Conjugates, compositions thereof, and related methods

Abstract

Some embodiments of the invention include compounds of the invention (e.g., compounds of Formula (I)). Other embodiments include compositions (e.g., pharmaceutical compositions) that include compounds of the invention. Still other embodiments of the invention include compositions using compounds of the invention to treat, for example, certain diseases. Some embodiments include methods of using compounds of the invention (e.g., in compositions or pharmaceutical compositions) for administration and treatment. Further embodiments include methods for making compounds of the invention. Additional embodiments of the invention are also discussed herein.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 265,421, filed December 15, 2021, entitled “CONJUGATES, THEIR COMPOSITIONS, AND THEIR RELATED METHODS,” which is incorporated by reference in its entirety. [Background technology]

[0002] Some compounds (e.g., inhibitors and / or macrocyclic compounds) have potential anti-cancer and / or anti-tumor activity. Some of these compounds can be toxic to normal (e.g., non-cancerous) animal cells and are sometimes ineffective at tolerated doses. There remains a broad need to develop new compounds to treat cancer in animals. Certain embodiments of the present invention may address one or more of the deficiencies discussed above.

[0003] Some embodiments of the invention include compounds of the invention (e.g., compounds of Formula (I)). Other embodiments include compositions (e.g., pharmaceutical compositions) that include compounds of the invention. Still other embodiments of the invention include compositions using compounds of the invention to treat, for example, certain diseases. Some embodiments include methods of using compounds of the invention (e.g., in compositions or pharmaceutical compositions) for administration and treatment. Further embodiments include methods for making compounds of the invention. Additional embodiments of the invention are also discussed herein. Summary of the Invention

[0004] Some embodiments of the present invention are of formula (I) [ka] (I), In certain embodiments, R 1is selected from H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), and phenyl-(C1-C4 alkyl), wherein the -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, C1-C4 alkylsulfonyl, or phenyl-(C1-C4 alkyl) is optionally selected from halogen, hydroxy (-OH), meta In other embodiments, R may be substituted with one or more of the following: noyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, phenyl, perfluorinated methyl, perfluorinated ethyl, amino, C1-C4 alkanoylamino (-NH-CO-(C1-C4 alkyl)), C1-C4 alkoxy, benzyloxy (-O-CH2-phenyl), oxo (=O), C2-C5 alkoxycarbonyl (-CO-O-(C2-C5 alkyl)), methylenedioxy (-O-CH2-O- with one or two bonded carbons), or C1-C4 alkylthio (-S-(C1-C4 alkyl)). 2 is selected from H, allyl, vinyl, hydroxyl, Cl, Br, F, I, thiol, amino, nitro, cyano, C1-C4 alkyl, C1-C4 alkylnoic, phenyl, C1-C2 perfluorinated alkyl, alkylamino, oxo, carboxy, acetyl, amido, and C1-C3 alkoxy. 3 , H, C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, cycloalkyl, and -COR 4 C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20Alkynyl, methyl, ethyl, aryl, or cycloalkyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbo In yet other embodiments, R may be substituted with one or more of the following: nyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 4 is C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, cycloalkyl, and [ka] C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20Alkynyl, methyl, ethyl, aryl, or cycloalkyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbo In yet other embodiments, R may be substituted with one or more of the following: nyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 5 , R 6 , and R 7 are the same or different, and each independently represents C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, and cycloalkyl; 18 Alkyl, C2-C 20 Alkenyl, C2-C 20Alkynyl, methyl, ethyl, aryl, or cycloalkyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbo nyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). In certain embodiments, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, Z is selected from albumin, human serum albumin (HSA), bovine serum albumin (BSA), canine serum albumin (CSA), feline serum albumin (FSA), equine serum albumin (ESA), domain I of HSA, domain II of HSA, domain III of HSA, engineered albumins, variants thereof, and fragments thereof.

[0005] In other embodiments, R 1is selected from H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl, where the -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, or ethyl may be optionally substituted with one or more of halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, amino, C1-C4 alkoxy, or oxo. 1 is selected from the group consisting of H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl.

[0006] In yet another embodiment, R 2 is selected from the group consisting of H, Cl, Br, F, I, allyl, ethyl, methyl, and OH. 2 is H or Cl.

[0007] In other embodiments, R 3 is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and nonyl. 3 teeth, [ka] In some embodiments, R 3 teeth, [ka] In yet another embodiment, R 4 is C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, and cycloalkyl; 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 Alkynyl, methyl, ethyl, aryl, or cycloalkyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbo In some embodiments, R may be substituted with one or more of the following: nyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 4 is C1-C 10 Alkyl, C2-C 10 Alkenyl, C2-C 10 alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; 10 Alkyl, C2-C 10 Alkenyl, C2-C 10Alkynyl, methyl, or ethyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbonyl (-O-C O-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). In certain embodiments, R 4 is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, and hexyl. 4 teeth, [ka] and R 5 , R 6 , and R 7 are the same or different, and each independently represents C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; 18 Alkyl, C2-C 20 Alkenyl, C2-C 20Alkynyl, methyl, or ethyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbonyl (-O-C O-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 5 , R 6 , and R 7 are the same or different and each is independently selected from methyl, ethyl, propyl, butyl, pentyl, and hexyl. 4 teeth, [ka] In some embodiments, m is 1, 2, 3, 4, or 5. In still other embodiments, m is 1 or 2. In still other embodiments, Z is BSA, CSA, or HSA. In certain embodiments, Z is HSA.

[0008] In some embodiments, the compound is I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, or I-10. In other embodiments, the compound is I-1 or I-3.

[0009] Some embodiments of the present invention include compositions comprising any of the compounds disclosed herein. In certain embodiments, the amount of the compound is from about 0.0001% (by weight of the total composition) to about 99%. In other embodiments, the composition comprises a lyophilized compound.

[0010] Some embodiments of the present invention include a pharmaceutical composition comprising any of the compounds disclosed herein. In yet other embodiments, the amount of the compound is from about 0.0001% (by weight of the total composition) to about 50%. In certain embodiments, the pharmaceutical composition further comprises a compounding ingredient. In other embodiments, the pharmaceutical composition comprises a lyophilized compound.

[0011] Some embodiments of the present invention include a method for providing any compound disclosed herein to an animal, the method comprising one or more administrations of one or more compositions comprising any compound disclosed herein, the compositions, if there are two or more administrations, may be the same or different. In yet other embodiments, at least one of the one or more compositions further comprises a formulation component. In other embodiments, at least one of the one or more compositions comprises any composition disclosed herein or any pharmaceutical composition disclosed herein. In certain embodiments, at least one of the one or more administrations comprises parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration. In some embodiments, if there are two or more administrations, at least one composition used in at least one administration is different from the composition of at least one other administration. In certain embodiments, at least one compound of the one or more compositions is administered to the animal in an amount of about 0.01 mg / kg animal body weight to about 500 mg / kg animal body weight. In still other embodiments, the animal is a human, a dog, or a primate.

[0012] Some embodiments of the present invention include a method for treating an animal for a disease, the method comprising one or more administrations of one or more compositions comprising any compound disclosed herein, the compositions, if there are two or more administrations, may be the same or different. In certain embodiments, at least one of the one or more compositions further comprises a formulation component. In other embodiments, at least one of the one or more compositions comprises any composition disclosed herein or any pharmaceutical composition disclosed herein. In some embodiments, at least one of the one or more administrations comprises parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration. In still other embodiments, if there are two or more administrations, at least one composition used in at least one administration is different from the composition of at least one other administration. In certain embodiments, at least one compound of the one or more compositions is administered to the animal in an amount of about 0.01 mg / kg animal body weight to about 500 mg / kg animal body weight. In still other embodiments, the animal is a human, a dog, or a primate. In still other embodiments, the animal is in need of treatment. In certain embodiments, the method is for treating cancer. In some embodiments, the method is for treating acute lymphoblastic leukemia, astrocytoma, basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, chronic lymphocytic leukemia (CLL), CNS cancer, colon cancer, colorectal cancer, endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma, gastric cancer, thyroid cancer, uterine cancer, cancer that may lead to metastasis, cancer resulting from metastasis, or cancerous tumors thereof.In other embodiments, the method is for treating basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, CNS cancer, colon cancer, colorectal cancer, endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma, gastric cancer, thyroid cancer, uterine cancer, or cancerous tumors thereof. In yet other embodiments, the method is for treating a cancerous tumor. In yet other embodiments, the method is for treating breast cancer, head and neck cancer, lung cancer, non-small cell lung cancer, lymphoma, ovarian cancer, or kidney cancer.

[0013] Some embodiments of the present invention include a method for preparing any of the compounds disclosed herein, comprising: (a) reacting Z with a disulfide to provide an activated Z; (b) reacting the activated Z with Formula (II) to provide a compound; and (c) optionally recovering the compound. In certain embodiments, Formula (II) is [ka] (II) In some embodiments, the disulfide is formamidine disulfide, aldrithiol, or 5,5'-dithiobis(2-nitrobenzoic acid). In yet other embodiments, the disulfide is formamidine disulfide. In other embodiments, the disulfide is at a concentration that is in molar equivalent excess of Z. In still other embodiments, prior to step (a), one or more thiol reducing agents are added to Z, and optionally, at least a portion of the unreacted one or more thiol reducing agents is then removed. In some embodiments, the pH of the reaction solution in step (b) is about 1.0 to about 7.0 (e.g., about 1.0 to about 6.0). In still other embodiments, Formula (II) is dissolved in a polar solvent prior to its addition to activated Z. In certain embodiments, the reaction in step (b) is for about 4 hours to about 48 hours. In other embodiments, the temperature of the reaction in step (b) is about -5°C to about 20°C. In still other embodiments, step (c) is not optional.In some embodiments, the method further comprises lyophilization after step (b) or after step (c).

[0014] Other embodiments of the present invention are also disclosed herein.

[0015] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the invention. The invention may be better understood by reference to one or more of these drawings in combination with the description of specific embodiments presented herein. [Brief description of the drawings]

[0016] [Figure 1] Comparison of the effects of AP3 and compound I-1 on the viability of human Daudi lymphoma tumor cells after 72 hours of culture. Similar potency suggests the bioavailability of compound I-1.

[0017] [Diagram 2]Repeated experiments comparing the effects of AP3 and compound I-1 on the viability of human Daudi lymphoma tumor cells after 72 hours of culture. Similar efficacy also suggests the bioavailability of compound I-1.

[0018] [Diagram 3] Comparison of the effects of AP3 and compound I-1 on the viability of cultures of human SGC-7901 gastric tumor cells after 72 hours of culture. Similar potency suggests the bioavailability of compound I-1.

[0019] [Figure 4] Repeated experiments comparing the effects of AP3 and compound I-1 on the viability of cultures of human SGC-7901 gastric tumor cells after 72 hours of incubation. Similar efficacy also suggests the bioavailability of compound I-1.

[0020] [Diagram 5] Comparison of the effects of AP3 and compound I-1 on the viability of human U-937 lymphoma tumor cells after 72 hours of culture. Similar potency suggests the bioavailability of compound I-1.

[0021] [Figure 6] Repeated experiments comparing the effects of AP3 and compound I-1 on the viability of human U-937 lymphoma tumor cells after 72 hours of culture. Similar potency also suggests the bioavailability of compound I-1.

[0022] [Figure 7] Comparison of the effects of AP3 and compound I-3 on the viability of human U-937 lymphoma tumor cells after 72 hours of culture. Similar potency suggests the bioavailability of compound I-3.

[0023] [Figure 8]Repeated experiments comparing the effects of AP3 and compound I-3 on the viability of human U-937 lymphoma tumor cells after 72 hours of culture. Similar potency also suggests the bioavailability of compound I-3.

[0024] [Figure 9] Effects of AP3 and compound I-1 on the proliferation of human gastric tumor SGC-7901 cells subcutaneously injected into nude mice on day 0. Compounds were injected into the tail vein on days 0 (200 mg / kg) and 14 (150 mg / kg) for compound I-1, and on days 0 and 14 for AP3. AP3 was only marginally effective at this dose level compared to the control, whereas compound I-1 was effective throughout the study period.

[0025] [Figure 10] The effect of AP3 and compound I-1 on the body weight of nude mice treated under the experimental protocol described in Figure 9, in which compounds were injected into the tail vein on days 0 (200 mg / kg) and 14 (150 mg / kg) for compound I-1, and on days 0 and 14 for AP3. AP3 was only slightly effective at this dose level compared to the control, while compound I-1 was effective throughout the study period. A decrease in body weight was observed in the compound I-1 treatment group.

[0026] [Figure 11] Effects of AP3 and compound I-1 on the growth of human U-937 lymphoma tumor cells subcutaneously injected into SCID mice on day 0. AP3 (0.5 mg / kg) was injected into the tail vein on days 0 and 15, and compound I-1 (300 mg / kg) was injected on days 0, 7, 15, and 21. Compound I-1 was more effective than AP3. Compound I-1 caused almost complete tumor growth inhibition up to 30 days.

[0027] [Figure 12]The effect of AP3 and compound I-1 on the body weight of SCID mice treated under the experimental protocol described in Figure 11. During the period, mice in both treatment groups were observed to lose weight. The body weight returned to normal approximately one week after the last injection.

[0028] [Figure 13] Affinity of recombinant HSA for recombinant human FcRn receptor using surface plasmon resonance microscopy indicates that recombinant HSA binds in a pH-dependent manner.

[0029] [Figure 14] The affinity of compound I-1 to recombinant human FcRn receptor using surface plasmon resonance microscopy shows that compound I-1 binds in a pH-dependent manner. HSA (from Figure 13) and compound I-1 exhibited similar concentration-dependent binding and response rates. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0030] Although embodiments embodying the general inventive concept may take a variety of forms, various embodiments are described herein with the understanding that the disclosure should be considered merely as illustrative and that the general inventive concept is not intended to be limited to the disclosed embodiments.

[0031] Some embodiments of the invention include compounds of the invention (e.g., compounds of Formula (I)). Other embodiments include compositions (e.g., pharmaceutical compositions) that include compounds of the invention. Still other embodiments of the invention include compositions using compounds of the invention to treat, for example, certain diseases. Some embodiments include methods of using compounds of the invention (e.g., in compositions or pharmaceutical compositions) for administration and treatment. Further embodiments include methods for making compounds of the invention. Additional embodiments of the invention are also discussed herein.

[0032] As defined herein, "amino acid" includes, but is not limited to, any naturally occurring amino acid (including the standard 20 amino acids, which are Gly, Ala, Val, Leu, Ile, Met, Pro, Phe, Trp, Ser, Thr, Asn, Gln, Tyr, Cys, Lys, Arg, His, Asp, and Glu) and unusual amino acids. As defined herein, an "unusual amino acid" is an amino acid that is not one of the standard 20 amino acids. Some examples of unusual amino acids are listed in Table A, but are not limited to those listed in Table A. [Table 1]

[0033] Amino acids may be in the L-form, D-form, or neither (e.g., glycine). As used herein, unless otherwise indicated, when the three letter or one letter designation of an amino acid is not given, the amino acid is in the L-form. When not within a sequence, the form of the amino acid when designated includes a hyphen (e.g., L-Lys). When within a sequence, the hyphen is omitted.

[0034] As used herein (unless otherwise specified), a "fragment" of a protein or amino acid sequence comprises at least 3 consecutive amino acids. For example, a fragment may comprise 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, 30, 35, 40, 45, 50, 75, 100, 125, 150, 175, 200, 250, 500, 1000, 1500, 2000, 50 or less, 100 or less, 500 or less, 1000 or less, 1500 or less, 2000 or less, at least 10, at least 15, at least 20, at least 25, at least 35, at least 40, at least 45, at least 50, at least 100, at least 150, at least 175, at least 200, at least 500, at least 1000, at least 1500, or at least 2000 consecutive amino acids of a protein or amino acid sequence. In some embodiments, the fragment may be at least 150 contiguous amino acids. In some embodiments, the fragment may be at least 175 contiguous amino acids. The length of the fragment may be varied appropriately depending on the desired properties or functions.

[0035] As used herein (unless otherwise specified), a "variant" (or similar language such as mutated or mutation) of a protein or amino acid sequence includes truncations, additions, deletions, substitutions, and other modifications of a protein or amino acid sequence, provided that some desired property or desired function remains. In some embodiments, a mutation may be a combination of two or more truncations, deletions, additions, substitutions, or other modifications. In some embodiments, one or more substitutions may be conservative substitutions. In some embodiments, conservative substitutions may be based on the hydropathy index of Kyte and Doolittle J. Mol. Biol. 1982, Vol. 157, pp. 105-132 (e.g., substitutions within ±2, ±1, or ±0.5), the hydrophilicity value of U.S. Pat. No. 4,554,101 (e.g., substitutions within ±2, ±1, or ±0.5), or the size of the amino acid (e.g., side group size). In certain embodiments, the following substitutions are considered conservative when one amino acid is substituted for another amino acid in the same group: Group 1 is Ile, Leu, Val, Ala, Gly, Group 2 is Trp, Tyr, Phe, Group 3 is Asp, Glu, Asn, Gln, Group 4 is Cys, Ser, Thr, Met, and Group 5 is His, Lys, Arg. In some cases, conservative substitutions minimally disrupt (or may enhance) one or more desired properties or functions.

[0036] As used herein (unless otherwise specified), the term "alkyl" refers to a monovalent, straight or branched hydrocarbon chain (e.g., C-C 24). For example, the term "C1-C7 alkyl" or "C1-C4 alkyl" refers to a straight-chain or branched saturated hydrocarbon group having 1 to 7 (e.g., 1, 2, 3, 4, 5, 6, or 7) or 1 to 4 (e.g., 1, 2, 3, or 4) carbon atoms, respectively. Examples of C1-C7 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, t-butyl, n-pentyl, s-pentyl, n-hexyl, and n-septyl. Examples of C1-C4 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, and t-butyl.

[0037] As used herein (unless otherwise specified), the term “alkenyl” refers to an alkyl group having one or more (e.g., 1, 2, 3, or 4) double bonds (e.g., C2-C 24 ) means a monovalent, straight or branched hydrocarbon chain containing the alkyl group, aryl ...

[0038] As used herein (unless otherwise specified), the term "alkoxy" refers to an alkyl group as defined above attached to the remainder of the molecule by an oxygen atom (alkyl-O-) (e.g., C1-C 23 Examples of alkoxy groups include, but are not limited to, methoxy (sometimes referred to as MeO-), ethoxy, isopropoxy, propoxy, and butyloxy.

[0039] As used herein (unless otherwise specified), the term "alkynyl" refers to an alkyl group that includes one or more (e.g., 1, 2, 3, or 4) triple bonds, and optionally one or more (e.g., 1, 2, 3, or 4) double bonds (e.g., C2-C 24) means a monovalent, straight or branched hydrocarbon chain, which may also include an alkynyl group. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, and 5-hexynyl.

[0040] As used herein (unless otherwise specified), the term "aryl" refers to a monovalent, monocyclic or bicyclic, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered aromatic hydrocarbon group when unsubstituted. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, tolyl, and xylyl. In the case of bicyclic aryls, which are designated as substituted, one or both rings can be substituted.

[0041] As used herein (unless otherwise specified), the term "cycloalkyl" refers to a monovalent, monocyclic or bicyclic, 3-, 4-, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered aromatic hydrocarbon group. The ring can be saturated or partially unsaturated. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and bicycloalkyl (e.g., bicyclooctanes, such as [2.2.2]bicyclooctane or [3.3.0]bicyclooctane, bicyclononanes, such as [4.3.0]bicyclononane, and bicyclodecanes, such as [4.4.0]bicyclodecane (decalin), or spiro compounds), and adamantane. In the case of monocyclic cycloalkyls, the ring is not aromatic. For bicyclic cycloalkyls, if one ring is aromatic, the other is not aromatic. For bicyclic cycloalkyls designated as substituted, one or both rings can be substituted.

[0042] As used herein (unless otherwise specified), the term "halogen" means monovalent Cl, F, Br, or I.

[0043] As used herein, the term "heteroaryl" refers to a monovalent, monocyclic or bicyclic, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered hydrocarbon group in which 1, 2, 3, 4, 5, or 6 carbon atoms are replaced by heteroatoms independently selected from nitrogen, oxygen, or sulfur atoms, and the monocyclic or bicyclic ring system is aromatic. Examples of heteroaryl groups include, but are not limited to, thienyl (or thiophenyl), furyl, indolyl, pyrrolyl, pyridinyl, pyrazinyl, oxazolyl, thiaxolyl, quinolinyl, pyrimidinyl, imidazolyl, triazolyl, tetrazolyl, 1H-pyrazol-4-yl, 1-Me-pyrazol-4-yl, pyridin-3-yl, pyridin-4-yl, 3,5-dimethylisoxazolyl, 1H-pyrrol-3-yl, 3,5-di-Me-pyrazolyl, and 1H-pyrazol-4-yl. For bicyclic heteroaryls, if one ring is aryl, the other is heteroaryl. For bicyclic heteroaryls, one or both rings can have one or more heteroatoms. For bicyclic heteroaryls designated as substituted, one or both rings can be substituted.

[0044] As used herein, the term "heterocyclyl" means a monovalent, monocyclic or bicyclic, 5-, 6-, 7-, 8-, 9-, 10-, 11-, or 12-membered hydrocarbon in which 1, 2, 3, 4, 5, or 6 carbon atoms are replaced by heteroatoms independently selected from nitrogen, oxygen, or sulfur atoms, and the monocyclic or bicyclic ring system is not aromatic. Examples of heterocyclyl groups include, but are not limited to, tetrahydropyran, pyrrolidinyl (e.g., pyrrolidin-1-yl, pyrrolidin-2-yl, pyrrolidin-3-yl, or pyrrolidin-4-yl), piperazinyl (e.g., piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, or piperazin-4-yl), piperidinyl (e.g., piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, or piperazin-4-yl), and morpholinyl (e.g., morpholin-1-yl, morpholin-2-yl, morpholin-3-yl, or morpholin-4-yl). For bicyclic heterocyclyls, when one ring is aromatic (e.g., a monocyclic aryl or heteroaryl), the other ring is not aromatic. For bicyclic heterocyclyls, either or both rings may have one or more heteroatoms. For bicyclic heterocyclyls designated as substituted, either or both rings may be substituted.

[0045] As used herein (unless otherwise specified), the term "heteroatom" means an atom selected from a nitrogen atom, an oxygen atom, or a sulfur atom.

[0046] As used herein (unless otherwise specified), the term "hydroxy" or "hydroxyl" denotes the presence of a monovalent --OH group.

[0047] As used herein (unless otherwise specified), the term "substituted" (e.g., as in substituted alkyl) means that one or more hydrogen atoms of a chemical group (having one or more hydrogen atoms) may be replaced by one or more non-hydrogen substituents selected from a specified selection. The substitution may occur at one or more positions. The term "optionally substituted" means that one or more hydrogen atoms of a chemical group (having one or more hydrogen atoms) may be, but need not be, replaced.

[0048] Some compounds of the present invention may have one or more chiral centers, and can exist and be isolated in optically active and racemic forms for any of the one or more chiral centers.Some compounds may exhibit polymorphism.The compounds of the present invention (e.g., Formula I) encompass any optically active, racemic, stereoisomeric form, polymorph, or mixture thereof.If a chiral center does not provide an indication of its configuration (i.e., R or S) in the chemical structure, it should be considered as representing R, S, or racemic.

[0049] Some embodiments of the present invention include a compound of formula (I). [ka] (I)

[0050] In some embodiments, the compound of formula (I) may be in the form of salts, optical and geometric isomers, and salts of isomers. In other embodiments, the compound may be in various forms, such as uncharged molecules, components of molecular complexes, or non-irritating pharmacologically acceptable salts, including but not limited to hydrochlorides, hydrobromides, sulfates, phosphates, nitrates, borates, acetates, maleates, tartrates, and salicylates. In some cases, for acidic compounds, salts may include metals, amines, or organic cations (e.g., quaternary ammonium). In still other embodiments, simple derivatives of compounds (e.g., ethers, esters, or amides) may be used that have the desired retention and release properties, but are easily hydrolyzed by body pH, enzymes, or other suitable means.

[0051] In some embodiments, R 1is H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, or C8 alkyl), C2-C8 alkenyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkenyl), C2-C8 alkynyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkynyl), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkyl and phenyl-(C1-C4 alkyl), including -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, or C8 alkyl), C2-C8 alkenyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkenyl), C2-C8 alkyn ... , C7 or C8 alkynyl), methyl, ethyl, C1-C4 alkylsulfonyl, or phenyl-(C1-C4 alkyl) are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, phenyl, perfluorinated methyl, perfluorinated ethyl, amino, C1-C4 alkano The arylamino group may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of ylamino (-NH-CO-(C1-C4 alkyl)), C1-C4 alkoxy, benzyloxy (-O-CH2-phenyl), oxo (=O), C2-C5 alkoxycarbonyl (-CO-O-(C2-C5 alkyl)), methylenedioxy (-O-CH2-O- having one or two bonded carbons), or C1-C4 alkylthio (-S-(C1-C4 alkyl)).

[0052] In other embodiments, R 1may be selected from H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, or C8 alkyl), C2-C8 alkenyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkenyl), C2-C8 alkynyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkynyl), methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl, wherein -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C8 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, or C8 alkyl), C2-C8 alkenyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkynyl), For example, C2, C3, C4, C5, C6, C7, or C8 alkenyl), C2-C8 alkynyl (e.g., C2, C3, C4, C5, C6, C7, or C8 alkynyl), methyl, or ethyl can be optionally substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, phenyl, perfluorinated methyl, perfluorinated ethyl, amino, C1-C4 alkoxy, benzyloxy, oxo, or methylenedioxy. In other embodiments, R 1may be selected from H, -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C4 alkyl (e.g., C1, C2, C3, or C4 alkyl), C2-C4 alkenyl (e.g., C2, C3, or C4 alkenyl), C2-C4 alkynyl (e.g., C2, C3, or C4 alkynyl), methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl, wherein -COCH3, carboxy (-CO2H), ethynyl (-CCH), C1-C4 alkyl (e.g., C1, C2, C3, or C4 alkyl), C2-C4 alkenyl (e.g., C2, C3, or C4 alkynyl), , C2, C3, or C4 alkenyl), C2-C4 alkynyl (e.g., C2, C3, or C4 alkynyl), methyl, or ethyl can be optionally substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, amino, C1-C4 alkoxy, or oxo. In certain embodiments, R 1 may be selected from H, -COCH, carboxy (-COH), ethynyl (-CCH), methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl. In certain embodiments, R 1 may be selected from H, methyl, and perfluorinated methyl. In some embodiments, R 1 may be identical to Y in US 4,424,219 to Hashimoto et al., which is incorporated herein by reference in its entirety.

[0053] In some embodiments, R 2may be selected from H, allyl, vinyl, hydroxyl, Cl, Br, F, I, thiol, amino, nitro, cyano, C1-C4 alkyl (e.g., C1, C2, C3, or C4 alkyl), C1-C4 alkylnoic (e.g., C1, C2, C3, or C4 alkylnoic), phenyl, C1-C2 perfluorinated alkyl, alkylamino, oxo, carboxy, acetyl, amido, and C1-C3 alkoxy (e.g., C1, C2, or C3 alkoxy). In some embodiments, R 2 can be H, Cl, Br, F, I, allyl, ethyl, methyl, or OH. In some embodiments, R 2 can be H or Cl. In some embodiments, R 2 may be identical to X in US 4,424,219 to Hashimoto et al., which is incorporated herein by reference in its entirety.

[0054] In some embodiments, R 3 , H, C1-C 18 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkynyl), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), and -COR 4 and C1-C 18 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20alkynyl), methyl, ethyl, aryl (e.g., phenyl or naphthyl), or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-( In some embodiments, R may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of C2-C4 alkyl), C2-C4 alkoxycarbonyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 3 can be methyl, ethyl, propyl (e.g., isopropyl), butyl (e.g., isobutyl), pentyl (e.g., isopentyl), hexyl (e.g., isohexyl), heptyl (e.g., isoheptyl), octyl (e.g., isooctyl), or nonyl (e.g., isononyl). In some embodiments, R 3 can be isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, or isononyl. 3 teeth, [ka] In certain embodiments, R 3 teeth, [ka] In some embodiments, R 3 may be identical to R in US 4,424,219 to Hashimoto et al., which is incorporated herein by reference in its entirety.

[0055] In other embodiments, R 4 is C1-C 18 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkynyl), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), and [ka] and C1-C 18Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20alkynyl), methyl, ethyl, aryl (e.g., phenyl or naphthyl), or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-( In other embodiments, R may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of C2-C4 alkyl), C2-C4 alkoxycarbonyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 4 is C1-C 10 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkenyl), C2-C 10 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkynyl), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), and [ka] and C1-C 10Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkenyl), C2-C 10 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkynyl), methyl, ethyl, aryl (e.g., phenyl or naphthyl), or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-( In some embodiments, R may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of C2-C4 alkyl), C2-C4 alkoxycarbonyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 4 can be methyl, ethyl, propyl (e.g., isopropyl), butyl (e.g., isobutyl), pentyl (e.g., isopentyl), hexyl (e.g., isohexyl), heptyl (e.g., isoheptyl), octyl (e.g., isooctyl), or nonyl (e.g., isononyl). In some embodiments, R 4can be isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, or isononyl. 4 can be methyl, ethyl, propyl (e.g., isopropyl), butyl, pentyl, or hexyl. In some embodiments, R 4 can be isopropyl.

[0056] In certain embodiments, R 4 teeth, [ka] It could be.

[0057] In yet another embodiment, R 5 , R 6 , and R 7 can be the same or different, and each independently represents C1-C 18 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C16 , C 17 , C 18 , C 19 , or C 20 alkynyl), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl (e.g., phenyl or naphthyl), and cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), 18 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , or C 18 Alkyl), C2-C 20 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20 alkenyl), C2-C 20 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , C 12 , C 13 , C 14 , C 15 , C 16 , C 17 , C 18 , C 19 , or C 20alkynyl), methyl, ethyl, aryl (e.g., phenyl or naphthyl), or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-( In yet other embodiments, R may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of C2-C4 alkyl), C2-C4 alkoxycarbonyl (-O-CO-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 5 , R 6 , and R 7 can be the same or different, and each independently represents C1-C 18 Alkyl, C2-C 20 Alkenyl, C2-C 20 alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; 18 Alkyl, C2-C 20 Alkenyl, C2-C 20Alkynyl, methyl, or ethyl are optionally selected from halogen, hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbonyl (-O-C O-(C2-C4 alkyl)), nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). In certain embodiments, R 5 , R 6 , and R 7 can be the same or different, and each independently represents C1-C 10 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkenyl), C2-C 10 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkynyl), methyl, ethyl, perfluorinated methyl, or perfluorinated ethyl, 10 Alkyl (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C 10 alkenyl), C2-C 10 Alkynyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9, or C10 alkynyl), methyl, or ethyl are optionally substituted with halogen (e.g., F, Cl, Br, or I), hydroxy (-OH), methanoyl (-COH), -COCH3, carboxy (-CO2H), ethynyl (-CCH), cyano (-CN), sulfo (-SO3H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkoxycarbonyl (-O-CO- In some embodiments, R may be substituted with one or more (e.g., 0, 1, 2, 3, 4, 5, or 6) of: nitro, amino, mono(C1-C4 alkyl)amino, di(C1-C4 alkyl)amino, C1-C4 alkylthio (-S-(C1-C4 alkyl)), C1-C4 alkylsulfinyl (-SO-(C1-C4 alkyl)), C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)), oxo (=O), thioxo (=S), or C1-C4 alkanoylamino (-CO-NH-(C1-C4 alkyl)). 5 , R 6 , and R 7 can be the same or different and each can be independently selected from methyl, ethyl, propyl (e.g., isopropyl), butyl (e.g., isobutyl), pentyl (e.g., isopentyl), hexyl (e.g., isohexyl), heptyl (e.g., isoheptyl), octyl (e.g., isooctyl), and nonyl (e.g., isononyl). In some embodiments, R 5 , R 6 , and R 7 can be the same or different and each can be independently selected from isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, and isononyl. In some embodiments, R 5 , R 6 , and R 7can be the same or different and each can be independently selected from methyl, ethyl, propyl (e.g., isopropyl), butyl, pentyl, and hexyl. In some embodiments, R 5 , R 6 , and R 7 may be the same and are methyl.

[0058] In other embodiments, R 4 teeth, [ka] It could be.

[0059] In some embodiments, R 4 , R 5 , R 6 , and R 7 and R in Hashimoto et al., US Pat. No. 4,424,219, which is incorporated herein by reference in its entirety. 1 , R 2 , R 3 , and R 4 may be identical to

[0060] In some embodiments, m can be 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, m can be 1, 2, 3, 4, or 5. In certain embodiments, m is 1, or m is 2. In some embodiments, a solution comprising Formula (I) can have a mixture of formulas with different values ​​of m, resulting in a solution from which an average m value can be obtained. In some examples, the average m value of the solution can be any rational number between 1 and 10, such as, but not limited to, about 1, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3, about 3.5, about 4, about 4.5, about 5, about 5.5, about 6, about 6.5, about 7, about 7.5, about 8, about 8.5, about 9, about 9.5, or about 10.

[0061] In some embodiments, Z can be, but is not limited to, albumin, human serum albumin (HSA), bovine serum albumin (BSA), canine serum albumin (CSA), feline serum albumin (FSA), equine serum albumin (ESA), domain I of HSA, domain II of HSA, domain III of HSA, engineered albumin (e.g., Veltis® from Novozymes), variants thereof, or fragments thereof (including variant fragments). In other embodiments, Z can be BSA, CSA, or HSA. In other embodiments, Z can be BSA or HSA. In other embodiments, Z can be CSA or HSA. In other embodiments, Z can be HSA.

[0062] In other embodiments, Z stabilizes the compound. Stabilizing a compound can include, for example, maintaining or modulating (e.g., increasing or decreasing) any suitable desired property or function of the compound, including, but not limited to, water solubility (e.g., increasing), blood half-life (e.g., increasing), half-life in the body before reaching the target (e.g., increasing), circulation stability (e.g., increasing), preventing or reducing absorption by the kidney, or preventing or reducing absorption by the brain.

[0063] In certain embodiments, Z is substituted or unsubstituted in the compounds of the invention (e.g., in formula (I)) via a free thiol on Z. [ka] In another embodiment, Z is HSA and Z is connected via cys-34 to [ka] In another embodiment, Z is CSA and Z is connected to the compound of the invention (e.g., formula (I)) via cys-34. [ka] In another embodiment, Z is BSA and Z is connected to the compound of the invention (e.g., formula (I)) via cys-34. [ka] In yet other embodiments, Z (e.g., HSA or domain I of HSA) is connected to a compound of the invention (e.g., formula (I)) via a free thiol on activated Z (e.g., using any suitable activating compound, such as, but not limited to, formamidine disulfide, as described herein). [ka] In yet other embodiments, Z (e.g., canine, feline, equine, or bovine albumin) is connected to a compound of the invention (e.g., Formula (I)) via a free thiol on Z that has been activated (e.g., using any suitable activating compound, such as, but not limited to, formamidine disulfide, as described herein) after reacting Z with a reducing compound (e.g., using any suitable reducing compound, such as, but not limited to, mercaptoethanol or dithiothreitol). [ka] is connected to.

[0064] In some embodiments, the compound of formula (I) can be selected from those specified in Table 1. [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4] [Table 2-5] [Table 2-6]

[0065] In some embodiments, the compound of the invention can be I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, or I-10. In some embodiments, the compound of the invention can be I-1, I-2, I-3, I-4, or I-5. In other embodiments, the compound of the invention can be I-1, I-2, I-3, I-6, or I-7. In yet other embodiments, the compound of the invention can be I-1 or I-3. In yet other embodiments, the compound of the invention can be I-1.

[0066] Compositions including pharmaceutical compositions One or more compounds of the present invention (e.g., Formula (I)) may be part of the composition and may be at least about 0.0001%, at least about 0.001%, at least about 0.10%, at least about 0.15%, at least about 0.20%, at least about 0.25%, at least about 0.50%, at least about 0.75%, at least about 1%, at least about 10%, at least about 25%, at least about 50%, at least The amount may be about 75%, at least about 90%, at least about 95%, at least about 99%, at least about 99.99%, up to about 75%, up to about 90%, up to about 95%, up to about 99%, or up to about 99.99%, from about 0.0001% to about 99%, from about 0.0001% to about 50%, from about 0.01% to about 95%, from about 1% to about 95%, from about 10% to about 90%, or from about 25% to about 75% (by weight of the total composition).

[0067] 0.0001 or more compounds of the invention (e.g., Formula (I)) are at least about 0.0001%, at least about 0.001%, at least about 0.10%, at least about 0.15%, at least about 0.20%, at least about 0.25%, at least about 0.50%, at least about 0.75%, at least about 1%, at least about 10%, at least about 25%, at least about 50%, at least about 75% , at least about 90%, at least about 95%, at least about 99%, at least about 99.99%, about 75% or less, about 90% or less, about 95% or less, about 99% or less, about 99.99% or less, about 0.0001% to about 99%, about 0.0001% to about 50%, about 0.01% to about 95%, about 1% to about 95%, about 10% to about 90%, or about 25% to about 75% (by weight of the total composition).

[0068] Some embodiments of the present invention include compositions comprising a compound of the present invention (e.g., Formula (I)). In certain embodiments, the composition is a pharmaceutical composition, such as a composition suitable for administration to an animal (e.g., a mammal, a primate, a monkey, a human, a dog, a cat, a horse, a cow, a pig, a mouse, a rabbit, or a rat). In some cases, the pharmaceutical composition is non-toxic, does not cause side effects, or both. In some embodiments, there may be inherent side effects (e.g., it may be harmful to the patient, or it may be toxic or harmful to some degree in some patients).

[0069] In some embodiments, a compound of the invention (e.g., Formula (I)) may be part of a pharmaceutical composition and may be in an amount of at least about 0.0001%, at least about 0.001%, at least about 0.10%, at least about 0.15%, at least about 0.20%, at least about 0.25%, at least about 0.50%, at least about 0.75%, at least about 1%, at least about 10%, at least about 25%, at least about 50%, at least about 75%, at least about 90%, at least about 95%, at least about 99%, at least about 99.99%, up to about 75%, up to about 90%, up to about 95%, up to about 99% or up to about 99.99%, from about 0.001% to about 99%, from about 0.001% to about 50%, from about 0.1% to about 99%, from about 1% to about 95%, from about 10% to about 90%, or from about 25% to about 75%. In some embodiments, the pharmaceutical composition can be presented in a dosage form suitable for topical, subcutaneous, intrathecal, intraperitoneal, oral, parenteral, rectal, dermal, nasal, vaginal, or ocular routes of administration. In other embodiments, the pharmaceutical composition can be presented in a dosage form suitable for parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration. The pharmaceutical composition can be in the form of, for example, a tablet, capsule, pill, powder granule, suspension, emulsion, solution, gel (including hydrogel), paste, ointment, cream, plaster, drench, delivery device, suppository, enema, injection, implant injection, spray, aerosol, or other suitable form.

[0070] In some embodiments, the pharmaceutical composition may include one or more compounding ingredients. "Compounding ingredients" include, but are not limited to, water (e.g., boiled water, distilled water, filtered water, pyrogen-free water, or water containing chloroform), sugar (e.g., sucrose, glucose, mannitol, sorbitol, xylitol, or syrups made therefrom), ethanol, glycerol, glycol (e.g., propylene glycol), acetone, ether, DMSO, surfactants (e.g., anionic surfactants, cationic surfactants, zwitterionic surfactants, or nonionic surfactants (e.g., polysorbates)), oils (e.g., animal oils, vegetable oils (e.g., coconut oil or peanut oil), or mineral oils), oil derivatives (e.g., oleic acid, ethylhexyl ether ... The composition may contain any suitable ingredients (e.g., suitable for the compound of the invention (e.g., Formula (I)), the dosage of the compound of the invention (e.g., Formula (I)), the timing of release of the compound of the invention (e.g., Formula (I)), the disease state, the organ affected, or the delivery route), including an additive, a preservative (e.g., cysteine, methionine, an antioxidant (e.g., a vitamin (e.g., A, E, or C), selenium, retinol palmitate, sodium citrate, citric acid, chloroform, or a paraben (e.g., methylparaben or propylparaben)), or a combination thereof.

[0071] In certain embodiments, pharmaceutical compositions can be formulated to release a compound of the invention (e.g., one or more compounds of formula (I)) immediately after administration or at any substantially predetermined or any time after administration. Such formulations can include controlled release formulations, such as, for example, various controlled release compositions and coatings.

[0072] Other formulations (e.g., formulations of pharmaceutical compositions) may, in certain embodiments, include incorporating a compound of the invention (e.g., Formula (I)) (or a controlled release formulation) into a food, foodstuff, feed, or beverage.

[0073] Still other formulations (e.g., formulations of pharmaceutical compositions) may, in certain embodiments, include lyophilized compounds of the invention (e.g., Formula (I)), which may (in some instances) include any suitable substance for stabilizing one or more of the compounds of the invention, such as, but not limited to, one or more simple carbohydrates (e.g., sucrose, mannitol, trehalose, or combinations thereof).

[0074] Other embodiments of the present invention may include methods of administering or treating an animal (e.g., a human or a dog), which may involve treatment with at least one compound of the present invention (e.g., Formula (I)) in an amount effective to treat a cancer that the animal has, is suspected of having, or is susceptible to, or to produce a desired physiological effect. In some embodiments, the composition or pharmaceutical composition comprises at least one compound of the present invention (e.g., Formula (I)) that can be administered to an animal (e.g., a mammal, primate, monkey, or human) in an amount of about 0.01 to about 15 mg / kg body weight, about 0.1 to about 10 mg / kg body weight, about 0.5 to about 7 mg / kg body weight, about 0.01 mg / kg, about 0.05 mg / kg, about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 3 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.5 mg / kg, about 7 mg / kg, about 7.5 mg / kg, about 8 mg / kg, about 10 mg / kg, about 12 mg / kg, or about 15 mg / kg. For some conditions, the dosage can be about 0.5 mg / kg human body weight or about 6.5 mg / kg human body weight. In some examples, some animals (e.g., mammals, mice, rabbits, cats, pigs, or dogs) may be administered a dosage of about 0.01 to about 15 mg / kg body weight, about 0.1 to about 10 mg / kg body weight, about 0.5 to about 7 mg / kg body weight, about 0.01 mg / kg, about 0.05 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 80 mg / kg, about 100 mg / kg, or about 150 mg / kg. Of course, one of skill in the art will appreciate that many concentrations can be used in the methods of the invention, and that, using in part the guidance provided herein, any number of concentrations can be adjusted and tested to find a concentration that achieves the desired result in a given situation. In other embodiments, the compounds of the invention can be administered in combination with one or more other therapeutic agents for cancer.

[0075] In some embodiments, the composition may include a dose (e.g., a unit dose) of one or more compounds of the present invention (e.g., Formula (I)) in combination with a pharma- ceutically acceptable carrier, and may additionally include other medicinal agents, pharmaceutical agents, carriers, adjuvants, diluents, additives, or combinations thereof. In certain embodiments, the carrier, vehicle, or additive may facilitate administration, delivery, and / or storage of the composition. In other embodiments, the one or more carriers include, but are not limited to, saline solutions such as saline, Ringer's solution, PBS (phosphate buffered saline), and mixtures of various salts, including potassium salts and phosphate salts, generally with or without sugar additives such as glucose. Carriers further include aqueous and non-aqueous sterile injection solutions that may contain antioxidants, buffers, bacteriostatic agents, bactericidal antibiotics, and solutes that render the formulation isotonic with the body fluids of the intended recipient, as well as aqueous and non-aqueous sterile suspensions that may include suspending agents and thickening agents. In other embodiments, the one or more additives may include, but are not limited to, water, saline, dextrose, glycerol, ethanol, and the like, and combinations thereof. Non-toxic auxiliary substances, such as wetting agents, buffering agents, or emulsifying agents, may also be added to the compositions. Oral formulations can include such commonly employed additives as, for example, pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc.

[0076] Parenteral administration, if used, is generally characterized by injection. Sterile injectables can be prepared in conventional forms, either as liquid solutions or suspensions, solid forms suitable for solution of suspension in liquid prior to injection, or as emulsions.

[0077] Methods for Administration and Treatment The compounds of the present invention (e.g., Formula (I)) can be administered to animals by any number of suitable routes of administration or formulations. The compounds of the present invention (e.g., Formula (I)) can also be used to treat animals for various diseases. Animals include, but are not limited to, mammals, primates, monkeys (e.g., macaques, rhesus monkeys, or pig-tailed macaques), humans, dogs, cats, horses, cows, pigs, birds (e.g., chickens), mice, rabbits, and rats. As used herein, the term "subject" refers to both human and animal subjects.

[0078] The route of administration of the compound of the present invention (e.g., Formula (I)) may be any suitable route. The route of administration may be, but is not limited to, oral, parenteral, dermal, nasal, rectal, vaginal, and ocular. In other embodiments, the route of administration may be parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular. The choice of the route of administration may depend, for example, on the identity of the compound of the present invention (e.g., Formula (I)) (e.g., the physical and chemical properties of the compound of the present invention (e.g., Formula (I))), as well as the age and weight of the animal, the particular disease (e.g., cancer), and the severity of the disease (e.g., the stage of the cancer). Of course, a combination of routes of administration may be administered if desired.

[0079] Some embodiments of the invention include methods for providing a subject with a composition (e.g., a pharmaceutical composition) comprising a compound of the invention described herein (e.g., Formula (I)), comprising one or more administrations of one or more such compositions, which, if two or more administrations are administered, may be the same or different, and which, if two or more administrations are administered, may be the same or different.

[0080] Some embodiments of the invention include methods for treating a subject with a composition (e.g., a pharmaceutical composition) comprising a compound of the invention described herein (e.g., Formula (I)), comprising one or more administrations of one or more such compositions, which, if two or more administrations are administered, may be the same or different, and which, if two or more administrations are administered, may be the same or different.

[0081] Animals that may be treated include, but are not limited to, mammals, primates, monkeys (e.g., macaques, rhesus monkeys, pig-tailed macaques), humans, dogs, cats, horses, pigs, birds (e.g., chickens), cows, mice, rabbits, and rats. As used herein, the term "subject" refers to both human and animal subjects. A subject susceptible to a disease (e.g., cancer) may be a human subject or an animal subject. In some cases, an animal (e.g., a human or a dog) is in need of treatment for cancer.

[0082] Diseases that may be treated in animals (e.g., mammals, pigs, dogs, birds (e.g., chickens), cows, cats, primates, rodents, monkeys, rabbits, mice, rats, and humans) using the compounds of the invention (e.g., Formula (I) or (I-1)) include, but are not limited to, cancer.

[0083] In some embodiments, cancers that may be treated in animals (e.g., mammals, pigs, dogs, birds (e.g., chickens), cattle, cats, primates, rodents, monkeys, rabbits, mice, rats, and humans) using the compounds of the invention (e.g., Formula (I) or (I-1)) include, but are not limited to, acute lymphoblastic leukemia, astrocytoma, basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, chronic lymphocytic leukemia (CLL), CNS cancer (e.g., glioblastoma, glioblastoma multiforme, gliosarcoma, or astrocytoma), colon cancer, colorectal cancer (e.g., colon cancer or rectal cancer), endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer (e.g., kidney cancer ... cancer), leukemia, liver cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma neoplasia), malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma (e.g., diffuse large B-cell lymphoma), non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer (e.g., pancreatic ductal adenocarcinoma), prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma (e.g., head and neck squamous cell carcinoma), gastric cancer, thyroid cancer, uterine cancer, cancer that may lead to metastasis, cancer resulting from metastasis, or cancerous tumors thereof.In some embodiments, cancers that may be treated include, but are not limited to, basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, CNS cancer (e.g., glioblastoma, glioblastoma multiforme, gliosarcoma, or astrocytoma), colon cancer, colorectal cancer (e.g., colon or rectal cancer), endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer (e.g., kidney cancer, cancer), leukemia, liver cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma neoplasia), malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma (e.g., diffuse large B-cell lymphoma), non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer (e.g., pancreatic ductal adenocarcinoma), prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma (e.g., head and neck squamous cell carcinoma), gastric cancer, thyroid cancer, uterine cancer, or cancerous tumors thereof. In some embodiments, cancers that may be treated include, but are not limited to, leukemia, lung cancer (e.g., non-small cell lung cancer), head and neck cancer, colorectal cancer (e.g., colon or rectal cancer), CNS cancer (e.g., glioblastoma, glioblastoma multiforme, gliosarcoma, or astrocytoma), lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma neoplasia), ovarian cancer, renal cancer, kidney cancer, prostate cancer, breast cancer, or cancerous tumors thereof. In some embodiments, cancers that may be treated include, but are not limited to, breast cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof. In some embodiments, cancers that may be treated include, but are not limited to, breast cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, or renal cancer. In some embodiments, cancers that may be treated include, but are not limited to, cancerous tumors. Animals that may be treated include, but are not limited to, mammals, rodents, primates, monkeys (e.g., macaques, rhesus monkeys, pig-tailed macaques), humans, dogs, cats, pigs, horses, birds (e.g., chickens), cows, mice, rabbits, and rats.As used herein, the term "subject" refers to both human and animal subjects. In some cases, the animal is in need of treatment (e.g., by exhibiting symptoms of disease or cancer, or by having a cancerous tumor).

[0084] In some embodiments, cancers that may be treated in animals (e.g., mammals, pigs, dogs, horses, birds (e.g., chickens), cows, cats, primates, rodents, monkeys, rabbits, mice, rats, and humans) using the compounds of the invention (e.g., Formula (I) or (I-1)) include, but are not limited to, cancers that are affected (e.g., reduced tumor size) by tubulin inhibitors.

[0085] As used herein, the term "treating" (and its variants, such as "treatment") should be considered in its broadest context. In particular, the term "treating" does not necessarily mean that an animal is treated to complete recovery. Thus, "treating" includes amelioration of symptoms, alleviation of symptoms or effects associated with a condition, reduction in the severity of a condition, or prevention, prophylactic alleviation of symptoms, or otherwise reducing the risk of developing a particular condition. As used herein, reference to "treating" an animal includes, but is not limited to, prophylactic and therapeutic treatment. Any of the compositions (e.g., pharmaceutical compositions) described herein can be used to treat an animal.

[0086] In relation to the treatment of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof), treatment can include, but is not limited to, prophylactic and therapeutic treatment. Thus, treatment includes, but is not limited to, preventing cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof), reducing the risk of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof), improving or alleviating the symptoms of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof), eliciting the body's response to cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof), inhibiting the development or progression of cancer (e.g., breast cancer, The present invention may include inhibiting or preventing the onset of symptoms associated with cancer (e.g., lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof), reducing the severity of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof), causing regression of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof) or causing one or more symptoms associated with cancer (e.g., reduction in tumor size), causing remission of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof), or preventing recurrence of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof). In some embodiments, the treatment does not include prophylactic treatment of cancer (e.g., preventing or ameliorating future cancer).

[0087] Treatment of an animal may be carried out using any suitable method of administration (such as those disclosed herein) and using any suitable amount of a compound of the invention (e.g., Formula (I) or (I-1)). In some embodiments, the treatment method includes treating an animal for cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof). Some embodiments of the invention include methods for treating a subject (e.g., an animal such as a human or a primate) with a composition (e.g., a pharmaceutical composition) comprising a compound of the invention (e.g., Formula (I) or (I-1)) comprising one or more administrations of one or more such compositions, which compositions, if there are two or more administrations, may be the same or different.

[0088] In some embodiments, the method of treatment includes administering an effective amount of a composition comprising a compound of the present invention (e.g., Formula (I) or (I-1)). As used herein, the term "effective amount" refers to a dose or series of doses sufficient to effect treatment in an animal (e.g., treat cancer, such as, but not limited to, breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, renal cancer, or cancerous tumors thereof). In some embodiments, the effective amount can include a therapeutically effective amount as disclosed herein. In certain embodiments, the effective amount can vary depending on the subject and the particular treatment being effected. The exact amount required can vary from subject to subject, depending, for example, on the age and general condition of the subject, the particular adjuvant (if any) being used, the administration protocol, etc. Thus, the effective amount can vary, for example, based on the particular circumstances, and an appropriate effective amount can be determined in a particular case. The effective amount can include, for example, any dosage or amount of the composition disclosed herein. In some embodiments, an effective amount of at least one compound of the present invention (e.g., Formula (I) or (I-1)), which may be administered to an animal, such as a mammal, a primate, a monkey, or a human, may be an amount of about 0.005 to about 50 mg / kg body weight, about 0.01 to about 15 mg / kg body weight, about 0.1 to about 10 mg / kg body weight, about 0.5 to about 7 mg / kg body weight, about 0.005 mg / kg, about 0.01 mg / kg, about 0.05 mg / kg, about 0.1 mg / kg, about 0.5 mg / kg, about 1 mg / kg, about 3 mg / kg, about 5 mg / kg, about 5.5 mg / kg, about 6 mg / kg, about 6.5 mg / kg, about 7 mg / kg, about 7.5 mg / kg, about 8 mg / kg, about 10 mg / kg, about 12 mg / kg, or about 15 mg / kg. For some embodiments, the dosage may be about 0.5 mg / kg human body weight or about 6.5 mg / kg human body weight.In some examples, an effective amount of at least one compound of the present invention (e.g., Formula (I) or (I-1)) (which may be administered to an animal, such as a mammal, rodent, mouse, rabbit, cat, horse, pig, or dog) may be an amount of about 0.005 to about 50 mg / kg body weight, about 0.01 to about 15 mg / kg body weight, about 0.1 to about 10 mg / kg body weight, about 0.5 to about 7 mg / kg body weight, about 0.005 mg / kg, about 0.01 mg / kg, about 0.05 mg / kg, about 0.1 mg / kg, about 1 mg / kg, about 5 mg / kg, about 10 mg / kg, about 20 mg / kg, about 30 mg / kg, about 40 mg / kg, about 50 mg / kg, about 80 mg / kg, about 100 mg / kg, or about 150 mg / kg. In some embodiments, an effective amount of at least one compound of the present invention (e.g., Formula (I) or (I-1)), which may be administered to an animal, such as a mammal, a primate, a monkey, or a human, may be in an amount of about 1 to about 1000 mg / kg body weight, about 5 to about 500 mg / kg body weight, about 10 to about 200 mg / kg body weight, about 25 to about 100 mg / kg body weight, about 1 mg / kg, about 2 mg / kg, about 5 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 100 mg / kg, about 150 mg / kg, about 200 mg / kg, about 300 mg / kg, about 400 mg / kg, about 500 mg / kg, about 600 mg / kg, about 700 mg / kg, about 800 mg / kg, about 900 mg / kg, or about 1000 mg / kg. For some conditions, the dosage may be about 20 mg / kg human body weight or about 100 mg / kg human body weight. In some examples, an effective amount of at least one compound of the present invention (e.g., Formula (I) or (I-1)) (which may be administered to an animal, such as a mammal, rodent, mouse, rabbit, cat, horse, pig, or dog) may be an amount of about 1 to about 1000 mg / kg body weight, about 5 to about 500 mg / kg body weight, about 10 to about 200 mg / kg body weight, about 25 to about 100 mg / kg body weight, about 1 mg / kg, about 2 mg / kg, about 5 mg / kg, about 10 mg / kg, about 25 mg / kg, about 50 mg / kg, about 100 mg / kg, about 150 mg / kg, about 200 mg / kg, about 300 mg / kg, about 400 mg / kg, about 500 mg / kg, about 600 mg / kg, about 700 mg / kg, about 800 mg / kg, about 900 mg / kg, or about 1000 mg / kg.

[0089] "Therapeutically effective amount" means an amount effective to achieve a desired and / or beneficial effect (e.g., reduction in tumor size). A therapeutically effective amount can be administered in one or more administrations. For some purposes of the present invention, a therapeutically effective amount is an amount appropriate to treat an indication (e.g., to treat cancer). Treating an indication means achieving any desired effect, such as one or more of mitigating, alleviating, stabilizing, reversing, slowing, or delaying the progression of a disease (e.g., cancer), improving quality of life, or extending lifespan. Such achievement can be measured by any suitable method, such as, but not limited to, measuring tumor size.

[0090] In some embodiments, treatment can also include one or more of surgical intervention, chemotherapy, radiation therapy, hormone therapy, immunotherapy, and adjuvant systemic therapy.Adjuvants can include, but are not limited to, chemotherapy (e.g., temozolomide), radiation therapy, antiangiogenic therapy (e.g., bevacizumab), and hormone therapy such as administration of LHRH agonists, antiestrogens such as tamoxifen, high-dose progestogens, aromatase inhibitors, and / or adrenalectomy.Chemotherapy can be used as a single agent or in combination with known or new therapies.

[0091] In some embodiments, administration of at least one compound of the present invention (e.g., Formula (I) or (I-1)) is an adjuvant cancer therapy or part of an adjuvant cancer therapy. Adjuvant therapy includes treatment by the mechanisms disclosed herein, as well as treatment of cancers disclosed herein, including but not limited to tumors. The corresponding primary therapy may include, but is not limited to, surgery, chemotherapy, or radiation therapy. In some cases, adjuvant therapy may be a combination of a chemokine receptor antagonist with a conventional chemotoxic agent or immunotherapy, which increases the specificity of the treatment to the cancer and potentially limits additional systemic side effects. In yet other embodiments, the compounds of the present invention (e.g., Formula (I) or (I-1)) can be used as an adjuvant with other chemotherapeutic agents. The use of the compounds of the present invention (e.g., Formula (I) or (I-1)) can, in some cases, reduce the duration of the dose of both the drug and the drug combination, which reduces side effects.

[0092] In some embodiments, the treatments disclosed herein may include the use of other drugs (e.g., antibiotics) or therapies to treat the disease. For example, antibiotics may be used to treat an infection and may be combined with the compounds of the present invention to treat the disease (e.g., infections associated with cancer). In other embodiments, intravenous immunoglobulin (IVIG) therapy may be used as part of a treatment regimen (e.g., in addition to administering the compound(s) of the present invention).

[0093] Methods for preparing compounds of the present invention (e.g., Formula (I)) Some embodiments of the present invention include methods for the preparation of compounds of the present invention (e.g., Formula (I)). In certain embodiments, compounds of the present invention (e.g., Formula (I)) can be prepared including a step of reacting Z (such as HSA or BSA, as described herein) with a disulfide (e.g., formamidine disulfide) to provide an activated Z. In other embodiments, the disulfide may be, but is not limited to, formamidine disulfide, aldrithiol, 5,5'-dithiobis(2-nitrobenzoic acid), disulfides disclosed in SADOWSKY et al. (2017) "Development of Efficient Chemistry to Generate Site-Specific Disulfide-Linked Protein- and Peptide-Payload Conjugates: Application to THIOMAB Antibody-Drug Conjugates" Bioconjugate Chem., Vol. 28, pp. 2086-2098, which is incorporated herein in its entirety, and ANDREU et al. (1994) "Formation of Disulfide Bonds in Synthetic Peptides and Proteins" from Methods in Molecular Biology, Vol. 35: Peptide Synthesis Protocols, Edited by: MWPennington and BMDunn, Humana Press Inc., Totowa, NJ The disulfide may be any suitable disulfide for the reaction (e.g., a disulfide that is suitable for linking Z to formula (II) by a disulfide bond), including the disulfides disclosed in (incorporated herein in its entirety). In yet other embodiments, the disulfide is formamidine disulfide or 5,5'-dithiobis(2-nitrobenzoic acid). In yet other embodiments, the disulfide is formamidine disulfide.In certain embodiments, the pH of the reaction solution can be about 1.0 to about 6.0, about 2.0 to about 5.0, about 3.0 to about 4.0, about 1.0, about 1.5, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.5, or about 6.0. In other embodiments, the reaction solution comprises a buffer system (e.g., 0.1 M sodium acetate or 0.1 M sodium acetate with 150 mM sodium chloride). In some embodiments, the disulfide (e.g., formamidine disulfide) is at a concentration that is in molar excess of Z, e.g., the disulfide (e.g., formamidine disulfide) is at a concentration that is in molar excess of Z, e.g., about 1.0 to about 20.0, about 5.0 to about 15.0, about 8.0 to about 12.0, about 1.0, about 1.05, about 1.1, about 1.15, about 1.2, about 1.25, about 1.3, about 1.35 , about 1.4, about 1.45, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, about 2.0, about 3.0, about 4.0, about 5.0, about 6.0, about 7.0, about 8.0, about 9.0, about 10.0, about 11.0, about 12.0, about 13.0, about 14.0, about 15.0, about 16.0, about 17.0, about 18.0, about 19.0, or about 20.0 molar equivalents in excess of Z. In some embodiments, following this reaction step (e.g., after the reaction has begun, when the reaction is partially complete (e.g., >50% complete), when the reaction is nearly complete (e.g., >80% complete), or when the reaction is complete (e.g., >95% complete)), some or all of the unreacted disulfide (e.g., formamidine disulfide) can be removed, for example, using any suitable method including, but not limited to, HPLC (e.g., reverse phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography, Sephadex G-25 size exclusion chromatography, or ion exchange chromatography), use of silica gel, or a combination thereof.In some embodiments, activated Z comprises, as a result of the reaction, an S-substituted thiosothiourea structure.

[0094] In some embodiments, it may be optional to add one or more thiol reducing agents (e.g., any suitable thiol reducing agent, such as, but not limited to, mercaptoethanol or dithiothreitol) to Z (e.g., BSA, canine albumin, or non-human albumin) at any suitable concentration (e.g., about 1 mM, about 5 mM, about 10 mM, about 15 mM, or about 20 mM) prior to reacting Z with the disulfide (e.g., formamidine disulfide). It is then further optional to remove some or all of the unreacted thiol reducing agent, which may be performed using any suitable method, including, for example, but not limited to, HPLC (e.g., reverse phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography, Sephadex G-25 size exclusion chromatography, or ion exchange chromatography), the use of silica gel, or a combination thereof.

[0095] Formula (II) is [ka] (II) wherein R 1 , R 2 , and R 3is the same as disclosed herein. Formula (II) may be purchased or synthesized using any suitable method, including but not limited to those disclosed herein and in US 4,424,219 to Hashimoto et al., which is incorporated herein by reference in its entirety. In some embodiments, Formula (II) may be synthesized by contacting a starting compound (e.g., any compound of Formula (II) where -SH is replaced by -OH, such as ansamitocin P3 (AP3) or maytansine) with a solution containing P2S5 (e.g., a solution containing P2S5 and pyridine) for any suitable period of time (e.g., 0.25 hours to 8 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, or 8 hours), at any suitable temperature or set of temperatures (e.g., 15°C to 100°C, 15°C to 80°C, room temperature, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, or 100°C; room temperature for 30 minutes and then at 65°C for 3 hours). In other embodiments, formula (II) can be synthesized by contacting a solution comprising the starting compound (e.g., AP3 or maytansine) (e.g., a solution comprising the starting compound dissolved in methylene chloride) with H2S or a solution comprising H2S (e.g., by bubbling H2S gas into the starting compound solution), which in certain embodiments comprises a solution comprising trifluoroacetic acid (TFA) (e.g., TFA in methylene chloride) at any suitable temperature (e.g., 15°C to 100°C, 15°C to 80°C) with the starting compound (e.g., AP3 or maytansine) / H2S solution. The mixture may be contacted at a temperature of 0.25 to 72 hours (e.g., room temperature, 15°C, 20°C, 25°C, 30°C, 35°C, 40°C, 45°C, 50°C, 55°C, 60°C, 65°C, 70°C, 75°C, 80°C, 85°C, 90°C, 95°C, or 100°C) for any suitable period of time (e.g., 0.25 hours to 72 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 18 hours, 20 hours, 24 hours, 32 hours, 40 hours, 48 ​​hours, 56 hours, 64 hours, or 72 hours).In certain embodiments, recovery of Formula (II) is optional. In other embodiments, recovery of Formula (II) may optionally be performed using any suitable method, including HPLC (e.g., reverse phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion or ion exchange chromatography), use of silica gel, or a combination thereof. In some embodiments, recovery of Formula (II) may be achieved using HPLC with a linear solvent gradient (e.g., as disclosed herein). In some embodiments of the synthesis of Formula (II), water may optionally be removed in the starting compound (e.g., AP3 or maytansine) powder (e.g., by drying under vacuum at 40° C.) and / or P2S5 (e.g., by drying under vacuum at 40° C.).

[0096] In other embodiments (e.g., for the synthesis of Formula (II)), it is optional to reflux pyridine with calcium hydride and distill (e.g., prior to the synthesis of Formula (II)). In yet other embodiments, glassware and spin bars are optionally dried (e.g., prior to the synthesis of Formula (II)).

[0097] In some embodiments, activated Z can be reacted with Formula (II) to provide a compound of the present invention (eg, Formula (I)). In certain embodiments, the pH of the reaction solution is about 1.0 to about 7.0, about 2.0 to about 5.0, about 3.0 to about 4.0, about 1.0, about 2.0, about 2.1, about 2.2, about 2.3, about 2.4, about 2.5, about 2.6, about 2.7, about 2.8, about 2.9, about 3.0, about 3.1, about 3.2, about 3.3, about 3.4, about 3.5, about 3.6, about 3.7, about 3.8, about 3.9, about 4.0, about 4.1, about 4.2, about 4.3, about 4.4, about 4.5, about 4.6, about 4.7, about 4.8, about 4.9, about 5.0, about 5.5, about 6.0, about 6.5, or about 7.0. In other embodiments, the reaction solution comprises a buffer system (e.g., 0.1 M sodium acetate or 0.1 M sodium acetate with 150 mM sodium chloride). In certain embodiments, activated Z is in a solution that is colder than room temperature (e.g., in an ice bath). In other embodiments, Formula (II) is dissolved in a polar solvent prior to its addition to activated Z, and dissolution of Formula (II) can be achieved with any suitable polar solvent, such as, but not limited to, polar aprotic solvents, acetonitrile, acetone, THF, polar protic solvents, methanol, or ethanol. In yet other embodiments, dissolved Formula (II) is added in no more than about 5%, about 10%, about 15%, about 20%, about 25%, or about 30% volume / volume of the total reaction solution. In certain embodiments, after Formula (II) (e.g., dissolved Formula (II)) is added to activated Z, the reaction is allowed to proceed for about 4 hours to about 48 hours, about 4 hours, about 8 hours, about 12 hours, about 16 hours, about 24 hours, about 36 hours, or about 48 hours. In some embodiments, the reaction is carried out at a temperature of about -5°C to about 20°C, about -5°C, about 0°C, about 5°C, about 10°C, about 15°C, or about 20°C.

[0098] In certain embodiments, the compounds of the present invention (e.g., Formula (I)) can be optionally recovered. Recovery can be performed using any suitable method, including, but not limited to, HPLC (e.g., reverse phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion or ion exchange chromatography), the use of silica gel, or a combination thereof. In some embodiments, recovery of the compounds of the present invention (e.g., Formula (I)) can be achieved using a butyl sepharose column with reduced concentrations of ammonium sulfate, followed by size exclusion chromatography (e.g., sephadex G-25) using water, dilute acetic acid, ammonium acetate, or a combination thereof.

[0099] In other embodiments, the compounds of the invention (e.g., Formula (I)) can be lyophilized. In yet other embodiments, the lyophilized compositions comprising the compounds of the invention (e.g., Formula (I)) can further comprise any suitable substance for stabilizing the compounds of the invention (e.g., Formula (I)), including, but not limited to, one or more simple carbohydrates (e.g., sucrose, mannitol, trehalose, or combinations thereof).

[0100] The subject matter of the present disclosure is further illustrated by the following specific, but non-limiting, examples. The following examples may include compilations of data representative of data collected at various times during the course of development and experimentation related to the present invention. EXAMPLES

[0101] Example 1: Synthesis of Ansamitocin P3 Thiol (AP3-SH) - Method A [ka] Ansamitocin P3 (AP3 or AP-3) (MedChemExpress, LLC, Junction, NJ US) (19 mg) was weighed into a glass screw-cap vial. Separately, phosphorus pentasulfide (P2S5) (7.3 mg) was weighed into a screw-cap vial equipped with a small spin bar. The top of the vial was secured with filter paper and both vials were dried by warming to 40°C under high vacuum for 1 hour. Meanwhile, pyridine was dried in a Stark apparatus using calcium hydride for several hours. The dissolved phosphorus pentasulfide was then dissolved in about 2 mL of dry pyridine with a screw cap (15 min). AP3 was dissolved in about 1 mL of dry pyridine and added to the P2S5 solution, which was mixed at room temperature with the screw cap on for 30 minutes and then placed in a heated oil bath at 65°C for 3 hours. The solution was transferred to a round-bottom flask and evaporated on a rotovap at 60°C. The residue was dissolved in a mixture of HPLC solvents A and B (1:1) (HPLC solvent A-0.01% TFA, solvent B-80% acetonitrile in 0.01% TFA) and purified by preparative HPLC using a linear gradient from 10% B to 100% B. The product eluted at approximately 74% B. Pure fractions were pooled and lyophilized.

[0102] Example 2: Synthesis of Ansamitocin P3 Thiol (AP3-SH) - Method B [ka] Ansamitocin P3 (AP3 or AP-3) (MedChemExpress, LLC, Junction, NJ US) (10 mg) was weighed into a glass screw-cap vial, the top was secured with filter paper, and the vial was warmed to 40° C. under high vacuum for 1 hour to dry. The dried AP3 was then dissolved in 50 mL of dry methylene chloride and added to a dry two-neck round-bottom flask equipped with a dropping funnel assembly under a flow of argon and a bubbler to remove moisture. H2S gas was then bubbled into the solution while cooling on an ice bath. Dry methylene chloride (5 ml) and TFA (1 ml) were added to the dropping funnel and the mixture was slowly added dropwise to the AP3 solution and mixed overnight. The solution was transferred to a round-bottom flask and evaporated to dryness at room temperature. The residue was dissolved in a mixture of HPLC solvents A and B (1:1) (HPLC solvent A-0.01% TFA, solvent B-80% acetonitrile (ACN) in 0.01% TFA) and purified by preparative HPLC using a linear gradient from 10% B to 100% B. The product eluted at approximately 74% B. Pure fractions were pooled and lyophilized.

[0103] Example 3: Synthesis of Human Serum Albumin (HSA)-AP3 Conjugate [ka] Recombinant human serum albumin (100 mg) (Wuhan Healthgen Biotechnology Corp., Wuhan, China) was dissolved in 3 mL of ice-cold 0.1 M acetate buffer (pH 3.5 with 150 mM NaCl) in a glass culture tube and placed in an ice bath. Formamidine disulfide (FDS) (3 mg) was placed in the culture tube, dissolved in 500 μL of ice-cold 0.1 M acetate buffer and placed on ice. The FDS solution was then added to the albumin with mixing. After 5 min, excess FDS was separated from the activated albumin using Sephadex G-25 eluted with 0.1 M acetate buffer. The purified activated albumin was cooled to 0 °C in an ice bath. Ansamitocin P3 (AP3) (1.05 equivalents of thiol) was dissolved in 200 μL of acetonitrile and added to the ice-cold isothiourea-albumin with mixing, which was then placed in the refrigerator overnight. The reaction was monitored using HPLC. The reaction reached approximately 60-80% completeness and the product was purified using Sephadex G-25 size exclusion chromatography using distilled water as the eluent.

[0104] Example 4: Synthesis of Canine Serum Albumin (CSA)-AP3 Conjugate Canine serum albumin (Animal Blood Resources International, Stockbridge, MI US) (100 mg) was dissolved in 4 mL of ice-cold PBS (pH 7.4) in a 50 mL Falcon tube and placed in an ice bath. In a separate tube, 3.5 μL of mercaptoethanol was added to 1 mL of PBS, cooled in an ice bath, then added to the solution of canine serum albumin, mixed, and placed in the refrigerator overnight. The reaction was purified on Sephadex G-25 using 0.1 M acetate buffer. Formamidine disulfide (FDS) (4 mg) was weighed into a culture tube, dissolved in 500 μL of ice-cold 0.1 M acetate buffer, placed on ice, and added to the albumin in one go with mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. AP3 (1.05 equiv.) dissolved in 200 μL of acetonitrile was then added to the ice-cold isothiourea-albumin with mixing and placed in the refrigerator overnight. The reaction was monitored using HPLC. The reaction reached approximately 60-80% completeness and the product was purified using Sephadex G-25 size-exclusion chromatography using distilled water as the solvent.

[0105] Example 5: Synthesis of Feline Serum Albumin (FSA)-AP3 Conjugate Feline serum albumin (BioWorld, Dublin, OH US) (100 mg) was dissolved in 4 mL of ice-cold PBS (pH 7.4) in a 50 mL Falcon tube and placed in an ice bath. In a separate tube, 3.5 μL of mercaptoethanol was added to 1 mL of PBS, cooled in an ice bath, then added to the solution of feline serum albumin, mixed, and placed in the refrigerator overnight. The reaction was purified on Sephadex G-25 using 0.1 M acetate buffer. Formamidine disulfide (FDS) (4 mg) was weighed in a culture tube, dissolved in 500 μL of ice-cold 0.1 M acetate buffer, placed on ice, and added to the albumin in one go with mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. AP3 thiol (1.05 equiv.) dissolved in 200 μL of acetonitrile was then added to the ice-cold isothiourea-albumin with mixing and placed in the refrigerator overnight. The reaction was monitored using HPLC. The reaction reached approximately 60-80% completeness and the product was purified using Sephadex G-25 size-exclusion chromatography using distilled water as the solvent.

[0106] Example 6: Synthesis of Equine Serum Albumin (ESA)-AP3 Conjugate Horse serum albumin (Abcom, Boston, MA US) (100 mg) was dissolved in 4 mL of ice-cold PBS (pH 7.4) in a 50 mL Falcon tube and placed in an ice bath. In a separate tube, 3.5 μL of mercaptoethanol was added to 1 mL of PBS, cooled in an ice bath, then added to the horse serum albumin solution, mixed, and placed in the refrigerator overnight. The reaction was purified on Sephadex G-25 using 0.1 M acetate buffer. Formamidine disulfide (FDS) (4 mg) was weighed into a culture tube, dissolved in 500 μL of ice-cold 0.1 M acetate buffer, placed on ice, and added to the albumin in one go with mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. AP3 thiol (1.05 equiv.) dissolved in 200 μL of acetonitrile was then added to the ice-cold isothiourea-albumin with mixing and placed in the refrigerator overnight. The reaction was monitored using HPLC. The reaction reached approximately 60-80% completeness and the product was purified using Sephadex G-25 size-exclusion chromatography using distilled water as the solvent.

[0107] Example 7: Synthesis of bovine serum albumin (BSA)-AP3 conjugate Bovine serum albumin (Sigma Aldrich, St. Louis, MO US) (100 mg) was dissolved in 4 mL of ice-cold PBS (pH 7.4) in a 50 mL Falcon tube and placed in an ice bath. In a separate tube, 3.5 μL of mercaptoethanol was added to 1 mL of PBS, cooled in an ice bath, then added to the solution of bovine serum albumin, mixed, and placed in the refrigerator overnight. The reaction was purified on Sephadex G-25 using 0.1 M acetate buffer. Formamidine disulfide (FDS) (4 mg) was weighed in a culture tube, dissolved in 500 μL of ice-cold 0.1 M acetate buffer, placed on ice, and added to the albumin in one go with mixing. After 5 min, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. AP3 thiol (1.05 equiv.) dissolved in 200 μL of acetonitrile was then added to the ice-cold isothiourea-albumin with mixing and placed in the refrigerator overnight. The reaction was monitored using HPLC. The reaction reached approximately 60-80% completeness and the product was purified using Sephadex G-25 size-exclusion chromatography using distilled water as the solvent.

[0108] Example 8: Lyophilization of AP3-BSA conjugate Mannitol (approximately 150 mg) and 100 mg sucrose were added to the purified AP3-BSA conjugate and mixed gently until everything was dissolved. The solution was then placed in a culture vial, which was placed in a bath of liquid nitrogen until it became a frozen solid. The vial was then placed in a freeze dryer for 2 days, resulting in a fluffy white powder.

[0109] Example 9: CCK8 Assay Method (In Vitro Cell Proliferation Assay) Cell viability assays were performed using Cell Counting Kit-8 (CCK-8; Dojindo Laboratories, Kumamoto, Japan) according to the manufacturer's protocol. Briefly, cells were seeded into 96-well cell culture plates at 8 × 10 cells per well. 3 The cells were seeded at different cell densities (10 -13 , 10-12 , 10 -11 , 10 -10 , 10 -9 , 10 -8 , 10 -7 , 10 -6 Compounds (100 μL, 100 μL each) were added to each well. The contents of the wells were mixed thoroughly. The plates were incubated at 37°C for 72 h. Then, 10 μL of CCK-8 solution was added and the plates were incubated for an additional 1–4 h. OD values ​​were measured at 450 nm using a microplate reader (BioTek, USA).

[0110] Figures 1-8 show that compounds of the invention (e.g., compounds I-1 and I-3) provide similar potency compared to AP3 when tested according to this method in several human and animal cancer cell lines.

[0111] Example 10: Xenograft Mouse Model (In Vivo Antitumor Assay) - Lymphoma Tumors Female BALB / c nude mice aged 4 to 6 weeks were purchased (Tyercan, Ltd, Shenzhen) and kept in the laboratory animal facility for 1 week to adapt to the new environment. A total of 5 × 10 6 Human gastric tumor SGC-7901 cells (200 μl) were inoculated by subcutaneous injection. 3 When the tumor volume reaches 0.5×length×width, the tumor volume becomes 2 ), tumor-bearing mice in the experimental group received (via tail vein injection) 100 μl of AP3-HSA at a first dose of 200 mg / kg (on day 0) and a second dose of 150 mg / kg (on day 14). The control group was injected in a similar manner with 100 μl of AP-3 containing 0.4 mg of AP-3 on days 0 and 14. On the same days, in the second control group, 100 μl of saline was injected via the tail vein. Tumor volumes were measured and mice were weighed twice a week from the first dose.

[0112] Figures 9 and 10 show that compound I-1 was more effective in treating gastric tumor growth compared to AP3, but compound I-1 caused a statistically insignificant weight loss suggesting an acceptable level of toxicity.

[0113] Example 11: Xenograft Mouse Model (In Vivo Antitumor Assay) - Gastric Tumor Four to six-week-old female SCID mice were purchased (Tyercan, Ltd, Shenzhen) and housed in the laboratory animal facility for one week to adapt to the new environment. A total of 5 × 10 6 Human U-937 lymphoma tumor cells (200 μl) were inoculated by subcutaneous injection. 3 When the tumor volume reaches 0.5×length×width, the tumor volume becomes 2 ), tumor-bearing mice in the experimental group received 100 μl of AP3-HSA (via tail vein injection) at a dose of 300 mg / kg on days 0, 7, 15, and 21. The control group was injected in a similar manner with 100 μl of AP-3 containing 0.5 mg of AP-3 on days 0, 7, 15, and 21. On the same days, in a second control group, 100 μl of saline was injected via the tail vein. Tumor volumes were measured and mice were weighed twice a week from the first dose.

[0114] Figures 11 and 12 show that compound I-1 was more effective in treating lymphoma tumor growth compared to AP3, but both AP3 and compound I-1 resulted in weight loss using this higher dosing protocol.

[0115] Example 12: Surface plasmon resonance affinity measurement of Thio-AP-3-HSA with human FcRn protein receptor Recombinant HSA was purchased from Wuhan Heyuan Biotechnology Co.,Ltd. Recombinant human FcRn (6his-tagged) was purchased from Novoprotein. FcRn protein was immobilized on flow channel 4 (FC4) of a CM5 sensor chip via amine coupling. First, the carboxymethylated dextran layers on FC3 and FC4 were activated by a freshly prepared mixture of 0.4 M EDC and 0.1 M NHS (1:1, v / v). 170 μL of the mixture was introduced to FC4 at a flow rate of 20 μL / min. The ligand was diluted to a final concentration of 100 μg / mL in sodium acetate buffer at pH 4.4 and coupled to the activated FC4 surface by its free amine at a flow rate of 10 μL / min for 100 μL. After coupling, 1 M ethanolamine was added to block the unreacted sites of FC3 and FC4 at a flow rate of 20 μL / min for 170 μL. HSA as a control was observed in a range of concentrations (1485nM, 742nM, 371nM, 1485nM, 2970nM, and 5940nM), and AP-3-HSA as an experimental sample was injected in a range of concentrations (448nM, 224nM, 3582nM, 1791nM, 1791nM, and 896nM) at a flow rate of 20μL / min into FC4 and FC3 over 2min. The dissociation time was set to 2min by the curve fitting process.

[0116] 13 and 14 show that HSA and compound I-1 exhibited similar concentration-dependent binding and kinetics for binding to the recombinant human FcRn receptor, suggesting no adverse structural effects on the HSA protein.

[0117] Example 13: Synthesis of Maytansine-SH Reaction Equation [ka]

[0118] process Pyridine was dried with CaH2 at room temperature for 16 hours. When the moisture was less than 0.1%, the drying was stopped and the pyridine was filtered for the next step. Otherwise, the drying time should be extended or CaH2 should be added until the moisture meets the standard.

[0119] 300 mg of maytansine (MTX) (1.0 equiv.) was placed in a 50 mL single-necked bottle and dried in vacuum at 55° C. for 4.0 hours. 360 mg of P2S5 (3.4 equiv.) was placed in a 50 mL single-necked bottle and dried in vacuum at 58° C. for 4.0 hours.

[0120] P2S5 was cooled to 20°C-30°C, 10 mL of pyridine was added, and the mixture was stirred at room temperature for 30 min.

[0121] Maytansine was dissolved in 10 mL of pyridine and the resulting solution was injected into the P2S5 solution and stirring was continued at room temperature for 30 min.

[0122] The reaction solution was placed under nitrogen and then heated to 60° C. for 2-4 hours. In-process control was performed until the maytansine level was less than 1%.

[0123] The solution was concentrated under reduced pressure and the residue was dissolved in methanol and purified by preparative HPLC.

[0124] The purified product was lyophilized to give maytansine-SH. [Table 3]

[0125] Analytical procedures Column: Agilent ZORBAX SB-C18 (150 x 4.6 mm 5 μm) Injection volume: 10 μL Flow rate: 1.0mL / min Column temperature: 30℃ Detection wavelength: 254 nm Mobile phase A: 0.01% trifluoroacetic acid (TFA)-HO Mobile phase B: 0.01% TFA-H2O:acetonitrile (CAN) = 2:8 [Table 4] Diluent: Methanol Sample solution: Place 3 drops of the reaction solution in a 10 mL centrifuge tube, add 1 mL of methanol to dissolve, shake well, and filter.

[0126] Example 14: Coupling and lyophilization of maytansine-HSA Solution preparation: 100 mM PBS (Buffer 1) Weigh out 2.6218 g of sodium dihydrogen phosphate, 5.5177 g of disodium hydrogen phosphate, and 2.922 g of sodium chloride and dissolve them in 400 mL of ultrapure water. Adjust the pH to 7.4 with 2 M NaOH, then fill to 500 mL using ultrapure water.

[0127] 0.1 M acetate, 0.15 M NaCl (Buffer 2) Weigh out 3.319 g of sodium acetate, 58.585 g of glacial acetic acid, and 87.660 g of sodium chloride and dissolve in 8000 mL of ultrapure water. Adjust the pH to 3 with 20% AcOH and fill to 10000 mL using ultrapure water.

[0128] 20 mM histidine, 4% mannitol (Buffer 3) Weigh out 15.515 g of histidine and 200.00 g of mannitol and dissolve in 4000 mL of ultrapure water, adjust the pH to 6.5 with dilute hydrochloric acid, then fill to 5000 mL using ultrapure water.

[0129] 20 mM histidine, 4% mannitol, 30% sucrose (Buffer 4) Weigh out 0.3103 g of histidine, 4.00 g of mannitol, and 30.00 g of sucrose, dissolve in 80 mL of ultrapure water, adjust the pH to 6.5 with dilute hydrochloric acid, then fill to 100 mL using ultrapure water.

[0130] 20 mM histidine, 4% mannitol, 1% sucrose (Buffer 5) 3. Weigh out 103 g of histidine, 40.000 g of mannitol, and 10.00 g of sucrose, dissolve in 800 mL of ultrapure water, then fill to 1000 mL with ultrapure water to obtain an alkaline solution of histidine. 4. Weigh out 193 g of histidine hydrochloride, 40.000 g of mannitol, and 10.00 g of sucrose, dissolve in 800 mL of ultrapure water, then fill to 1000 mL with ultrapure water to obtain an acidic solution of histidine. Slowly add the acidic solution of histidine to the alkaline solution of histidine until the pH is 6.5 to obtain buffer 5.

[0131] procedure Take an appropriate amount of liquid HSA, dissolve it in buffer 1, and measure its concentration.

[0132] reduction The reduction was carried out according to the conditions in Table 14A, but note that the buffer was added first, followed by the addition of DTT (dithiothreitol). [Table 5]

[0133] Add 197.260 mL of HSA (3809.091 mg, 57.334 μmol, 1 equiv., 19.310 mg / mL) to a 1 L Schott bottle, control the temperature at 20-24 °C, add 166.27 mL of buffer 1 with magnetic stirring, slowly add 17.374 mL of DTT (5 mM, 86.001 μmol, 1.5 equiv.) aqueous solution, then transfer to a shaker, maintain at 22 °C, and react for 1 h at 90 rpm. At the end of the reaction, weigh the net weight of the reaction solution, adjust the pH to 3.5-4.0 with 1 / 8 of the net weight of the reaction solution in 20% AcOH (w / w) aqueous solution, then perform buffer exchange.

[0134] Buffer exchange: a) The pump speed was set to 160 rpm (320 mL / min) and the mixture was washed with water, then with an alkaline solution and allowed to circulate for 30 minutes. b) Rinse with water and then wash the membrane with Buffer 2 until the pH of the solution passing through the end matches the pH of the buffer. c) Set the pump speed to 160 rpm (320 mL / min), transmembrane pressure (TMP): 0.25 bar, and exchange the solution for 10 DV (Diavolume). d) Buffer 2 was used for buffer exchange, pump speed was set at 160 rpm (320 mL / min), TMP: 0.25 bar, and the solution was exchanged for 10 DV. e) 298.495 mL was obtained, the concentration was 12.712 mg / mL and it contained 3,794.468 mg of protein.

[0135] Qualification: The modification was carried out according to the conditions in Table 14B, but note that Buffer 2 was added first, followed by the addition of formamide disulfide (FDS). [Table 6]

[0136] 298.75 mL of HSA-red (3797.610 mg, 57.161 μmol, 1 equivalent, 12.712 mg / mL) was added to a 1 L Schott bottle, the temperature was controlled at 5-10 °C, 51.547 mL of buffer 2 was added with magnetic stirring, 29.464 mL of FDS (25 mM, 714.512 μmol, 12.5 equivalent) aqueous solution was quickly added, then transferred to a shaker, maintained at 10 °C, and reacted at 90 rpm for 0.5 h. Buffer exchange was then performed.

[0137] buffer exchange a) The pump speed was set to 160 rpm (320 mL / min) and the mixture was washed with water, then with an alkaline solution and allowed to circulate for 30 minutes. b) Rinse with water and then wash the membrane with Buffer 2 until the pH of the solution passing through the end matches the pH of the buffer. c) Set pump speed to 160 rpm (320 mL / min), TMP: 0.25 bar, and exchange the solution for 10 DV (Diavolume). d) Buffer 2 was used for buffer exchange, pump speed was set at 160 rpm (320 mL / min), TMP: 0.25 bar, and the solution was exchanged for 10 DV. e) 257.96 mL was obtained, the concentration was 14.663 mg / mL and it contained 3782.553 mg of protein.

[0138] Coupling The coupling was carried out according to the conditions in Table 14C, but note that acetonitrile (ACN) was added first, followed by maytansine-SH. [Table 7]

[0139] 256.160 mL of HSA-mod (3756.159 mg, 56.473 μmol, 1 equiv., 14.663 mg / mL) was added to a 1 L Schott bottle, the temperature was controlled at 0-10 °C, 81.864 mL of buffer 2 was added with magnetic stirring, 26.176 mL of ACN was slowly added, and 11.386 mL of ACN containing maytansine-SH (10 mM, 112.947 μmol, 2 equiv.) was slowly added, then transferred to a shaker, maintained at 10 °C, and reacted at 90 rpm for 16 h. At the end of the reaction, the net weight of the reaction solution was weighed, and the pH was adjusted to 6.0-6.5 with 1 / 10 of the net weight of the reaction solution in 1 M Tris aqueous solution, and then buffer exchange was performed.

[0140] buffer exchange a) The pump speed was set to 160 rpm (320 mL / min) and the mixture was washed with water, then with an alkaline solution and allowed to circulate for 30 minutes. b) Rinse with water and then wash the membrane with Buffer 3 until the pH of the solution passing through the end matches the pH of the buffer. c) Set pump speed to 160 rpm (320 mL / min), TMP: 0.25 bar, and exchange the solution for 10 DV (Diavolume). d) Buffer 3 was used for buffer exchange, pump speed was set at 160 rpm (320 mL / min), TMP: 0.25 bar, and the solution was exchanged for 10 DV.

[0141] Mixing with additives The sample obtained after buffer exchange was diluted with buffer 3 to a concentration of 10.345 mg / mL, and 1 / 29 of the volume of buffer 4 was added, followed by filtration to obtain the drug substance. [Table 8]

[0142] Freeze drying Drug substance (5.3mL, density 1.020g / cm 3 ) and Buffer 5 were added to the cleaned and sterile vial, then the rubber plug was pushed halfway through the plug and the freeze dryer was turned on.

[0143] After 3 days, the freeze dryer was turned off and the vials were sealed with aluminum caps. [Table 9]

[0144] A heading used in this disclosure is not meant to imply that all disclosure relating to the heading is found in the section beginning with that heading, but rather that disclosure of any subject matter may be found throughout the entire specification.

[0145] It should be noted that terms such as "preferably," "generally," and "typically" are not used herein to limit the scope of the claimed invention or to imply that a particular feature is critical, essential, or essential to the structure or function of the claimed invention. Rather, these terms are merely intended to highlight alternative or additional features that may or may not be utilized in a particular embodiment of the invention.

[0146] As used in this disclosure, "a" or "an" means one or more unless otherwise specified. When used in the claims, when used with the word "comprising", the word "a" or "an" can mean one or more than one unless otherwise specified. When used in this disclosure or claims, "another" means at least a second or more unless otherwise specified. When used in this disclosure, the words "such as", "for example", and "eg" mean "for example, but not limited to", in that the list following that term ("such as", "for example", or "eg") provides some examples, but the list is not necessarily an entirely exhaustive list. The word "comprising" means that the items following the word "comprising" may include additional unrecited elements or steps, i.e., "comprising" does not exclude additional unrecited steps or elements.

[0147] In certain cases, the sequences disclosed herein are included in publicly available databases such as GENBANK® and SWISSPROT. Unless otherwise indicated or apparent, references to such publicly available databases are to the most current versions of the databases as of the filing date of this application.

[0148] Unless otherwise indicated, all numbers expressing properties such as amounts of ingredients, reaction conditions, and the like used in the specification and claims are to be understood as being modified in all instances by the term "about." Accordingly, unless otherwise indicated to the contrary, the numerical parameters set forth in the specification and claims are approximations that may vary depending upon the desired properties or functions sought by the subject matter of the present disclosure.

[0149] As used herein, the term "about," when referring to a value or amount of mass, weight, time, volume, concentration, or percentage, is meant to encompass variations in some embodiments of ±20%, in some embodiments of ±10%, in some embodiments of ±5%, in some embodiments of ±1%, in some embodiments of ±0.5%, and in some embodiments of ±0.1%, as appropriate for carrying out the disclosed methods.

[0150] A detailed description of one or more embodiments is provided herein. However, it should be understood that the present invention can be embodied in various forms. Therefore, the specific details disclosed herein (even if specified as preferred or advantageous) should not be construed as limiting, but rather should be used as an exemplary basis for the claims and as a representative basis for teaching a person skilled in the art to use the present invention in any suitable manner. Indeed, various modifications of the present invention in addition to those described herein will become apparent to a person skilled in the art from the foregoing description and the accompanying drawings. Such modifications are intended to fall within the scope of the appended claims.

Claims

1. Formula (I) 【Chemistry 1】 (I), A compound selected from the salts, optical isomers, geometric isomers, salts of isomers, and derivatives thereof, During the ceremony, -R １ H, -COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), C １ -C ８ Alkyl, C ２ -C ８ Alkenyl, C ２ -C ８ Alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl)), and phenyl-(C １ -C ４ alkyl), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), C １ -C ８ Alkyl, C ２ -C ８ Alkenyl, C ２ -C ８ Alkynyl, methyl, ethyl, C １ -C ４ alkylsulfonyl, or phenyl-(C １ -C ４ alkyl) is optionally selected from halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, phenyl, perfluorinated methyl, perfluorinated ethyl, amino, C １ -C ４ Alkanoylamino (-NH-CO-(C １ -C ４ alkyl), C １ -C ４ Alkoxy, benzyloxy (—O—CH ２ -phenyl), oxo (=O), C ２ -C ５ Alkoxycarbonyl (-CO-O-(C ２ -C ５ alkyl), methylenedioxy (having one or two bonded carbons, —O—CH ２ -O-), or C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl)), -R ２ H, allyl, vinyl, hydroxyl, Cl, Br, F, I, thiol, amino, nitro, cyano, C １ -C ４ Alkyl, C １ -C ４ Alkylnoic, phenyl, C １ -C ２ Perfluorinated alkyl, alkylamino, oxo, carboxy, acetyl, amido, and C １ -C ３ alkoxy; -R ３ But H, C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, cycloalkyl, and —COR ４ is selected from C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, aryl, or cycloalkyl may optionally be selected from the group consisting of halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ alkyl)), -R ４ But C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, cycloalkyl, and 【Chemistry 2】 is selected from C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, aryl, or cycloalkyl may optionally be selected from the group consisting of halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ alkyl)), -R ５ , R ６ , and R ７ are the same or different, and each independently represents C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ selected from alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, and cycloalkyl; １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, aryl, or cycloalkyl may optionally be selected from the group consisting of halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ alkyl)), - m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; - Z is selected from albumin, human serum albumin (HSA), bovine serum albumin (BSA), canine serum albumin (CSA), feline serum albumin (FSA), equine serum albumin (ESA), domain I of HSA, domain II of HSA, domain III of HSA, engineered albumin, variants thereof, and fragments thereof.

2. R １ H, -COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), C １ -C ４ Alkyl, C ２ -C ４ Alkenyl, C ２ -C ４ selected from alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), C １ -C ４ Alkyl, C ２ -C ４ Alkenyl, C ２ -C ４ Alkynyl, methyl, or ethyl may optionally be substituted with halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, amino, C １ -C ４ The compound of claim 1 , which may be substituted with one or more of alkoxy or oxo.

3. R １ H, -COCH ３ , carboxy (-CO ２ 2. The compound of claim 1, wherein the alkyl group is selected from the group consisting of aryl (-H), ethynyl (-CCH), methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl.

4. R ２ 2. The compound of claim 1, wherein is selected from the group consisting of H, Cl, Br, F, I, allyl, ethyl, methyl, and OH.

5. R ２ The compound of claim 1 , wherein is H or Cl.

6. R ３ 2. The compound of claim 1, wherein is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and nonyl.

7. R ３ but, 【Transformation 3】 2. The compound of claim 1, wherein:

8. R ３ but, 【Chemistry 4】 2. The compound of claim 1, wherein:

9. R ４ But C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ selected from alkynyl, methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, aryl, and cycloalkyl; １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, ethyl, aryl, or cycloalkyl may optionally be selected from the group consisting of halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ 10. The compound of claim 1, wherein the compound is substituted with one or more of the following:

10. R ４ But C １ -C １０ Alkyl, C ２ -C １０ Alkenyl, C ２ -C １０ selected from alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; １ -C １０ Alkyl, C ２ -C １０ Alkenyl, C ２ -C １０ Alkynyl, methyl, or ethyl may optionally be substituted with halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ 10. The compound of claim 1, wherein the compound is substituted with one or more of the following:

11. R ４ 2. The compound of claim 1, wherein is selected from the group consisting of methyl, ethyl, propyl, butyl, pentyl, and hexyl.

12. R ４ but, 【Transformation 5】 and R ５ , R ６ , and R ７ are the same or different, and each independently represents C １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ selected from alkynyl, methyl, ethyl, perfluorinated methyl, and perfluorinated ethyl; １ -C １８ Alkyl, C ２ -C ２０ Alkenyl, C ２ -C ２０ Alkynyl, methyl, or ethyl may optionally be substituted with halogen, hydroxy (—OH), methanoyl (—COH), —COCH ３ , carboxy (-CO ２ H), ethynyl (-CCH), cyano (-CN), sulfo (-SO ３ H), methyl, ethyl, perfluorinated methyl, perfluorinated ethyl, C １ -C ４ Alkoxy, C ２ -C ４ Alkanoyl (-CO-(C ２ -C ４ alkyl), C ２ -C ４ Alkanoyloxy (—CO—O—(C ２ -C ４ alkyl), C ２ -C ４ Alkoxycarbonyl (—O—CO—(C ２ -C ４ alkyl), nitro, amino, mono(C １ -C ４ alkyl)amino, di(C １ -C ４ alkyl)amino, C １ -C ４ Alkylthio (-S-(C １ -C ４ alkyl), C １ -C ４ Alkyl sulfinyl (-SO-(C １ -C ４ alkyl), C １ -C ４ Alkylsulfonyl (-S(=O) ２ -(C １ -C ４ alkyl), oxo (=O), thioxo (=S), or C １ -C ４ Alkanoylamino (—CO—NH—(C １ -C ４ 10. The compound of claim 1, wherein the compound is substituted with one or more of the following:

13. R ５ , R ６ , and R ７ are the same or different and each is independently selected from methyl, ethyl, propyl, butyl, pentyl, and hexyl.

14. R ４ but, 【Transformation 6】 2. The compound of claim 1, wherein:

15. 2. The compound of claim 1, wherein m is 1, 2, 3, 4, or 5.

16. 2. The compound of claim 1, wherein m is 1 or 2.

17. 2. The compound of claim 1, wherein Z is BSA, CSA, HSA, FSA, or ESA (e.g., BSA, CSA, or HSA).

18. 2. The compound of claim 1, wherein Z is HSA.

19. 2. The compound of claim 1, wherein the compound is I-1, I-2, I-3, I-4, I-5, I-6, I-7, I-8, I-9, or I-10.

20. The compound of claim 1, wherein the compound is I-1 or I-3.

21. A composition comprising a compound according to any one of claims 1 to 20.

22. 22. The composition of claim 21, wherein the amount of the compound is from about 0.0001% (by weight of the total composition) to about 99%.

23. 22. The composition of claim 21, wherein the composition comprises a lyophilized compound.

24. A pharmaceutical composition comprising a compound according to any one of claims 1 to 20.

25. 25. The pharmaceutical composition of claim 24, wherein the amount of the compound is from about 0.0001% (by weight of the total composition) to about 50%.

26. 25. The pharmaceutical composition of claim 24, wherein the pharmaceutical composition further comprises a compounding ingredient.

27. 25. The pharmaceutical composition of claim 24, wherein the pharmaceutical composition comprises a lyophilized compound.

28. 21. An agent for providing a compound according to any one of claims 1 to 20 to an animal, comprising the compound according to any one of claims 1 to 20, wherein one or more compositions comprising the compound according to any one of claims 1 to 20 are administered one or more times, said compositions being the same or different if there are two or more administrations.

29. 30. The method of claim 28, wherein at least one of the one or more compositions further comprises a formulation ingredient.

30. A method for providing a compound according to any one of claims 1 to 20 to an animal, the method comprising administering one or more compositions comprising the compound according to any one of claims 1 to 20 one or more times, the compositions being the same or different if administered two or more times, and at least one of the one or more compositions comprising a compound according to any one of claims 1 to 20 or a pharmaceutical composition comprising a compound according to any one of claims 1 to 20.

31. 29. The method of claim 28, wherein at least one of the one or more administrations comprises parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration.

32. 29. The method of claim 28, wherein, when there is more than one administration, at least one composition used in at least one administration is different from the composition in at least one other administration.

33. 30. The method of claim 28, wherein at least one compound of the one or more compositions is administered to the animal in an amount of about 0.01 mg / kg to about 500 mg / kg of animal body weight.

34. 29. The method of claim 28, wherein the animal is a human, dog, primate, cat, or horse (e.g., a human, dog, or primate).

35. 21. A method for treating an animal for a disease, comprising administering one or more compositions comprising a compound according to any one of claims 1 to 20 one or more times, said compositions being the same or different when there are two or more administrations.

36. 36. The method of claim 35, wherein at least one of the one or more compositions further comprises a formulation ingredient.

37. 21. A method for treating an animal for a disease, comprising administering one or more compositions comprising the compound of any one of claims 1 to 20 one or more times, said compositions, if administered two or more times, may be the same or different, and at least one of said one or more compositions comprises a composition comprising the compound of any one of claims 1 to 20 or a pharmaceutical composition comprising the compound of any one of claims 1 to 20.

38. 36. The method of claim 35, wherein at least one of the one or more administrations comprises parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration.

39. 36. The method of claim 35, wherein, when there is more than one administration, at least one composition used in at least one administration is different from the composition in at least one other administration.

40. 36. The method of claim 35, wherein at least one compound of the one or more compositions is administered to the animal in an amount of from about 0.01 mg / kg to about 500 mg / kg of animal body weight.

41. 36. The method of claim 35, wherein the animal is a human, dog, primate, cat, or horse (e.g., a human, dog, or primate).

42. The method of claim 35, wherein the animal is in need of treatment.

43. The agent according to claim 35, which is for treating cancer.

44. 36. The agent according to claim 35, which is for treating acute lymphoblastic leukemia, astrocytoma, basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, chronic lymphocytic leukemia (CLL), CNS cancer, colon cancer, colorectal cancer, endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma, gastric cancer, thyroid cancer, uterine cancer, cancer that can lead to metastasis, cancer resulting from metastasis, or cancerous tumors thereof.

45. 36. The agent according to claim 35, which is for treating basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, CNS cancer, colon cancer, colorectal cancer, endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer, leukemia, liver cancer, lung cancer, lymphoma, melanoma, malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, renal cell carcinoma, rhabdomyosarcoma, squamous cell carcinoma, gastric cancer, thyroid cancer, uterine cancer, or cancerous tumors thereof.

46. The agent according to claim 35, which is for treating cancerous tumors.

47. The agent according to claim 35, which is for treating breast cancer, head and neck cancer, lung cancer, non-small cell lung cancer, lymphoma, ovarian cancer, or kidney cancer.

48. A process for preparing a compound according to any one of claims 1 to 20, said process comprising: (a) reacting Z with a disulfide to provide an activated Z; (b) reacting activated Z with formula (II) to provide a compound; (c) optionally recovering the compound; During the ceremony, Formula (II) 【Transformation 7】 (II) That's the method.

49. 49. The method of claim 48, wherein the disulfide is formamidine disulfide, aldrithiol, or 5,5'-dithiobis(2-nitrobenzoic acid).

50. 49. The method of claim 48, wherein the disulfide is formamidine disulfide.

51. 49. The method of claim 48, wherein the disulfide is at a concentration that is in molar equivalent excess of Z.

52. 49. The method of claim 48, wherein prior to step (a), one or more thiol reducing agents are added to Z, and optionally, at least a portion of the unreacted one or more thiol reducing agents is then removed.

53. 49. The method of claim 48, wherein the pH of the reaction solution in step (b) is from about 1.0 to about 7.0 (e.g., from about 1.0 to about 6.0).

54. 49. The method of claim 48, wherein formula (II) is dissolved in a polar solvent prior to its addition to activated Z.

55. 49. The method of claim 48, wherein the reaction in step (b) is for about 4 hours to about 48 hours.

56. 49. The method of claim 48, wherein the temperature of the reaction in step (b) is from about -5°C to about 20°C.

57. 49. The method of claim 48, wherein step (c) is not optional.

58. 49. The method of claim 48, wherein the method further comprises lyophilization after step (b) or after step (c). 【Request Item 59】 【Transformation 8】 wherein HSA is human serum albumin.

60. A pharmaceutical composition comprising the compound of claim 59.

61. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition comprises (a) a compound and (b) sucrose, mannitol, or both.

62. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition comprises a freeze-dried composition comprising (a) the compound and (b) sucrose, mannitol, or both.

63. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition comprises a compound and sucrose.

64. The pharmaceutical composition of claim 60, wherein the pharmaceutical composition comprises a freeze-dried composition comprising the compound and sucrose.

65. A drug comprising the pharmaceutical composition of any one of claims 60 to 64 for treating a disease in a human, wherein the disease is breast cancer, non-small cell lung cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, ovarian cancer, glioblastoma multiforme, hepatocellular carcinoma, squamous cell carcinoma of the head and neck, gastric cancer, melanoma, lymphoma, or non-Hodgkin's lymphoma.

66. The agent described in claim 65, wherein the disease is breast cancer, non-small cell lung cancer, lymphoma, gastric cancer or ovarian cancer.

67. The agent described in claim 65, wherein the disease is lymphoma or gastric cancer.

68. The agent described in claim 65, wherein the disease is lymphoma.

69. A method for treating a human for lymphoma, comprising administering the pharmaceutical composition of any one of claims 60 to 64 parenterally. 【Request Item 70】 【Chemistry 9】 【Chemistry 10】 wherein BSA is bovine serum albumin, CSA is canine serum albumin, FSA is feline serum albumin, and ESA is equine serum albumin.

71. A pharmaceutical composition comprising the compound described in claim 70.

72. The pharmaceutical composition of claim 71, wherein the pharmaceutical composition comprises a freeze-dried composition comprising (a) the compound and sucrose, or (b) the compound and sucrose.

73. The pharmaceutical composition of claim 71, wherein the pharmaceutical composition comprises a freeze-dried composition comprising (a) the compound, sucrose and mannitol, or (b) the compound, sucrose and mannitol.

74. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, wherein the animal is a cow, dog, cat or horse, and the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma or a cancerous tumor.

75. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a dog; The compound 【Chemistry 11】 and The agent, wherein the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma or a cancerous tumor.

76. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a dog; The compound 【Chemistry 12】 and The agent, wherein the disease is lymphoma, gastric cancer, or melanoma.

77. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a cat, The compound 【Chemistry 13】 and The agent, wherein the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma or a cancerous tumor.

78. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a cat, The compound 【Chemistry 14】 and The agent, wherein the disease is lymphoma or gastric cancer.

79. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a horse, The compound 【Chemistry 15】 and The agent, wherein the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma or a cancerous tumor.

80. An agent for treating an animal for a disease, comprising the pharmaceutical composition of any one of claims 71 to 73, The animal is a horse, The compound 【Chemistry 16】 and The agent, wherein the disease is lymphoma, gastric cancer, or melanoma.

81. A method for preparing a lyophilized composition comprising one or more of compounds I-1 to I-10, the method comprising: (a) providing one or more of compounds I-1 to I-10; (b) contacting one or more of compounds I-1 to I-10 of (a) with sucrose, mannitol, or both to obtain a mixture; (c) freezing the mixture of (b); and (d) removing water to obtain a lyophilized composition comprising one or more of compounds I-1 to I-10. Including, Compounds I-1 to I-10 are 【Chemistry 17】 wherein human serum albumin (HSA) is attached via its cys-34 thiol (I-1), [Chemistry 18] wherein bovine serum albumin (BSA) is attached via its cys-34 thiol (I-2). 【Chemistry 19】 where canine serum albumin (CSA) is attached via its cys-34 thiol (I-3). 【Chemistry 20】 where feline serum albumin (FSA) is attached via its cys-34 thiol (I-4). 【Chemistry 21】 where horse serum albumin (ESA) is attached via its cys-34 thiol (I-5). 【Chemistry 22】 wherein human serum albumin (HSA) is attached via its cys-34 thiol (I-6). 【Chemistry 23】 where bovine serum albumin (BSA) is attached via its cys-34 thiol (I-7). 【Chemistry 24】 where canine serum albumin (CSA) is attached via its cys-34 thiol (I-8). 【Chemistry 25】 (I-9), or 【Chemistry 26】 (I-10) That's the method.

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