a variable slope. The samples were normalized to the average of the two media controls on each plate. By acquiring IC calculated at each combination of values of ₅₀ and this equivalent single agent ABBV-621 IC ₅₀ determined by comparing IC ₅₀ fold. If the single-agent therapy ABBV-621 IC _50> 20 nM, ABBV-621 treatment of the IC ₅₀ was set to be 20 nM to avoid extrapolation data. If all combinations produce an ABBV-621 IC ₅₀ value of >20 nM, the multiple IC ₅₀ is set to zero. Result: Single agent activity :

The single agent activity of ABBV-621 as defined by IC _{50 of} <20 nM was observed in 9 of the 14 cell lines. Among the 9 sensitive cell lines, 6 cell lines exhibited subnemol sensitivity. The IC ₅₀ of ABBV-621 and the% growth inhibition of ABBV-621 alone or in combination with each test compound are shown in Table 5.

表6：24小時培育後之所有測試化合物的單藥劑活性(µM)。

組合活性 ： The single agent activity of each test compound during 24 hours of treatment was also determined (Table 6). If the IC _{50 is} less than the maximum concentration used, the compound is considered to have single agent activity. Compound (I) and SN-38 exhibited single agent activity in 5 or 6 pancreatic cancer cell lines out of 14 pancreatic cancer cell lines, respectively. Navitork demonstrated single agent activity in 3 of the 14 cell lines. 5-fluorouracil (5FU) and erlotinib exhibited weak activity in cell lines DAN- and BXPC3, respectively. No single agent activity of gemcitabine, venetoc or paclitaxel was observed in any cell lines. 5:24 ABBV-621 of Table IC ₅₀ hours of incubation and after ABBV-621 alone or in combination with the growth of the percent inhibition of each test compound

Table 6: Single-agent activity (µM) of all test compounds after 24 hours of incubation.

Combination activity :

The percentage values of growth inhibition resulting from the combination of ABBV-621 treatment and each test compound are listed in Table 5. Although the change in growth inhibition is suitable for understanding the parameter of combined activity, it is difficult to use this parameter alone to assess synergy because many cell lines have extremely high growth inhibition (>90%) when only ABBV-621 is used . In this case, the change in growth inhibition may be minimal, but the amount of ABBV-621 required to achieve growth inhibition may be very different when used in combination. Thus, based on the maximum score BLISS each combination of the IC / cell lines of the ₅₀ fold change is defined synergistic combination. The cutoff value was assigned to both the BLISS score and the IC _50- fold change (Table 7). Assign a point according to the cutoff value, and add the final objective judgment that proposes strong synergy (Table 8), which is defined as strong synergy, weak synergy, and no synergy. Using these guidelines, of the 112 total combinations/cell lines tested, 59 tests showed weak or strong synergy, while 53 tests did not show significant synergy. Table 9 shows the final evaluation of the synergy of each cell line and combination.

In standard care agents, the combination of ABBV-621 and SN-38 exhibited strong synergy in 11 of 14 cell lines and weak synergy in 2 of the remaining cell lines. Only one cell line (HPAC) used objective criteria to show no synergy with SN-38. This particular cell line is sensitive to ABBV-621 as a single agent. The representative dose-response curves of the combination of ABBV-621 and SN-38 in cell lines BXPC-3 and SU.86.86 are shown in Figures 7 and 8, respectively, which show weak and strong synergy, respectively.

The strong synergy of gemcitabine/ABBV-621 combination was observed in DAN-G and CFPAC-1, while the weak synergy was observed in AsPC-1 and Su.86.86. Among these cell lines, two cell lines (AsPC-1, DAN-G) are relatively resistant to ABBV-621 as a single agent, and sensitive to the other two (Su.86.86, CFPAC-1) . In addition, although according to the criterion of IC ₅₀ <maximum dose (1 µM) within 24 hours, no cell line is sensitive to gemcitabine, but the obvious line is three of the four cell lines (SU.86.86, DAN-G And CFPAC-1) some single agent activity appears. The representative dose-response curves of the combination of ABBV-621 and gemcitabine in cell lines SU.86.86 and DAN-G are shown in Figure 3 and Figure 4, respectively, which show weak and strong synergy.

Paclitaxel showed strong synergy with ABBV-621 in two cell lines (PA-TU-8688T and HPAF-II), and in three cell lines (DAN-G, MIA PaCa-2 and Panc-1) Shows a weak synergy. Interestingly, all these cell lines except PA-TU-8688T are relatively resistant to ABBV-621 as a single agent (IC ₅₀ >1 nM). In addition, although none of these cell lines are considered to be sensitive to paclitaxel (IC ₅₀ <1 µM in the 24-hour treatment), there are markers of single agent activity in several cell lines. The representative dose-response curves of the combination of ABBV-621 and paclitaxel in the cell lines Mia PaCa-2 and PA-Tu-8988T are shown in Figures 5 and 6, respectively, where weak synergy and strong synergy are shown separately .

The cell lines did not show strong synergy with ABBV-621 in combination with 5-fluorouracil or erlotinib, but a small number of cell lines showed weak synergy with either combination. For 5-fluorouracil, two cell lines (BXPC-3 and SU.86.86) exhibited single agent activity markers, while three cell lines (DAN-G) exhibited minimal single agent activity. In contrast, none of the four cell lines (BXPC-3, AsPC-1, Capan-2, and HPAF-II) exhibiting synergy with ABBV-621 and erlotinib exhibited Single agent activity sign.

For the anti-apoptotic agent, the combination of Navitok and ABBV-621 and the combination of Bcl-xL inhibitor (Compound (I)) and ABBV-621 showed synergy in 12 of 14 cell lines effect. Not surprisingly, the two agents exhibited a synergistic effect in 11 cell lines of the same cell line. Using objective criteria, PA-TU-8988T and PA-TU-8988S exhibited synergy with Navitork and showed no synergy with Bcl-xL inhibitor (Compound (I)). In contrast, the Bcl-xL inhibitor (Compound (I)) exhibited a weak synergy in HPAC, while Navitork did not. This cell line shows a single agent activity with a Bcl-xL inhibitor (Compound (I)), and the observed synergy seems to be driven by the single agent activity. The representative dose-response curves of the combination of ABBV-621 and Navitoc in the cell lines BXPC-3 and MIA PaCa-2 are shown in Figures 1 and 2, respectively, where strong synergy and weak synergy are shown separately.

表8：用於判定協同作用之客觀準則

表9：判定胰臟癌細胞株之活體外組合協同作用(最大BLISS/倍數IC₅₀ )。

結論 Of the specific cell lines tested, none of them showed strong synergy with the combination of ABBV-621 and Venetoc, as defined herein, but several cell lines did show weak synergy. Among the 5 cell lines that exhibited weak synergy with Venetoc, only the highest concentration (10 µM) exhibited a different dose response than the ABBV-621 control alone. The higher concentration of venetoc does indeed seem to be cytotoxic in multiple cell lines in general, so it is difficult to determine whether the observed synergistic effect is only the cytotoxic function from venetoc. Table 7: Cutoff value and cutoff point calculated by BLISS and multiple IC ₅₀

Table 8: Objective criteria used to determine synergy

Table 9: Judgment of in vitro combination synergy of pancreatic cancer cell lines (maximum BLISS/fold IC ₅₀ ).

in conclusion

Cytotoxicity analysis was performed on 14 pancreatic cancer cell lines using a combination of ABBV-621 and 6 standard care agents for pancreatic cancer and 3 anti-cancer agents that induce apoptosis. Surprisingly, substantial synergy with several tested combinations was observed in several cell lines. Obviously, SN-38, the active form of irinotecan, showed synergistic activity with ABBV-621 in 13 of the 14 cell lines tested. Both paclitaxel and gemcitabine also demonstrated strong synergy with ABBV-621 in several cell lines. Interesting systems should note that two of these agents provide at least two cell lines with substantially synergistic activity. The remaining standard care agents (5-fluorouracil and erlotinib) will not show substantial synergy in any cell line, but will show weak synergy in several cell lines.

Since ABBV-621 induces apoptosis via foreign and internal apoptosis pathways, the inventors aimed to evaluate the combination of ABBV-621 and agents that interfere with internal anti-apoptotic proteins. The apoptosis inducing agents tested include Navitork, which is an inhibitor of both Bcl-xL and Bcl-2; Venetoc, which is a specific inhibitor of Bcl-2; and Compound (I), that is Specific inhibitor of Bcl-xL. Among them, the agent that inhibits Bcl-xL alone (Compound (I)) or the agent that inhibits the combination of Bcl-xL and Bcl-2 (Navitock) showed strong in 9 of 14 cell lines Synergy. It will show that when the internal pathway is activated by ABBV-621, many pancreatic cancer cell lines are partially dependent on Bcl-xL.

After 24 hours of incubation using the combined treatment, the combined activity was tested. For most test compounds, very little single agent activity was observed under these conditions. Since the activity of several of these compounds (5-fluorouracil, gemcitabine, paclitaxel, and SN-38) is cell cycle-dependent, it will be possible to observe other single-agent activities with longer incubation. Therefore, it is possible that other synergistic or additive effects can be observed in the case of pre-incubating the cells with the test compound before adding ABBV-621 or in the case of continuing the combined treatment for a longer period of time. The fact that the synergistic effects of these compounds are observed at 24 hours will indicate that cell cycle-independent changes occur, which promote apoptosis or block the inhibition of apoptosis after being treated with ABBV-621 (ie, sensitive cells Agents for apoptosis). Example 2: composition by ABBV-621 and docetaxel or gemcitabine the xenograft from the human patient of pancreatic cancer tumor growth inhibition in mice and breeding materials and methods

Conduct xenotransplantation (PDX) studies derived from patients with pancreatic cancer. Female nude mice were obtained from Harlan Laboratories (Indianapolis, IN). The weight on arrival is 18 to 20 g. Food and water are available at will. Before the start of the experiment, the mice were acclimated to the animal facility for at least one week. Animals were tested in the bright phase of 12 hours light: 12 hours dark time period (lights turned on at 06:00 hours). In a facility certified by the Laboratory Animal Care Assessment and Certification Association, all experiments were conducted in accordance with the laboratory animal care and use guidelines of the AbbVie Institute Animal Care and Use Committee and the National Institutes of Health. Efficacy parameter

The parameters of treatment response amplitude (tumor growth inhibition, TGI) and persistence (tumor growth retardation, TGD) are used to refer to drug efficacy. TGI indicates the difference between the average tumor volume of the drug-treated group and the average tumor volume of the control, and is expressed as a percentage of the average volume of the control group. The TGI value was determined at the last time point before removing the first animal from a group because the value reached its tumor burden limit. TGD indicates the difference between the median time for the drug-treated group to reach the defined tumor volume (mm ³ ) and the median time for the vehicle-treated control group to reach the same volume. The difference is expressed as the percentage of the median time for the control group to reach the specified tumor volume. Compounds and formulations

ABBV-621 is a TRAIL receptor agonist composed of human immunoglobulin G1 (IgG1)-Fc fused with natural single-chain TRAIL receptor binding domain (RBD) monomers. These monomers are glycosylated The linkers are covalently linked to produce a dimer consisting of two sets of trimeric RBD. The raw material ABBV-621 was kept in a solution (pH 7.2) of 20 mm Tris, 70 g/L sucrose, 1.0 g/L polysorbate 80 and stored at 80°C. Dilute the appropriate amount of raw materials in phosphate buffered saline before administration. Docetaxel and gemcitabine were obtained and formulated in phosphate buffered saline at pH 7.2. statistics

The Student's t-test was used to assess the statistically significant difference in the average tumor volume between compound-treated and vehicle-treated. Statistical analysis of survival was performed using log rank and Wilkerson comparison using JMP software (SAS). Generation of tumor-bearing mice and measurement of tumor volume of subcutaneous flank tumors

Fresh tumor tissue is cut into fragments and implanted. Unless otherwise specified, the size of all tumors matches approximately 200 to 250 mm ³ . Therapy starts within 24 hours after tumor size matches. At the beginning of therapy, the mice were weighed at approximately 22 grams. The tumor volume is estimated two to three times a week. The measurement values of the length (L) and width (W) of the tumor are obtained through an electronic caliper, and the volume is calculated according to the following equation: V = L × W ² /2. When the tumor volume reached at most 2,000 mm ³ or skin ulcers appeared, the mice were euthanized. Experimental conditions

CTG-0282, CTG-0289, CTG-0314 or CTG-0492 tumor-bearing mice are used to provide tumor tissue for reproduction into the study mice. The tumor was excised and cut into 2 mm × 2 mm fragments. Use a trocar to place the fragment under the skin. Calculate the data and save it with Excel (Microsoft Office). The details of treatment and administration routes are described in Table 10. Results and discussion

The efficacy of the combination of ABBV-621 with docetaxel and gemcitabine was measured in a xenograft model derived from CTG-0282 pancreatic cancer patients. The medicament was administered as shown in Table 10. ABBV-621, which was administered with 3 mg/kg Q7D×3 as monotherapy, was effective, with TGI of 64% (p<0.05). Compared with ABBV-621 alone, docetaxel at 50 mg/kg QD×1 as monotherapy or in combination with ABBV-621 administration was effective in this model, as shown in FIG. 9. Except for CTG-0282 mice treated with a combination of ABBV-621 and gemcitabine, all treatment groups were well tolerated and no significant weight loss was observed. For the mice administered the combination of ABBV-621 and gemcitabine in the CTG-0282 model, 4 of the 5 mice had died at a time point early in the study. Therefore, only the surviving mice of one of the specific treatment groups are plotted in FIG. 9.

The efficacy of the combination of ABBV-621 with docetaxel and gemcitabine was measured in a xenograft model derived from CTG-0289 pancreatic cancer patients. The medicament was administered as shown in Table 10. ABBV-621 administered with 3 mg/kg Q7D×3 as monotherapy was effective, with a TGI of 41% (NS). Docetaxel administered with 50 mg/kg QD×1 as monotherapy was effective, with a TGI of 91% (p<0.005). As shown in Figure 10, the combination of ABBV-621 and docetaxel showed an increased efficacy compared to docetaxel alone by TGI (66%; p<0.01). Gemcitabine administered at 80 mg/kg Q3D×4 as monotherapy is effective, with a TGI of 59% (NS). As shown in Figure 10, the combination of ABBV-621 and gemcitabine showed an increased efficacy compared to gemcitabine alone by TGI (46%; p<0.05). All treatment groups were well tolerated and no significant weight loss was observed.

The efficacy of the combination of ABBV-621 with docetaxel and gemcitabine was measured in a xenograft model derived from CTG-0314 pancreatic cancer patients. The medicament was administered as shown in Table 10. ABBV-621 administered with 3 mg/kg Q7D×3 as monotherapy is not effective in this model. Docetaxel administered with 50 mg/kg QD×1 as monotherapy was effective, with TGI of 33% (p<0.05). As shown in FIG. 11, the combination of ABBV-621 and docetaxel showed synergistic efficacy and lack of TGI (74%; p<0.05) compared to docetaxel alone. The lack of ABBV-621 observed in this model Effect. Gemcitabine administered with 80 mg/kg Q3D×4 as monotherapy is not effective in this model. Compared with gemcitabine alone, the combination of ABBV-621 and gemcitabine did not show increased efficacy. All treatment groups were well tolerated and no significant weight loss was observed.

The efficacy of the combination of ABBV-621 with docetaxel and gemcitabine was measured in a xenograft model derived from CTG-0492 pancreatic cancer patients. The medicament was administered as shown in Table 10. ABBV-621 administered with 3 mg/kg Q7D×3 as monotherapy is not effective in this model. Docetaxel administered at 50 mg/kg QD×1 as monotherapy is effective, with a TGI of 42% (NS). As shown in FIG. 12, the combination of ABBV-621 and docetaxel showed a trend of increased efficacy when compared with docetaxel alone by TGI (39%; NS) and TGD (16%; NS). Gemcitabine administered with 80 mg/kg Q3D×4 as monotherapy is not effective in this model. As shown in Figure 12, the combination of ABBV-621 and gemcitabine showed a synergistic trend of increased efficacy when compared to gemcitabine alone through TGI (53%; NS) and TGD (29%; NS), in view of ABBV- The fact that neither 621 nor gemcitabine is effective in this model is further emphasized. All treatment groups were well tolerated and no significant weight loss was observed. Table 10: Research design

Figure 1 shows the dose-response curve of the combined anti-BXPC-3 pancreatic cancer cell line of ABBV-621 and Navitok. The maximum BLISS score is 46 and the maximum multiple IC ₅₀ is 131. It represents a strong collaborative response.

Figure 2 shows the dose-response curve of the combined anti-MIA PaCa-2 pancreatic cancer cell line of ABBV-621 and Navitok. The maximum BLISS score is 19 and the maximum multiple IC ₅₀ is 4. It does not represent a synergistic reaction.

Figure 3 shows the dose response curve of the combination of ABBV-621 and gemcitabine against the SU.86.86 pancreatic cancer cell line. The maximum BLISS score is 21 and the maximum multiple IC ₅₀ is 74. It represents a weak cooperative response.

Figure 4 shows the dose response curve of the combination of ABBV-621 and gemcitabine against DAN-G pancreatic cancer cell line. The maximum BLISS score is 51 and the maximum multiple IC ₅₀ is 351. It represents a strong collaborative response.

Figure 5 shows the dose response curve of the combination of ABBV-621 and paclitaxel against MIA PaCa-2 pancreatic cancer cell line. The maximum BLISS score is 27 and the maximum multiple IC ₅₀ is 25. It represents a weak cooperative response.

Figure 6 shows the dose response curve of the combination of ABBV-621 and paclitaxel against PA-TU-8988T pancreatic cancer cell line. The maximum BLISS score is 58 and the maximum multiple IC ₅₀ is 531. It represents a strong collaborative response.

Figure 7 shows the dose-response curve of the combination of ABBV-621 and SN-38 against the BXPC-3 pancreatic cancer cell line. The maximum BLISS score is 29 and the maximum multiple IC ₅₀ is 6290. It represents a weak cooperative response.

Figure 8 shows the dose response curve of the combination of ABBV-621 and SN-38 against the SU.86.86 pancreatic cancer cell line. The maximum BLISS score is 42 and the maximum multiple IC ₅₀ is 257. It represents a strong collaborative response.

Figure 9 shows xenotransplanted human pancreatic cancer tumors (derived from patients) with ABBV-621, gemcitabine or docetaxel as a single agent or with the combination of ABBV-621 and gemcitabine or docetaxel as described in Example 2 Xenograft model) CTG-0282 growth inhibition curve. As shown in the figure, ABBV-621 administered as monotherapy is effective. Compared to ABBV-621 alone, docetaxel administered as monotherapy or in combination with ABBV-621 is not effective in this model. As shown in the figure, gemcitabine or the combination of ABBV-621 and gemcitabine as monotherapy caused enhanced tumor growth inhibition.

Figure 10 shows xenograft human pancreatic cancer tumors (derived from patients) with ABBV-621, gemcitabine or docetaxel as a single agent or with the combination of ABBV-621 and gemcitabine or docetaxel as described in Example 2 Xenotransplantation model) CTG-0289 growth inhibition curve. As shown in Figure 10, the combination of ABBV-621 and docetaxel showed increased efficacy compared to docetaxel alone. Gemcitabine administered as a monotherapy is effective. As shown in Figure 10, the combination of ABBV-621 and gemcitabine showed increased efficacy compared to gemcitabine alone.

Figure 11 shows xenograft human pancreatic cancer tumors (derived from patients) with ABBV-621, gemcitabine or docetaxel as a single agent or with the combination of ABBV-621 and gemcitabine or docetaxel as described in Example 2 Xenograft model) CTG-0314 growth inhibition curve. As shown in FIG. 11, the combination of ABBV-621 and docetaxel demonstrated synergistic efficacy compared to docetaxel alone and lacked the efficacy observed for ABBV-621 in this model. Gemcitabine administered as monotherapy is not effective in this model. Compared with gemcitabine alone, the combination of ABBV-621 and gemcitabine did not show increased efficacy.

Figure 12 shows xenograft human pancreatic cancer tumors (derived from patients) with ABBV-621, gemcitabine or docetaxel as a single agent or with the combination of ABBV-621 and gemcitabine or docetaxel as described in Example 2 Xenograft model) CTG-0492 growth inhibition curve. As shown in Figure 12, ABBV-621 administered as monotherapy is not effective in this model. Docetaxel administered as monotherapy is effective. As shown in FIG. 12, the combination of ABBV-621 and docetaxel showed a trend of increased efficacy compared with docetaxel alone. Gemcitabine administered as monotherapy is not effective in this model. As shown in Figure 12, the combination of ABBV-621 and gemcitabine showed a synergistic trend of increased efficacy compared to gemcitabine alone, which was obtained in view of the fact that neither ABBV-621 nor gemcitabine alone were effective in this model Further emphasis.

Claims (20)

A method for treating pancreatic cancer in an individual in need thereof, comprising administering to the individual a combination of a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and one or more anti-pancreatic cancer agents. The method of claim 1, wherein the anti-pancreatic cancer agent is selected from the group consisting of gemcitabine, 5-fluorouracil, erlotinib, paclitaxel, and more Docetaxel and irinotecan. The method of claim 2, wherein the anti-pancreatic cancer agent is administered before, after, or at the same time as ABBV-621. The method of claim 2, wherein the anti-pancreatic cancer agent is gemcitabine. The method of claim 2, wherein the anti-pancreatic cancer agent is 5-fluorouracil. The method of claim 2, wherein the anti-pancreatic cancer agent is erlotinib. The method of claim 2, wherein the anti-pancreatic cancer agent is paclitaxel. The method of claim 2, wherein the anti-pancreatic cancer agent is irinotecan. The method of claim 1, wherein ABBV-621 is administered in combination with an anti-cancer agent that induces apoptosis, the anti-cancer agent that induces apoptosis is selected from the group consisting of: navitoclax, vitamin Netecolax and compound (I). The method of claim 9, wherein the anti-cancer agent that induces apoptosis is Navitok. The method of claim 9, wherein the anti-cancer agent that induces apoptosis is Venetoc. The method according to claim 9, wherein the anti-cancer agent that induces apoptosis is compound (I). The method of claim 2, further comprising administering albumin-bound paclitaxel (nab-paclitaxel). The method according to claim 2, wherein the paclitaxel is albumin-bound paclitaxel. The method of claim 2, wherein the anti-pancreatic cancer agent is docetaxel. A method of treating pancreatic cancer in an individual in need thereof, comprising administering to the individual a combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine). A method for treating pancreatic cancer in an individual in need thereof, comprising administering to the individual a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (gemcitabine and albumin-bound paclitaxel) Of the combination. A method for treating pancreatic cancer in an individual in need thereof, comprising administering to the individual a combination of a therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent, the anti-pancreatic cancer agent is selected Free from the group consisting of paclitaxel and albumin-bound paclitaxel. A method of treating pancreatic cancer in an individual in need thereof, which comprises administering to the individual a combination of a therapeutically effective amount of a combination of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (SN-38). A method for treating pancreatic cancer in an individual in need thereof, comprising administering to the individual a combination of a synergistically therapeutically effective amount of TRAIL receptor agonist ABBV-621 and an anti-pancreatic cancer agent (docetaxel).

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