Lyophilized compositions comprising (S)-2-((S)-2-acetylamino-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester for subcutaneous administration and uses thereof

Abstract

The present disclosure provides a lyophilisate comprising (S)-2-((S)-2-acetylamino-3-(1H-indol-3-yl)propanoylamino)-6-diazo-5-oxohexanoic acid isopropyl ester for subcutaneous administration to a subject, and pharmaceutical compositions, pharmaceutical formulations and uses thereof.

Description

Background of the Invention

[0002] 6-Diazo-5-oxo-L-ortholeucine (DON) is a glutamine antagonist that has shown promising activity in preclinical models for treating a variety of diseases, such as cancer. See, for example, Ahluwalia et al., Pharmac. The. 46:243-371 (1990). However, the clinical development of DON has been hampered by its dose-limiting toxicity in humans, particularly in the intestinal epithelium. See, for example, Rosenfeld and Roberts, Cancer Research 41:1324-1328 (1981) and Lynch et al., Am J Clin Oncol (CCT) 5:541-543 (1982). Administration of DON as a prodrug may help mitigate this toxicity.

[0003] US 10,336,778B2 discloses isopropyl (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoate (“Compound 1”) and other prodrugs for the treatment of cancer and other diseases. A pharmaceutical composition comprising Compound 1 is desired for subcutaneous administration to a subject in need of such treatment.

[0004] Invention Summary

[0005] In one aspect, this disclosure provides a lyophilized form comprising compound 1 for subcutaneous administration to a subject in need.

[0006] In another aspect, this disclosure provides a lyophilized product comprising compound 1 and a filler, such as mannitol.

[0007] In another aspect, this disclosure provides a lyophilized product comprising compound 1 and a cellulose-based suspension modifier, such as sodium carboxymethyl cellulose.

[0008] In another aspect, this disclosure provides a lyophilized product comprising compound 1 and a poloxamer-based suspension modifier, such as poloxamer 188.

[0009] In another aspect, this disclosure provides a lyophilized product comprising compound 1 and a lecithin-based suspension modifier, such as soy lecithin.

[0010] In another aspect, this disclosure provides a lyophilized product comprising compound 1 and a buffer, such as L-histidine.

[0011] In another aspect, this disclosure provides a pharmaceutical composition comprising a lyophilized material already suspended in a pharmaceutically acceptable excipient (e.g., a solvent, such as water), said lyophilized material comprising compound 1.

[0012] In another aspect, this disclosure provides a method for treating cancer in a subject with this need, the method comprising subcutaneously administering a therapeutically effective amount of a pharmaceutical composition comprising a lyophilized compound 1.

[0013] In another aspect, this disclosure provides a method for preparing a lyophilized product comprising compound 1.

[0014] In another aspect, this disclosure provides a method for preparing a pharmaceutical composition comprising a lyophilized compound 1.

[0015] In another aspect, this disclosure provides a kit comprising a lyophilized compound 1 packaged as a single unit dose in a tubular vial. Brief description of the attached diagram

[0017] Figure 1 The images show the appearance of freeze-dried cakes containing five different lyophilized compounds of compound 1.

[0018] Figure 2 The image shows the appearance of a pharmaceutical composition containing compound 1 suspended in 2.5 mL of water for injection. Invention Details

[0020] I. Lyophilized products of this disclosure

[0021] In one embodiment, this disclosure provides a lyophilized form comprising compound 1 for subcutaneous administration to a subject in need.

[0022] In another embodiment, this disclosure provides a lyophilized product comprising compound 1 and a filler. In another embodiment, the weight ratio of compound 1 to filler is about 5 to about 0.05. In another embodiment, the weight ratio of compound 1 to filler is about 3 to about 0.1. In another embodiment, the weight ratio of compound 1 to filler is about 1 to about 0.2. In another embodiment, the weight ratio of compound 1 to filler is about 3, about 2, about 1, about 0.9, about 0.8, about 0.7, about 0.6, about 0.5, about 0.4, about 0.3, about 0.2, or about 0.1. In another embodiment, the filler is mannitol.

[0023] In another embodiment, this disclosure provides a lyophilized product comprising compound 1 and a cellulose-based suspension modifier. In another embodiment, the weight ratio of compound 1 to the cellulose-based suspension modifier is about 25 to about 0.3. In another embodiment, the weight ratio of compound 1 to the cellulose-based suspension modifier is about 15 to about 0.5. In another embodiment, the weight ratio of compound 1 to the cellulose-based suspension modifier is about 10 to about 1. In another embodiment, the weight ratio of compound 1 to the cellulose-based suspension modifier is about 5 to about 2. In another embodiment, the weight ratio of compound 1 to the cellulose-based suspension modifier is about 10, about 9, about 8, about 7.5, about 7, about 6, about 5, about 4, about 3, about 2.7, about 2, or about 1. In another embodiment, the cellulose-based suspension modifier is sodium carboxymethyl cellulose.

[0024] In another embodiment, this disclosure provides a lyophilized product comprising compound 1 and a poloxamer-based suspension modifier. In another embodiment, the weight ratio of compound 1 to the poloxamer-based suspension modifier is about 80 to about 1. In another embodiment, the weight ratio of compound 1 to the poloxamer-based suspension modifier is about 40 to about 2. In another embodiment, the weight ratio of compound 1 to the poloxamer-based suspension modifier is about 20, about 19, about 18, about 17, about 16, about 15, about 14, about 13, about 12, about 11, about 10, about 9, about 8.8, about 7, about 6, about 5, about 4, about 3, or about 2. In another embodiment, the poloxamer-based suspension modifier is poloxamer 188 (a copolymer of polyethylene oxide and polypropylene oxide).

[0025] In another embodiment, this disclosure provides a lyophilized product comprising compound 1 and a lecithin-based suspension modifier. In another embodiment, the weight ratio of compound 1 to the lecithin-based suspension modifier is about 15 to about 0.15. In another embodiment, the weight ratio of compound 1 to the lecithin-based suspension modifier is about 8 to about 0.3. In another embodiment, the weight ratio of compound 1 to the lecithin-based suspension modifier is about 5 to about 0.6. In another embodiment, the weight ratio of compound 1 to the lecithin-based suspension modifier is about 3 to about 1. In another embodiment, the weight ratio of compound 1 to the lecithin-based suspension modifier is about 5, about 4, about 3, about 2, about 1.5, about 1, or about 0.6. In another embodiment, the lecithin-based suspension modifier is soybean lecithin.

[0026] In another embodiment, this disclosure provides a lyophilized product comprising compound 1 and a buffer. In another embodiment, the weight ratio of compound 1 to buffer is about 50 to about 1. In another embodiment, the weight ratio of compound 1 to buffer is about 25 to about 10. In another embodiment, the weight ratio of compound 1 to buffer is about 20 to about 15. In another embodiment, the weight ratio of compound 1 to buffer is about 25, about 24, about 23, about 22, about 21, about 20, about 21, about 20, about 19, about 18, about 17.4, about 17, about 16, about 15, about 14, about 13, about 12, about 11, or about 10. In another embodiment, the buffer is L-histidine.

[0027] In another embodiment, this disclosure provides a lyophilized product comprising: compound 1, a filler, at least one of a cellulose-based suspension regulator, a poloxamer-based suspension regulator, or a lecithin-based suspension regulator, and optionally a buffer.

[0028] In another embodiment, this disclosure provides a lyophilized product comprising: compound 1, a filler, and at least two of a cellulose-based suspension regulator, a poloxamer-based suspension regulator, or a lecithin-based suspension regulator, and optionally a buffer.

[0029] In another embodiment, this disclosure provides a lyophilized product comprising: compound 1, a filler, a cellulose-based suspension regulator, a poloxamer-based suspension regulator, and a lecithin-based suspension regulator, and optionally a buffer.

[0030] In another embodiment, this disclosure provides a lyophilized product comprising about 20% wt / wt to about 45% wt / wt of compound 1 and about 35% wt / wt to about 55% wt / wt of filler.

[0031] In another embodiment, this disclosure provides a lyophilized product comprising compound 1, mannitol, sodium carboxymethyl cellulose, poloxamer 188, and soybean lecithin, and optionally L-histidine.

[0032] In another embodiment, this disclosure provides a lyophilized product comprising about 10 mg to about 120 mg of compound 1. In another embodiment, this disclosure provides a lyophilized product comprising about 20 mg to about 110 mg of compound 1. In another embodiment, this disclosure provides a lyophilized product comprising about 30 mg to about 100 mg of compound 1. In another embodiment, this disclosure provides a lyophilized product comprising about 40 mg to about 90 mg of compound 1. In another embodiment, this disclosure provides a lyophilized product comprising about 50 mg to about 80 mg of compound 1. In another embodiment, this disclosure provides a lyophilized product comprising about 60 mg to about 70 mg of compound 1. In another embodiment, this disclosure provides a lyophilized compound comprising about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 42 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 62.5 mg, about 65 mg, about 67.5 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, or about 120 mg of compound 1.

[0033] In another embodiment, this disclosure provides a lyophilized product comprising about 50 mg to about 250 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 60 mg to about 210 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 70 mg to about 200 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 80 mg to about 190 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 90 mg to about 180 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 100 mg to about 170 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 110 mg to about 160 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 120 mg to about 150 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 125 mg to about 140 mg of filler. In another embodiment, this disclosure provides a lyophilized product comprising about 50 mg, about 55 mg, about 60 mg, about 65 mg, about 70 mg, about 75 mg, about 80 mg, about 85 mg, about 90 mg, about 95 mg, about 100 mg, about 105 mg, about 110 mg, about 115 mg, about 120 mg, about 125 mg, about 130 mg, about 135 mg, about 140 mg, about 145 mg, about 150 mg, about 155 mg, about 160 mg, about 165 mg, about 170 mg, about 175 mg, about 180 mg, about 185 mg, about 190 mg, about 195 mg, about 200 mg, about 205 mg, about 210 mg, about 215 mg, or about 220 mg of filler. In another embodiment, the filler is mannitol.

[0034] In another embodiment, this disclosure provides a lyophilized product comprising about 1 mg to about 100 mg of a cellulose-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 5 mg to about 75 mg of a cellulose-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 5 mg to about 50 mg of a cellulose-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 5 mg to about 10 mg of a cellulose-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 20 mg to about 30 mg of a cellulose-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 5 mg, about 6 mg, about 7 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, about 20 mg, about 21 mg, about 22 mg, about 23 mg, about 24 mg, about 25 mg, about 25.2 mg, about 2... 6 mg, about 27 mg, about 28 mg, about 29 mg, about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, or about 50 mg of a cellulose-based suspension modifier. In another embodiment, the cellulose-based suspension modifier is sodium carboxymethyl cellulose.

[0035] In another embodiment, this disclosure provides a lyophilized product comprising about 0.5 mg to about 70 mg of a poloxamer-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 1 mg to about 40 mg of a poloxamer-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 2 mg to about 30 mg of a poloxamer-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 3 mg to about 20 mg of a poloxamer-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 4 mg to about 10 mg of a poloxamer-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 3 mg, about 4 mg, about 5 mg, about 6 mg, about 7 mg, about 7.65 mg, about 8 mg, about 9 mg, about 10 mg, about 11 mg, about 12 mg, about 13 mg, about 14 mg, about 15 mg, about 16 mg, about 17 mg, about 18 mg, about 19 mg, or about 20 mg of a poloxamer-based suspension modifier. In another embodiment, the poloxamer-based suspension modifier is poloxamer 188.

[0036] In another embodiment, this disclosure provides a lyophilized product comprising about 1 mg to about 100 mg of a lecithin-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 10 mg to about 80 mg of a lecithin-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 20 mg to about 70 mg of a lecithin-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 30 mg to about 60 mg of a lecithin-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 40 mg to about 50 mg of a lecithin-based suspension modifier. In another embodiment, this disclosure provides a lyophilized product comprising about 30 mg, about 31 mg, about 32 mg, about 33 mg, about 34 mg, about 35 mg, about 36 mg, about 37 mg, about 38 mg, about 39 mg, about 40 mg, about 41 mg, about 42 mg, about 43 mg, about 44 mg, about 45 mg, about 46 mg, about 47 mg, about 48 mg, about 49 mg, about 50 mg, about 51 mg, about 52 mg, about 53 mg, about 54 mg, about 55 mg, about 56 mg, about 57 mg, about 58 mg, about 59 mg, or about 60 mg of a lecithin-based suspension modifier. In another embodiment, the lecithin-based suspension modifier is soybean lecithin.

[0037] In another embodiment, this disclosure provides a lyophilized product containing about 0.1 mg to about 20 mg of buffer. In another embodiment, this disclosure provides a lyophilized product containing about 0.5 mg to about 15 mg of buffer. In another embodiment, this disclosure provides a lyophilized product containing about 1 mg to about 10 mg of buffer. In another embodiment, this disclosure provides a lyophilized product containing about 2 mg to about 8 mg of buffer. In another embodiment, this disclosure provides a lyophilized product containing about 3 mg to about 5 mg of buffer. In another embodiment, this disclosure provides a lyophilized product comprising about 0.1 mg, about 0.5 mg, about 1 mg, about 1.5 mg, about 2 mg, about 2.5 mg, about 3 mg, about 3.5 mg, about 3.9 mg, about 4 mg, about 4.5 mg, about 5 mg, about 5.5 mg, about 6 mg, about 6.5 mg, about 7 mg, about 7.5 mg, about 8 mg, about 8.5 mg, about 9 mg, about 9.5 mg, about 10 mg, about 10.5 mg, about 11 mg, about 11.5 mg, about 12 mg, about 12.5 mg, about 13 mg, about 13.5 mg, about 14 mg, about 14.5 mg, or about 15 mg of buffer. In another embodiment, the buffer is L-histidine.

[0038] This disclosure provides the following specific implementation methods:

[0039] Implementation scheme I. A lyophilized product containing about 60 mg to about 70 mg of compound 1 for subcutaneous administration to a subject.

[0040] Implementation Scheme II. The lyophilized product of Implementation Scheme I, which contains about 125 mg to about 140 mg of filler, such as mannitol.

[0041] Implementation Scheme III. The lyophilized product of Implementation Scheme I or II, which contains about 8 mg to about 30 mg of a cellulose-based suspension regulator, such as sodium carboxymethyl cellulose.

[0042] Implementation Scheme IV. A lyophilized product of any one of Implementation Schemes I-III, comprising about 5 mg to about 10 mg of a poloxamer-based suspension modifier, such as poloxamer 188.

[0043] Implementation scheme V. The lyophilized product of any one of implementation schemes I-IV, which contains about 40 mg to about 50 mg of a lecithin-based suspension regulator, such as soy lecithin.

[0044] Implementation scheme VI. The lyophilized product of any of implementation scheme IV, which contains about 2 mg to about 5 mg of buffer, such as L-histidine.

[0045] This disclosure provides the following specific implementation methods:

[0046] Implementation scheme IA. A lyophilized product containing about 110 mg to about 130 mg of compound 1 for subcutaneous administration to a subject.

[0047] Implementation scheme II-A. The lyophilized product of implementation scheme IA contains about 110 mg to about 250 mg of filler, such as mannitol.

[0048] Implementation scheme III-A. The lyophilized product of implementation scheme IA or II-A contains about 5 mg to about 30 mg of a cellulose-based suspension regulator, such as sodium carboxymethyl cellulose.

[0049] Implementation scheme IV-A. The lyophilized product of any one of implementation schemes IA to III-A contains about 5 mg to about 15 mg of a poloxamer-based suspension modifier, such as poloxamer 188.

[0050] Implementation scheme VA. The lyophilized product of any one of implementation schemes IA to IV-A contains about 40 mg to about 90 mg of a lecithin-based suspension modifier, such as soy lecithin.

[0051] Implementation scheme VI-A. The lyophilized product of any one of implementation schemes IA to VA contains about 2 mg to about 10 mg of buffer, such as L-histidine.

[0052] The representative lyophilized products of this disclosure (designated L1-L5) contain the following amounts of compound 1, mannitol, NaCMC, poloxamer 188, NF, histidine, and soybean lecithin:

[0053]

[0054]

[0055] In another embodiment, the moisture content of the lyophilized product is determined by the Karl Fischer method to be about 4% or less. In another embodiment, the moisture content of the lyophilized product is about 3% or less. In another embodiment, the moisture content of the lyophilized product is about 2% or less. In another embodiment, the moisture content of the lyophilized product is about 1% or less. In another embodiment, the moisture content of the lyophilized product is about 0.9%. In another embodiment, the moisture content of the lyophilized product is about 0.8%. In another embodiment, the moisture content of the lyophilized product is about 0.7%. In another embodiment, the moisture content of the lyophilized product is about 0.6%. In another embodiment, the moisture content of the lyophilized product is about 0.5%. In another embodiment, the moisture content of the lyophilized product is about 0.4%. In another embodiment, the moisture content of the lyophilized product is about 0.3%. In another embodiment, the moisture content of the lyophilized product is about 0.2%. In another embodiment, the moisture content of the lyophilized product is about 0.1%.

[0056] In another embodiment, the chemical purity of compound 1 in the lyophilized product, as determined by HPLC, is about 96% to about 99.9%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 96%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 96.5%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 97%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 97.5%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 98%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 98.5%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 99.0%. In another embodiment, the chemical purity of compound 1 in the lyophilized product is about 99.5%.

[0057] In another embodiment, this disclosure provides a lyophilized product comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0058] In another embodiment, this disclosure provides a lyophilized product that is substantially composed of about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0059] In another embodiment, this disclosure provides a lyophilized product comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0060] In another embodiment, this disclosure provides a lyophilized product comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0061] In another embodiment, this disclosure provides a lyophilized product that is substantially composed of about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0062] In another embodiment, this disclosure provides a lyophilized product comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine and about 45 mg of soybean lecithin.

[0063] In another embodiment, this disclosure provides a lyophilized product comprising about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine and about 41.7 mg of soybean lecithin.

[0064] In another embodiment, this disclosure provides a lyophilized product that is substantially composed of about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine and about 41.7 mg of soybean lecithin.

[0065] In another embodiment, this disclosure provides a lyophilized product comprising about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine and about 41.7 mg of soybean lecithin.

[0066] In another embodiment, this disclosure provides a lyophilized product comprising about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, and about 80.2 mg of soybean lecithin.

[0067] In another embodiment, this disclosure provides a lyophilized product that is substantially composed of about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, and about 80.2 mg of soybean lecithin.

[0068] In another embodiment, this disclosure provides a lyophilized product comprising about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, and about 80.2 mg of soybean lecithin.

[0069] In another embodiment, this disclosure provides a lyophilized product comprising about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, and about 40.1 mg of soybean lecithin.

[0070] In another embodiment, this disclosure provides a lyophilized product that is substantially composed of about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, and about 40.1 mg of soybean lecithin.

[0071] In another embodiment, this disclosure provides a lyophilized product comprising about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, and about 40.1 mg of soybean lecithin.

[0072] The lyophilized products described in this section are collectively referred to as "lyophilized products of this disclosure".

[0073] II. Pharmaceutical compositions disclosed herein

[0074] In another embodiment, this disclosure provides a pharmaceutical composition comprising a lyophilized product of this disclosure suspended in a solvent. In one embodiment, the solvent comprises water. In another embodiment, the solvent is water, for example, sterile water for injection from the United States Pharmacopeia.

[0075] In another embodiment, this disclosure provides a pharmaceutical composition comprising a lyophilized product of this disclosure suspended in a solvent, wherein the concentration of compound 1 is from about 20 mg / mL to about 60 mg / mL, for example, from about 24 mg / mL to about 35 mg / mL. In another embodiment, the concentration of compound 1 is from about 25 mg / mL to about 30 mg / mL. In another embodiment, the concentration of compound 1 is about 20 mg / mL. In another embodiment, the concentration of compound 1 is about 21 mg / mL. In another embodiment, the concentration of compound 1 is about 22 mg / mL. In another embodiment, the concentration of compound 1 is about 23 mg / mL. In another embodiment, the concentration of compound 1 is about 24 mg / mL. In another embodiment, the concentration of compound 1 is about 25 mg / mL. In another embodiment, the concentration of compound 1 is about 26 mg / mL. In another embodiment, the concentration of compound 1 is about 27 mg / mL. In another embodiment, the concentration of compound 1 is about 28 mg / mL. In another embodiment, the concentration of compound 1 is about 29 mg / mL. In another embodiment, the concentration of compound 1 is about 30 mg / mL. In another embodiment, the concentration of compound 1 is about 31 mg / mL. In another embodiment, the concentration of compound 1 is about 32 mg / mL. In another embodiment, the concentration of compound 1 is about 33 mg / mL. In another embodiment, the concentration of compound 1 is about 34 mg / mL. In another embodiment, the concentration of compound 1 is about 35 mg / mL. In another embodiment, the concentration of compound 1 is about 40 mg / mL. In another embodiment, the concentration of compound 1 is about 45 mg / mL. In another embodiment, the concentration of compound 1 is about 50 mg / mL. In another embodiment, the concentration of compound 1 is about 55 mg / mL. In another embodiment, the concentration of compound 1 is about 60 mg / mL.

[0076] In another embodiment, this disclosure provides a pharmaceutical composition comprising a lyophilized product of this disclosure suspended in a solvent, wherein, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is from about 0.1 μm to about 50 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is from about 1 μm to about 40 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is from about 10 μm to about 30 μm. In another embodiment, the average particle size distribution of compound 1 in the suspension is from about 10 μm to about 30 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is about 10 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is about 15 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is about 20 μm. In another embodiment, after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is about 25 μm.

[0077] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soy lecithin, and about 2.5 mL of sterile water for injection (WFI USP).

[0078] In another embodiment, this disclosure provides a pharmaceutical composition comprising essentially about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0079] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 25.2 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0080] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soy lecithin, and about 2.25 mL of sterile water for injection from the United States Pharmacopeia.

[0081] In another embodiment, this disclosure provides a pharmaceutical composition comprising essentially about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soy lecithin, and about 2.25 mL of sterile water for injection from the United States Pharmacopeia.

[0082] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 67.5 mg of compound 1, about 135 mg of mannitol, about 9.0 mg of sodium carboxymethyl cellulose, about 7.65 mg of poloxamer 188, about 3.88 mg of L-histidine, about 45 mg of soybean lecithin, and about 2.25 mL of sterile water for injection from the United States Pharmacopeia.

[0083] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine, about 41.7 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0084] In another embodiment, this disclosure provides a pharmaceutical composition comprising essentially about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine, about 41.7 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0085] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 62.5 mg of compound 1, about 125 mg of mannitol, about 8.33 mg of sodium carboxymethyl cellulose, about 7.08 mg of poloxamer 188, about 4.31 mg of L-histidine, about 41.7 mg of soybean lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0086] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, about 80.2 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0087] In another embodiment, this disclosure provides a pharmaceutical composition comprising essentially about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, about 80.2 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0088] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 120.2 mg of compound 1, about 240.4 mg of mannitol, about 16 mg of sodium carboxymethyl cellulose, about 13.6 mg of poloxamer 188, about 8.3 mg of L-histidine, about 80.2 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0089] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8.0 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, about 40.1 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0090] In another embodiment, this disclosure provides a pharmaceutical composition comprising essentially about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8.0 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, about 40.1 mg of soy lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0091] In another embodiment, this disclosure provides a pharmaceutical composition comprising about 120.2 mg of compound 1, about 120.2 mg of mannitol, about 8.0 mg of sodium carboxymethyl cellulose, about 6.8 mg of poloxamer 188, about 4.1 mg of L-histidine, about 40.1 mg of soybean lecithin, and about 2.5 mL of sterile water for injection from the United States Pharmacopeia.

[0092] In another embodiment, this disclosure provides a pharmaceutical composition comprising lyophilized products L1, L2, L3, L4, or L5 (see table above) and 1 mL WFI USP, 1.5 mL WFI USP, 2.0 mL WFI USP, 2.5 mL WFI USP, 3.0 mL WFI USP, 3.5 mL WFI USP, 4.0 mL WFI USP, 4.5 mL WFI USP, or 5 mL WFI USP.

[0093] The pharmaceutical compositions described in this section are collectively referred to as "the pharmaceutical compositions of this disclosure".

[0094] III. Treatment Methods

[0095] In another embodiment, this disclosure provides a method for treating cancer in a subject in need, the method comprising subcutaneously administering a therapeutically effective amount of the pharmaceutical composition of this disclosure to the subject.

[0096] In another embodiment, this disclosure provides a method for treating cancer in a subject in need, the method comprising subcutaneously administering a therapeutically effective amount of the pharmaceutical composition of this disclosure, and one or more optional therapeutic agents, to the subject.

[0097] In another embodiment, this disclosure provides a pharmaceutical composition of this disclosure for treating cancer in a subject.

[0098] In another embodiment, this disclosure provides a pharmaceutical composition of this disclosure for treating cancer in a subject, wherein the pharmaceutical composition of this disclosure, along with one or more optional therapeutic agents, is administered subcutaneously.

[0099] In another embodiment, the pharmaceutical composition of this disclosure is administered subcutaneously to the subject according to an intermittent dosing regimen. For example, the pharmaceutical composition of this disclosure may be administered to the subject on three non-consecutive days per week, such as Monday-Wednesday-Friday, or two non-consecutive days per week, such as Monday and Thursday.

[0100] In one implementation, the cancer is a solid tumor.

[0101] In another embodiment, the cancer is a hematologic cancer. In yet another embodiment, the hematologic cancer is acute lymphoblastic leukemia, chronic lymphocytic leukemia (including B-cell chronic lymphocytic leukemia), or acute myeloid leukemia.

[0102] In another embodiment, the cancer is any one or more of the cancers listed in Table 1.

[0103] Table 1

[0104]

[0105]

[0106]

[0107] In another implementation, the cancer is any one or more of the cancers listed in Table 2.

[0108] Table 2

[0109] Acute lymphoblastic leukemia (ALL) Acute eosinophilic leukemia Acute myeloid leukemia (AML) Acute erythroleukemia Chronic lymphocytic leukemia (CLL) Acute lymphoblastic leukemia Small lymphocytic lymphoma (SLL) Acute megakaryoblastic leukemia Multiple myeloma (MM) Acute monocytic leukemia Hodgkin's lymphoma (HL) Acute promyelocytic leukemia Non-Hodgkin's lymphoma (NHL) Acute myeloid leukemia Mantle cell lymphoma (MCL) B-cell prolymphocytic leukemia Marginal zone B-cell lymphoma B-cell lymphoma Splenic marginal zone lymphoma MALT lymphoma Follicular lymphoma (FL) Precursor T-lymphoblastic lymphoma Waldenström macroglobulinemia (WM) T-cell lymphoma Diffuse large B-cell lymphoma (DLBCL) Mast cell leukemia Marginal zone lymphoma (MZL) Adult T-cell leukemia / lymphoma Hairy cell leukemia (HCL) Invasive NK cell leukemia Burkitt's lymphoma (BL) Angioimmunoblastic T-cell lymphoma Richter's transformation

[0110] In another embodiment, the cancer is selected from the group consisting of: squamous cell carcinoma of the head and neck (SCCHN), adenocarcinoma of the esophagus, adenocarcinoma of the stomach, adenocarcinoma of the colon, hepatocellular carcinoma, bile duct carcinoma of the biliary system, adenocarcinoma of the gallbladder, adenocarcinoma of the pancreas, ductal carcinoma in situ of the breast, adenocarcinoma of the breast, adenocarcinoma of the lung, squamous cell carcinoma of the lung, transitional cell carcinoma of the bladder, squamous cell carcinoma of the bladder, squamous cell carcinoma of the cervix, adenocarcinoma of the cervix, endometrial cancer, squamous cell carcinoma of the penis, and squamous cell carcinoma of the skin.

[0111] In another implementation, the precancerous tumor is selected from the set of the following members: leukoplakia of the head and neck, Barrett's esophagus, gastric tissue transformation, colonic adenoma, chronic hepatitis, bile duct hyperplasia, pancreatic intraepithelial neoplasia, atypical adenomatous hyperplasia of the lungs, bladder developmental abnormalities, cervical intraepithelial neoplasia, penile intraepithelial neoplasia, and actinic keratosis of the skin.

[0112] In another embodiment, the cancer is selected from the set of the following members: hepatocellular carcinoma, glioblastoma, lung cancer, breast cancer, head and neck cancer, prostate cancer, melanoma, and colorectal cancer.

[0113] In another implementation, the cancer is selected from the set of the following members: colorectal cancer, breast cancer, lymphoma, melanoma, kidney cancer, and lung cancer.

[0114] In another embodiment, the cancer has become resistant to conventional cancer treatments. As used herein, the term "conventional cancer treatment" means any cancer drug, biologic, or radiation therapy, or a combination of cancer drugs and / or biologics and / or radiation therapy, that has been tested and / or approved by the U.S. Food and Drug Administration, the European Medicines Agency, or a similar regulatory body for therapeutic use in humans.

[0115] IV. Optional treatments

[0116] In some treatment methods and uses of this disclosure, the pharmaceutical composition of this disclosure is administered as a single agent to a subject suffering from cancer. In other treatment methods and uses of this disclosure, the pharmaceutical composition of this disclosure is administered in combination with one or more optional therapeutic agents to a subject suffering from cancer. In one embodiment, the pharmaceutical composition of this disclosure is administered in combination with one optional therapeutic agent. In another embodiment, the pharmaceutical composition of this disclosure is administered in combination with two optional therapeutic agents. In yet another embodiment, the pharmaceutical composition of this disclosure is administered in combination with three optional therapeutic agents. Optional therapeutic agents that can be used to treat cancer patients include those known in the art and those developed in the future.

[0117] The optional therapeutic agent is administered in an amount that provides the desired therapeutic effect. The effective dose range of each optional therapeutic agent is known in the art, and the optional therapeutic agent is administered to the individual in need within such a defined range.

[0118] The pharmaceutical compositions and optional therapeutic agents of this disclosure can be administered separately as multiple unit doses in any order, for example, wherein the pharmaceutical compositions of this disclosure are administered before the optional therapeutic agents, or vice versa. One or more doses of the pharmaceutical compositions and optional therapeutic agents of this disclosure may be administered to a subject.

[0119] In one embodiment, the optional therapeutic agent comprises one or more immune checkpoint inhibitors. Examples of immune checkpoint inhibitors include PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, cd47 inhibitors, TIGIT inhibitors, and B7-H1 inhibitors. Therefore, in one embodiment, the immune checkpoint inhibitor is selected from the set of members: PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, and cd47 inhibitors.

[0120] In another embodiment, the immune checkpoint inhibitor is a programmed cell death (PD-1) inhibitor. PD-1 is a T-cell co-inhibitory receptor that plays a crucial role in the ability of tumor cells to evade the host's immune system. Blocking the interaction between PD-1 and PD-L1 (a ligand of PD-1) enhances immune function and mediates anti-tumor activity. Examples of PD-1 inhibitors include antibodies that specifically bind to PD-1. Specific anti-PD-1 antibodies include, but are not limited to, nivolumab, pembrolizumab, STI-A1014, pidilzumab, and cimipril (cemiplimab-rwlc). For a general discussion of the availability, manufacturing methods, mechanism of action, and clinical studies of anti-PD-1 antibodies, see US2013 / 0309250, US6,808,710, US7,595,048, US8,008,449, US8,728,474, US8,779,105, US8,952,136, US8,900,587, US9,073,994, US9,084,776, and Naido et al., British Journal of Cancer 111:2214-19 (2014).

[0121] In another embodiment, the immune checkpoint inhibitor is a PD-L1 (also known as B7-H1 or CD274) inhibitor. Examples of PD-L1 inhibitors include antibodies that specifically bind to PD-L1. Specific anti-PD-L1 antibodies include, but are not limited to, avelumab, atezolizumab, durvalumab, and BMS-936559. For a general discussion of availability, manufacturing methods, mechanism of action, and clinical studies, see US8,217,149, US2014 / 0341917, US2013 / 0071403, WO2015036499, and Naido et al., British Journal of Cancer 111:2214-19 (2014).

[0122] In another embodiment, the immune checkpoint inhibitor is a CTLA-4 inhibitor. CTLA-4, also known as cytotoxic T-lymphocyte antigen 4, is a protein receptor that downregulates the immune system. CTLA-4 is characterized as a “brake” that binds to co-stimulatory molecules on antigen-presenting cells, preventing interaction with CD28 on T cells and also producing a significant inhibitory signal that limits T cell activation. Examples of CTLA-4 inhibitors include antibodies that specifically bind to CTLA-4. Specific anti-CTLA-4 antibodies include, but are not limited to, ipilimumab and trimemumab. For a general discussion of availability, manufacturing methods, mechanism of action, and clinical studies, see US6,984,720, US6,207,156, and Naido et al., British Journal of Cancer 111:2214-19 (2014).

[0123] In another embodiment, the immune checkpoint inhibitor is a LAG3 inhibitor. LAG3 (Lymphocyte Activation Gene 3) is a negative co-stimulatory receptor that regulates T cell homeostasis, proliferation, and activation. Furthermore, LAG3 has been reported to participate in the suppressive function of regulatory T cells (Tregs). Most LAG3 molecules are retained in cells near the microtubule center and are induced only upon antigen-specific T cell activation. (US2014 / 0286935). Examples of LAG3 inhibitors include antibodies that specifically bind to LAG3. Specific anti-LAG3 antibodies include, but are not limited to, GSK2831781. For a general discussion of availability, manufacturing methods, mechanism of action, and studies, see US2011 / 0150892, US2014 / 0093511, US20150259420, and Huang et al., Immunity 21:503-13 (2004).

[0124] In another embodiment, the immune checkpoint inhibitor is a TIM3 inhibitor. TIM3 (T-cell immunoglobulin and mucin domain 3) is an immune checkpoint receptor that restricts T cells... H 1 and T C 1. Duration and intensity of T-cell responses. Due to TIM3 in dysfunctional CD8... +The TIM3 pathway, expressed on T cells and Tregs (two populations of immune cells reported to constitute immunosuppression in tumor tissues), is considered a target for anticancer immunotherapy. (Anderson, Cancer Immunology Research 2:393-98 (2014)). Examples of TIM3 inhibitors include antibodies that specifically bind to TIM3. For a general discussion of the availability, manufacturing methods, mechanisms of action, and research on TIM3 inhibitors, see US20150225457, US20130022623, US8,522,156, Ngiow et al., Cancer Res 71:6567-71 (2011), Ngiow et al., Cancer Res 71:3540-51 (2011), and Anderson, Cancer Immunology Research 2:393-98 (2014).

[0125] In another embodiment, the immune checkpoint inhibitor is a CD47 inhibitor. See Unanue, ER, PNAS 110:10886-87 (2013).

[0126] In another embodiment, the immune checkpoint inhibitor is a TIGIT inhibitor. See also and Guillerey, Clin Exp Immunol 200:108-119 (2019).

[0127] The one or more immune checkpoint inhibitors are administered to the subject in a therapeutically effective dose. These doses are guided by standard clinical practice, for example, based on prescribing information associated with the immune checkpoint inhibitor.

[0128] For example, when the immune checkpoint inhibitor is a monoclonal antibody, it can be administered as an intravenous infusion of about 1 to about 2000 mg every 2-4 weeks. For example, antibodies can be administered in doses of approximately 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1100 mg, 1200 mg, 1300 mg, 1400 mg, 1500 mg, 1600 mg, 1700 mg, 1800 mg, 1900 mg, or 2000 mg.

[0129] For example, when the immune checkpoint inhibitor is nivolumab, 240 mg can be administered to the subject via intravenous infusion every 2 weeks, or 480 mg can be administered via intravenous infusion every 4 weeks.

[0130] For example, when the immune checkpoint inhibitor is pembrolizumab, it can be administered to the subject at a dose of 200 mg via intravenous infusion every 3 weeks, or at a dose of 400 mg via intravenous infusion every 6 weeks.

[0131] For example, when the immune checkpoint inhibitor is avelumab, 800 mg can be administered to the subject via intravenous infusion every 2 weeks.

[0132] For example, when the immune checkpoint inhibitor is atezolizumab, it can be administered by intravenous infusion of 840 mg every 2 weeks, or by intravenous infusion of 1200 mg every 3 weeks, or by intravenous infusion of 1680 mg every 4 weeks.

[0133] For example, when the immune checkpoint inhibitor is ipilimumab, it can be administered to the subject at 3 mg / kg via intravenous infusion every 3 weeks.

[0134] For example, when the immune checkpoint inhibitor is trimemumab, it can be administered to the subject every 4 weeks via intravenous infusion at a dose of 3-20 mg / kg.

[0135] For example, when the immune checkpoint inhibitor is GSK2831781, it can be administered to the subject via intravenous infusion at a dose of 1.5 to 5 mg / kg every 2 to 4 weeks.

[0136] Representative dosing regimens for certain immune checkpoint inhibitors used to treat certain cancers are provided in Table 3.

[0137] Table 3

[0138]

[0139]

[0140] In one embodiment, the immune checkpoint inhibitor is an antibody, and is administered to the subject via intravenous infusion at a dose of 1-20 mg / kg every 2-4 weeks. In another embodiment, the antibody is administered to the subject via intravenous infusion at a dose of 20-2000 mg every 2-4 weeks. In another embodiment, compound 1 is administered before the administration of the antibody. In another embodiment, compound 1 is administered to the subject 1, 2, 3, 4, 5, 6, or 7 days prior to the day of antibody administration. In another embodiment, compound 1 is administered to the subject on the day of antibody administration. In another embodiment, compound 1 is administered to the subject 1, 2, 3, 4, 5, 6, or 7 days after the day of antibody administration.

[0141] For example, the subject received pembrolizumab administered intravenously every 3 weeks and compound 1 administered subcutaneously twice weekly, wherein the first dose of compound 1 was administered before the first dose of pembrolizumab, on the same day as the first dose of pembrolizumab, or after the first dose of pembrolizumab, for example, until disease progression or until no treatment benefit was found.

[0142] For example, the subject received nivolumab administered intravenously every 2 weeks and compound 1 administered subcutaneously twice weekly, wherein the first dose of compound 1 was administered before the first dose of nivolumab, on the same day as the first dose of nivolumab, or after the first dose of nivolumab, for example, until disease progression or until no treatment benefit was found.

[0143] In another implementation, treatment of cancer patients with compound 1 and one or more immune checkpoint inhibitors induces an antiproliferative response more quickly than when immune checkpoint inhibitors are administered alone.

[0144] The term "antibody" is intended to include complete monoclonal antibodies, polyclonal antibodies, multispecific antibodies formed from at least two complete antibodies, and antibody fragments, provided they exhibit the desired biological activity. In another embodiment, "antibody" is intended to include a soluble receptor that does not possess the Fc portion of the antibody. In one embodiment, the antibody is a humanized monoclonal antibody or fragment thereof prepared by recombinant genetic engineering.

[0145] Another class of immune checkpoint inhibitors includes peptides that bind to and block the PD-1 receptor on T cells without triggering inhibitor signal transduction. Such peptides include B7-DC peptide, B7-H1 peptide, B7-1 peptide, and B7-2 peptide, and their soluble fragments, as disclosed in U.S. Patent 8,114,845.

[0146] Another class of immune checkpoint inhibitors includes compounds having a peptide moiety that inhibits PD-1 signaling. Examples of such compounds are disclosed in U.S. Patent 8,907,053.

[0147] Another class of immune checkpoint inhibitors includes inhibitors of certain metabolic enzymes, such as indoleamine 2,3 dioxygenase (IDO), expressed by infiltrating myeloid cells and tumor cells, and isocitrate dehydrogenase (IDH), which is mutated in leukemia cells. Mutants of the IDH enzyme lead to elevated levels of 2-hydroxyglutarate (2-HG), which inhibits myeloid differentiation. Stein et al., Blood 130:722-31 (2017); Wouters, Blood 130:693-94 (2017). Specific mutant IDH blockers include, but are not limited to, evanixifen and ensidipin mesylate. Dalle and DiNardo, TherAdv Hematol 9(7):163-73 (2018); Nassereddine et al., Onco Targets Ther 12:303-08 (2018). IDO enzymes suppress immune responses by depleting amino acids essential for anabolic functions in T cells or by synthesizing specific natural ligands of cytosol receptors that can alter lymphocyte function. (Pardoll, Nature Reviews. Cancer 12:252-64 (2012)) Cancer Immunol Immunother 58:153-57 (2009). Specific IDO blockers include, but are not limited to, L-1-methyltryptophan (L-1MT) and 1-methyltryptophan (1MT). Qian et al., Cancer Res 69:5498-504 (2009); and et al., Cancer Immunol Immunother 58:153-7 (2009).

[0148] In one embodiment, the immune checkpoint inhibitor is nivolumab, pembrolizumab, pildizumab, STI-A1110, avelumab, atezolizumab, durvalumab, STI-A1014, ipilimumab, trimemumab, GSK2831781, BMS-936559, or MED14736.

[0149] In another embodiment, the optional therapeutic agent is an epigenetic drug. As used herein, the term "epigenetic drug" refers to a therapeutic agent that targets an epigenetic regulator. Examples of epigenetic regulators include histone lysine methyltransferases, histone arginine methyltransferases, histone demethylases, histone deacetylases, histone acetylesterases, and DNA methyltransferases. Histone deacetylase inhibitors include, but are not limited to, vorinostat and lactate parbistat.

[0150] Other examples of conventional therapies and anticancer agents that can be used in combination with the pharmaceutical compositions of this disclosure include surgery, radiation therapy (e.g., gamma radiation, neutron beam radiation therapy, electron beam radiation therapy, proton therapy, brachytherapy, and systemic radioisotopes), endocrine therapy, biological response modulators (e.g., interferon, interleukin, tumor necrosis factor (TNF)), hyperthermia and cryotherapy, agents to mitigate any adverse effects (e.g., antiemetics), and any other approved biological or chemotherapeutic therapies (e.g., treatment regimens that use drugs to stop cancer cell growth by killing cancer cells or by preventing them from dividing). Chemotherapy can be administered orally, by injection or infusion, or over the skin, depending on the type and stage of the cancer being treated.

[0151] Non-limiting exemplary antiproliferative compounds include: aromatase inhibitors; antiestrogens; antiandrogens; gonadotropin-releasing hormone agonists; topoisomerase I inhibitors; topoisomerase II inhibitors; microtubule activators; alkylating agents, such as temozolomide; retinoids, carotenoids, or tocopherols; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antimetabolites; platinum compounds; methionine aminopeptidase inhibitors; bisphosphonates; antiproliferative antibodies; heparinase inhibitors; Ras oncogenic isoform inhibitors; telomere-terminal transferase inhibitors; proteasome inhibitors; compounds for the treatment of hematologic malignancies; Flt-3 inhibitors; Hsp90 inhibitors; kinin spindle protein inhibitors; MEK inhibitors; antitumor antibiotics; nitrosoureas; compounds that target / reduce the activity of protein or lipid kinases, compounds that target / reduce the activity of protein or lipid phosphatases, or any other anti-angiogenic compounds.

[0152] Non-limiting exemplary aromatase inhibitors include steroids such as atamitan, exemestane, and formestane, and non-steroids such as aminoglutethimide, lotamethasone, pyridoglutethimide, tralostertan, testosterone, ketoconazole, vorcicloazole, fazodazole, anastrozole, and letrozole.

[0153] Non-restrictive anti-estrogens include tamoxifen, fulvestrant, raloxifene, and raloxifene hydrochloride. Anti-androgens include, but are not limited to, bicalutamide and apalutamide. Gonadotropin-releasing hormone agonists include, but are not limited to, abareg, goserelin, and goserelin acetate.

[0154] Non-limiting exemplary topoisomerase I inhibitors include topotecan, gemmatocan, irinotecan, camptothecin and its analogues, 9-nitrocamptothecin, and the macromolecular camptothecin conjugate PNU-166148. Topoisomerase II inhibitors include, but are not limited to, anthracycline antibiotics such as doxorubicin, daunorubicin, epirubicin, idarubicin, and nemorubicin; anthraquinones such as mitoxantrone and loxoantrone; and podophyllotoxins such as etoposide and teniposide.

[0155] Microtubule activators include microtubule stabilizing compounds, microtubule destabilizing compounds, and microtubule polymerization inhibitors, including, but not limited to, taxanes such as paclitaxel and docetaxel; sponge lactone; colchicine and epothilone and their derivatives.

[0156] Non-limiting exemplary alkylating agents include cyclophosphamide, ifosfamide, melphalan, trabectedin, and nitrosoureas such as carmustine and lomustine.

[0157] Non-limiting exemplary matrix metalloproteinase inhibitors (“MMP inhibitors”) include collagen peptide mimics and non-peptide mimics inhibitors, tetracycline derivatives, palmastat, malimasatana, prrinstat, metastat, BMS-279251, BAY 12-9566, TAA211, MMI270B, and AAJ996.

[0158] Non-limiting exemplary mTOR inhibitors include compounds that inhibit mammalian target of rapamycin (mTOR) and have antiproliferative activity, such as sirolimus, everolimus, CCI-779, and ABT578.

[0159] Non-limiting exemplary antimetabolites include 5-fluorouracil (5-FU), capecitabine, gemcitabine, DNA demethylating compounds (such as 5-azacytidine and decitabine), methotrexate and edarax, and folic acid antagonists (such as pemetrexed).

[0160] Non-limiting exemplary platinum compounds include carboplatin, cisplatin, cisplatinum, and oxaliplatin.

[0161] Non-limiting exemplary methionine aminopeptidase inhibitors include bengamide or derivatives thereof and PPI-2458.

[0162] Non-limiting exemplary bisphosphonates include hydroxyethyl phosphate, chloromethyl bisphosphonate, teludromic acid, pamidromic acid, alendromic acid, ibandronic acid, risedromic acid, and zoledronic acid.

[0163] Non-limiting exemplary heparinase inhibitors include compounds that target, reduce, or inhibit the degradation of heparan sulfate, such as PI-88 and OGT2115.

[0164] Non-limiting exemplary compounds that target, reduce, or inhibit the carcinogenic activity of Ras include farnesyltransferase inhibitors, such as L-744832, DK8G557, tepifanabine, and lonafabine.

[0165] Non-limiting exemplary telomere-terminal transferase inhibitors include compounds that target, reduce, or inhibit the activity of telomere-terminal transferases, such as compounds that inhibit telomere-terminal transferase receptors, such as telomerostin.

[0166] Non-limiting exemplary proteasome inhibitors include compounds that target, reduce, or inhibit the activity of the proteasome, including, but not limited to, bortezomib. In some embodiments, the proteasome inhibitor is carfilzomib or ixazomib.

[0167] Non-limiting exemplary FMS-like tyrosine kinase inhibitors, as compounds that target, reduce or inhibit the activity of FMS-like tyrosine kinase receptor (Flt-3R), include giretinib, interferon, 1-β-D-arabinofuranylcytosine (cytarabine), and busulfan; and ALK inhibitors, as compounds that target, reduce or inhibit anaplastic lymphoma kinases, include alectinib, brigatinib, and lorlatinib.

[0168] Non-limiting exemplary Flt-3 inhibitors include PKC412, midotaurin, astrocytocin derivatives, SU11248, MLN518, and giretinib.

[0169] Non-limiting exemplary HSP90 inhibitors include compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90; or compounds that degrade, target, reduce, or inhibit HSP90 client proteins via the ubiquitin-proteasome pathway. Compounds that target, reduce, or inhibit the intrinsic ATPase activity of HSP90 are particularly compounds, proteins, or antibodies that inhibit the ATPase activity of HSP90, such as 17-allylamino, 17-demethoxygeldmycin (17AAG), geldmycin derivatives; other geldmycin-related compounds; rhizocarpine and HDAC inhibitors.

[0170] Non-limiting exemplary protein tyrosine kinase and / or serine and / or threonine kinase inhibitors or lipid kinase inhibitors include: a) compounds that target, reduce, or inhibit the activity of platelet-derived growth factor receptors (PDGFRs), such as compounds that target, reduce, or inhibit the activity of PDGFRs, including olarumab and N-phenyl-2-pyrimidinylamine derivatives, such as imatinib, SU101, SU6668, and GFB-111; b) compounds that target, reduce, or inhibit the activity of fibroblast growth factor receptors (FGFRs), such as erdatinib and lenvatinib; c) Compounds that target, reduce, or inhibit the activity of insulin-like growth factor receptor I (IGF-IR), such as brigatinib; d) Compounds that target, reduce, or inhibit the activity of vascular endothelial growth factor receptor (VEGFR), such as lenvatinib; e) Compounds that target, reduce, or inhibit the activity of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors, such as larotrectinib; f) Compounds that target, reduce, or inhibit the activity of the Axl receptor tyrosine kinase family; g) Compounds that target, reduce, or inhibit the activity of Ret receptor tyrosine kinase, such as alectinib. h) Compounds that target, reduce, or inhibit the activity of Kit / SCFR receptor tyrosine kinases, such as imatinib; i) Compounds that target, reduce, or inhibit the activity of c-Kit receptor tyrosine kinases, such as imatinib; j) Compounds that target, reduce, or inhibit the activity of members of the c-Abl family, their gene fusion products (e.g., Bcr-Abl kinases), and mutants, such as N-phenyl-2-pyrimidinylamine derivatives, such as imatinib or nilotinib; PD180970; AG957; NSC680410; PD173955; or dasatinib; k) Compounds that target, reduce, or inhibit the activity of serine / threonine kinases, such as astronein derivatives disclosed in U.S. Patent No. 5,093,330, such as midotulin; other examples include UCN-01, safingo, BAY43-9006, lichenstatin 1, perifoxin; imofoxin; RO 318220 and RO 320432; GO 6976; Isis 3521; LY333531 / LY379196; isoquinoline compounds; farnesyltransferase inhibitors; PD184352 or QAN697 or AT7519; abecicilline; bimetinib; cobimetinib; cannefenib; neratinib; palbociclib; rebociclib;l) Compounds that target, reduce, or inhibit the activity of protein-tyrosine kinases, such as acalatinib, imatinib mesylate, or tyrosine phosphorylation inhibitors, such as tyrosine phosphorylation inhibitors A23 / RG-50810; AG 99; tyrosine phosphorylation inhibitor AG 213; tyrosine phosphorylation inhibitor AG 1748; tyrosine phosphorylation inhibitor AG 490; tyrosine phosphorylation inhibitor B44; tyrosine phosphorylation inhibitor B44(+) enantiomer; tyrosine phosphorylation inhibitor AG 555; AG 494; Tyrosine phosphorylation inhibitors AG556, AG957 and adaphostin (4-{[(2,5-dihydroxyphenyl)methyl]amino}-adaphostin; NSC680410, adaphostin); m) compounds that target, reduce or inhibit the activity of receptor tyrosine kinases (EGFR, ErbB2, ErbB3, ErbB4 as homodimers or heterodimers) and their mutants in the epidermal growth factor family, such as brigatinib, CP358774, ZD 1839, ZM 105180; Trastuzumab, Cetuximab, Gefitinib, Erlotinib, Osimertinib, Dacomitinib, Nexilimumab, Neratinib, OSI-774, Cl-1033, EKB-569, GW-2016, Antibody E1.1, E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 and E7.6.3 and 7H-pyrrolo-[2,3-d]pyrimidine derivatives; n) Compounds that target, reduce or inhibit the activity of phosphatidylinositol 3-kinase (PI3K), such as apeliximab, cupannixi, and davidicilxi; and o) Compounds that target, reduce or inhibit the activity of c-Met receptors.

[0171] Non-limiting exemplary compounds that target, reduce, or inhibit the activity of protein or lipid phosphatases include inhibitors of phosphatase 1, phosphatase 2A, or CDC25, such as okadaic acid or derivatives thereof.

[0172] Other angiogenic compounds include those with an alternative mechanism of activity unrelated to protein or lipid kinase inhibition, such as thalidomide and TNP-470.

[0173] Other, non-limiting, exemplary chemotherapy compounds (one or more of which may be used in combination with the pharmaceutical compositions of this disclosure) include: Avastin, daunorubicin, doxorubicin, cytarabine, VP-16, teniposide, mitoxantrone, idarubicin, carboplatin, PKC412, 6-mercaptopurine (6-MP), fludarabine phosphate, octreotide, SOM230, FTY720, 6-thioguanine, cladribine, 6-mercaptopurine, pentostatin, hydroxyurea, 2-hydroxy-1H-isoindole-1,3- Diketone derivatives, l-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceutically acceptable salt thereof, 1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate, angiostatin, endostatin, anthranilamide, ZD4190, ZD6474, SU5416, SU6668, bevacizumab, rhuMAb, rhuFab, macuganon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgG1 antibody, RPI 4610, porphyrin sodium, anechodine, triamcinolone, hydrocortisone, 11-a-epihydrocotisol, corticosterone, 17a-hydroxyprogesterone, corticosterone, deoxycorticosterone, testosterone, estrone, dexamethasone, fluocinolone, phytoalkaloids, hormone compounds and / or antagonists, biological response modulators such as lymphokines or interferons, antisense oligonucleotides or oligonucleotide derivatives, shRNA and siRNA.

[0174] Many suitable alternative therapeutic agents (e.g., anticancer agents) have been considered for use in the treatment methods provided herein. In fact, the methods provided herein may include, but are not limited to, the administration of a wide range of alternative therapeutic agents, such as: agents that induce apoptosis; polynucleotides (e.g., antisense, ribozymes, siRNA); peptides (e.g., enzymes and antibodies); biomimics (e.g., gossypol or BH3 mimics); agents that bind to Bcl-2 family proteins such as Bax (e.g., oligomerization or complex formation); alkaloids; alkylating agents; antitumor antibiotics; antimetabolites; hormones; platinum compounds; monoclonal or polyclonal antibodies (e.g., those binding to anti-…). Cancer drugs, toxins, antibodies conjugated with defenseins; radionuclides; biological response modulators (e.g., interferons (e.g., IFN-α) and interleukins (e.g., IL-2)); adoptive immunotherapy agents; hematopoietic growth factors; agents that induce tumor cell differentiation (e.g., all-trans-retinoic acid); gene therapy agents (e.g., antisense agents and nucleotides); tumor vaccines; angiogenesis inhibitors; proteasome inhibitors: NF-κB modulators; anti-CDK compounds; HDAC inhibitors; etc. Numerous other examples of optional therapeutic agents (such as chemotherapy compounds and anticancer therapies) suitable for co-administration with the disclosed compounds are known to those skilled in the art.

[0175] In some embodiments, the optional therapeutic agent comprises an agent that induces or stimulates apoptosis. Agents that induce or stimulate apoptosis include, for example, agents that interact with or modify DNA (such as by intercalation, cross-linking, alkylation, or other methods of disrupting or chemically modifying DNA). Agents that induce apoptosis include, but are not limited to, radiation (e.g., X-rays, gamma rays, UV); tumor necrosis factor (TNF)-related factors (e.g., TNF family receptor proteins, TNF family ligands, TRAIL, antibodies against TRAIL-R1 or TRAIL-R2); and kinase inhibitors (e.g., epidermal growth factor receptor (EGFR) kinase inhibitors). Other anticancer agents include: angiogenesis factor receptor (VGFR) kinase inhibitors, fibroblast growth factor receptor (FGFR) kinase inhibitors, platelet-derived growth factor receptor (PDGFR) kinase inhibitors, and Bcr-Abl kinase inhibitors (such as Gleevec); antisense molecules; antibodies (e.g., Herceptin, RITUXAN, Zyvalil, and Avastin); anti-estrogens (e.g., raloxifene and tamoxifen); anti-androgens (e.g., flutamide, apalutamide, bicalutamide, finasteride, amglutide, ketoconazole, and corticosteroids); BCL-2 inhibitors (e.g., venetoclax); cyclooxygenase 2 (COX-2) inhibitors (e.g., celecoxib, meloxicam, NS-398, and nonsteroidal anti-inflammatory drugs (NSAIDs)); and anti-inflammatory drugs (e.g., phenylbutazone, decatetron, DELTASONE, dexamethasone, dexamethasone). Intensol, DEXONE, HEXADROL, hydroxychloroquine, METICORTEN, ORADEXON, ORASONE, hydroxybutanol, PEDIAPRED, phenylbutazone, PLAQUENIL, prednisolone, prednisone, PRELONE, and TANDEARIL; and cancer chemotherapy drugs (e.g., irinotecan (CAMPTOSAR), CPT-11, fludarabine (Fodahua), dacarbazine (DTIC), dexamethasone, mitoxantrone, merotazone, VP-16, cisplatin, carboplatin, oxaliplatin, 5-FU, doxorubicin, gemcitabine, bortezomib, gefitinib, bevacizumab, taxostatin, or paclitaxel); cell signaling molecules; ceramides and cytokines; astrococci, etc.

[0176] In other embodiments, the treatment methods provided herein include administering a therapeutically effective amount of the pharmaceutical composition of this disclosure, an immune checkpoint inhibitor, and at least one additional optional therapeutic agent, such as an alkylating agent, antimetabolite, and an antiproliferator or antitumor agent selected from natural products (e.g., herbs and other plant and / or animal-derived compounds).

[0177] Alkylating agents suitable for the method of the present invention include, but are not limited to: 1) nitrogen mustard (e.g., dichloromethyldiethylamine, cyclophosphamide, ifosfamide, melphalan (L-saccharin); and chlorambucil); 2) ethyleneimine and methylmelamine (e.g., hexamethylmelamine and thiotepa); 3) alkyl sulfonates (e.g., busulfan); 4) nitrosoureas (e.g., carmustine (BCNU); lomustine (CCNU); semustine (methylcyclohexanenitrosourea); and streptozotocin (streptozomycin)); and 5) triazines (e.g., dacarbazine (DTIC; dimethyltriazine imidazole carboxamide).

[0178] In some embodiments, antimetabolites suitable for the methods of the present invention include, but are not limited to: 1) folic acid analogs (e.g., methotrexate); 2) pyrimidine analogs (e.g., fluorouracil (5-fluorouracil; 5-FU), fluorouridine (fluorodeoxyuridine; FudR), and cytarabine (cytosine cytarabine)); and 3) purine analogs (e.g., mercaptopurine (6-mercaptopurine; 6-MP), thioguanine (6-thioguanine; TG), and pentostatin (2'-deoxymyostatin)).

[0179] In other embodiments, chemotherapeutic agents suitable for the methods of this disclosure include, but are not limited to: 1) vinca alkaloids (e.g., vincristine (VLB), vinblastine); 2) epipodophyllotoxin (e.g., etoposide and teniposide); 3) antibiotics (e.g., daunorubicin (actinomycin D), doxorubicin (daunorubicin; erythromycin), doxorubicin, bleomycin, procainomycin (procainomycin), and mitomycin (mitomycin C)); 4) enzymes (e.g., L-asparaginase); 5) biological response modulators (e.g., interferon-α); 6) platinum coordination complexes (e.g., cisplatin (cis-DDP) and carboplatin); 7) anthraquinones (e.g., mitoxantrone). 8) Substituted ureas (e.g., hydroxyurea); 9) Methylhydrazine derivatives (e.g., procarbazine (N-methylhydrazine; MIH)); 10) Adrenocortical inhibitors (e.g., mitotane (o,p'-DDD) and aminoglutethimide); 11) Adrenocortical steroids (e.g., prednisone); 12) Progestins (e.g., hydroxyprogesterone caproate, medroxyprogesterone acetate and megestrol acetate); 13) Estrogens (e.g., diethylstilbestrol and ethinylestradiol); 14) Antiestrogens (e.g., tamoxifen); 15) Androgens (e.g., testosterone propionate and flumethasone); 16) Antiandrogens (e.g., flutamide); and 17) Gonadotropin-releasing hormone analogs (e.g., leuprorelin).

[0180] Any oncolytic agent routinely used in the context of cancer therapy may be used in the treatments described in this disclosure. For example, the U.S. Food and Drug Administration (FDA) maintains a formulary of oncolytic agents approved for use in the United States. The FDA's international counterparts possess similar formularies. Those skilled in the art will understand that the required "product label" for all U.S.-approved chemotherapy agents describes the approved indications, dosing information, toxicity data, etc., of the exemplary agent.

[0181] Anticancer agents further include compounds that have been identified as having anticancer activity. Examples include, but are not limited to, 3-AP, 12-O-tetradecanoylphorbol-13-acetate, 17AAG, 852A, ABI-007, ABR-217620, ABT-751, ADI-PEG20, AE-941, AG-013736, AGRO100, Aranoxin, AMG 706, Antibody G250, Antitumor ketone, AP23573, Apazolidin, APC8015, Atemod, ATN-161, Atrasentan, Azacitidine, BB-10901, BCX-1777, Bevacizumab, BG00001, Bicalutamide, BMS 247550, Bortezomib, Lichenstatin-1, Buserelin, Long-acting PEGylated Asparaginase, Calcitriol, CCI-779, CDB-2914, Cefixime, Cetuximab, CG0070, Silengidate, Clofarabine, Cobustatin A4 Phosphate, CP-675, 206, CP-724, 714, CpG 7909, Curcumin, Daratumumab, Decitabine, DENSPM, Denutoximab, Ducalciferol, E7070, E7389, Seaweed Extract 743, Efaxiro, Eflunomide, EKB-569, Elotuzumab, Enzatolin, Erlotinib, Exisulin, Fenivel A, Vrappine, Fludarabine, Flutamide, Formusine, FR901228, G17DT, Garliximab, Gefitinib, Gentamicin, Graziebrion, Glufosinate, GTI-2040, Histamine Relin, HKI-272, Homoharringtonine, HSPPC-96, Hu14.18-Interleukin-2 fusion protein, HuMax-CD4, iloprost, imiquimod, infliximab, olibutrazumab, interleukin-12, IPI-504, iroflavone, ixaprilone, lapatinib, lenalidomide, leuprolide, LMB-9 immunotoxin, lonafaxine, ruximab, lutetium 177 dotatate, masphosphatamide, MB07133, MDX-010, MLN2704, mogliflozin, monoclonal antibody 3F8, monoclonal antibody J591, moxetumomab pasudotox-tdfk, MS-275, MVA-MUC1-IL2, nilumet, nirapanib, nitrocamptothecin, noratrexate hydrochloride, Novadex, NS-9, O6-benzylguanine, omeprazole sodium, ONYX-015, ozovozab, OSI-774, palumumab, berberine, PD-0325901, pemetrexed, PHY906, pioglitazone, pirfenidone, picotinib, polotuzumab, PS-341, PSC 833, PXD101, pyrazoloacridine, R115777, RAD001, leopard frog enzyme, davidimycin analogue, recombinant human angiostatin, rhuMab 2C4, Rosiglitazone, Rubotecan, Rucapapanib, S-1, S-8184, Saplatin, SB-, 15992, SGN-0010, SGN-40, Sonidazole, Sorafenib, SR31747A, ST1571, SU011248, Sulfamethoxazole, Sulfamethoxazole, Tagraxofusp-erzs, Talapopanib, Taliquid Tadalafil, Tamromoxetine, TGFa-PE38 immunotoxin, Thalidomide, Thymofasin, Tilpifab, Tirazamine, TLK286, Trabectin, Trifluridine and Tipyrimidine Hydrochloride, Trimethoprim Glucuronide, TroVax, UCN-1, Valproic acid, Vinflunidine, VNP40101M, Voloximab, Vorinostat, VX-680, ZD1839, ZD6474, Ziloutong and Zosquidone Trihydrochloride.

[0182] In one implementation, the optional therapeutic agent includes one of the anticancer drugs or combinations of anticancer drugs listed in Table 5.

[0183] Table 5

[0184]

[0185]

[0186]

[0187]

[0188]

[0189]

[0190]

[0191]

[0192] For a more detailed description of anticancer agents and other alternative treatments, those skilled in the art can refer to any number of guidance manuals, including, but not limited to, the Physician's Desk Reference and Goodman and Gilman, Pharmaceutical Basis of Therapeutics, 10th Edition, Hardman et al., 2002.

[0193] In another embodiment, the method of treating cancer provided herein includes administering the pharmaceutical composition of this disclosure in combination with radiation therapy and optionally an immune checkpoint inhibitor to a subject. The method provided herein is not limited by the type, amount, or delivery and administration system of radiation used to deliver the therapeutic dose to the patient. For example, the patient may receive photon radiation therapy, particle beam radiation therapy, other types of radiation therapy, or combinations thereof. In some embodiments, a linear accelerator is used to deliver radiation to the patient. In still other embodiments, a gamma knife is used to deliver radiation.

[0194] Radiation sources can be external to or internal to the patient. External radiation therapy is the most common and involves using, for example, a linear accelerator to guide a high-energy radiation beam through the skin to the tumor site. While the radiation beam is targeted to the tumor site, it is almost impossible to avoid exposure to normal healthy tissue. However, patients generally tolerate external radiation well. Internal radiation therapy involves implanting radiation-emitting sources, such as beads, threads, pellets, capsules, or granules, into the body at or near the tumor site, including the use of delivery systems that specifically target cancer cells (e.g., using ligands that bind to cancer cells). Such implants can be removed after treatment or remain inactive in the body. Types of internal radiation therapy include, but are not limited to, brachytherapy, interstitial irradiation, intracavitary irradiation, and radioimmunotherapy.

[0195] Patients may optionally receive radiosensitizers (e.g., metronidazole, misonitroazole, intra-arterial Budr, intravenous iodide-1H-imidazole (IudR), nitroimidazole, 5-substituted 4-nitroimidazole, 2H-isoindoledione, [[(2-bromoethyl)-amino]methyl]-nitro-1H-imidazole-1-ethanol, nitroaniline derivatives, DNA-affinity hypoxia-selective cytotoxins, halogenated DNA ligands, 1,2,4-benzotriazine oxides, 2-nitroimidazole derivatives, fluorinated nitroazole derivatives, benzoyl... Amines, nicotinamide, acridine-intercalating agents, 5-thiotetrazole derivatives, 3-nitro-1,2,4-triazole, 4,5-dinitroimidazole derivatives, hydroxylated tetraphrins, cisplatin, mitomycin, teirazamine, nitrosourea, mercaptopurine, methotrexate, fluorouracil, bleomycin, vincristine, carboplatin, epirubicin, doxorubicin, cyclophosphamide, vinblastine, etoposide, paclitaxel, heat (high heat), etc.), and radiation protectants (e.g., cysteine, aminoalkyl dihydrothiophosphate, amifostine (WR 2721), IL-1, IL-6, etc.). Radiosensitizers enhance the killing of tumor cells. Radiation protectants protect healthy tissues from the harmful effects of radiation.

[0196] Any type of radiation can be administered to a patient, provided the patient tolerates the radiation dose and there are no unacceptable negative side effects. Suitable types of radiation therapy include, for example, ionizing (electromagnetic) radiation therapy (e.g., X-rays or gamma rays) or particle beam radiation therapy (e.g., high linear energy radiation). Ionizing radiation is defined as radiation containing particles or photons with sufficient energy to produce ionization, i.e., an increase or loss of electrons (as described, for example, in US 5,770,581, which is incorporated herein by reference in its entirety). Clinicians can at least partially control the effects of radiation. In one embodiment, the radiation dose is graded to achieve maximum target cell exposure and reduced toxicity.

[0197] In one embodiment, the total dose of radiation administered to the patient is from about 0.01 Gy to about 100 Gy. In another embodiment, about 10 Gy to about 65 Gy is administered during treatment (e.g., about 15 Gy, 20 Gy, 25 Gy, 30 Gy, 35 Gy, 40 Gy, 45 Gy, 50 Gy, 55 Gy, or 60 Gy). While the full radiation dose may be administered over a single day in some embodiments, it is ideal to divide the total dose and administer it over several days. Ideally, radiation therapy is administered over a period of at least about 3 days, for example, at least 5, 7, 10, 14, 17, 21, 25, 28, 32, 35, 38, 42, 46, 52, or 56 days (about 1–8 weeks). Therefore, the daily radiation dose will consist of approximately 1-5 Gy (e.g., about 1 Gy, 1.5 Gy, 1.8 Gy, 2 Gy, 2.5 Gy, 2.8 Gy, 3 Gy, 3.2 Gy, 3.5 Gy, 3.8 Gy, 4 Gy, 4.2 Gy, or 4.5 Gy) or 1-2 Gy (e.g., 1.5-2 Gy). The daily radiation dose should be sufficient to induce the destruction of target cells. In one implementation, if the treatment is extended, radiation is not administered daily, allowing the animal to rest and achieve the therapeutic effect. For example, for weekly treatment, ideally, radiation is administered for 5 consecutive days with 2 days of no administration, allowing for 2 days of rest per week. However, radiation may be administered 1 day / week, 2 days / week, 3 days / week, 4 days / week, 5 days / week, 6 days / week, or all 7 days / week, depending on the animal's responsiveness and any potential side effects. Radiation therapy can be initiated at any time during the treatment phase. In one implementation, radiation is initiated in week 1 or week 2 and administered for the remainder of the treatment phase. For example, for the treatment of solid tumors, radiation is administered during weeks 1-6 or 2-6 of a 6-week treatment phase. Alternatively, radiation is administered during weeks 1-5 or 2-5 of a 5-week treatment phase. However, these exemplary radiotherapy administration schedules are not intended to limit the methods provided herein.

[0198] V. Method for preparing the lyophilized product of this disclosure

[0199] In another embodiment, this disclosure provides a method for preparing the lyophilized product of this disclosure, the method comprising:

[0200] (i) Prepare a pre-lyophilization solution containing compound 1, filler and water;

[0201] (ii) Cool the solution prior to lyophilization until it is frozen; and

[0202] (iii) Apply a vacuum to the frozen pre-lyophilized solution to produce the lyophilized product.

[0203] In another embodiment, the filler is mannitol.

[0204] In another embodiment, the pre-lyophilization solution further comprises ethanol. In yet another embodiment, the pre-lyophilization solution further comprises tert-butanol.

[0205] In another embodiment, this disclosure provides a method for preparing the lyophilized product of this disclosure, the method comprising:

[0206] (i) Dissolve compound 1 and a filler (e.g., mannitol) in a mixture of tert-butanol, ethanol and water at a temperature of about 20°C to about 50°C to produce a solution before lyophilization;

[0207] (ii) Cool the solution prior to lyophilization until it is frozen or partially frozen; and

[0208] (iii) Apply a vacuum to the frozen or partially frozen pre-lyophilized solution to produce the lyophilized product.

[0209] In another embodiment, compound 1 is dissolved in a mixture of tert-butanol, ethanol, and water at a temperature of about 25°C to about 35°C. In another embodiment, the temperature is about 25°C to about 30°C. In another embodiment, the temperature is about 25°C. In another embodiment, the temperature is about 30°C.

[0210] In another embodiment, the pre-lyophilization solution further comprises sodium carboxymethyl cellulose. In another embodiment, the pre-lyophilization solution further comprises poloxamer 188. In another embodiment, the pre-lyophilization solution further comprises soy lecithin. In another embodiment, the pre-lyophilization solution further comprises L-histidine. In another embodiment, the pre-lyophilization solution further comprises ethanol, tert-butanol, sodium carboxymethyl cellulose, poloxamer 188, soy lecithin, and L-histidine.

[0211] In another embodiment, the concentration of compound 1 in the solution before lyophilization is about 1 mg / mL to about 15 mg / mL. In another embodiment, the concentration of compound 1 in the solution before lyophilization is about 10 mg / mL.

[0212] VI. Methods for preparing pharmaceutical compositions

[0213] In another embodiment, this disclosure provides a method for preparing a pharmaceutical composition of this disclosure, the method comprising suspending a lyophilized product of this disclosure in a solvent. In another embodiment, the solvent comprises water.

[0214] VII. Reagent Kit

[0215] In another embodiment, this disclosure provides a kit comprising a lyophilized product of this disclosure packaged as a single unit dose in a tubular vial. In another embodiment, the tubular vial has a stopper and a cap. In yet another embodiment, the tubular vial is a glass bottle.

[0216] In another embodiment, this disclosure provides a kit for treating cancer in a subject, comprising a lyophilized version of this disclosure packaged as a single unit dose in a tubular vial.

[0217] In another embodiment, the kit further includes instructions on suspending the lyophilized material in a solvent to produce the pharmaceutical composition of this disclosure.

[0218] In another embodiment, the kit further includes instructions for administering the pharmaceutical composition of this disclosure to a subject.

[0219] In another embodiment, the kit may further comprise an optional therapeutic agent.

[0220] In another embodiment, the kit further includes a device suitable for subcutaneous administration of the pharmaceutical composition of this disclosure to a subject.

[0221] VIII. Definition

[0222] The terms “(S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester” and “compound 1” represent a prodrug of 6-diazo-5-oxo-L-leucine (DON) having the following structure:

[0223]

[0224] Compound 1 is described in US 10,336,778B2.

[0225] As used in this article, the term "lyophilized product" refers to powder obtained by lyophilization.

[0226] As used herein, the terms "lyophilization," "lyophilizing," and "lyophilized" refer to the lyophilization process by which compound 1 is frozen and, while still frozen, water and other solvents (if present) are removed by sublimation under vacuum. Compound 1 may be lyophilized in the presence of other reagents (e.g., fillers, cellulose-based suspension modifiers, poloxamer-based suspension modifiers, lecithin-based suspension modifiers, buffers) to enhance the properties of the resulting lyophilized product and / or the properties of any pharmaceutical composition containing the lyophilized product.

[0227] As used herein, the term "solvent" refers to a liquid (e.g., water) or a mixture of liquids (e.g., water and ethanol) suitable for administration to a subject as part of a pharmaceutical composition or formulation. In one embodiment, the solvent comprises a combination of water and one, two, three, or four other pharmaceutically acceptable water-miscible solvents (e.g., dioxane, dimethylacetamide, butanediol, polyethylene glycol, glycerol, ethanol, etc., or combinations thereof). In another embodiment, the solvent is water. In another embodiment, the solvent is a combination of water and one other pharmaceutically acceptable water-miscible solvent. In another embodiment, the solvent is a combination of water and ethanol. In another embodiment, the solvent comprises a combination of water and one, two, three, or four other pharmaceutically acceptable water-immiscible solvents (e.g., peanut oil, ethyl oleate, etc.). In another embodiment, the solvent comprises about 10% to about 30% water and about 70% to about 90% a water-miscible solvent, such as ethanol. In another embodiment, the solvent comprises about 15% to about 25% water and about 75% to about 85% a water-miscible solvent. In another embodiment, the solvent consists essentially of about 20% water and about 80% a water-miscible solvent.

[0228] As used herein, the term "filler" refers to a pharmaceutically acceptable excipient that provides structure to a freeze-dried cake to prevent shrinkage and collapse. Exemplary, non-limiting fillers include mannitol, lactose, sucrose, polyethylene glycol, and glycine.

[0229] As used herein, the term "cellulose-based suspension regulator" refers to a polysaccharide composed of linear β(1→4)-linked D-glucose units. Exemplary, non-limiting cellulose-based suspension regulators include hydroxypropyl cellulose, hydroxymethyl cellulose, and sodium carboxymethyl cellulose.

[0230] As used herein, the term "poloxam-based suspension modifier" refers to a nonionic triblock copolymer comprising a central hydrophobic chain of polyoxypropylene flanked by two hydrophilic chains of polyethylene oxide. Exemplary, non-limiting poloxamer-based suspension modifiers include poloxamer 188,NF (…). F-68) and Polosham 407 F-127).

[0231] As used herein, the term "lecithin-based suspension regulator" refers to a mixture containing glycerophospholipids, such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and phosphatidic acid. An exemplary, non-limiting lecithin-based suspension regulator is soybean lecithin.

[0232] As used herein, the term "buffer" refers to a pharmaceutically acceptable excipient that helps maintain pH during lyophilization and after remodeling of the resulting lyophilized product. Exemplary, non-limiting buffers include glycine, L-histidine, phosphate, acetic acid, lactic acid, citric acid, and Tris.

[0233] As used herein, the term "diluent" refers to a liquid used to dilute a pharmaceutical composition prior to parenteral administration to a subject. In one embodiment, the diluent is physiological saline, 5% dextran, lactate Ringer's solution, or any other sterile liquid designed to be compatible with human administration (e.g., via intravenous infusion). In another embodiment, the diluent is physiological saline.

[0234] As used herein, the term "weight ratio" refers to the mass of Compound 1 in the lyophilized product divided by the mass of another reagent (e.g., a filler or buffer). For example, in a lyophilized product containing 67.5 mg of Compound 1 and 135 mg of mannitol, the weight ratio of Compound 1 to filler is 0.5. In a lyophilized product containing 67.5 mg of Compound 1, 135 mg of mannitol, and 3.88 mg of L-histidine, the weight ratio of Compound 1 to buffer is 17.4.

[0235] The term “%wt / wt” as used herein refers to the mass of a lyophilized ingredient divided by the combined mass of all lyophilized ingredients, multiplied by 100. For example, in a lyophilized product containing 62.5 mg of Compound 1, 125 mg of mannitol, 8.33 mg of sodium carboxymethyl cellulose (NaCMC), 7.08 mg of poloxamer 188,NF, 4.31 mg of histidine, and 41.7 mg of soybean lecithin, the %wt / wt of Compound 1 is 25.1% (62.5 mg / 248.92 mg = 0.251 x 100 = 25.1% wt / wt).

[0236] The terms "average particle size distribution" or "D" used in this article 50"The diameter is determined by laser diffraction, where 50% by mass of the particles have a larger equivalent diameter, while the other 50% by mass of the particles have a smaller equivalent diameter."

[0237] As used herein, the terms "intermittent dosing," "intermittent dosing regimen," and similar terms refer to the non-continuous administration of the pharmaceutical composition of this disclosure to a subject. Intermittent dosing regimens available in this disclosure cover any non-continuous administration regimen that provides a therapeutically effective amount of the pharmaceutical composition of this disclosure to a subject in need. Intermittent dosing regimens may use doses of the pharmaceutical composition of this disclosure equal to, lower than, or higher than those used in continuous dosing regimens. The advantages of intermittent dosing include, but are not limited to, improved safety, reduced toxicity (e.g., reduced weight loss), increased exposure, increased potency, and / or increased subject compliance. These advantages can be achieved when the pharmaceutical composition of this disclosure is administered as a single agent or when administered in combination with one or more other therapeutic agents (e.g., immune checkpoint inhibitors).

[0238] In one embodiment, the pharmaceutical composition of this disclosure is administered to a subject according to an intermittent dosing regimen to treat cancer. In another embodiment, the intermittent dosing regimen increases the therapeutic index of the pharmaceutical composition of this disclosure. The therapeutic index is a comparison of the amount of the pharmaceutical composition of this disclosure that causes a therapeutic effect (e.g., tumor shrinkage, increased time to tumor progression, and / or increased subject survival) to the amount that causes toxicity (e.g., weight loss).

[0239] In one embodiment, the pharmaceutical composition of this disclosure is administered to a subject every other day.

[0240] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject once a week.

[0241] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject twice a week on consecutive days (e.g., on Monday and Tuesday).

[0242] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject twice a week on non-consecutive days (e.g., on Monday and Wednesday, on Monday and Thursday, or on Tuesday and Friday).

[0243] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject twice a week, on Mondays and Thursdays.

[0244] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject twice a week, on Tuesdays and Fridays.

[0245] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject three times a week on consecutive days (e.g., on Monday, Tuesday, and Wednesday).

[0246] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject three times a week on non-consecutive days (e.g., on Mondays, Wednesdays, and Fridays).

[0247] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject four times a week on consecutive days (e.g., on Monday, Tuesday, Wednesday, and Thursday).

[0248] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject four times a week on non-consecutive days (e.g., Monday, Tuesday, Thursday, and Friday).

[0249] In another embodiment, the pharmaceutical composition of this disclosure is administered to the subject five times a week on consecutive days (e.g., on Monday, Tuesday, Wednesday, Thursday, and Friday).

[0250] The terms “parallel administration,” “combined administration,” “simultaneous administration,” and similar phrases refer to the administration of two or more pharmaceutical agents in parallel to a subject being treated. “Parallel” means that each agent is administered simultaneously or sequentially at different time points, either at the same time or in any order. However, if not administered simultaneously, it means that they are administered to the individual sequentially and sufficiently close in time to provide the desired therapeutic effect and to act synergistically. For example, the pharmaceutical compositions of this disclosure may be administered to a subject simultaneously or sequentially at different time points, either with optional therapeutic agents. The pharmaceutical compositions and optional therapeutic agents of this disclosure may be administered separately in any suitable form and via any suitable route. When the pharmaceutical compositions and optional therapeutic agents of this disclosure are not administered in parallel, it should be understood that they may be administered to a subject in need in any order. For example, the pharmaceutical composition of this disclosure may be administered before (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week or longer before), concurrently with or after (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours, 1 week or longer after) an optional therapeutic agent.

[0251] The terms “a” and “a kind” mean one or more kinds.

[0252] The term “about” as used in this article includes ±10% of the listed numbers. Therefore, “about 10” refers to 9-11.

[0253] As used herein, the terms “treat,” “treating,” “treatment,” “method of treatment,” etc., mean the elimination, reduction, or improvement of a disease or condition and / or its associated symptoms. While not excluded, treating a disease or condition does not necessarily require the complete elimination of the disease, condition, or its associated symptoms. However, in one embodiment, administration of compound 1, along with optionally one or more optional therapeutic agents, results in remission of cancer.

[0254] As used herein, the term "therapeutic effective amount" refers to an amount of therapeutic agent sufficient to cause improvement in one or more symptoms of the disorder, or to prevent the progression of the disorder, or to cause the remission of the disorder. For example, in relation to cancer treatment, in one embodiment, the therapeutic effective amount would refer to an amount of therapeutic agent that elicits a therapeutic response, such as normalization of blood cell counts, reduction in tumor growth rate, reduction in tumor mass, reduction in the number of metastases, increase in time to tumor progression, and / or increase in subject survival by at least about 2%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or at least about 100% or more.

[0255] The terms “pharmaceutically acceptable carrier” or “pharmaceutically acceptable medium” encompass any standard drug carrier, solvent, surfactant, or medium. Suitable pharmaceutically acceptable media include aqueous and non-aqueous media. Standard drug carriers and their formulations are described in Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton, PA, 1995, 19th edition.

[0256] The term "container" refers to any reservoir and enclosure suitable for storing, transporting, dispensing, and / or handling pharmaceutical products. In one embodiment, the container is a tubular bottle.

[0257] The term "insert" refers to information accompanying a pharmaceutical product that provides a description of how to administer the product, as well as safety and efficacy data necessary to allow physicians, pharmacists, and patients to make informed decisions about its use. Inserts on packaging are often considered the "label" of a pharmaceutical product.

[0258] In some embodiments, when administered in combination, two or more agents may have a synergistic effect. The terms “synergistic,” “synergistic,” “synergistically,” and their derivatives, as used herein, such as in “synergistic effect,” “synergistic combination,” or “synergistic composition,” indicate a situation where the biological activity of the agent in combination with at least one other therapeutic agent is greater than the sum of the biological activities of the individual agents when administered alone. For example, the term “synergistically effective” as used herein refers to an interaction between compound 1 and one or more immune checkpoint inhibitors, resulting in a total effect of said drug that is greater than the sum of the individual effects of each drug. See, for example, Berenbaum, Pharmacological Reviews 41:93-141 (1989).

[0259] IX. Specific Implementation Plans

[0260] In another aspect, this disclosure provides the following specific implementation schemes.

[0261] Implementation Scheme 1. A freeze-dried product comprising compound 1 and a filler.

[0262] Implementation Scheme 2. The lyophilized product of Implementation Scheme 1, which contains approximately 67.5 mg of compound 1.

[0263] Implementation Scheme 3. The lyophilized product of Implementation Scheme 1 or 2, wherein the weight ratio of compound 1 to filler is about 5 to about 0.05.

[0264] Implementation Scheme 4. The lyophilized product of Implementation Scheme 3, wherein the weight ratio of compound 1 to filler is about 1 to about 0.2.

[0265] Implementation Scheme 5. The lyophilized product of Implementation Scheme 5, wherein the weight ratio of compound 1 to filler is about 0.5.

[0266] Implementation Scheme 6. The freeze-dried product of any one of Implementation Schemes 1-5, wherein the filler is mannitol.

[0267] Implementation Scheme 7. The freeze-dried product of Implementation Scheme 6 contains approximately 135 mg of mannitol.

[0268] Implementation Scheme 8. The freeze-dried product of any one of Implementation Schemes 1-7, further comprising a cellulose-based suspension regulator.

[0269] Implementation Scheme 9. The freeze-dried product of Implementation Scheme 8, wherein the ratio of compound 1 to cellulose-based suspension regulator is about 25 to about 0.3.

[0270] Implementation Scheme 10. The freeze-dried product of Implementation Scheme 9, wherein the weight ratio of compound 1 to cellulose-based suspension regulator is about 5 to about 2.

[0271] Implementation Scheme 11. The lyophilized product of Implementation Scheme 10, wherein the weight ratio of compound 1 to cellulose-based suspension regulator is about 2.7 or about 7.5.

[0272] Implementation Scheme 12. The method of any one of Implementation Schemes 8-11, wherein the cellulose-based suspension regulator is sodium carboxymethyl cellulose.

[0273] Implementation Scheme 13. The lyophilized product of Implementation Scheme 12, which contains about 25.2 mg or about 9.0 mg of sodium carboxymethyl cellulose.

[0274] Implementation Scheme 14. The lyophilized product of any one of Implementation Schemes 1-13, further comprising a poloxamer-based suspension modifier.

[0275] Implementation Scheme 15. The lyophilized product of Implementation Scheme 14, wherein the ratio of compound 1 to poloxamer-based suspension modifier is about 80 to about 1.

[0276] Implementation Scheme 16. The lyophilized product of Implementation Scheme 15, wherein the weight ratio of compound 1 to the poloxamer-based suspension modifier is about 10 to about 6.

[0277] Implementation Scheme 17. The lyophilized product of Implementation Scheme 16, wherein the weight ratio of compound 1 to the poloxamer-based suspension modifier is about 8.8.

[0278] Implementation Scheme 18. The freeze-dried product of any one of Implementation Schemes 14-17, wherein the poloxamer-based suspension modifier is poloxamer 188.

[0279] Implementation Scheme 19. The lyophilized product of Implementation Scheme 18, which contains approximately 7.65 mg of poloxamer 188.

[0280] Implementation Scheme 20. The lyophilized product of any one of Implementation Schemes 1-19, further comprising a lecithin-based suspension regulator.

[0281] Implementation Scheme 21. The lyophilized product of Implementation Scheme 20, wherein the ratio of compound 1 to lecithin-based suspension regulator is about 20 to about 0.1.

[0282] Implementation Scheme 22. The lyophilized product of Implementation Scheme 21, wherein the weight ratio of compound 1 to the lecithin-based suspension regulator is about 10 to about 0.5.

[0283] Implementation Scheme 23. The lyophilized product of Implementation Scheme 22, wherein the weight ratio of compound 1 to the lecithin-based suspension regulator is about 1.5.

[0284] Implementation Scheme 24. The freeze-dried product of any one of Implementation Schemes 20-23, wherein the lecithin-based suspension regulator is soybean lecithin.

[0285] Implementation Scheme 25. The freeze-dried product of Implementation Scheme 24 contains approximately 45 mg of soybean lecithin.

[0286] Implementation Scheme 26. The freeze-dried product of any one of Implementation Schemes 1-25, further comprising a buffer.

[0287] Implementation Scheme 27. The lyophilized product of Implementation Scheme 26, wherein the weight ratio of compound 1 to buffer is about 50 to about 1.

[0288] Implementation Scheme 28. The lyophilized product of Implementation Scheme 27, wherein the weight ratio of compound 1 to buffer is about 20 to about 10.

[0289] Implementation Scheme 29. The lyophilized product of Implementation Scheme 28, wherein the weight ratio of compound 1 to buffer is about 17.4.

[0290] Implementation scheme 30. The lyophilized product of any one of implementation schemes 26-29, wherein the buffer is L-histidine.

[0291] Implementation Scheme 31. The lyophilized product of Implementation Scheme 30 contains approximately 3.88 mg of L-histidine.

[0292] Implementation Scheme 32. A pharmaceutical composition comprising a lyophilized product of any one of Implementation Schemes 1-31, wherein the lyophilized product is suspended in a solvent.

[0293] Implementation Scheme 33. The pharmaceutical composition of Implementation Scheme 32, wherein the solvent comprises water.

[0294] Implementation Scheme 34. The pharmaceutical composition of Implementation Scheme 32 or 33, wherein the concentration of said compound 1 is from about 25 mg / mL to about 30 mg / mL.

[0295] Implementation Scheme 35. The pharmaceutical composition of any one of Implementation Schemes 32-34, wherein after 3 days at 2-8°C, the average particle size distribution of compound 1 in the suspension is about 1 μm to about 30 μm.

[0296] Implementation Scheme 36. A method for treating cancer in a subject with such need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of any one of Implementation Schemes 32-35.

[0297] Implementation Scheme 37. The method of Implementation Scheme 36, wherein the pharmaceutical composition is administered subcutaneously to the subject.

[0298] Implementation scheme 38. The method of implementation scheme 36 or 37, the method further comprising administering an optional therapeutic agent to the subject.

[0299] Implementation Scheme 39. A method for preparing the lyophilized product of Implementation Scheme 1, the method comprising:

[0300] (i) Prepare a solution containing compound 1, filler and water before lyophilization;

[0301] (ii) Cool the solution prior to lyophilization until it is frozen or partially frozen; and

[0302] (iii) Apply a vacuum to the frozen or partially frozen pre-lyophilized solution to produce the lyophilized product.

[0303] Implementation Scheme 40. The method of Implementation Scheme 39, wherein the filler is mannitol.

[0304] Implementation Scheme 41. The method of Implementation Scheme 39 or 40, wherein the solution prior to lyophilization further comprises ethanol.

[0305] Implementation Scheme 42. The method of any one of Implementation Schemes 39-41, wherein the solution before lyophilization further comprises tert-butanol.

[0306] Implementation Scheme 43. The method of any one of Implementation Schemes 39-42, wherein the solution prior to lyophilization further comprises sodium carboxymethyl cellulose.

[0307] Implementation Scheme 44. The method of any one of Implementation Schemes 39-43, wherein the solution prior to lyophilization further comprises poloxamer 188.

[0308] Implementation Scheme 45. The method of any one of Implementation Schemes 39-44, wherein the pre-lyophilized solution further comprises soybean lecithin.

[0309] Implementation Scheme 46. The method of any one of Implementation Schemes 39-45, wherein the solution before lyophilization further comprises L-histidine.

[0310] Implementation Scheme 47. A method for preparing the pharmaceutical composition of Implementation Scheme 32, the method comprising suspending the lyophilized material in a solvent.

[0311] Implementation Scheme 48. The method of Implementation Scheme 47, wherein the solvent comprises water.

[0312] Implementation Scheme 49. A reagent kit comprising a lyophilized product of any one of Implementation Schemes 1-31 packaged as a single unit dose in a tubular vial.

[0313] Implementation scheme 50. A kit for treating cancer in subjects with this need, according to implementation scheme 49.

[0314] Implementation scheme 51. The kit of implementation scheme 49 or 50, wherein the kit further comprises instructions on suspending the lyophilized material in a solvent to produce a pharmaceutical composition.

[0315] Implementation Scheme 52. The kit of Implementation Schemes 49-51, wherein the kit further comprises instructions for administering the pharmaceutical composition to the subject.

[0316] In another aspect, this disclosure provides the following specific implementation schemes.

[0317] Implementation Scheme A1. A lyophilized product comprising about 20% wt / wt to about 45% wt / wt of compound 1 and about 35% wt / wt to about 55% wt / wt of filler.

[0318] Implementation Scheme A2. The lyophilized product of Implementation Scheme A1 further comprises about 1% wt / wt to about 10% wt / wt of a cellulose-based suspension regulator.

[0319] Implementation Scheme A3. The lyophilized product of Implementation Scheme A1 or A2 further comprises about 1% wt / wt to about 5% wt / wt of a poloxamer-based suspension modifier.

[0320] Implementation scheme A4. The lyophilized product of any one of implementation schemes A1-A3, further comprising about 10% wt / wt to about 20% wt / wt of a lecithin-based suspension regulator.

[0321] Implementation Scheme A5. The lyophilized product of any one of Implementation Schemes A1-A5 further comprises a buffer of about 0.5% wt / wt to about 2.5% wt / wt.

[0322] Implementation Scheme A6. The freeze-dried product of any one of Implementation Schemes A1-A5, comprising:

[0323] (i) Compound 1, approximately 20% wt / wt to approximately 30% wt / wt;

[0324] (ii) Fillers of about 45% wt / wt to about 55% wt / wt;

[0325] (iii) Cellulose-based suspension modifiers of about 1% wt / wt to about 9% wt / wt;

[0326] (iv) Poloxamer-based suspension modifiers of about 1% wt / wt to about 4% wt / wt;

[0327] (v) a lecithin-based suspension modifier of about 12% wt / wt to about 20% wt / wt; and

[0328] (vi) Buffer of about 1% wt / wt to about 2% wt / wt.

[0329] Implementation scheme A7. A lyophilized product of any one of implementation schemes A1-A6, comprising about 25.1% wt / wt of compound 1.

[0330] Implementation scheme A8. The lyophilized product of any one of implementation schemes A1-A7, which contains about 50.2% wt / wt of filler.

[0331] Implementation scheme A9. The lyophilized product of any one of implementation schemes A1-A8, which contains about 3.3% wt / wt of a cellulose-based suspension regulator.

[0332] Implementation scheme A10. The lyophilized product of any one of implementation schemes A1-A9, which contains about 2.8% wt / wt of a poloxamer-based suspension modifier.

[0333] Implementation scheme A11. A lyophilized product of any one of implementation schemes A1-A10, comprising about 16.8% wt / wt of a lecithin-based suspension modifier.

[0334] Implementation scheme A12. The lyophilized product of any one of implementation schemes A1-A11, which contains about 1.7% wt / wt of buffer.

[0335] Implementation scheme A13. The lyophilized product of any one of implementation schemes A1-A12, wherein the filler is mannitol.

[0336] Implementation scheme A14. The freeze-dried product of any one of implementation schemes A1-A13, wherein the cellulose-based suspension regulator is sodium carboxymethyl cellulose.

[0337] Implementation Scheme A15. The lyophilized product of any one of Implementation Schemes A1-A14, wherein the poloxamer-based suspension modifier is poloxamer 188 (a copolymer of ethylene oxide and propylene oxide).

[0338] Implementation scheme A16. The freeze-dried product of any one of implementation schemes A1-A15, wherein the lecithin-based suspension regulator is soybean lecithin.

[0339] Implementation scheme A17. A lyophilized product of any one of implementation schemes A1-A16, wherein the buffer is histidine.

[0340] Implementation Scheme A18. The lyophilized product of Implementation Scheme A1, comprising:

[0341] (i) Approximately 62.5 mg of compound 1;

[0342] (ii) Approximately 125 mg of mannitol;

[0343] (iii) Approximately 8.3 mg of sodium carboxymethyl cellulose;

[0344] (iv) Approximately 7.1 mg poloxamer 188, NF;

[0345] (v) Approximately 41.7 mg of soy lecithin; and

[0346] (vi) Approximately 4.3 mg of histidine.

[0347] Implementation Scheme A19. A pharmaceutical composition comprising a lyophilized product of any one of Implementation Schemes A1-A18, wherein the lyophilized product is suspended in a pharmaceutically acceptable carrier.

[0348] Implementation scheme A20. Pharmaceutical composition of implementation scheme A19, wherein the pharmaceutically acceptable carrier comprises water.

[0349] Implementation Scheme A21. The pharmaceutical composition of Implementation Scheme A19 or A20, wherein the concentration of said compound 1 is from about 20 mg / mL to about 60 mg / mL.

[0350] Implementation Scheme A22. The pharmaceutical composition of Implementation Scheme A21, wherein the concentration of compound 1 is about 25 mg / mL.

[0351] Implementation scheme A23. A pharmaceutical composition of any one of implementation schemes A19-22, wherein the average particle size distribution of compound 1 in the suspension is about 1 μm to about 30 μm.

[0352] Implementation Scheme A24. A method for treating cancer in a subject with such need, the method comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition of any one of Implementation Schemes A19-A23.

[0353] Implementation scheme A25. The method of implementation scheme A24, wherein the cancer is a solid tumor.

[0354] Implementation scheme A26. The method of implementation scheme A24, wherein the cancer is a hematologic cancer.

[0355] Implementation scheme A27. The method of implementation scheme A24, wherein the cancer is one or more of the cancers listed in Table 1.

[0356] Implementation scheme A28. The method of implementation scheme A24, wherein the cancer is hepatocellular carcinoma, glioblastoma, lung cancer, breast cancer, head and neck cancer, prostate cancer, melanoma, or colorectal cancer.

[0357] Implementation scheme A29. The method of implementation scheme A24, wherein the cancer is colorectal cancer, breast cancer, lymphoma, melanoma, kidney cancer or lung cancer.

[0358] Implementation scheme A30. The method of implementation scheme A24, wherein the cancer is an advanced solid tumor, non-small cell lung cancer, or squamous cell carcinoma of the head and neck.

[0359] Implementation scheme A31. The method of any one of implementation schemes A24-30, wherein the pharmaceutical composition is administered subcutaneously to the subject.

[0360] Implementation scheme A32. The method of implementation scheme A31, wherein the pharmaceutical composition is administered to the subject according to an intermittent dosing regimen.

[0361] Implementation scheme A33. The method of implementation scheme A32, wherein the pharmaceutical composition is administered to the subject twice a week on non-consecutive days.

[0362] Implementation scheme A34. The method of any one of implementation schemes A24-3A3, the method further comprising administering one or more optional therapeutic agents to the subject.

[0363] Implementation scheme A35. The method of implementation scheme A34, wherein the one or more optional therapeutic agents comprise one or more immune checkpoint inhibitors.

[0364] Implementation scheme A36. The method of implementation scheme A35, wherein the one or more immune checkpoint inhibitors comprise PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors, or TIGIT inhibitors, or combinations thereof.

[0365] Implementation scheme A37. The method of implementation scheme A36, wherein the one or more immune checkpoint inhibitors comprises an anti-PD-1 antibody.

[0366] Implementation Scheme A38. The method of Implementation Scheme A37, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, pildizumab, STI-A1110, spartalizumab, JTX-4014, sintilimab, MEDI0680, AGEN2034, BGB-A317, AB122, dostarlimab, PF-06801591, cimiprimab, tislelizumab, toripalimab, camrelizumab, SYM021, JNJ 63723283, HLX10, LZM009, or MGA012.

[0367] Implementation scheme A39. The method of any one of implementation schemes A35-A38, wherein the one or more immune checkpoint inhibitors comprises an anti-PD-L1 antibody.

[0368] Implementation scheme A40. The method of implementation scheme A39, wherein the anti-PD-L1 antibody is averrucumab, atezolizumab, durvalumab, KN035 or STI-A1014.

[0369] Implementation scheme A41. The method of any one of implementation schemes A35-A40, wherein the one or more immune checkpoint inhibitors comprises an anti-CTLA-4 antibody.

[0370] Implementation scheme A42. The method of implementation scheme A41, wherein the anti-CTLA-4 antibody is ipilimumab or trimemumab.

[0371] Implementation scheme A43. The method of any one of implementation schemes A35-A42, wherein the one or more immune checkpoint inhibitors comprises an anti-LAG3 antibody.

[0372] Implementation scheme A44. The method of implementation scheme A43, wherein the anti-LAG3 antibody is GSK2831781.

[0373] Implementation scheme A45. The method of any one of implementation schemes A35-A44, wherein the one or more immune checkpoint inhibitors comprises an anti-TIM3 antibody.

[0374] Implementation scheme A46. The method of any one of implementation schemes A35-A45, wherein the one or more immune checkpoint inhibitors comprises an anti-TIGIT antibody.

[0375] Implementation Scheme A47. The method of Implementation Scheme A46, wherein the anti-TIGIT antibody is vimbrolizumab (MK-7684), tirelimumab (RG6058), EOS-448, BMS-986207, BGB-A1217, MTIG7192A, AB154, ASP8374 or MK-7684.

[0376] Implementation scheme A48. The method of any one of implementation schemes A35-A47, wherein the cancer is resistant to or has become resistant to treatment with at least one immune checkpoint inhibitor.

[0377] Implementation scheme A49. A kit comprising a lyophilized product of any one of implementation schemes A1-A18 packaged as a single unit dose in a container.

[0378] The kit of implementation scheme A50 and implementation scheme A49 further includes instructions on suspending the lyophilized material in a solvent to produce a pharmaceutical composition.

[0379] Implementation scheme A51. The kit according to implementation scheme A49 or A50, the kit further comprising a label having instructions on how to use the kit according to any one of implementation schemes A24-48.

[0380] Implementation scheme A52. The kit according to implementation scheme A51, wherein the label is approved by the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), the China Food and Drug Administration (CFDA), or the Japanese Ministry of Health, Labor and Welfare (MHLW).

[0381] Implementation Scheme A53. A method for preparing the lyophilized product of Implementation Scheme A1, the method comprising:

[0382] (i) Prepare a solution containing compound 1, filler and water before lyophilization;

[0383] (ii) Cool the solution prior to lyophilization until it is frozen or partially frozen; and

[0384] (iii) Apply a vacuum to the frozen or partially frozen pre-lyophilized solution to produce the lyophilized product.

[0385] Implementation scheme A54. The method of implementation scheme A54, wherein the filler is mannitol.

[0386] Implementation scheme A55. The method of implementation scheme A53 or A54, wherein the solution before lyophilization further comprises ethanol.

[0387] Implementation scheme A56. The method of any one of implementation schemes A53-A55, wherein the solution before lyophilization further comprises tert-butanol.

[0388] Implementation scheme A57. The method of any one of implementation schemes A53-A56, wherein the solution before lyophilization further comprises sodium carboxymethyl cellulose.

[0389] Implementation scheme A58. The method of any one of implementation schemes A53-A57, wherein the solution before lyophilization further comprises poloxamer 188.

[0390] Implementation scheme A59. The method of any one of implementation schemes A53-A58, wherein the solution before freeze-drying further comprises soybean lecithin.

[0391] Implementation scheme A60. The method of any one of implementation schemes A53-A59, wherein the solution before lyophilization further comprises L-histidine.

[0392] Implementation Scheme A61. A method for preparing a pharmaceutical composition according to Implementation Scheme A19, the method comprising suspending the lyophilized material in a solvent.

[0393] Implementation scheme A62. The method of implementation scheme A61, wherein the solvent comprises water.

[0394] Implementation Scheme A63. A kit for implementing the method of any one of Implementation Schemes A24-A48, said kit comprising a single unit dose of the lyophilized product of any one of Implementation Schemes A1-18 packaged in a container.

[0395] Implementation scheme A64. The kit of implementation scheme A63, wherein the kit further comprises instructions on suspending the lyophilized material in a solvent to produce a pharmaceutical composition.

[0396] Implementation scheme A65. The kit according to implementation scheme A63 or A64, wherein the kit further comprises a label approved by the U.S. Food and Drug Administration (FDA), the European Medicines Agency (EMA), the China Food and Drug Administration (CFDA), or the Japanese Ministry of Health, Labour and Welfare (MHLW).

[0397] Implementation scheme A66. A pharmaceutical composition of any one of implementation schemes A19-A23 for treating cancer in subjects in need of such treatment.

[0398] Implementation scheme A67. A pharmaceutical composition for use in implementation scheme A66, wherein the cancer is a solid tumor.

[0399] Implementation scheme A68. A pharmaceutical composition for use in implementation scheme A66, wherein the cancer is a hematologic cancer.

[0400] Implementation scheme A69. A pharmaceutical composition for use in implementation scheme A66, wherein the cancer is one or more of the cancers listed in Table 1.

[0401] Implementation scheme A70. A pharmaceutical composition for use in implementation scheme A66, wherein the cancer is hepatocellular carcinoma, glioblastoma, lung cancer, breast cancer, head and neck cancer, prostate cancer, melanoma, or colorectal cancer.

[0402] Implementation Scheme A71. A pharmaceutical composition for use in Implementation Scheme A66, wherein the cancer is colorectal cancer, breast cancer, lymphoma, melanoma, kidney cancer, or lung cancer.

[0403] Implementation Scheme A72. A pharmaceutical composition for use in Implementation Scheme A66, wherein the cancer is an advanced solid tumor, non-small cell lung cancer, or squamous cell carcinoma of the head and neck.

[0404] Implementation scheme A73. A pharmaceutical composition for use in any one of implementation schemes A66-A72, wherein the pharmaceutical composition is administered subcutaneously to the subject.

[0405] Implementation scheme A74. A pharmaceutical composition for use in implementation scheme A73, wherein the pharmaceutical composition is administered to the subject according to an intermittent dosing regimen.

[0406] Implementation scheme A75. A pharmaceutical composition for use in implementation scheme A74, wherein the pharmaceutical composition is administered to the subject twice a week on non-consecutive days.

[0407] Implementation Scheme A76. A pharmaceutical composition for use in any of Implementation Schemes A66-A75, wherein the pharmaceutical composition is administered to the subject together with one or more optional therapeutic agents.

[0408] Implementation Scheme A77. A pharmaceutical composition for use in Implementation Scheme A76, wherein the one or more optional therapeutic agents comprise one or more immune checkpoint inhibitors.

[0409] Implementation Scheme A78. A pharmaceutical composition for use in Implementation Scheme A77, wherein the one or more immune checkpoint inhibitors comprise a PD-1 inhibitor, a PD-L1 inhibitor, a CTLA-4 inhibitor, a LAG3 inhibitor, a TIM3 inhibitor, or a TIGIT inhibitor, or a combination thereof.

[0410] Implementation Scheme A79. A pharmaceutical composition for use in Implementation Scheme A78, wherein the one or more immune checkpoint inhibitors comprise an anti-PD-1 antibody.

[0411] Implementation Scheme A80. A pharmaceutical composition for use in Implementation Scheme A79, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, pildizumab, STI-A1110, (spartazumab), JTX-4014, sintilimab, MEDI 0680, AGEN2034, BGB-A317, AB122, dotalimab, PF-06801591, cimiprimab, tislelizumab, toripalimab, camrelizumab, SYM021, JNJ 63723283, HLX10, LZM009, or MGA012.

[0412] Implementation Scheme A81. A pharmaceutical composition for use in any of Implementation Schemes A77-A80, wherein the one or more immune checkpoint inhibitors comprise an anti-PD-L1 antibody.

[0413] Implementation Scheme A82. A pharmaceutical composition for use in Implementation Scheme A81, wherein the anti-PD-L1 antibody is avelumab, atezolizumab, durvalumab, KN035, or STI-A1014.

[0414] Implementation Scheme A83. A pharmaceutical composition for use in any of Implementation Schemes A77-A82, wherein the one or more immune checkpoint inhibitors comprise an anti-CTLA-4 antibody.

[0415] Implementation Scheme A84. A pharmaceutical composition for use in Implementation Scheme A83, wherein the anti-CTLA-4 antibody is ipilimumab or trimemumab.

[0416] Implementation Scheme A85. A pharmaceutical composition for use in any of Implementation Schemes A77-A84, wherein the one or more immune checkpoint inhibitors comprise an anti-LAG3 antibody.

[0417] Implementation Scheme A86. A pharmaceutical composition for use in Implementation Scheme A85, wherein the anti-LAG3 antibody is GSK2831781.

[0418] Implementation scheme A87. A pharmaceutical composition for use in any of implementation schemes A77-A86, wherein the one or more immune checkpoint inhibitors comprise an anti-TIM3 antibody.

[0419] Implementation scheme A88. A pharmaceutical composition for use in any of implementation schemes A77-A87, wherein the one or more immune checkpoint inhibitors comprise an anti-TIGIT antibody.

[0420] Implementation Scheme A89. A pharmaceutical composition for use in Implementation Scheme A88, wherein the anti-TIGIT antibody is vimbrolizumab (MK-7684), tirelinumab (RG6058), EOS-448, BMS-986207, BGB-A1217, MTIG7192A, AB154, ASP8374, or MK-7684.

[0421] Implementation Scheme A90. A pharmaceutical composition for use in any of Implementation Schemes A77-A89, wherein the cancer is resistant to or has become resistant to treatment with at least one immune checkpoint inhibitor.

[0422] Implementation Scheme A91. Use of the pharmaceutical composition of any one of Implementation Schemes A19-A23 in the preparation of a medicament for treating cancer in a subject in need of such treatment.

[0423] Implementation scheme A92. Use of implementation scheme A91, wherein the cancer is a solid tumor.

[0424] Implementation scheme A93. Use of implementation scheme A91, wherein the cancer is a hematologic cancer.

[0425] Implementation scheme A94. Use of implementation scheme A91, wherein the cancer is one or more of the cancers listed in Table 1.

[0426] Implementation scheme A95. Use of implementation scheme A91, wherein the cancer is hepatocellular carcinoma, glioblastoma, lung cancer, breast cancer, head and neck cancer, prostate cancer, melanoma, or colorectal cancer.

[0427] Implementation scheme A96. Use of implementation scheme A91, wherein the cancer is colorectal cancer, breast cancer, lymphoma, melanoma, kidney cancer or lung cancer.

[0428] Implementation scheme A97. Use of implementation scheme A91, wherein the cancer is an advanced solid tumor, non-small cell lung cancer, or squamous cell carcinoma of the head and neck.

[0429] Implementation scheme A98. Use of any one of implementation schemes A91-A97, wherein the pharmaceutical composition is administered subcutaneously to the subject.

[0430] Implementation scheme A99. Use of implementation scheme A98, wherein the pharmaceutical composition is administered to the subject according to an intermittent dosing regimen.

[0431] Implementation scheme A100. Use of implementation scheme A99, wherein the pharmaceutical composition is administered to the subject twice a week on non-consecutive days.

[0432] Implementation scheme A101. Use of any one of implementation schemes A91-A100, wherein the pharmaceutical composition is administered to the subject together with one or more optional therapeutic agents.

[0433] Implementation scheme A102. Use of implementation scheme A101, wherein the one or more optional therapeutic agents comprise one or more immune checkpoint inhibitors.

[0434] Implementation scheme A103. Use of implementation scheme A102, wherein the one or more immune checkpoint inhibitors comprise PD-1 inhibitors, PD-L1 inhibitors, CTLA-4 inhibitors, LAG3 inhibitors, TIM3 inhibitors or TIGIT inhibitors or combinations thereof.

[0435] Implementation scheme A104. Use of implementation scheme A103, wherein the one or more immune checkpoint inhibitors comprises an anti-PD-1 antibody.

[0436] Implementation Scheme A105. Use of Implementation Scheme A104, wherein the anti-PD-1 antibody is nivolumab, pembrolizumab, pildizumab, STI-A1110, (spartazumab), JTX-4014, sintilimab, MEDI 0680, AGEN2034, BGB-A317, AB122, dotalimab, PF-06801591, cimiprimab, tislelizumab, toripalimab, camrelizumab, SYM021, JNJ 63723283, HLX10, LZM009, or MGA012.

[0437] Implementation scheme A106. Use of any one of implementation schemes A102-A105, wherein the one or more immune checkpoint inhibitors comprises an anti-PD-L1 antibody.

[0438] Implementation scheme A107. Use of implementation scheme A106, wherein the anti-PD-L1 antibody is averrucumab, atezolizumab, durvalumab, KN035 or STI-A1014.

[0439] Implementation scheme A108. Use of any one of implementation schemes A102-A107, wherein the one or more immune checkpoint inhibitors comprise an anti-CTLA-4 antibody.

[0440] Implementation scheme A109. Use of implementation scheme A108, wherein the anti-CTLA-4 antibody is ipilimumab or trimemumab.

[0441] Implementation scheme A110. Use of any one of implementation schemes A102-A109, wherein the one or more immune checkpoint inhibitors comprises an anti-LAG3 antibody.

[0442] Implementation scheme A111. Use of implementation scheme A110, wherein the anti-LAG3 antibody is GSK2831781.

[0443] Implementation scheme A112. Use of any one of implementation schemes A102-A111, wherein the one or more immune checkpoint inhibitors comprise an anti-TIM3 antibody.

[0444] Implementation scheme A113. Use of any one of implementation schemes A102-A112, wherein the one or more immune checkpoint inhibitors comprise an anti-TIGIT antibody.

[0445] Implementation Scheme A114. Use of Implementation Scheme A113, wherein the anti-TIGIT antibody is vimbrolizumab (MK-7684), tirelimumab (RG6058), EOS-448, BMS-986207, BGB-A1217, MTIG7192A, AB154, ASP8374 or MK-7684.

[0446] Implementation scheme A115. Use of any one of implementation schemes A102-A114, wherein the cancer is resistant to or has become resistant to treatment with at least one immune checkpoint inhibitor.

[0447] Example 1

[0448] Screening study of lyophilized products

[0449] Study No. 1

[0450] The lyophilized products were prepared from the ingredients provided in Table 4. In each case, 13.4 mg of compound 1 was used.

[0451] Table 4

[0452]

[0453]

[0454] The freeze-dried cakes of P3, P4, P7, P8, and P9 have different appearances, with P7 producing the best-shaped cake. See also Figure 1 .

[0455] The results of the screening performance of the lyophilized cakes are presented in Tables 5 and 6. The solvent used to suspend the lyophilized products in Table 6 was 1 mL of water for injection (WFI).

[0456] Table 5

[0457]

[0458]

[0459] *Time includes the time used for hovering + an additional 1 minute.

[0460] Table 6

[0461]

[0462] Study No. 2

[0463] Table 7 provides lyophilized products containing Compound 1 and other components. In each case, 15 mg / mL of Compound 1 was used.

[0464] Table 7

[0465]

[0466]

[0467] Example 2

[0468] Preparation of a unit dose of a pharmaceutical composition comprising compound 1 for subcutaneous administration to a subject.

[0469] Table 8 shows the amounts of compound 1, mannitol, sodium carboxymethyl cellulose, poloxamer 188, histidine, and other components used to prepare lyophilized products L1, L2, and L3.

[0470] Table 8

[0471] Freeze-dried product ID L1 L2 L3 Element Quantity (g) Quantity (g) Quantity (g) Compound 1 (g) 0.06750 0.06750 0.0625 Mannitol (g) 0.135 0.135 0.125 Sodium carboxymethyl cellulose*(g) 0.0252 0.009 0.00833 Polosham 188, NF*** 0.00765 0.00765 0.00708 Soy lecithin** 0.0450 0.0450 0.04167 Histidine (g) 0.00388 0.00388 0.00431 Ethanol (g) 0.355 0.355 0.3945 Ethanol (L) (d=0.789) 0.000450 0.000450 0.0005 tert-Butanol (g) 1.299 1.2285 1.365 tert-Butanol (L) (d=0.780) 0.00158 0.00158 0.00175 QS to (using WFI)(L) 0.0045 0.0045 0.0050 1M HCl (titrate to pH 6.6-6.8) (mL) 0.0042 0.00416 0.00462

[0472] *CMC Sodium 7LF (Ashland Part No. 891158); **Lipoid S 100; ***Spectrum Chemical MFG Corp. Product Code P1169

[0473] 1. Dissolve to prepare a solution before lyophilization:

[0474] First, dissolve mannitol, sodium carboxymethyl cellulose, poloxamer 188, and histidine in water at 40°C. After dissolution, the solution can be cooled to 25°C without precipitation.

[0475] Next, titrate the solution to pH 6.6-6.8 (target 6.7) using 1M HCl.

[0476] Third, mix the solution with warm tert-butanol and ethanol.

[0477] Fourth, dissolve soybean lecithin in a solution at 30°C.

[0478] Fifth, completely dissolve compound 1 in a solution at 30°C (this may take several hours). Optionally, protect the solution from light.

[0479] Sixth, lower the temperature to 20-25℃.

[0480] Seventh, use the target QS value to bring the solution to the correct mass (or volume), and add room temperature WFI until the target is reached.

[0481] After 12 hours at room temperature, the L2 solution before lyophilization contained precipitated solids, which could be redissolved by mixing at 30°C.

[0482] 2. Aseptic filtration

[0483] The drug product solution from step 1 is aseptically filtered using redundant 1) Millipore Polysep II pre-filters and 2) redundant Millipore Durapore 0.22-micron capsule filters. The pressure should be maintained at 20 PSI or lower.

[0484] Precipitation occurs during the filtration of the solution before L2 lyophilization.

[0485] 3. Freeze-drying

[0486] The initial rack temperature is room temperature. The rack temperature is lowered to -40°C and the pressure reduced to 45 mTorr. These conditions are maintained for 44.5 hours. Then, the rack temperature is raised to 0°C over 5 hours and maintained at that temperature for 12 hours. Next, the rack temperature is raised to 35°C over 1 hour and maintained at that temperature for 28 hours. The rack temperature is then lowered back to room temperature over 1 hour, followed by nitrogen equilibration, and then the stopper is inserted. Freeze-dry according to one of the following procedures.

[0487] Program 1

[0488]

[0489] Program 2

[0490]

[0491]

[0492] The chemical purity of compound 1 in the lyophilized product obtained in this way is greater than or equal to 97%.

[0493] 4. Preparation of pharmaceutical compositions

[0494] For L1 and L3, add 2.5 mL of WFI to the lyophilized material and shake the container (e.g., a tubular bottle) to create a suspension. For L2, add 2.25 mL of WFI to the lyophilized material and shake the container (e.g., a tubular bottle) to create a suspension.

[0495] Example 3

[0496] Optimization study of freeze-dried products

[0497] Study No. 1

[0498] Table 9 provides the mixed components for preparing 100 mL scale (100 mL of each formulation), and Table 10 provides the observations of these lyophilized products.

[0499]

[0500]

[0501] Table 10

[0502]

[0503] Study No. 2

[0504] Table 11 provides the mixed composition of lyophilized products L12-L17 for preparing 100 mL scale (100 mL of each formulation), and Tables 12 and 13 provide the observation results of these lyophilized products.

[0505] Table 11

[0506]

[0507]

[0508] Table 12

[0509]

[0510] Table 13

[0511]

[0512] Example 4

[0513] Suspension Research

[0514] The lyophilized product L2 (see Example 2) was suspended in 1.5, 2.0, 2.25, and 2.5 mL of WFI. The test results for this pharmaceutical composition are provided in Table 14.

[0515] Table 14

[0516]

[0517] *The suspension thickens approximately 15-20 minutes after reconstruction, making PSD testing impossible.

[0518] Injection force was tested using an Instron instrument (BD 3mL syringe, 27G 1 / 2-inch needle, injection rate 1mm / s). The results are presented in Table 15.

[0519] Table 15

[0520] WFI volume 2.25mL 2.5mL Force (N) 6.8 5.6 Syringe ID (mm) 8.1 8.1 Area (mm2) 51.2 51.2 Manual injection ability light easy

[0521] Example 5

[0522] Subcutaneous drug composition research

[0523] To evaluate the stability of the L1 pharmaceutical composition (also referred to as E8h) prepared according to Example 2 at 2–8°C for 5 days, a study was conducted to determine the chemical and colloidal stability of suspensions of the lyophilized material at two concentrations—2.5 mL WFI and 10.0 mL WFI. Five vials were used for each concentration. All were suspended at time zero and their performance was subsequently determined after different holding times. Data regarding suspension (reconstitution) time and appearance are summarized in Table 16 below.

[0524] Table 16

[0525]

[0526] In the first 3 days, the tubular vial can be easily resuspended (shaked again to obtain a uniform drug product), but on the 5th day, the appearance is no longer uniform.

[0527] Particle size distribution (PSD) was measured throughout the study. Each test vial was suspended on day 0 and held until resuspended and sampled at its predetermined test time. Data on PSD are summarized in Table 17.

[0528] Table 17

[0529]

[0530] PSD was homogeneous until day 3, but shifted upwards by day 5. No changes in impurities were observed by HPLC during the 5-day study. A reduction in compound 1 was observed in 10 mL of resuspended compound 1 on day 5. This data is summarized in Table 18, which shows the percentage of label claims.

[0531] Table 18

[0532]

[0533] After resuspension, the accuracy of compound 1 delivery from the pharmaceutical composition was tested using a 0.5 mL tuberculin syringe. 150 μL of suspension was drawn into the syringe and injected into a volumetric flask for determination. Repeated tests were performed using fresh syringes to confirm the relative standard deviation (RSD), with HPLC determinations performed in duplicate. The data are presented in the table below.

[0534]

[0535] Average %LC-10 of E8h reconstructed using 10 mL

[0536]

[0537] Precision and accuracy of reconstructed E8h using 2.5 mL

[0538]

[0539]

[0540] Average % LC of E8 reconstructed with 2.5 mL

[0541]

[0542] Example 6

[0543] Research on the use of L3 drug compositions

[0544] In this study, simulated injections were delivered at doses of 3, 6, and 50 mg to evaluate the stability and absorption of compound 1 during resuspension and administration of the L3 pharmaceutical composition. Assays were performed at 0, 4, and 24 hours. Results are presented in Tables 19, 20, 21, and 22.

[0545] Table 19

[0546] Stability Study of Tubular Bottles

[0547]

[0548]

[0549] *Below the limit of quantitation 2.48 μg / mL

[0550] Table 20

[0551] Instron Injection Force Measurement

[0552] <![CDATA[T0]]> <![CDATA[T 4小时 ]]> <![CDATA[T 24小时 ]]> Force (N)* 5.5 6.5 8.0

[0553] * Syringe ID: 8.1mm; Syringe area: 51.2mm² 2

[0554] Table 21

[0555] Syringe stability study (BD 3mL syringe, 27G 1 / 2-inch needle, injection rate 1mm / s)

[0556]

[0557]

[0558] Table 22

[0559] Syringe delivery simulation injection study

[0560]

[0561] Example 7

[0562] Pharmacokinetic studies

[0563] The L1 and L2 drug compositions described in Example 2 were administered to dogs via subcutaneous (SC) injection at a dose of 0.83 mg / kg. In the subcutaneous group, blood samples were collected from all animals via the saphenous or cephalic vein at pre-administration and at 0.25, 0.5, 1, 2, 4, 6, 8, 12, 24, 36, 48, 60, and 72 hours post-administration in tubes containing potassium ethylenediaminetetraacetate (K2EDTA). Plasma was separated from blood by centrifugation at 4°C and stored at -80°C until analysis. The concentrations of compound 1 and DON in plasma were quantified using liquid chromatography-tandem mass spectrometry (LC-MS / MS).

[0564] In dogs, following a single subcutaneous administration of the L1 drug composition at 0.83 mg / kg, at 0.500 hours post-administration (median T... max The average peak plasma concentration of compound 1 (C) was reached. max =120±33.5nM). Mean terminal T 1 / 2 The duration was 8.71 ± 3.90 hours. AUC 最后 and AUC inf The results are calculated as 1270±469 and 1430±468 hours * nM, respectively. MRT 最后 and MRT INF The durations were 9.31 ± 2.30 hours and 13.0 ± 4.98 hours, respectively.

[0565] In dogs, following a single subcutaneous administration of the L2 drug composition at 0.83 mg / kg, at 4.00 hours post-administration (median T... max The average peak plasma concentration of compound 1 (C) was reached. max =70.7±18.9nM). Mean terminal T 1 / 2 The duration was 12.1 ± 12.3 hours. AUC 最后 and AUC inf The results were calculated as 759±230 and 1030±478 hours * nM, respectively. MRT 最后 and MRT INF The durations were 7.61 ± 2.09 hours and 17.0 ± 16.5 hours, respectively.

[0566] Conversely, in dogs, after a single subcutaneous administration of a solution formulation of compound 1 (mediates: 5% ethanol, 45% PEG 400, and 0.2% poloxamer 188 in phosphate buffer) at a dose of 0.15 mg / kg, the median T value was [missing data]. max In dogs, the mean peak plasma concentration (C) of compound 1 was reached.max =170nM). Mean terminal T 1 / 2 It is 0.71 hours. AUC 最后 The result is calculated to be 0.249 hours * μM.

[0567] It should be understood that the embodiments and examples described above are not intended to limit the scope of this disclosure in any way, and the claims set forth herein are intended to cover all embodiments and examples, whether or not they are expressly stated herein.

[0568] All patents and publications cited in this article are incorporated in their entirety by reference.

Claims

1. A lyophilized product, said lyophilized product comprising: (i) 20% wt / wt to 30% wt / wt of (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester; (ii) Mannitol, 45% wt / wt to 55% wt / wt; (iii) Sodium carboxymethyl cellulose, ranging from 1% wt / wt to 9% wt / wt; (iv) 1% wt / wt to 4% wt / wt of poloxamer 188; (v) 12% wt / wt to 20% wt / wt soybean lecithin; and (vi) 1% wt / wt to 2% wt / wt histidine.

2. The lyophilized product according to claim 1, wherein the lyophilized product comprises 25.1% wt / wt of isopropyl (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoate.

3. The lyophilized product according to claim 1, wherein the lyophilized product comprises 50.2% wt / wt of mannitol.

4. The lyophilized product according to claim 1, wherein the lyophilized product comprises 3.3% wt / wt sodium carboxymethyl cellulose.

5. The lyophilized product according to claim 1, wherein the lyophilized product comprises 2.8% wt / wt of poloxamer 188.

6. The freeze-dried product according to claim 1, wherein the freeze-dried product contains 16.8% wt / wt of soybean lecithin.

7. The lyophilized product according to claim 1, wherein the lyophilized product contains 1.7% wt / wt histidine.

8. The lyophilized product according to claim 1, wherein the lyophilized product comprises: (i) 62.5 mg (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester; (ii) 125 mg mannitol; (iii) 8.3 mg sodium carboxymethyl cellulose; (iv) 7.1 mg poloxamer 188; (v) 41.7 mg soy lecithin; and (vi) 4.3 mg histidine.

9. A pharmaceutical composition comprising the lyophilized product according to claim 1, wherein the lyophilized product is suspended in a pharmaceutically acceptable carrier.

10. The pharmaceutical composition of claim 9, wherein the pharmaceutically acceptable carrier comprises water.

11. The pharmaceutical composition according to claim 9, wherein the concentration of (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester is from 20 mg / mL to 60 mg / mL.

12. The pharmaceutical composition according to claim 11, wherein the concentration of (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoic acid isopropyl ester is 25 mg / mL.

13. The pharmaceutical composition according to claim 9, wherein the average particle size distribution of isopropyl (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoate in the suspension is from 1 μm to 30 μm.

14. A kit comprising a lyophilized product according to any one of claims 1-8, packaged as a single unit dose in a container.

15. The kit of claim 14, further comprising instructions for suspending the lyophilized material in a solvent to produce a pharmaceutical composition.

16. The kit of claim 14, wherein the kit further comprises a label.

17. A method for preparing the lyophilized product according to claim 1, the method comprising: (i) Prepare a solution before lyophilization, which contains isopropyl (S)-2-((S)-2-acetamido-3-(1H-indol-3-yl)propionylamino)-6-diazo-5-oxohexanoate, mannitol, sodium carboxymethyl cellulose, poloxamer 188, soybean lecithin, histidine and water. (ii) Cool the solution prior to lyophilization until it is frozen or partially frozen; and (iii) Apply a vacuum to the frozen or partially frozen pre-lyophilized solution to produce the lyophilized product.

18. The method of claim 17, wherein the solution prior to lyophilization further comprises ethanol.

19. The method of claim 17, wherein the solution prior to lyophilization further comprises tert-butanol.

20. A method for preparing a pharmaceutical composition according to claim 9, the method comprising suspending the lyophilized material in a solvent.

21. The method of claim 20, wherein the solvent comprises water.

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