Conjugates, compositions thereof, and related methods

Abstract

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

Description

[0001] This application is a divisional application of Chinese Patent Application No. 202280082516.8 (filed on December 14, 2022, entitled "Conjugates, Compositions Thereof and Related Methods").

[0002] 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 associated methods,” which is incorporated herein by reference in its entirety. Background Technology

[0003] Some compounds (e.g., inhibitors and / or macrocyclic compounds) possess potential anticancer and / or antitumor activities. Some of these compounds may be toxic to normal (e.g., non-cancer) animal cells and are sometimes ineffective at tolerable doses. There remains a significant need to develop new compounds for treating cancers in animals. Certain embodiments of the present invention address one or more of the aforementioned deficiencies.

[0004] Some embodiments of the present invention comprise compounds of the present invention (e.g., compounds of formula (I)). Other embodiments comprise compositions comprising compounds of the present invention (e.g., pharmaceutical compositions). Still other embodiments of the present invention comprise compositions for treating, for example, certain diseases using compounds of the present invention. Some embodiments comprise methods of administering and treating with compounds of the present invention (e.g., compounds of the present invention in a composition or in a pharmaceutical composition). Further embodiments comprise methods for preparing compounds of the present invention. Additional embodiments of the present invention are also discussed herein. Summary of the Invention

[0005] Some embodiments of the present invention include a compound selected from formula (I). (I) Its salts, optical isomers, geometric isomers, salts of isomers, and derivatives. In some embodiments, R 1The group selected from H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)) and phenyl-(C1-C4 alkyl), wherein the -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, C1-C4 alkylsulfonyl or phenyl-(C1-C4 alkyl) may optionally be substituted by one or more of the following: halogen, hydroxyl (- OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, phenyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkylamino (-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 linked carbons) or C1-C4 alkylthio (-S-(C1-C4 alkyl)). In other embodiments, R 2 The group is selected from H, allyl, vinyl, hydroxy, Cl, Br, F, I, thiol, amino, nitro, cyano, C1-C4 alkyl, C1-C4 alkylinoic, phenyl, C1-C2 perfluoroalkyl, alkylamino, oxo, carboxyl, acetyl, amide, and C1-C3 alkoxy. In some embodiments, R 3 Selected from H, C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, cycloalkyl and -COR 4 The C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In yet other embodiments, R 4 Selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, cycloalkyl and The C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In still other embodiments, R 5 R 6 and R 7 The same or different and each independently selected from C1-C 18 Alkyl, C2-C20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, and cycloalkyl, wherein C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S), or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some implementations, 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), HSA domain I, HSA domain II, HSA domain III, engineered albumin, mutants thereof, and fragments thereof.

[0006] In other implementations, R 1 The group is selected from H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, ethyl, perfluoromethyl, and perfluoroethyl. The -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, or ethyl group may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkoxy, or oxo. In some embodiments, R... 1Choose from the following groups: H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), methyl, ethyl, perfluoromethyl, and perfluoroethyl.

[0007] In other implementation schemes, R 2 The group consisting of H, Cl, Br, F, I, allyl, ethyl, methyl, and OH is selected. In other embodiments, R 2 It can be H or Cl.

[0008] In other implementations, R 3 The group consisting of methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and nonyl is selected. In some embodiments, R 3 for In some implementations, R 3 for In other implementation schemes, R 4 Selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, and cycloalkyl, wherein C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S), or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 4 Selected from C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne, methyl, ethyl, perfluoromethyl and perfluoroethyl, the C1-C 10 Alkyl, C2-C 10alkenyl, C2-C 10 The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkyl Oxycarbonyl (-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), thio (=S), or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 4 The group consisting of methyl, ethyl, propyl, butyl, pentyl, and hexyl is selected. In other embodiments, R 4 for And R 5 R 6 and R 7 The same or different and each independently selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl and perfluoroethyl, the C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkyl Oxycarbonyl (-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), thio (=S), or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 5 R 6 and R 7 The same or different and each independently selected from methyl, ethyl, propyl, butyl, pentyl, and hexyl. In other embodiments, R 4 for In some embodiments, m is 1, 2, 3, 4, or 5. In still other embodiments, m is 1 or 2. In yet another embodiment, Z is BSA, CSA, or HSA. In some embodiments, Z is HSA.

[0009] 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.

[0010] Some embodiments of the present invention comprise compositions including any of the compounds disclosed herein. In some 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.

[0011] Some embodiments of the present invention comprise pharmaceutical compositions including any of the compounds disclosed herein. In other embodiments, the amount of the compound is from about 0.0001% (by weight of the total composition) to about 50%. In some embodiments, the pharmaceutical composition further comprises formulation ingredients. In other embodiments, the pharmaceutical composition comprises lyophilized compounds.

[0012] Some embodiments of the invention include a method for administering any of the compounds disclosed herein to an animal, comprising one or more administrations of one or more compositions comprising any of the compounds disclosed herein, wherein if more than one administration is performed, the compositions may be the same or different. In yet another embodiment, at least one of the one or more compositions further comprises a formulation ingredient. 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 administration, mucosal administration, intravenous administration, subcutaneous administration, topical administration, intradermal administration, oral administration, sublingual administration, intranasal administration, or intramuscular administration. In some embodiments, if more than one administration is performed, at least one composition used for at least one administration is different from the composition used for at least one other administration. In some embodiments, at least one of the compounds in one or more compositions is administered to an animal at 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.

[0013] Some embodiments of the present invention include a method for treating a disease in animals, comprising administering one or more compositions comprising any of the compounds disclosed herein, once or multiple times, wherein if more than one administration is performed, the compositions may be the same or different. In some embodiments, at least one of the one or more compositions further comprises a formulation ingredient. 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 administration, mucosal administration, intravenous administration, subcutaneous administration, topical administration, intradermal administration, oral administration, sublingual administration, intranasal administration, or intramuscular administration. In still other embodiments, if more than one administration is performed, at least one composition used for at least one administration is different from the composition used for at least one other administration. In some embodiments, at least one compound of one or more compositions is administered to an animal at 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, dog, or primate. In yet another embodiment, the animal requires treatment. In some embodiments, the method is used to treat cancer. In some implementations, the method is used to treat 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 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, metastatic cancer, cancer resulting from metastasis, or cancerous tumors thereof. In other embodiments, the method is used to treat basal cell carcinoma, bladder cancer, myeloma, 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 other cancerous tumors thereof. In yet another embodiment, the method is used to treat cancerous tumors. In still another embodiment, the method is used to treat breast cancer, head and neck cancer, lung cancer, non-small cell lung cancer, lymphoma, ovarian cancer, or kidney cancer.

[0014] 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 activated Z; (b) reacting the activated Z with formula (II) to provide said compound; and (c) optionally, recovering said compound. In some embodiments, formula (II) is (II).

[0015] In some embodiments, the disulfide is formamidinium disulfide, dithiopyridine, or 5,5'-dithiobis(2-nitrobenzoic acid). In other embodiments, the disulfide is formamidinium disulfide. In other embodiments, the concentration of the disulfide exceeds Z in molar equivalents. In other embodiments, prior to step (a), one or more thiol reducing agents are added to Z, and optionally, at least some of the unreacted one or more thiol reducing agents are then removed. In some embodiments, 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). In other embodiments, formula (II) is dissolved in a polar solvent before being added to the activated Z. In some embodiments, the reaction in step (b) is carried out for about 4 hours to about 48 hours. In other embodiments, the temperature of the reaction in step (b) is from about -5°C to about 20°C. In yet other embodiments, step (c) is not optional. In some embodiments, the method further includes freeze-drying after step (b) or after step (c).

[0016] Other embodiments of the present invention are also disclosed herein. Attached Figure Description

[0017] The following figures form part of this specification and are included to further illustrate certain aspects of the invention. A better understanding of the invention can be achieved by referring to one or more of these figures in conjunction with the description of the specific embodiments presented herein.

[0018] Figure 1 A comparison of the effects of AP3 and the compound on the viability of I-1 human Daudi lymphoma tumor cells after 72 h of culture. Similar potency indicates the bioavailability of compound I-1.

[0019] Figure 2 The experiment comparing the effects of AP3 and compound I-1 on the viability of human Daudi lymphoma tumor cells after 72 h of culture was repeated. Similar potency again demonstrated the bioavailability of compound I-1.

[0020] Figure 3A comparison of the effects of AP3 and compound I-1 on the viability of human SGC-7901 gastric tumor cell cultures after 72 h of culture. Similar potency indicates the bioavailability of compound I-1.

[0021] Figure 4 The experiment comparing the effects of AP3 and compound I-1 on the viability of human SGC-7901 gastric tumor cell cultures after 72 h of culture was repeated. Similar potency again demonstrated the bioavailability of compound I-1.

[0022] Figure 5 A comparison of the effects of AP3 and compound I-1 on the viability of human U-937 lymphoma tumor cells after 72 h of culture. Similar potency indicates the bioavailability of compound I-1.

[0023] Figure 6 The experiment comparing the effects of AP3 and compound I-1 on the viability of human U-937 lymphoma tumor cells after 72 h of culture was repeated. Similar potency again demonstrated the bioavailability of compound I-1.

[0024] Figure 7 A comparison of the effects of AP3 and compound I-3 on the viability of human U-937 lymphoma tumor cells after 72 h of culture. Similar potency indicates the bioavailability of compound I-3.

[0025] Figure 8 The experiment comparing the effects of AP3 and compound I-3 on the viability of human U-937 lymphoma tumor cells after 72 h of culture was repeated. Similar potency again demonstrated the bioavailability of compound I-3.

[0026] Figure 9 Effects of AP3 and compound I-1 on the growth of human gastric tumor SGC-7901 cells subcutaneously injected into nude mice on day 0. For compound I-1, the compound was injected via tail vein on day 0 (200 mg / kg) and day 14 (150 mg / kg), and for AP3, the compound was injected via tail vein on day 0 and day 14. Compared to the control, AP3 was only slightly effective at these dose levels, while compound I-1 was effective throughout the study period.

[0027] Figure 10 AP3 and compound I-1 according to Figure 9The experimental protocol described herein was used to study the effects on body weight in nude mice, where compound I-1 was injected into the tail vein on days 0 (200 mg / kg) and 14 (150 mg / kg), and AP3 was injected into the tail vein on days 0 and 14. Compared to the control, AP3 was only slightly effective at this dose level, while compound I-1 was effective throughout the study period. A decrease in body weight was observed in the compound I-1 treatment group.

[0028] Figure 11 Effects of AP3 and Compound I-1 on the growth of human U-937 lymphoma tumor cells injected subcutaneously 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 into the tail vein on days 0, 7, 15, and 21. Compound I-1 was more effective than AP3. Compound I-1 almost completely inhibited tumor growth up to day 30.

[0029] Figure 12 AP3 and compound I-1 according to Figure 11 The experimental protocol described herein was used to study the effect on the body weight of SCID mice. Weight loss was observed in both treatment groups during this period. Body weight returned to normal approximately one week after the last injection.

[0030] Figure 13 By using surface plasmon resonance microscopy, the affinity of recombinant HSA for recombinant human FcRn receptor was shown to bind in a pH-dependent manner.

[0031] Figure 14 Using surface plasmon resonance microscopy, the affinity of compound I-1 for recombinant human FcRn receptor was shown to be pH-dependent. HSA (from Figure 13 Compound I-1 and compound I-1 exhibited similar concentration-dependent binding and response rates. Detailed Implementation

[0032] While embodiments covering the overall inventive concept may take different forms, various embodiments will be described herein, and it should be understood that this disclosure is to be considered exemplary only, and the overall inventive concept is not intended to be limited to the disclosed embodiments.

[0033] Some embodiments of the present invention comprise compounds of the present invention (e.g., compounds of formula (I)). Other embodiments comprise compositions comprising compounds of the present invention (e.g., pharmaceutical compositions). Still other embodiments of the present invention comprise compositions for treating, for example, certain diseases using compounds of the present invention. Some embodiments comprise methods of administering and treating with compounds of the present invention (e.g., compounds of the present invention in a composition or in a pharmaceutical composition). Further embodiments comprise methods for preparing compounds of the present invention. Additional embodiments of the present invention are also discussed herein.

[0034] As defined herein, “amino acid” includes, but is not limited to, any naturally occurring amino acid (including, but not limited to, the standard 20 amino acids: 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. Table A lists some examples of unusual amino acids, but is not limited to those listed in Table A. Amino acids can be in L form, D form, or neither (e.g., glycine). As used herein, if the three-letter or one-letter name of an amino acid is not indicated, the amino acid is in L form unless otherwise indicated. When not in a sequence, the form of an amino acid is specified with a hyphen (e.g., L-Lys). When in a sequence, the hyphen is removed.

[0035] As used herein (unless otherwise stated), a “fragment” of a protein or amino acid sequence contains at least three consecutive amino acids. For example, a fragment may contain 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, not exceeding 50, not exceeding 100, not exceeding 500, not exceeding 1000, not exceeding 1500, not exceeding 2000, 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 consecutive amino acids. In some implementations, the fragment may be at least 175 consecutive amino acids. The length of the fragment can be appropriately varied depending on the desired properties or functions.

[0036] As used herein (unless otherwise stated), a “mutant” of a protein or amino acid sequence (or similar terms such as mutant or mutated) includes truncation, addition, deletion, substitution, and other alterations to a protein or amino acid sequence, provided that a certain degree of desired property or desired function is maintained. In some embodiments, a mutation may be a combination of two or more truncations, deletions, additions, substitutions, or other alterations. In some embodiments, one or more substitutions may be conserved substitutions. In some embodiments, conserved substitutions may be based on the hydrophilicity index (e.g., within ±2, ±1, or ±0.5) as specified in Kyte and Doolittle's *Journal of Molecular Biology*, 1982, Vol. 157, pp. 105–132, U.S. Patent No. 4,554,101, or based on the hydrophilicity value (e.g., within ±2, ±1, or ±0.5) as specified in U.S. Patent No. 4,554,101, or based on the size of the amino acid (e.g., side group size). In some implementations, if one amino acid in the same group is substituted by another, the following substitutions are considered conserved: Group 1: Ile, Leu, Val, Ala, Gly; Group 2: Trp, Tyr, Phe; Group 3: Asp, Glu, Asn, Gln; Group 4: Cys, Ser, Thr, Met; Group 5: His, Lys, Arg. In some cases, conserved substitutions minimally disrupt (or may enhance) one or more desired properties or functions.

[0037] As used herein (unless otherwise stated), the term "alkyl" means a monovalent straight-chain or branched hydrocarbon chain (e.g., C1-C1). 24 For example, the terms "C1-C7 alkyl" or "C1-C4 alkyl" refer to straight-chain or branched saturated hydrocarbon groups 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, isopropyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, sec-pentyl, n-hexyl, and n-heptyl. Examples of C1-C4 alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, and tert-butyl.

[0038] As used herein (unless otherwise stated), the term "alkenyl" means containing one or more (e.g., 1, 2, 3, or 4) double bonds (e.g., C2-C). 24 Alkenyl groups are monovalent straight-chain or branched hydrocarbon chains. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, and 5-hexenyl.

[0039] As used herein (unless otherwise stated), the term "alkoxy" means any of the aforementioned alkyl groups (alkyl-O-) attached to the remainder of the molecule via an oxygen atom (e.g., C1-C). 23 Examples of alkoxy groups include, but are not limited to, methoxy (sometimes shown as MeO-), ethoxy, isopropoxy, propoxy, and butoxy.

[0040] As used herein (unless otherwise stated), the term "alkynyl" means a monovalent straight-chain or branched hydrocarbon chain containing one or more (e.g., 1, 2, 3, or 4) triple bonds, and optionally may also contain one or more (e.g., 1, 2, 3, or 4) double bonds (e.g., C2-C). 24 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.

[0041] As used herein (unless otherwise stated), the term "aryl" means a 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. For bicyclic aryl groups specified as substituted, one or both rings may be substituted.

[0042] As used herein (unless otherwise stated), the term "cycloalkyl" means a monocyclic or bicyclic 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12-membered hydrocarbon group. The ring may 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., bicyclooctane such as [2.2.2]bicyclooctane or [3.3.0]bicyclooctane, bicyclononane such as [4.3.0]bicyclononane, and bicyclodecane such as [4.4.0]bicyclodecane (decahydronaphthalene) or spiro compounds) and adamantane. For monocyclic cycloalkyl, the ring is not aromatic. For bicyclic cycloalkyl, if one ring is aromatic, the other ring is not aromatic. For bicyclic cycloalkyl specified as substituted, one or both rings may be substituted.

[0043] As used herein (unless otherwise stated), the term "halogen" means monovalent Cl, F, Br, or I.

[0044] As used herein (unless otherwise stated), the term "heteroaryl" means a monovalent monocyclic or bicyclic 5, 6, 7, 8, 9, 10, 11, or 12-membered hydrocarbon group in which one, two, three, four, five, or six carbon atoms are replaced by heteroatoms independently selected from nitrogen, oxygen, or sulfur atoms, and the monocyclic or bicyclic system is aromatic. Examples of heteroaryl groups include, but are not limited to, thienyl (or thiophenyl), furanyl, indolyl, pyrroleyl, pyridyl, pyrazinyl, oxazolyl, thiazolyl, quinolinyl, pyrimidinyl, imidazoleyl, triazolyl, tetrazolyl, 1H-pyrazol-4-yl, 1-Me-pyrazol-4-yl, pyridin-3-yl, pyridin-4-yl, 3,5-dimethylisooxazolyl, 1H-pyrrole-3-yl, 3,5-di-Me-pyrazolyl, and 1H-pyrazol-4-yl. For bicyclic heteroaryl groups, if one ring is aryl, then the other ring is heteroaryl. For bicyclic heteroaryl groups, one or both rings may have one or more heteroatoms. For bicyclic heteroaryl groups designated as substituted, one or both rings may be substituted.

[0045] As used herein (unless otherwise stated), the term "heterocyclic" means a monovalent monocyclic or bicyclic hydrocarbon of 5, 6, 7, 8, 9, 10, 11, or 12 members, wherein 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 system is not aromatic. Examples of heterocyclic groups include, but are not limited to, tetrahydropyran, pyrrolyl (e.g., pyrrolidine-1-yl, pyrrolidine-2-yl, pyrrolidine-3-yl, or pyrrolidine-4-yl), piperazinyl (e.g., piperazin-1-yl, piperazin-2-yl, piperazin-3-yl, or piperazin-4-yl), piperidinyl (e.g., piperidin-1-yl, piperidin-2-yl, piperidin-3-yl, or piperidin-4-yl), and morpholinyl (e.g., morpholin-1-yl, morpholin-2-yl, morpholin-3-yl, or morpholin-4-yl). For bicyclic heterocyclic groups, if one ring is aromatic (e.g., monocyclic aryl or heteroaryl), the other ring is not aromatic. For bicyclic heterocyclic groups, one or both rings may have one or more heteroatoms. For bicyclic heterocyclic groups specified as substituted, one or both rings may be substituted.

[0046] As used herein (unless otherwise stated), the term "heteroatom" means an atom selected from nitrogen, oxygen, or sulfur atoms.

[0047] As used herein (unless otherwise stated), the term "hydroxyl" or "hydroxyl" indicates the presence of a monovalent -OH group.

[0048] As used herein (unless otherwise stated), the term "substituted" (e.g., as in substituted alkyl groups) means that one or more hydrogen atoms in a chemical group (having one or more hydrogen atoms) may be replaced by one or more non-hydrogen substituents selected from the specified options. Substitution may occur at one or more positions. The term "optionally substituted" means that one or more hydrogen atoms in a chemical group (having one or more hydrogen atoms) may be substituted, but are not required to be substituted.

[0049] For any one or more chiral centers, some compounds of the present invention may have one or more chiral centers and may exist and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. The compounds of the present invention (e.g., Formula I) cover any optically active, racemic, stereoisomeric, polymorphic, or mixture thereof. If the chiral center does not provide an indication of its configuration (i.e., R or S) in the chemical structure, it should be considered to represent R, S, or a racemic form.

[0050] Some embodiments of the present invention include compounds of formula (I): (I).

[0051] In some embodiments, the compound of formula (I) may be in the form of a salt, an optical and geometric isomer, or a salt of an isomer. In other embodiments, the compound may be in various forms, such as an uncharged molecule, a component of a molecular complex, or a non-irritating, pharmacologically acceptable salt, including, but not limited to, hydrochlorides, hydrobromic acids, sulfates, phosphates, nitrates, borates, acetates, maleates, tartrates, and salicylates. In some cases, for acidic compounds, the salt may contain a metal, an amine, or an organic cation (e.g., a quaternary ammonium). In yet another embodiment, a simple derivative of the compound (e.g., an ether, ester, or amide) that has the desired retention and release properties but is readily hydrolyzed by body pH, enzymes, or other suitable means may be used.

[0052] In some implementation schemes, R 1It can be selected from H, -COCH3, carboxyl (-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 kynyl (e.g., C2, C3, C4, C5, C6, C7 or C8 kynyl), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C 4-alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)) and phenyl-(C1-C4 alkyl), wherein -COCH3, carboxyl (-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 alkenyl (e.g., C2, C3, C4, C5, C6, C7 or C8 alkenyl), C2-C8 alkenyl (e.g., C2, C3, C4, C5, C6, C7 or C8 alkenyl). 5. The group consisting of C6, C7, or C8 alkynyl, methyl, ethyl, C1-C4 alkylsulfonyl, or phenyl-(C1-C4 alkyl) may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), or sulfonyl (-SO3H). Methyl, ethyl, phenyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkylamino (-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 linked carbons) or C1-C4 alkylthio (-S-(C1-C4 alkyl)).

[0053] In other implementations, R 1The group may be selected from H, -COCH3, carboxyl (-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 alkenyl (e.g., C2, C3, C4, C5, C6, C7 or C8 alkenyl), methyl, ethyl, perfluoromethyl, and perfluoroethyl, wherein -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C8 alkyl (e.g., C1, C2, C3, C4, C5, C6, C7 or C8 alkyl), and C2-C8 alkenyl. (e.g., C2, C3, C4, C5, C6, C7, or C8 alkenyl), C2-C8 ynyl (e.g., C2, C3, C4, C5, C6, C7, or C8 ynyl), methyl or ethyl may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, phenyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkoxy, benzyloxy, oxo, or methylenedioxy. In other embodiments, R 1 The group can be selected from H, -COCH3, carboxyl (-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 alkenyl (e.g., C2, C3, or C4 alkenyl), methyl, ethyl, perfluoromethyl, and perfluoroethyl, wherein -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C4 alkyl (e.g., C1, C2, C3, or C4 alkyl), C2-C4 alkenyl (e.g., C1, C2, C3, or C4 alkenyl), methyl, ethyl, perfluoromethyl, and perfluoroethyl. 2. C3 or C4 alkenyl), C2-C4 ynyl (e.g., C2, C3, or C4 ynyl), methyl, or ethyl may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkoxy, or oxo. In some embodiments, R 1 It can be selected from H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), methyl, ethyl, perfluoromethyl, and perfluoroethyl. In some embodiments, R 1 It can be selected from H, methyl, and perfluoromethyl. In some embodiments, R 1It may be identical to Y in Hashimoto et al., US 4,424,219, which is incorporated herein by reference in its entirety.

[0054] In some implementation schemes, R 2 It can be selected from H, allyl, vinyl, hydroxy, 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 perfluoroalkyl, alkylamino, oxo, carboxyl, acetyl, amide, and C1-C3 alkoxy (e.g., C1, C2, or C3 alkoxy). In some embodiments, R 2 It can be H, Cl, Br, F, I, allyl, ethyl, methyl, or OH. In some embodiments, R 2 It can be H or Cl. In some implementations, R 2 It may be identical to X in Hashimoto et al., US 4,424,219, which is incorporated herein by reference in its entirety.

[0055] In some implementation schemes, R 3 Can be selected from H, C1-C 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 Or C 18 Alkyl), C2-C 20 Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15C 16 C 17 C 18 C 19 Or C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl) and -COR 4 The C1-C 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 Or C 18 Alkyl), C2-C 20 Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 Or C 20The alkynyl, methyl, ethyl, aryl (e.g., phenyl or naphthyl) or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) groups may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)) groups. C2-C4 alkoxy (-CO-O-(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), thio (=S) or C1-C4 alkoxyamino (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 3 It can be methyl, ethyl, propyl (e.g., isopropyl), butyl (e.g., isobutyl), pentyl (e.g., isopentyl), hexyl (e.g., isohexyl), heptyl (e.g., isohexyl), octyl (e.g., isooctyl), or nonyl (e.g., isononyl). In some embodiments, R 3 It can be isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, or isononyl. In other embodiments, R 3 It can be In some implementations, R 3 It can be In some implementations, R 3 It may be identical to R in Hashimoto et al., US 4,424,219, which is incorporated herein by reference in its entirety.

[0056] In other implementations, R 4 Can be selected from C1-C 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 Or C 18 Alkyl), C2-C 20Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 Or C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) and The C1-C 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 Or C 18 Alkyl), C2-C 20 Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C14 C 15 C 16 C 17 C 18 C 19 Or C 20 The alkynyl, methyl, ethyl, aryl (e.g., phenyl or naphthyl) or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) groups may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)) groups. C2-C4 alkoxy (-CO-O-(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), thio (=S) or C1-C4 alkoxyamino (-CO-NH-(C1-C4 alkyl)). In other embodiments, R 4 Can be selected from C1-C 10 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C6) 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 alkenyl), C2-C 10 Alkyne group (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl (e.g., phenyl or naphthyl), cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) and The C1-C 10 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C6) 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 alkenyl), C2-C 10 Alkyne group (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C10 The alkynyl, methyl, ethyl, aryl (e.g., phenyl or naphthyl) or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) groups may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)) groups. C2-C4 alkoxy (-CO-O-(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), thio (=S) or C1-C4 alkoxyamino (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 4 It can be methyl, ethyl, propyl (e.g., isopropyl), butyl (e.g., isobutyl), pentyl (e.g., isopentyl), hexyl (e.g., isohexyl), heptyl (e.g., isohexyl), octyl (e.g., isooctyl), or nonyl (e.g., isononyl). In some embodiments, R 4 It can be isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, or isononyl. In some embodiments, R 4 It can be methyl, ethyl, propyl (e.g., isopropyl), butyl, pentyl, or hexyl. In some embodiments, R 4 It can be isopropyl.

[0057] In some implementations, R 4 It can be .

[0058] In other implementation schemes, R 5 R 6 and R 7 They can be the same or different and each can be independently selected from C1-C. 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C17 Or C 18 Alkyl), C2-C 20 Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 Or C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl (e.g., phenyl or naphthyl), and cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), the C1-C 18 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, C6) 10 C 11 C 12 C 13 C 14 C 15 C 16 C 17 Or C 18 Alkyl), C2-C 20 Alkenyl groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9, C1) 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 Alkyne groups (e.g., C2, C3, C4, C5, C6, C7, C8, C9 ... 10 C 11 C12 C 13 C 14 C 15 C 16 C 17 C 18 C 19 Or C 20 The alkynyl, methyl, ethyl, aryl (e.g., phenyl or naphthyl) or cycloalkyl (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl) groups may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)) groups. C2-C4 alkoxy (-CO-O-(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), thio (=S) or C1-C4 alkoxyamino (-CO-NH-(C1-C4 alkyl)). In still other embodiments, R 5 R 6 and R 7 They can be the same or different and each can be independently selected from C1-C. 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl and perfluoroethyl, the C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkyl Oxycarbonyl (-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), thio (=S), or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 5 R 6 and R 7 They can be the same or different and each can be independently selected from C1-C. 10 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C6) 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 alkenyl), C2-C 10 Alkyne group (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 (alkynyl), methyl, ethyl, perfluoromethyl or perfluoroethyl, wherein C1-C 10 Alkyl groups (e.g., C1, C2, C3, C4, C5, C6, C7, C8, C9, or C6) 10 Alkyl), C2-C 10 Alkenyl (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10 alkenyl), C2-C 10 Alkyne group (e.g., C2, C3, C4, C5, C6, C7, C8, C9 or C 10The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following (e.g., 0, 1, 2, 3, 4, 5, or 6): halogen (e.g., F, Cl, Br, or I), hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-) (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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)). In some embodiments, R 5 R 6 and R 7 They can be the same or different and can each 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., isohexyl), octyl (e.g., isooctyl), and nonyl (e.g., isononyl). In some embodiments, R 5 R 6 and R 7 They can be the same or different and can each be independently selected from isopropyl, isobutyl, isopentyl, isohexyl, isoheptyl, isooctyl, and isononyl. In some embodiments, R 5 R 6 and R 7 They can be the same or different and can each be independently selected from methyl, ethyl, propyl (e.g., isopropyl), butyl, pentyl, and hexyl. In some embodiments, R 5 R 6 and R 7 They can be the same and both can be methyl.

[0059] In other implementations, R 4 It can be .

[0060] In some implementation schemes, R 4 R 5 R 6 and R 7 It can be compared with R in US 4,424,219 by Hashimoto et al.1 R 2 R 3 and R 4 The same applies; it is incorporated into this article in its entirety through reference.

[0061] 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 some embodiments, m is 1 or m is 2. In some embodiments, the solution comprising formula (I) can be a mixture of formulas with different m values, thereby producing a solution from which an average m value can be obtained. In some implementations, the solution average value of m can be any rational number from 1 to 10, such as 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.

[0062] 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, domain II, domain III of HSA, engineered albumin (e.g., Veltis® from Novozymes), mutants thereof, or fragments thereof (fragments containing mutants). 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.

[0063] In other embodiments, Z stabilizes the compound. A stabilizing compound may include, for example, maintaining or regulating (e.g., increasing or decreasing) any suitable desired property or function of the compound, including, but not limited to, water solubility (e.g., increased), half-life in blood (e.g., increased), half-life in vivo before reaching the target (e.g., increased), stability in circulation before reaching the target (e.g., increased), prevention or reduction of renal absorption, or prevention or reduction of brain absorption.

[0064] In some embodiments, Z is reacted with the free thiol on Z with the compound of the present invention (e.g., formula (I)). Connection. In other embodiments, Z is HSA and Z is connected via cys-34 to the compound of the present invention (e.g., formula (I)). Connection. In other embodiments, Z is CSA and Z is connected via cys-34 to the compound of the present invention (e.g., formula (I)). Connection. In other embodiments, Z is BSA and Z is connected to the compound of the present invention (e.g., formula (I)) via cys-34. Connection. In yet another embodiment, Z (e.g., HSA or HSA domain I) is connected via a free thiol on Z that has been activated (e.g., using any suitable activating compound, such as, but not limited to, formamidinium disulfide as described herein) to the compound of the invention (e.g., formula (I)). Connection. In other embodiments, Z (e.g., canine, cat, horse, or bovine albumin) is connected via a free thiol on Z that has been activated (e.g., using any suitable activating compound, such as, but not limited to, formamidinium disulfide) after a reaction of Z with a reducing compound (e.g., using any suitable activating compound, such as, but not limited to, formamidinium disulfide as described herein) and the compound of the present invention (e.g., formula (I)). connect.

[0065] In some implementations, the compound of formula (I) may be selected from those specified in Table 1. In some embodiments, the compounds of the present invention may 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 compounds of the present invention may be I-1, I-2, I-3, I-4, or I-5. In other embodiments, the compounds of the present invention may be I-1, I-2, I-3, I-6, or I-7. In still other embodiments, the compounds of the present invention may be I-1 or I-3. In still still other embodiments, the compounds of the present invention may be I-1.

[0066] Compositions containing pharmaceutical compositions One or more compounds of the present invention (e.g., formula (I)) may be part of the composition, and the amount thereof (by weight of the total composition) 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 about 75%, at least about 90%, at least about 95%, at least about 99%, at least about 99.99%, not more than about 75%, not more than about 90%, not more than about 95%, not more than about 99%, or not more than about 99.99%, 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%.

[0067] One or more compounds of the present invention (e.g., formula (I)) can be purified or isolated in an amount (by weight of the total composition) 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%, not more than about 75%, not more than about 90%, not more than about 95%, not more than about 99%, not more than about 99.99%, 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%.

[0068] Some embodiments of the present invention comprise compositions including compounds of the present invention (e.g., formula (I)). In some embodiments, the composition is a pharmaceutical composition, such as a composition suitable for administration to animals (e.g., mammals, primates, monkeys, humans, dogs, cats, horses, cattle, pigs, mice, rabbits, or rats). In some cases, the pharmaceutical composition is non-toxic, does not cause side effects, or both. In some embodiments, inherent side effects may be present (e.g., they may harm the patient or may be toxic or harmful to some extent in some patients).

[0069] In some embodiments, the compound of the present invention (e.g., formula (I)) may be a part of a pharmaceutical composition, and its amount 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 about 75%, at least about 90%, at least about 95%, at least about 99%, at least about 99.99%, not more than about 75%, not more than about 90%, not more than about 95%, not more than about 99%, not more than about 99.99%, about 0.001% to about 99%, about 0.001% to about 50%, about 0.1% to about 99%, about 1% to about 95%, about 10% to about 90%, or about 25% to about 75%. In some embodiments, the pharmaceutical composition may be in a dosage form suitable for topical, subcutaneous, intrathecal, intraperitoneal, oral, parenteral, rectal, skin, nasal, vaginal, or ocular administration. In other embodiments, the pharmaceutical composition may be in a dosage form suitable for parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular administration. The pharmaceutical composition may be in the form of, for example, tablets, capsules, pills, powders, suspensions, emulsions, solutions, gels (including hydrogels), pastes, ointments, creams, plasters, high-dose solutions, delivery devices, suppositories, enemas, injections, implants, sprays, aerosols, or other suitable forms.

[0070] In some embodiments, the pharmaceutical composition may comprise one or more formulation ingredients. A "formulation ingredient" can be any suitable (e.g., a dosage of a compound suitable for the present invention (e.g., formula (I)), a release timing of a compound suitable for the present invention (e.g., formula (I)), a disease suitable, a disease state suitable, a diseased organ suitable, or a route of delivery suitable) component, including but not limited to water (e.g., boiled water, distilled water, filtered water, pyrogen-free water, or chloroform-containing water), sugars (e.g., sucrose, glucose, mannitol, sorbitol, xylitol, or syrups made therefrom), ethanol, glycerol, diols (e.g., propylene glycol), acetone, ether, DMSO, surfactants The active ingredients include surfactants (e.g., anionic surfactants, cationic surfactants, amphoteric 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., ethyl oleate, glyceryl monostearate, or hydrogenated glyceryl esters), excipients, preservatives (e.g., cysteine, methionine), antioxidants (e.g., vitamins (e.g., A, E, or C), selenium, retinyl palmitate, sodium citrate, citric acid, chloroform, or parabens (e.g., methylparaben or propylparaben)) or combinations thereof.

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

[0072] In some embodiments, other formulations (e.g., formulations of pharmaceutical compositions) may include those incorporating the compounds of the present invention (e.g., formula (I)) (or controlled-release formulations) into food, food ingredients, feed, or beverages.

[0073] In some embodiments, other formulations (e.g., formulations of pharmaceutical compositions) may contain lyophilized compounds of the present invention (e.g., formula (I)), which may include (in some cases) any suitable substance to stabilize one or more compounds of the present 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 invention may include methods of administering or treating animals (e.g., humans or dogs) with an amount of at least one compound of the invention (e.g., formula (I)) that is effective in treating the animal with or suspected of having or being susceptible to cancer or producing the desired physiological effect. In some embodiments, the composition or pharmaceutical composition comprises at least one compound of the present invention (e.g., formula (I)) which may be administered to animals (e.g., mammals, primates, monkeys, or humans) in amounts 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. Under certain conditions, the dosage may be about 0.5 mg / kg human body weight or about 6.5 mg / kg human body weight. In some cases, certain animals (e.g., mammals, mice, rabbits, cats, pigs, or dogs) may be administered doses 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, those skilled in the art will understand that many concentrations can be used in the methods of the present invention, and that any number of concentrations will be able to be adjusted and tested in part using the guidance provided herein in order to find the concentration that achieves the desired results under given conditions. In other embodiments, the compounds of the present invention may be administered in combination with one or more other cancer therapeutic agents.

[0075] In some embodiments, the composition may comprise a dose (e.g., unit dose) of one or more compounds of the present invention (e.g., formula (I)) combined with a pharmaceutically acceptable carrier, and may additionally comprise other pharmaceutical reagents, agents, carriers, excipients, diluents, excipients, or combinations thereof. In some embodiments, the carrier, medium, or excipient may facilitate the administration, delivery, and / or improve the preservation of the composition. In other embodiments, one or more carriers comprise, but are not limited to, aqueous saline solutions, such as physiological saline, Ringer's solution, PBS (phosphate-buffered saline), and mixtures of various salts, including potassium and phosphate salts, with or without sugar additives such as glucose. The carrier may comprise aqueous and non-aqueous sterile injectable solutions, which may contain antioxidants, buffers, bacteriostatic agents, bactericidal antibiotics, and solutes that make the formulation isotonic with the body fluids of the intended recipient; and aqueous and non-aqueous sterile suspensions, which may contain suspending agents and thickeners. In other embodiments, one or more excipients may comprise, but are not limited to, water, saline, dextran, glycerol, ethanol, and combinations thereof. Non-toxic excipients, such as wetting agents, buffers, or emulsifiers, may also be added to the composition. Oral formulations may contain commonly used excipients, such as pharmaceutical-grade mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, and magnesium carbonate.

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

[0077] Methods of application and for treatment The compounds of the present invention (e.g., formula (I)) can be administered to animals via any suitable route of administration or formulation. The compounds of the present invention (e.g., formula (I)) can also be used to treat various diseases in animals. Animals include, but are not limited to, mammals, primates, monkeys (e.g., rhesus macaques, tamarins, or pig-tailed monkeys), humans, dogs, cats, horses, cattle, pigs, poultry (e.g., chickens), mice, rabbits, and rats. As used herein, the term "subject" refers to both human and animal subjects.

[0078] The compounds of the present invention (e.g., formula (I)) can be administered via any suitable route. Routes of administration can be, but are not limited to, oral, parenteral, skin, nasal, rectal, vaginal, and ocular routes. In other embodiments, the routes of administration can be parenteral, mucosal, intravenous, subcutaneous, topical, intradermal, oral, sublingual, intranasal, or intramuscular. The choice of route of administration can depend on, for example, 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))) and the age and weight of the animal, the specific disease (e.g., cancer), and the severity of the disease (e.g., the stage of cancer). Of course, combinations of routes of administration can be used as needed.

[0079] Some embodiments of the present invention include a method for administering to a subject a composition (e.g., a pharmaceutical composition) comprising the compounds of the present invention described herein (e.g., formula (I)), comprising administering one or more such compositions once or multiple times; if there are more than one administration, the compositions may be the same or different, and if there are more than one administration, the routes of administration may be the same or different.

[0080] Some embodiments of the present invention include a method for treating a subject with a composition (e.g., a pharmaceutical composition) comprising the compounds of the present invention described herein (e.g., formula (I)), comprising administering one or more such compositions once or multiple times; if there are more than one administration, the compositions may be the same or different, and if there are more than one administration, the routes of administration may be the same or different.

[0081] Animals that can be treated include, but are not limited to, mammals, primates, monkeys (e.g., macaques, rhesus monkeys, pig-tailed monkeys), humans, dogs, cats, horses, pigs, poultry (e.g., chickens), cattle, mice, rabbits, and rats. As used herein, the term "subject" refers to both human and animal subjects. Subjects with a predisposing disease (e.g., cancer) can be either human or animal subjects. In some cases, animals (e.g., humans or dogs) require treatment for cancer.

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

[0083] In some embodiments, the cancers that can be treated in animals (e.g., mammals, pigs, dogs, poultry (e.g., chickens), cattle, cats, primates, rodents, monkeys, rabbits, mice, rats, and humans) using the compounds of the present 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 cancers (e.g., glioblastoma, glioblastoma multiforme, glioma, or astrocytoma), colon cancer, colorectal cancer (e.g., colon cancer or rectal cancer), endometrial cancer, gastric cancer, glioblastoma, glioblastoma multiforme, glioma, and neurogenic cancer using the compounds of the present invention (e.g., formula (I) or (I-1)). Gliosarcoma, head and neck cancer, hepatocellular carcinoma, kidney cancer (e.g., renal cell carcinoma), leukemia, liver cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma tumor formation), malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin 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 can lead to metastasis, cancer resulting from metastasis, or cancerous tumors thereof. In some implementations, the treatable cancers include, but are not limited to, basal cell carcinoma, bladder cancer, bone marrow cancer, breast cancer, CNS cancers (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., renal cell carcinoma), leukemia, liver cancer, and lung cancer (e.g., non-small cell lung cancer). Lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma tumor), malignant nerve sheath tumor, medulloblastoma, meningioma, multiple myeloma, nasopharyngeal carcinoma, neuroblastoma, non-Hodgkin 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 other cancerous tumors thereof.In some implementations, treatable cancers 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 cancers (e.g., glioblastoma, glioblastoma multiforme, gliosarcoma, or astrocytoma), lymphoma, melanoma (e.g., cutaneous malignant melanoma or melanoma neoplasm), ovarian cancer, kidney cancer, prostate cancer, breast cancer, or cancerous tumors thereof. In some implementations, treatable cancers include, but are not limited to, breast cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof. In some implementations, treatable cancers 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 kidney cancer. In some implementations, treatable cancers include, but are not limited to, cancerous tumors. Animals that can be treated include, but are not limited to, mammals, rodents, primates, monkeys (e.g., rhesus monkeys, pig-tailed monkeys), humans, dogs, cats, pigs, horses, poultry (e.g., chickens), cattle, mice, rabbits, and rats. As used herein, the term "subject" refers to both human and animal subjects. In some cases, animals require treatment (e.g., by showing signs of disease or cancer, or by having a cancerous tumor).

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

[0085] As used herein, the term “treating” (and its variations, such as “treatment”) should be considered in its broadest context. Specifically, the term “treating” does not necessarily mean treating an animal until it is fully recovered. Therefore, “treatment” encompasses improving symptoms, alleviating symptoms or effects associated with a condition, reducing the severity of a condition, or preventing symptoms, preventively improving symptoms, or otherwise reducing the risk of developing a particular condition. As used herein, references to “treating” animals include, but are not limited to, preventative and therapeutic treatment. Any composition described herein (e.g., pharmaceutical compositions) may be used to treat animals.

[0086] As treatment is related to the treatment of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof), treatment may include, but is not limited to, preventive and therapeutic treatment. Therefore, treatment may include, 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 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); inducing a bodily 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, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof); inhibiting or preventing cancer-related complications. (For example, the onset of symptoms associated with breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney 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, kidney cancer, or cancerous tumors thereof); causing the resolution of one or more 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) or cancer-related symptoms (e.g., reducing tumor size); causing remission of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof); or preventing the recurrence of cancer (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof). In some implementations, the treatment does not include preventative treatment of cancer (e.g., preventing or improving future cancer).

[0087] Animal treatment can be performed using any suitable method of administration (such as those disclosed herein) and with any suitable amount of the compounds of the present invention (e.g., formula (I) or (I-1)). In some embodiments, the treatment methods include treating cancers in animals (e.g., breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof). Some embodiments of the present invention include methods for treating subjects (e.g., animals, such as humans or primates) with a composition (e.g., a pharmaceutical composition) comprising the compounds of the present invention (e.g., formula (I) or (I-1)), comprising administering one or more such compositions once or multiple times; if more than one administration is present, the compositions may be the same or different.

[0088] In some embodiments, the treatment method comprises administering an effective amount of a composition comprising the compounds 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 affect treatment (e.g., treatment of cancer, such as, but not limited to, breast cancer, lung cancer (e.g., non-small cell lung cancer), lymphoma, ovarian cancer, kidney cancer, or cancerous tumors thereof) in animals. In some embodiments, the effective amount may encompass the therapeutically effective amounts disclosed herein. In some embodiments, the effective amount may vary depending on the subject and the specific treatment affected. The exact amount required may vary among subjects, for example, based on the subject's age and general condition, the specific adjuvant used (if applicable), the administration regimen, etc. Therefore, the effective amount may vary, for example, based on specific circumstances, and an appropriate effective amount may be determined in specific circumstances. The effective amount may, for example, comprise any dose or amount of 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 can be administered to animals, such as mammals, primates, monkeys, or humans) may be from about 0.005 to about 50 mg / kg body weight, from about 0.01 to about 15 mg / kg body weight, from about 0.1 to about 10 mg / kg body weight, from about 0.5 to about 7 mg / kg body weight, from about 0.005 mg / kg, from about 0.01 mg / kg, from about 0.05 mg / kg, from about 0.1 mg / kg, from about 0.5 mg / kg, from about 1 mg / kg, from about 3 mg / kg, from about 5 mg / kg, from about 5.5 mg / kg, from about 6 mg / kg, from about 6.5 mg / kg, from about 7 mg / kg, from about 7.5 mg / kg, from about 8 mg / kg, from about 10 mg / kg, from about 12 mg / kg, or from about 15 mg / kg. In some embodiments, the dosage may be from about 0.5 mg / kg human body weight or from about 6.5 mg / kg human body weight. In some cases, the effective amount of at least one compound of the present invention (e.g., formula (I) or (I-1)) (which can be applied to animals, such as mammals, rodents, mice, rabbits, cats, horses, pigs or dogs) can be from about 0.005 to about 50 mg / kg body weight, from about 0.01 to about 15 mg / kg body weight, from about 0.1 to about 10 mg / kg body weight, from about 0.5 to about 7 mg / kg body weight, from about 0.005 mg / kg, from about 0.01 mg / kg, from about 0.05 mg / kg, from about 0.1 mg / kg, from about 1 mg / kg, from about 5 mg / kg, from about 10 mg / kg, from about 20 mg / kg, from about 30 mg / kg, from about 40 mg / kg, from about 50 mg / kg, from about 80 mg / kg, from about 100 mg / kg or from about 150 mg / kg.In some embodiments, the effective amount of at least one compound of the present invention (e.g., formula (I) or (I-1)) (which can be administered to animals, such as mammals, primates, monkeys, or humans) can be from about 1 to about 1000 mg / kg body weight, from about 5 to about 500 mg / kg body weight, from about 10 to about 200 mg / kg body weight, from about 25 to about 100 mg / kg body weight, from about 1 mg / kg, from about 2 mg / kg, from about 5 mg / kg, from about 10 mg / kg, from about 25 mg / kg, from about 50 mg / kg, from about 100 mg / kg, from about 150 mg / kg, from about 200 mg / kg, from about 300 mg / kg, from about 400 mg / kg, from about 500 mg / kg, from about 600 mg / kg, from about 700 mg / kg, from about 800 mg / kg, from about 900 mg / kg, or from about 1000 mg / kg. In some cases, the dose can be from about 20 mg / kg human body weight or from about 100 mg / kg human body weight. In some cases, the effective amount of at least one compound of the present invention (e.g., formula (I) or (I-1)) (which can be applied to animals, such as mammals, rodents, mice, rabbits, cats, horses, pigs or dogs) can be from about 1 to about 1000 mg / kg body weight, from about 5 to about 500 mg / kg body weight, from about 10 to about 200 mg / kg body weight, from about 25 to about 100 mg / kg body weight, from about 1 mg / kg, from about 2 mg / kg, from about 5 mg / kg, from about 10 mg / kg, from about 25 mg / kg, from about 50 mg / kg, from about 100 mg / kg, from about 150 mg / kg, from about 200 mg / kg, from about 300 mg / kg, from about 400 mg / kg, from about 500 mg / kg, from about 600 mg / kg, from about 700 mg / kg, from about 800 mg / kg, from about 900 mg / kg or from about 1000 mg / kg.

[0089] "Therapeutic effective amount" means the amount that effectively achieves the desired and / or beneficial effect (e.g., reduction of tumor size). A therapeutic effective amount may be administered in one or more applications. For some purposes of this invention, a therapeutic effective amount is an amount suitable for a therapeutic indication (e.g., treatment of cancer). A therapeutic indication means achieving any desired effect, such as alleviating, improving, stabilizing, reversing, slowing, or delaying disease (e.g., cancer) progression, improving quality of life, or prolonging lifespan, or one or more of these effects. Such achievement can be measured by any suitable method, such as, but not limited to, measurement of tumor size.

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

[0091] In some embodiments, the 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 via the mechanisms disclosed herein and treatment of cancers (including but not limited to tumors) as disclosed herein. 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 with immunotherapy, which increases the therapeutic specificity against cancer and potentially limits additional systemic side effects. In still other embodiments, the compounds of the present invention (e.g., formula (I) or (I-1)) may be used as adjuvants to other chemotherapeutic agents. In some cases, the use of the compounds of the present invention (e.g., formula (I) or (I-1)) may reduce the dose duration of both the drug and the drug combination, thereby reducing side effects.

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

[0093] Method for preparing the compounds of the present invention (e.g., formula (I)) Some embodiments of the present invention include methods for preparing the compounds of the present invention (e.g., formula (I)). In some embodiments, the preparation of the compounds of the present invention (e.g., formula (I)) may include the step of reacting Z (as described herein, such as HSA or BSA) with a disulfide (e.g., formamidinium disulfide) to produce activated Z. In other embodiments, the disulfide can be any suitable disulfide used for the reaction (e.g., a disulfide suitable for linking Z to formula (II) with a disulfide bond), including but not limited to formamidinium disulfide, dithiopyridine, 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 (in its entirety incorporated herein by reference), and ANDREU et al. (1994) "Formation of Disulfide Bonds in Synthetic Peptides and Proteins". "DisulfideBonds in Synthetic Peptides and Proteins," from Methods in Molecular Biology, Volume 35: Peptide Synthesis Protocols, edited by M.M. Pennington and BM. Dunn, Humana Press Inc., Totowa, NJ (in its entirety is incorporated herein). In still other embodiments, the disulfide is formamidinium disulfide or 5,5'-dithiobis(2-nitrobenzoic acid). In yet another embodiment, the disulfide is formamidinium disulfide.In some embodiments, the pH of the reaction solution may be from about 1.0 to about 6.0, from about 2.0 to about 5.0, from about 3.0 to about 4.0, from about 1.0, from about 1.5, from about 2.0, from about 2.1, from about 2.2, from about 2.3, from about 2.4, from about 2.5, from about 2.6, from about 2.7, from about 2.8, from about 2.9, from about 3.0, from about 3.1, from about 3.2, from about 3.3, from about 3.4, from about 3.5, from about 3.6, from about 3.7, from about 3.8, from about 3.9, from about 4.0, from about 4.1, from about 4.2, from about 4.3, from about 4.4, from about 4.5, from about 4.6, from about 4.7, from about 4.8, from about 4.9, from about 5.0, from about 5.5, or from about 6.0. In other embodiments, the reaction solution comprises a buffer system (e.g., comprising 0.1 M sodium acetate or comprising 0.1 M sodium acetate and 150 mM sodium chloride). In some embodiments, the concentration of the disulfide (e.g., formamidine disulfide) exceeds Z in molar equivalence; for example, the concentration of the disulfide (e.g., formamidine disulfide) exceeds Z in molar equivalence from 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, etc. Approximately 1.4, 1.45, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, 12.0, 13.0, 14.0, 15.0, 16.0, 17.0, 18.0, 19.0, or 20.0. In some embodiments, after this reaction step (e.g., after the reaction has started, when the reaction is partially completed (e.g., >50% complete), when the reaction is nearing completion (e.g., >80% complete), or when the reaction is complete (e.g., >95% complete)), some or all of the unreacted disulfide (e.g., formamidinium disulfide) may be removed; for example, this removal may be carried out using any suitable method, including but not limited to HPLC (e.g., reversed-phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography, Sephadex G-25 size exclusion chromatography, or ion exchange chromatography), using silica gel, or combinations thereof. In some embodiments, as a result of the reaction, the activated Z comprises an S-substituted thiothiourea structure.

[0094] In some embodiments, one or more thiol reducing agents (e.g., any suitable thiol reducing agent, such as, but not limited to, mercaptoethanol or dithiothreitol) may be added to Z at any suitable concentration (e.g., about 1 mM, about 5 mM, about 10 mM, about 15 mM, or about 20 mM) before reacting Z (e.g., BSA, canine albumin, or nonhuman albumin) with a disulfide (e.g., formamidinium disulfide). Then, optionally, some or all of the unreacted thiol reducing agent is removed; for example, this removal may be carried out using any suitable method, including, but not limited to, HPLC (e.g., reversed-phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography, Sephadex G-25 size exclusion chromatography, or ion exchange chromatography), using silica gel, or combinations thereof.

[0095] Equation (II) is provided. (II), where R 1 R 2 and R 3The same as those disclosed herein. Formula (II) may be purchased or synthesized by any suitable method, including but not limited to those disclosed herein and those disclosed in US 4,424,219 by Hashimoto et al., which are incorporated herein by reference in their entirety. In some embodiments, formula (II) can be synthesized by contacting a starting compound (e.g., any compound of formula (II) in which -SH is replaced by -OH, such as anserine P3 (AP3) or maytansine) with a solution comprising P2S5 (e.g., a solution comprising P2S5 and pyridine) at any suitable temperature or temperature group (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; 30 minutes at room temperature, then 3 hours at 65°C) for any suitable time period (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). In other embodiments, formula (II) can be synthesized by contacting a solution comprising a starting compound (e.g., AP3 or maytansine) (e.g., a solution comprising a starting compound dissolved in dichloromethane) with H2S or a solution comprising H2S (e.g., by bubbling H2S gas into a solution of the starting compound); in some embodiments, a solution comprising trifluoroacetic acid (TFA) (e.g., a dichloromethane solution of TFA) can be reacted with the starting compound (e.g., AP3 or maytansine) / H2S solution at any suitable temperature (e.g., 15°C to 100°C, 15°C to 80°C, room temperature, 15°C, 20°C, 25°C, 3°C). Exposure at 0°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 time period (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 some embodiments, recovery of formula (II) is optional. In other embodiments, the recovery of formula (II) may optionally be performed using any suitable method, including but not limited to HPLC (e.g., reversed-phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography or ion exchange chromatography), silica gel, or a combination thereof. In some embodiments, the recovery of formula (II) may be accomplished using HPLC with a linear solvent gradient (e.g., as disclosed herein).In some embodiments of the synthesis of formula (II), water in the starting compound (e.g., AP3 or maytansine) powder may be optionally removed (e.g., by drying under vacuum at 40°C) and / or water in P2S5 (e.g., by drying under vacuum at 40°C).

[0096] In other embodiments (e.g., in embodiments of the synthesis of formula (II)), reflux of pyridine with calcium hydride and distillation (e.g., prior to the synthesis of formula (II)) is optional. In yet another embodiment, the glassware and rotating rod are optionally dried (e.g., prior to the synthesis of formula (II)).

[0097] In some embodiments, the activated Z can react with formula (II) to provide the compound of the present invention (e.g., formula (I)). In some 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., comprising 0.1 M sodium acetate or comprising 0.1 M sodium acetate and 150 mM sodium chloride). In some embodiments, the activated Z is in a solution colder than room temperature (e.g., in an ice bath). In other embodiments, formula (II) is dissolved in a polar solvent before being added to the activated Z; the dissolution of formula (II) can be accomplished 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 another embodiment, the dissolved formula (II) is added so as not exceeding about 5%, about 10%, about 15%, about 20%, about 25%, or about 30% by volume / volume of the total reaction solution. In some embodiments, after formula (II) (e.g., dissolved formula (II)) is added to the 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 implementations, the reaction is carried out at temperatures 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 some embodiments, the compounds of the present invention (e.g., formula (I)) may optionally be recovered. Recovery may be performed using any suitable method, including but not limited to HPLC (e.g., reversed-phase), LC, precipitation, centrifugation, column chromatography (e.g., size exclusion chromatography or ion exchange chromatography), silica gel, or combinations thereof. In some embodiments, the recovery of the compounds of the present invention (e.g., formula (I)) may be performed using a butyl agarose column with a reduced concentration of ammonium sulfate, followed by size exclusion chromatography (e.g., Sephadex G-25) using water, dilute acetic acid, ammonium acetate, or combinations thereof.

[0099] In other embodiments, the compounds of the present invention (e.g., formula (I)) may be lyophilized. In yet another embodiment, the lyophilized composition comprising the compounds of the present invention (e.g., formula (I)) may further comprise any suitable substance for stabilizing the compounds of the present invention (e.g., formula (I)), such as, but not limited to, one or more simple carbohydrates (e.g., sucrose, mannitol, trehalose, or combinations thereof).

[0100] The subject matter disclosed herein is further illustrated by the following specific, but non-limiting, embodiments. These embodiments may include data compilations representing data collected at different times during the development and experimental processes related to the present invention.

[0101] Example Example 1: Synthesis of anserin P3 thiol (AP3-SH) - Method A Anserine P3 (AP3 or AP-3) (MedChemExpress, LLC, Monmouth Junction, NJ US) (19 mg) was weighed in a screw-capped glass vial. Separately, phosphorus pentasulfide (P2S5) (7.3 mg) was weighed in a screw-capped vial equipped with a small rotating rod. The tops of the vials were secured with filter paper, and both vials were dried under high vacuum and heated to 40°C for 1 hour. Simultaneously, pyridine was dried with calcium hydride in a Stark instrument for several hours. The dissolved phosphorus pentasulfide was then dissolved in approximately 2 mL of anhydrous pyridine and capped with a screw cap (15 min). AP3 was dissolved in approximately 1 mL of anhydrous pyridine and added to the P2S5 solution. The mixture was stirred at room temperature with the screw cap on for 30 min, 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 rotary evaporator at 60°C. The residue was dissolved in a 1:1 mixture of HPLC solvents A and B (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. The pure fractions were combined and lyophilized.

[0102] Example 2: Synthesis of anserin P3 thiol (AP3-SH) - Method B Anserine P3 (AP3 or AP-3) (MedChemExpress, LLC, Monmouth-Junxen, NJ, USA) (10 mg) was weighed in a glass screw-cap vial and secured with a filter paper top; the vial was dried under high vacuum and heated to 40°C for 1 hour. The dried AP3 was then dissolved in 50 mL of anhydrous dichloromethane and added to a dried two-necked round-bottom flask equipped with a dropping funnel assembly under an argon flow and a bubbler to remove moisture. H2S gas was then bubbled into the solution while cooling in an ice bath. Anhydrous dichloromethane (5 mL) and TFA (1 mL) were added to the dropping funnel, and the mixture was slowly added dropwise to the AP3 solution and allowed to mix overnight. The solution was transferred to a round-bottom flask and evaporated to dryness at room temperature. The residue was dissolved in a 1:1 mixture of HPLC solvents A and B (HPLC solvent A - 0.01% TFA, solvent B - 80% acetonitrile (ACN) in 0.01% TFA) and purified using a linear gradient of 10% B to 100% B on a preparative HPLC system. The product eluted at approximately 74% B. The pure fractions were combined and lyophilized.

[0103] Example 3: Synthesis of human serum albumin (HSA)-AP3 conjugate 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, containing 150 mM NaCl) in a glass culture tube and placed in an ice bath. Formamidinium disulfide (FDS) (3 mg) was placed in a culture tube and 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 while mixing. After 5 minutes, excess FDS was separated from the activated albumin using Sephadex G-25 eluted with 0.1 M acetate buffer. The purified and activated albumin was cooled to 0 °C in an ice bath. Anserin P3 (AP3) (1.05 equivalent thiols) was dissolved in 200 µL of acetonitrile and added to the ice-cold isothiourea-albumin while mixing; it was then placed in a refrigerator overnight. The reaction was monitored using HPLC. The reaction was completed at approximately 60%-80%, and the product was purified using Sephadex G-25 size exclusion chromatography with 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 and cooled in an ice bath, then added to the canine serum albumin solution, mixed, and placed in the refrigerator overnight. The reactants were purified using 0.1 M acetate buffer on a Sephadex G-25. Formamidinium disulfide (FDS) (4 mg) was weighed in a culture tube and dissolved in 500 µL of ice-cold 0.1 M acetate buffer and placed on ice, then immediately added to the albumin while mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. Then, while mixing, AP3 (1.05 equivalents) dissolved in 200 µL acetonitrile was added to ice-cold isothiourea-albumin and incubated overnight. The reaction was monitored using HPLC. The reaction was allowed to proceed to approximately 60%-80% completion, and the product was purified using Sephadex G-25 size exclusion chromatography with 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 and cooled in an ice bath, then added to the feline serum albumin solution, mixed, and placed in the refrigerator overnight. The reactants were purified using 0.1 M acetate buffer on a Sephadex G-25. Formamidinium disulfide (FDS) (4 mg) was weighed in a culture tube and dissolved in 500 µL of ice-cold 0.1 M acetate buffer and placed on ice, then immediately added to the albumin while mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. Then, while mixing, AP3 thiol (1.05 equivalents) dissolved in 200 µL acetonitrile was added to ice-cold isothiourea-albumin, and the mixture was placed in a refrigerator overnight. The reaction was monitored using HPLC. When the reaction reached approximately 60%–80% completion, the product was purified using Sephadex G-25 size exclusion chromatography with 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 and cooled in an ice bath, then added to the horse serum albumin solution, mixed, and placed in a refrigerator overnight. The reactants were purified using 0.1 M acetate buffer on a Sephadex G-25. Formamidinium disulfide (FDS) (4 mg) was weighed in a culture tube and dissolved in 500 µL of ice-cold 0.1 M acetate buffer and placed on ice, then immediately added to the albumin while mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. Then, while mixing, AP3 thiol (1.05 equivalents) dissolved in 200 µL acetonitrile was added to ice-cold isothiourea-albumin, and the mixture was placed in a refrigerator overnight. The reaction was monitored using HPLC. When the reaction reached approximately 60%–80% completion, the product was purified using Sephadex G-25 size exclusion chromatography with 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 and cooled in an ice bath, then added to the bovine serum albumin solution, mixed, and placed in a refrigerator overnight. The reactants were purified using 0.1 M acetate buffer on a Sephadex G-25. Formamidinium disulfide (FDS) (4 mg) was weighed in a culture tube and dissolved in 500 µL of ice-cold 0.1 M acetate buffer and placed on ice, then immediately added to the albumin while mixing. After 5 minutes, excess FDS was separated from the activated isothiourea-albumin using Sephadex G-25 and 0.1 M acetate buffer. Then, while mixing, AP3 thiol (1.05 equivalents) dissolved in 200 µL acetonitrile was added to ice-cold isothiourea-albumin, and the mixture was placed in a refrigerator overnight. The reaction was monitored using HPLC. When the reaction reached approximately 60%–80% completion, the product was purified using Sephadex G-25 size exclusion chromatography with distilled water as the solvent.

[0108] Example 8: Lyophilization of AP3-BSA conjugate Mannitol (approximately 150 mg) and 100 mg of sucrose were added to the purified AP3-BSA conjugate and gently mixed until all substances dissolved. The solution was then transferred to a culture vial and inverted in a liquid nitrogen bath until solid-state freezing. The vial was then placed in a lyophilizer for two days to obtain a fluffy white powder.

[0109] Example 9: CCK8 determination method ( in vitro Cell proliferation assay Cell viability was measured using the Cell Counting Kit-8 (CCK-8; Dojindo, Kumamoto, Japan) following the manufacturer's instructions. In short, cells were counted at 8 × 10⁶ cells / day. 3 Cells were seeded at a density of 10 cells / well in 96-well cell culture plates, with different concentrations (10... -13 10 -12 10 -11 10 -10 10 -9 10 -8 10 -7 10 -6Compound M was added to each well. The contents of the wells were thoroughly mixed. The plate was incubated at 37°C for 72 h. Then 10 μL of CCK-8 solution was added and the plate was incubated for another 1 to 4 h. The OD value was measured at 450 nm using a microplate reader (BioTek, USA).

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

[0111] Example 10: Xenograft Mouse Model ( in vivo Antitumor assay - Lymphoma tumor Female BALB / c nude mice aged 4-6 weeks (Tyercan, Ltd., Shenzhen) were purchased and housed in the laboratory animal facility for one week to acclimatize to the new environment. A total of 5 × 10⁶ mice were administered subcutaneously. 6 Individual gastric tumor SGC-7901 cells (200 µl) were inoculated into the right flank of each mouse. Tumors were inoculated when they reached 100–300 mm in size. 3 (Tumor volume = 0.5 × length × width) 2 Mice in the experimental group were given a first dose of 100 µl AP3-HSA 200 mg / kg (on day 0) and a second dose of 150 mg / kg (on day 14) via tail vein injection. The control group was similarly administered 100 µl of AP-3 containing 0.4 mg AP-3 via tail vein injection on days 0 and 14. On the same day, the second control group received 100 µl of saline via tail vein injection. Tumor volume was measured and mice were weighed twice weekly since the first administration.

[0112] Figure 9-10 The results showed that compound I-1 was more effective in treating gastric tumor growth than AP3, while compound I-1 caused statistically insignificant weight loss, indicating an acceptable level of toxicity.

[0113] Example 11: Xenograft Mouse Model ( in vivo Antitumor assay - gastric tumor Female SCID mice aged 4-6 weeks (Taircom Biotechnology, Shenzhen) were purchased and housed in the laboratory animal facility for one week to acclimatize to the new environment. A total of 5 × 10⁸ mice were administered subcutaneously. 6 Individual U-937 lymphoma tumor cells (200 µl) were inoculated into the right flank of each mouse. Tumors were inoculated when they reached a size of 100–300 mm. 3 (Tumor volume = 0.5 × length × width) 2On days 0, 7, 15, and 21, tumor-bearing mice in the experimental group were administered 100 µl of AP3-HSA at a dose of 300 mg / kg via tail vein injection. On days 0, 7, 15, and 21, the control group was administered 100 µl of AP-3 containing 0.5 mg AP-3 via tail vein injection in a similar manner. On the same day, the second control group received 100 µl of saline via tail vein injection. Tumor volume was measured and mice were weighed twice weekly since the first administration.

[0114] Figure 11-12 The results showed that compound I-1 was more effective in treating lymphoma tumor growth than AP3, and both AP3 and compound I-1 resulted in weight loss when using the higher dosing regimen.

[0115] Example 12: Measurement of surface plasmon resonance affinity between thio-AP-3-HSA and human FcRn protein receptor Recombinant HSA was purchased from Wuhan Heyuan Biotechnology Co., Ltd. Recombinant human FcRn (6 his-tagged) was derived from a novoprotein. The FcRn protein was immobilized on channel 4 (FC4) of the CM5 sensor chip via amine coupling. First, the carboxymethylated dextran layers on FC3 and FC4 were activated using a freshly prepared mixture of 0.4M EDC and 0.1M NHS (1:1, v / v). 170 μL of the mixture was introduced into FC4 at a flow rate of 20 μL / min. The ligand was diluted to a final concentration of 100 μg / mL with sodium acetate buffer at pH 4.4, and 100 μL was coupled to the activated FC4 surface via its free amine at a flow rate of 10 μL / min. After coupling, 170 μL of 1M ethanolamine was added to block unreacted sites on FC3 and FC4 at a flow rate of 20 μL / min. HSA was observed at concentrations of 1485 nM, 742 nM, 371 nM, 1485 nM, 2970 nM, and 5940 nM as controls, and 60 μL of AP-3-HSA at concentrations of 448 nM, 224 nM, 3582 nM, 1791 nM, 1791 nM, and 896 nM as experimental samples was injected into FC4 and FC3 at a flow rate of 20 μL / min for more than 2 minutes. The dissociation time was set to 2 minutes during curve fitting.

[0116] Figure 13-14 The results showed that HSA and compound I-1 exhibited similar concentration-dependent binding and response rates to the recombinant human FcRn receptor, indicating that they did not have a detrimental structural effect on the HSA protein.

[0117] Example 13: Synthesis of Maytansin-SH Reaction process Pyridine was dried with CaH2 at room temperature for 16 hours. Drying was stopped when the moisture content became less than 0.1%, and the pyridine was filtered for use in the next step. Otherwise, the drying time should be extended or CaH2 should be added until the moisture content met the standard.

[0118] 300 mg of maytansine (MTX) (1.0 equivalent) was placed in a 50 mL single-necked flask and dried under vacuum at 55 °C for 4.0 h. 360 mg of P2S5 (3.4 equivalent) was placed in a 50 mL single-necked flask and dried under vacuum at 58 °C for 4.0 h.

[0119] Cool P2S5 to 20°C to 30°C, add 10 mL of pyridine and stir the mixture at room temperature for 30 min.

[0120] Dissolve maytansine in 10 mL of pyridine, and inject the resulting solution into the P2S5 solution. Continue stirring at room temperature for 30 min.

[0121] The reaction solution was placed under nitrogen atmosphere and then heated to 60°C for 2 to 4 hours. During the process, the temperature was controlled until the maytansine level became less than 1%.

[0122] The solution was concentrated under reduced pressure, and the residue was dissolved in methanol and purified by preparative HPLC.

[0123] The purified product was lyophilized to obtain maytansine-SH. Analysis program Column: Agilent ZORBAX SB-C18 (150*4.6 mm 5 μm) Injection volume: 10 μL Flow rate: 1.0 mL / min Column temperature: 30℃ Detection wavelength: 254 nm Mobile phase A: 0.01% trifluoroacetic acid (TFA) - H2O Mobile phase B: 0.01% TFA-H2O: Acetonitrile (CAN) = 2:8 gradient procedure Diluent: Methanol Sample solution: Take 3 drops of the reaction solution into a 10 mL centrifuge tube, add 1 mL of methanol to dissolve, shake well and filter.

[0124] Example 14: Coupling and lyophilization of Maytansin-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 2M NaOH, and then fill the volume to 500 mL with ultrapure water.

[0125] 0.1M acetate, 0.15M NaCl (buffer 2) Weigh 3.319 g sodium acetate, 58.585 g glacial acetic acid, and 87.660 g sodium chloride, and dissolve them in 8000 mL of ultrapure water. Adjust the pH to 3 with 20% AcOH, and then fill the volume to 10000 mL with ultrapure water.

[0126] 20 mM histidine, 4% mannitol (buffer 3) Weigh 15.515 g histidine and 200.00 g mannitol, dissolve them in 4000 mL of ultrapure water, adjust the pH to 6.5 with dilute hydrochloric acid, and then fill the volume to 5000 mL with ultrapure water.

[0127] 20 mM histidine, 4% mannitol, 30% sucrose (buffer 4) Weigh 0.3103 g histidine, 4.00 g mannitol and 30.00 g sucrose, dissolve them in 80 mL of ultrapure water, adjust the pH to 6.5 with dilute hydrochloric acid, and then fill the volume to 100 mL with ultrapure water.

[0128] 20 mM histidine, 4% mannitol, 1% sucrose (buffer 5) Weigh 3.103 g histidine, 40.000 g mannitol, and 10.00 g sucrose, dissolve them in 800 mL of ultrapure water, and then fill the volume to 1000 mL with ultrapure water to obtain an alkaline solution of histidine. Weigh 4.193 g histidine hydrochloride, 40.000 g mannitol, and 10.00 g sucrose, dissolve them in 800 mL of ultrapure water, and then fill the volume 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 reaches 6.5 to obtain buffer 5.

[0129] program Take an appropriate amount of liquid HSA and dissolve it in buffer 1, then determine its concentration.

[0130] reduction Perform the reduction according to the conditions in Table 14A, noting that the buffer should be added first, followed by DTT (dithiothreitol).

[0131] Table 14A Add 197.260 mL of HSA (3809.091 mg, 57.334 μmol, 1 equivalent, 19.310 mg / mL) to a 1 L Schott flask, maintain the temperature between 20°C and 24°C, add 166.27 mL of buffer 1 under magnetic stirring, and slowly add 17.374 mL of DTT (5 mM, 86.001 μmol, 1.5 equivalent) aqueous solution. Then transfer to a shaker, maintain at 22°C, and react at 90 rpm for 1 h. At the end of the reaction, weigh the net weight of the reaction solution and 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, and then perform buffer exchange.

[0132] Buffer exchange: a) Set the pump speed to 160 rpm (320 mL / min), wash with water, then wash with alkaline solution, and circulate for 30 min.

[0133] b) Wash with water and rinse the membrane with buffer 2 until the pH of the solution passing through the end matches the pH of the buffer.

[0134] c) Set the pump speed to 160 rpm (320 mL / min), transmembrane pressure (TMP): 0.25 bar, and exchange solution to 10 DV (Diavolume).

[0135] d) Buffer 2 is used for buffer exchange, and the pump speed is set to 160 rpm (320 mL / min), TMP: 0.25 bar, and the exchange solution is 10 DV.

[0136] e) Provides 298.495 mL, with a concentration of 12.712 mg / mL, and contains 3794.468 mg of protein.

[0137] Modification: Modify according to the conditions in Table 14B, noting that buffer 2 should be added first, followed by formamidine disulfide (FDS).

[0138] Table 14B 298.75 mL of HSA-reducing agent (3797.610 mg, 57.161 μmol, 1 equivalent, 12.712 mg / mL) was added to a 1 L Schott flask. The temperature was maintained between 5°C and 10°C. 51.547 mL of buffer 2 was added with magnetic stirring, followed by the rapid addition of 29.464 mL of FDS (25 mM, 714.512 μmol, 12.5 equivalent) aqueous solution. The mixture was then transferred to a shaker and reacted at 10°C and 90 rpm for 0.5 h. Buffer exchange was then performed.

[0139] Buffer exchange a) Set the pump speed to 160 rpm (320 mL / min), wash with water, then wash with alkaline solution, and circulate for 30 min.

[0140] b) Wash with water and rinse the membrane with buffer 2 until the pH of the solution passing through the end matches the pH of the buffer.

[0141] c) Set the pump speed to 160 rpm (320 mL / min), TMP: 0.25 bar, and exchange solution to 10 DV (Diavolume).

[0142] d) Buffer 2 is used for buffer exchange, and the pump speed is set to 160 rpm (320 mL / min), TMP: 0.25 bar, and the exchange solution is 10 DV.

[0143] e) Provides 257.96 mL, with a concentration of 14.663 mg / mL, and contains 3782.553 mg of protein.

[0144] Couplet Perform coupling according to the conditions in Table 14C, noting that acetonitrile (ACN) should be added first, followed by maytansine-SH.

[0145] Table 14C Add 256.160 mL of HSA-modified solution (3756.159 mg, 56.473 μmol, 1 equivalent, 14.663 mg / mL) to a 1 L Schott flask, maintaining the temperature between 0°C and 10°C. Add 81.864 mL of buffer 2 under magnetic stirring, then slowly add 26.176 mL of ACN, followed by slowly adding 11.386 mL of ACN containing maytansin-SH (10 mM, 112.947 μmol, 2 equivalents). Transfer the solution to a shaker and maintain the temperature at 10°C and react at 90 rpm for 16 h. At the end of the reaction, weigh the net reaction solution and adjust the pH to 6.0–6.5 with 1 M Tris aqueous solution (1 / 10 of the net reaction solution weight). Then perform buffer exchange.

[0146] Buffer exchange a) Set the pump speed to 160 rpm (320 mL / min), wash with water, then wash with alkaline solution, and circulate for 30 min.

[0147] b) Wash with water and rinse the membrane with buffer 3 until the pH of the solution passing through the end matches the pH of the buffer.

[0148] c) Set the pump speed to 160 rpm (320 mL / min), TMP: 0.25 bar, and exchange solution to 10 DV (Diavolume).

[0149] d) Buffer 3 is used for buffer exchange, and the pump speed is set to 160 rpm (320 mL / min), TMP: 0.25 bar, and the exchange solution is 10 DV.

[0150] Mix with excipients The sample obtained after buffer exchange was diluted with buffer 3 to a concentration of 10.345 mg / mL, and 1 / 29 of its volume of buffer 4 was added, and then filtered to obtain the drug substance.

[0151] Table 14D - Drug Substance Results freeze-dried The drug substance (5.3 mL, density 1.020 g / cm³) was... 3 Add 5 and buffer 5 to the cleaned and sterilized vials, then press the rubber stopper halfway and turn on the freeze dryer.

[0152] Three days later, the freeze dryer was turned off, and the vials were sealed with aluminum caps.

[0153] Table 14E - Results of freeze-drying The headings used in this disclosure do not imply that all disclosures related to a given heading are found in the section beginning with that heading. Disclosures on any subject can be found throughout the specification.

[0154] It should be noted that terms such as “preferredly,” “usually,” and “typically” are not intended to limit the scope of the claimed invention or to imply that certain features are critical, necessary, or even important to the structure or function of the claimed invention. Rather, these terms are intended only to emphasize alternative or additional features that may or may not be used in specific embodiments of the invention.

[0155] As used in this disclosure, “an” or “a” means one or more than one, unless otherwise stated. As used in the claims, when combined with the word “comprising,” “an” or “a” means one or more, unless otherwise stated. As used in this disclosure or the claims, “another” means at least a second or more, unless otherwise stated. As used in this disclosure, the phrases “such as,” “for example,” and “e.g.” mean “for example, but not limited to,” because the list following the term (“such as,” “for example,” or “e.g.”) provides some embodiments, but the list is not necessarily an exhaustive list. The word “comprising” means that the item following the word “comprising” may include additional unlisted elements or steps; that is, “comprising” does not exclude additional unlisted steps or elements.

[0156] In some cases, the sequences disclosed in this paper are included in publicly available databases, such as GENBANK. ® And SWISSPROT. Unless otherwise stated or obvious, references to such publicly available databases are references to the most recent version of the database as of the filing date of this application.

[0157] Unless otherwise stated, all figures used in the specification and claims to indicate the amount of components, properties such as reaction conditions, etc., should in all cases be understood to be modified by the term "about". Therefore, unless indicated to the contrary, the numerical parameters set forth in this specification and claims are approximate values ​​that may vary depending on the desired properties or functions sought to be obtained from the subject matter of this disclosure.

[0158] As used herein, when referring to values ​​or amounts of mass, weight, time, volume, concentration, or percentage, the term “about” is intended to cover variations of ±20% of a specific amount in some embodiments, ±10% of a specific amount in some embodiments, ±5% of a specific amount in some embodiments, ±1% of a specific amount in some embodiments, ±0.5% of a specific amount in some embodiments, and ±0.1% of a specific amount in some embodiments, as such variations are suitable for implementing the disclosed methods.

[0159] This document provides a detailed description of one or more embodiments. However, it should be understood that the invention can be embodied in various forms. Therefore, the specific details disclosed herein (even those specified as preferred or advantageous) should not be construed as limiting, but rather serve as an illustrative basis for the claims and as a representative basis for teaching those skilled in the art to use the invention in any appropriate manner. Indeed, various modifications to the invention, in addition to those described herein, will become apparent to those skilled in the art from the foregoing description and drawings. Such modifications are intended to fall within the scope of the appended claims.

[0160] This disclosure relates to the following implementation plan: 1. A compound selected from formula (I). (I) Its salts, optical isomers, geometric isomers, salts of isomers, and derivatives thereof. in -R 1 The group selected from H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkylsulfonyl (-S(=O)2-(C1-C4 alkyl)) and phenyl-(C1-C4 alkyl), wherein the -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C8 alkyl, C2-C8 alkenyl, C2-C8 alkynyl, methyl, ethyl, C1-C4 alkylsulfonyl or phenyl-(C1-C4 alkyl) may optionally be substituted by one or more of the following: halogen, hydroxyl (- OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, phenyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkylamino (-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 linked carbons) or C1-C4 alkylthio (-S-(C1-C4 alkyl)); -R 2 Selected from H, allyl, vinyl, hydroxyl, Cl, Br, F, I, thiol, amino, nitro, cyano, C1-C4 alkyl, C1-C4 alkylnoic, phenyl, C1-C2 perfluoroalkyl, alkylamino, oxo, carboxyl, acetyl, amide and C1-C3 alkoxy; -R 3 Selected from H, C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, cycloalkyl and -COR 4 The C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)); -R 4 Selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, cycloalkyl and The C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)); -R 5 R 6 and R 7 The same or different and each independently selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, and cycloalkyl, wherein C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)); -m is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; and -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), HSA domain I, HSA domain II, HSA domain III, engineered albumin, its mutants and fragments.

[0161] 2. The compound according to claim 1, wherein R 1 The group is selected from H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, ethyl, perfluoromethyl, and perfluoroethyl, wherein the -COCH3, carboxyl (-CO2H), ethynyl (-CCH), C1-C4 alkyl, C2-C4 alkenyl, C2-C4 alkynyl, methyl, or ethyl may optionally be substituted by one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, amino, C1-C4 alkoxy, or oxo.

[0162] 3. The compound according to any one of claims 1 to 2, wherein R 1 Choose from the following groups: H, -COCH3, carboxyl (-CO2H), ethynyl (-CCH), methyl, ethyl, perfluoromethyl, and perfluoroethyl.

[0163] 4. The compound according to any one of claims 1 to 3, wherein R 2 Choose from the following groups: H, Cl, Br, F, I, allyl, ethyl, methyl, and OH.

[0164] 5. The compound according to any one of claims 1 to 4, wherein R 2 It can be H or Cl.

[0165] 6. The compound according to any one of claims 1 to 5, wherein R 3 Choose from the group consisting of: methyl, ethyl, propyl, butyl, pentyl, hexyl, heptyl, octyl, and nonyl.

[0166] 7. The compound according to any one of items 1 to 6, wherein R 3 for .

[0167] 8. The compound according to any one of items 1 to 6, wherein R 3 for .

[0168] 9. The compound according to any one of claims 1 to 8, wherein R4 Selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl, perfluoroethyl, aryl, and cycloalkyl, wherein C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 The alkynyl, methyl, ethyl, aryl, or cycloalkyl groups may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkylyl (-CO-(C2-C4 alkyl)), C2-C4 alkylyloxy (-CO-O-(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), thio (=S) or C1-C4 alkylamide (-CO-NH-(C1-C4 alkyl)).

[0169] 10. The compound according to any one of claims 1 to 9, wherein R 4 Selected from C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10 Alkyne, methyl, ethyl, perfluoromethyl and perfluoroethyl, the C1-C 10 Alkyl, C2-C 10 alkenyl, C2-C 10The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkyl Oxycarbonyl (-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), thio (=S) or C1-C4 alkylamino (-CO-NH-(C1-C4 alkyl)).

[0170] 11. The compound according to any one of items 1 to 10, wherein R 4 Choose from the group consisting of: methyl, ethyl, propyl, butyl, pentyl, and hexyl.

[0171] 12. The compound according to any one of items 1 to 11, wherein R 4 for And R 5 R 6 and R 7 The same or different and each independently selected from C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20 Alkyne, methyl, ethyl, perfluoromethyl and perfluoroethyl, the C1-C 18 Alkyl, C2-C 20 alkenyl, C2-C 20The alkynyl, methyl, or ethyl group may optionally be substituted with one or more of the following: halogen, hydroxyl (-OH), formyloxy (-COH), -COCH3, carboxyl (-CO2H), ethynyl (-CCH), cyano (-CN), sulfonyl (-SO3H), methyl, ethyl, perfluoromethyl, perfluoroethyl, C1-C4 alkoxy, C2-C4 alkanoyl (-CO-(C2-C4 alkyl)), C2-C4 alkanoyloxy (-CO-O-(C2-C4 alkyl)), C2-C4 alkyl Oxycarbonyl (-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), thio (=S) or C1-C4 alkylamino (-CO-NH-(C1-C4 alkyl)).

[0172] 13. The compound according to any one of items 1 to 12, wherein R 5 R 6 and R 7 They may be the same or different and are each independently selected from methyl, ethyl, propyl, butyl, pentyl and hexyl.

[0173] 14. The compound according to any one of items 1 to 13, wherein R 4 for .

[0174] 15. The compound according to any one of items 1 to 14, wherein m is 1, 2, 3, 4 or 5.

[0175] 16. The compound according to any one of items 1 to 15, wherein m is 1 or 2.

[0176] 17. The compound according to any one of items 1 to 16, wherein Z is BSA, CSA, HSA, FSA or ESA (e.g., BSA, CSA or HSA).

[0177] 18. The compound according to any one of items 1 to 17, wherein Z is HSA.

[0178] 19. The compound according to any one of claims 1 to 18, 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.

[0179] 20. The compound according to any one of claims 1 to 19, wherein the compound is I-1 or I-3.

[0180] 21. A composition comprising the compound according to any one of claims 1 to 20.

[0181] 22. The composition according to claim 21, wherein the amount of said compound is from about 0.0001% (by weight of the total composition) to about 99%.

[0182] 23. The composition according to any one of claims 21 to 22, wherein the composition comprises q lyophilized compounds.

[0183] 24. A pharmaceutical composition comprising a compound according to any one of claims 1 to 20.

[0184] 25. The pharmaceutical composition according to claim 24, wherein the amount of said compound is from about 0.0001% (by weight of the total composition) to about 50%.

[0185] 26. The pharmaceutical composition according to any one of claims 24 to 25, wherein the pharmaceutical composition further comprises a formulation ingredient.

[0186] 27. The pharmaceutical composition according to any one of claims 24 to 26, wherein the pharmaceutical composition comprises a lyophilized compound.

[0187] 28. A method for administering a compound according to any one of claims 1 to 20 to an animal, comprising administering one or more compositions comprising the compound according to any one of claims 1 to 20 once or multiple times, wherein if more than one administration is performed, the compositions may be the same or different.

[0188] 29. The method according to claim 28, wherein at least one of the one or more compositions further comprises a formulation ingredient.

[0189] 30. The method according to any one of items 28 to 29, wherein at least one of the one or more compositions comprises a composition according to any one of items 21 to 23 or a pharmaceutical composition according to any one of items 24 to 27.

[0190] 31. The method according to any one of claims 28 to 30, wherein at least one of the one or more administrations comprises parenteral administration, mucosal administration, intravenous administration, subcutaneous administration, local administration, intradermal administration, oral administration, sublingual administration, intranasal administration, or intramuscular administration.

[0191] 32. The method according to any one of items 28 to 31, wherein if there is more than one application, at least one composition used for at least one application is different from the composition used for at least one other application.

[0192] 33. The method according to any one of claims 28 to 32, wherein at least one of the compounds in 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.

[0193] 34. The method according to any one of items 28 to 33, wherein the animal is a human, dog, primate, cat, or horse (e.g., a human, dog, or primate).

[0194] 35. A method for treating a disease in an animal, comprising administering, once or multiple times, one or more compositions comprising a compound according to any one of claims 1 to 20, wherein if more than one administration is performed, the compositions may be the same or different.

[0195] 36. The method according to claim 35, wherein at least one of the one or more compositions further comprises a formulation ingredient.

[0196] 37. The method according to any one of items 35 to 36, wherein at least one of the one or more compositions comprises a composition according to any one of items 21 to 23 or a pharmaceutical composition according to any one of items 24 to 27.

[0197] 38. The method according to any one of claims 35 to 37, wherein at least one of the one or more administrations comprises parenteral administration, mucosal administration, intravenous administration, subcutaneous administration, local administration, intradermal administration, oral administration, sublingual administration, intranasal administration, or intramuscular administration.

[0198] 39. The method according to any one of items 35 to 38, wherein if there is more than one application, the at least one composition used for at least one application is different from the composition used for at least one other application.

[0199] 40. The method according to any one of claims 35 to 39, wherein at least one of the compounds in 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.

[0200] 41. The method according to any one of items 35 to 40, wherein the animal is a human, dog, primate, cat, or horse (e.g., a human, dog, or primate).

[0201] 42. The method according to any one of items 35 to 41, wherein the animal requires the treatment.

[0202] 43. The method according to any one of items 35 to 42, wherein the method is used to treat cancer.

[0203] 44. The method according to any one of claims 35 to 43, wherein the method is used to treat 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 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 metastasize, cancer arising from metastasis, or cancerous tumors thereof.

[0204] 45. The method according to any one of claims 35 to 44, wherein the method is used to treat basal cell carcinoma, bladder cancer, myeloma, 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 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.

[0205] 46. ​​The method according to any one of items 35 to 45, wherein the method is used to treat cancerous tumors.

[0206] 47. The method according to any one of items 35 to 46, wherein the method is used to treat breast cancer, head and neck cancer, lung cancer, non-small cell lung cancer, lymphoma, ovarian cancer, or kidney cancer.

[0207] 48. A method for preparing a compound according to any one of claims 1 to 20, comprising: (a) Reacting Z with disulfide to provide activated Z; (b) Reacting the activated Z with formula (II) to provide the compound; and (c) Optionally, the compound is recovered; in - Equation (II) is (II).

[0208] 49. The method according to claim 48, wherein the disulfide is formamidinium disulfide, dithiopyridine, or 5,5'-dithiobis(2-nitrobenzoic acid).

[0209] 50. The method according to any one of claims 48 to 49, wherein the disulfide is formamidinium disulfide.

[0210] 51. The method according to any one of items 48 to 50, wherein the concentration of said disulfide exceeds Z in molar equivalents.

[0211] 52. The method according to any one of items 48 to 51, wherein one or more thiol reducing agents are added to Z prior to step (a), and optionally, at least some of the unreacted one or more thiol reducing agents are then removed.

[0212] 53. The method according to any one of items 48 to 52, wherein 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).

[0213] 54. The method according to any one of items 48 to 53, wherein formula (II) is dissolved in a polar solvent before being added to the activated Z.

[0214] 55. The method according to any one of items 48 to 54, wherein the reaction in step (b) is carried out for about 4 hours to about 48 hours.

[0215] 56. The method according to any one of items 48 to 55, wherein the temperature of the reaction in step (b) is from about -5°C to about 20°C.

[0216] 57. The method according to any one of items 48 to 56, wherein step (c) is not optional.

[0217] 58. The method according to any one of items 48 to 57, wherein the method further comprises freeze-drying after step (b) or after step (c).

Claims

1. A compound, which is HSA is human serum albumin.

2. A pharmaceutical composition comprising the compound according to claim 1.

3. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition comprises (a) the compound and (b) sucrose, mannitol, or both.

4. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition comprises a lyophilized composition comprising (a) the compound and (b) sucrose, mannitol, or both.

5. The pharmaceutical composition according to claim 2, wherein the pharmaceutical composition comprises the compound and sucrose.

6. The pharmaceutical composition of claim 2, wherein the pharmaceutical composition comprises a lyophilized composition comprising the compound and sucrose.

7. A method for treating a human disease, comprising administering the pharmaceutical composition according to claim 2, wherein the disease is breast cancer, non-small cell lung cancer, pancreatic cancer, pancreatic ductal adenocarcinoma, ovarian cancer, glioblastoma multiforme, hepatocellular carcinoma, head and neck squamous cell carcinoma, gastric cancer, melanoma, lymphoma, or non-Hodgkin lymphoma.

8. The method according to claim 7, wherein the disease is breast cancer, non-small cell lung cancer, lymphoma, gastric cancer, or ovarian cancer.

9. The method according to claim 7, wherein the disease is lymphoma or gastric cancer.

10. The method of claim 7, wherein the disease is lymphoma.

11. A method for treating lymphoma in humans, comprising administering the pharmaceutical composition according to claim 2, wherein the administration includes parenteral administration.

12. A compound selected from the group consisting of: 、 、 and , BSA stands for bovine serum albumin, CSA for canine serum albumin, FSA for feline serum albumin, and ESA for equine serum albumin.

13. A pharmaceutical composition comprising the compound according to claim 12.

14. The pharmaceutical composition of claim 13, wherein the pharmaceutical composition comprises (a) the compound and sucrose or (b) a lyophilized composition comprising the compound and sucrose.

15. The pharmaceutical composition of claim 13, wherein the pharmaceutical composition comprises (a) the compound, sucrose, and mannitol or (b) a lyophilized composition comprising the compound, sucrose, and mannitol.

16. A method for treating a disease in an animal, comprising administering the pharmaceutical composition according to claim 13, wherein the animal is a cow, dog, cat, or horse, and wherein the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma, or cancerous tumor.

17. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a dog, the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma, or cancerous tumor, and the compound is... .

18. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a dog, the disease is lymphoma, gastric cancer, or melanoma, and the compound is... .

19. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a cat, the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma, or cancerous tumor, and the compound is... .

20. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a dog, the disease is lymphoma or gastric cancer, and the compound is... .

21. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a horse, the disease is lymphoma, gastric cancer, cancer, sarcoma, squamous cell carcinoma, melanoma, or cancerous tumor, and the compound is... .

22. A method for treating a disease in an animal, comprising administering a pharmaceutical composition according to claim 13, wherein the animal is a horse, the disease is lymphoma, gastric cancer, or melanoma, and the compound is... .

23. A method for preparing a lyophilized composition, said lyophilized composition comprising one or more compounds I-1 to I-10, said method comprising... (a) Providing one or more compounds I-1 to I-10; (b) Contact one or more of the compounds I-1 to I-10 of (a) with sucrose, mannitol or both to provide a mixture; (c) Freezing the mixture described in (b); and (d) Removing water to provide a lyophilized composition comprising one or more compounds I-1 to I-10. Compounds I-1 to I-10 are: Human serum albumin (HSA) is linked via its cys-34 thiol (I-1). Bovine serum albumin (BSA) is linked via its cys-34 thiol (I-2). Canine serum albumin (CSA) is linked via its cys-34 thiol (I-3). In this context, feline serum albumin (FSA) is linked via its cys-34 thiol (I-4). Horse serum albumin (ESA) is linked via its cys-34 thiol (I-5). Human serum albumin (HSA) is linked via its cys-34 thiol (I-6). Bovine serum albumin (BSA) is linked via its cys-34 thiol (I-7). Canine serum albumin (CSA) is linked via its cys-34 thiol linker (I-8). (I-9), or (I-10).

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