Certain pladienolide compounds and methods of use

Abstract

To provide compounds and pharmaceutical compositions for treating cancer.SOLUTION: The present invention provides a compound selected from the compound represented by formula (III) and a pharmaceutically acceptable salt thereof. The compound is useful for treating cancer, particularly cancers in which agents targeting the spliceosome and mutations therein are known to be useful.SELECTED DRAWING: None

Description

[Technical Field]

[0001] This application claims the benefit of priority to U.S. Provisional Patent Application No. 62 / 655,021, filed April 9, 2018; U.S. Provisional Patent Application No. 62 / 679,653, filed June 1, 2018; U.S. Provisional Patent Application No. 62 / 814,838, filed March 6, 2019; and U.S. Provisional Patent Application No. 62 / 814,843, filed March 6, 2019, all of which are incorporated herein by reference.

[0002] Disclosed herein are novel organic compounds and pharmaceutical compositions containing such compounds. The compounds are useful in the treatment of cancer, particularly cancers for which drugs targeting the spliceosome and mutations therein are known to be useful. The compounds are also useful in the treatment of cancer when administered in combination with at least one additional therapy. [Background technology]

[0003] In eukaryotes, newly synthesized messenger RNA typically contains multiple introns, which are excised to form mature mRNA. The spliceosome is a multisubunit complex that accomplishes this task. The spliceosome consists of five small nuclear RNAs (snRNAs; U1-U6) assembled with various proteins.

[0004] Mutations in spliceosomal splicing factor 3B subunit 1 (SF3B1) are found in several cancers and are targets for anticancer drugs. Compounds isolated from the bacterium Streptomyces platensis (Sakai, Takashi; Sameshima, Tomohiro; Matsufuji, Motoko; Kawamura, Naoto; Dobashi, Kazuyuki; Mizui, Yoshiharu. Pladienolides, New Substances from Culture of Streptomyces platensis Mer-11107. I. Taxonomy, Fermentation, Isolation and Screening. The Journal of Antibiotics. 2004, Vol. 57, No. 3.) were discovered during a screen for vascular endothelial growth factor (VEGF) promoter inhibitors, known as pladienolides. They inhibit the expression of reporter genes under the control of the human VEGF promoter, and this inhibition is known to be a useful mechanism of action for anticancer drugs.

[0005] The compounds also inhibit the growth of U251 human glioma cells in vitro. The most potent of these compounds, pladienolide B, inhibits VEGF-stimulated gene expression with an IC of 1.8 nM. 50 inhibited glioma cell proliferation with an IC of 3.5 nM 50The structure of pladienolide B is known (Sakai, Takashi; Sameshima, Tomohiro; Matsufuji, Motoko; Kawamura, Naoto; Dobashi, Kazuyuki; Mizui, Yoshiharu. Pladienolides, New Substances from Culture of Streptomyces platensis Mer-11107. II. Physico-chemical Properties and Structure Elucidation. The Journal of Antibiotics. Vol. 57, No. 3. (2004)). Pladienolide B is known to target the SF3b spliceosome, inhibit splicing, and alter gene expression patterns (Kotake et al., "Splicing factor SF3b as a target of the antitumor natural product"). "Splicing factor SF3b as a target of the anticancer natural product pladienolide", Nature Chemical Biology 2007,3,570-575.

[0006] Certain pladienolide B analogs are also known: WO 2002 / 060890; WO 2004 / 011459; WO 2004 / 011661; WO 2004 / 050890; WO 2005 / 052152; WO 2006 / 009276; WO 2008 / 126918; and WO 2015 / 175594. For example, the pladienolide compound, (8E,12E,14E)-7-((4-cycloheptylpiperazin-1-yl)carbonyl)oxy-3,6,16,21-tetrahydroxy-6,10,12,16,20-pentamethyl-18,19-epoxytricosa-8,12,14-trien-11-olide, also known as E7107, is a semisynthetic derivative of the natural product pladienolide D, and results of its phase I clinical trials have been reported. Another example is the pladienolide pyridine compound (2S,3S,6S,7R,10R,E)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-((R,2E,4E)-6-(pyridin-2-yl)hepta-2,4-dien-2-yl)oxacyclododec-4-en-6-yl 4-methylpiperazine-1-carboxylate (also known as "(2S,3S,4E, 6S,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-((2E,4E,6R)-6-(pyridin-2-yl)hepta-2,4-dien-2-yl)oxacyclododec-4-en-6-yl 4-methylpiperazine-1-carboxylate), also known as H3B-8800, has received orphan drug designation for the treatment of certain blood cancers. Summary of the Invention [Problem to be solved by the invention]

[0007] However, there is a need for additional agents useful in the treatment of cancer, particularly cancers where agents that target the spliceosome and mutations therein are known to be useful.

[0008] In recent years, immune checkpoint blockade (ICB) has emerged as a paradigm shift for the treatment of several different cancer types, however, not all patients demonstrate robust / durable responses to ICB. See, e.g., Zappasodi, R. et al. Emerging Concepts for Immune Checkpoint Blockade-Based Combination Therapies. Cancer Cell 33, 581-598, doi:10.1016 / j.ccell.2018.03.005 (2018); and Wolchok, J.D. et al. Overall Survival with Combined Nivolumab and Ipilimumab in Advanced Melanoma. N Engl J Med 377, 1345-1356, doi:10.1056 / NEJMoa1709684 (2017). Thus, there is also a need to discover complementary therapeutic agents or any other therapy to administer in combination with ICB to improve and / or maximize patient response. [Means for solving the problem]

[0009] Provided herein are compounds of formula I: [ka] and pharmaceutically acceptable salts thereof [In formula: n is selected from 0, 1, 2 and 3; R 1 represents a C1-C6 alkyl group, a C3-C8 cycloalkyl group, -NR 9 R 10 basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] selected from the group R 9 is hydrogen, -NR 11 R 12 a C1-C6 alkyl group, a -(C1-C6 alkyl)-CO2H group, a C3-C8 cycloalkyl group, and a C3-C8 heterocyclyl group, wherein the C3-C8 cycloalkyl group and the C3-C8 heterocyclyl group may be unsubstituted or substituted 1 to 3 times with groups independently selected from a C1-C6 alkyl group, a -(C1-C6 alkyl)-CO2H group, hydroxy, a halogen group, and a C1-C6 alkoxy group; R 10 is selected from hydrogen and C1-C6 alkyl groups; R 2 or R 3 one of which is selected from hydrogen and a C1-C6 alkyl group, and the other is hydrogen, -OR 10 , -OC(O)R 10 , -OC(O)R 1and a C1-C6 alkyl group; R 4 is hydrogen or hydroxy; R 5 and R 6 are each independently selected from a C1 to C6 alkyl group; R7 and R8 are each independently selected from hydrogen, hydroxy, a C1-C6 alkoxy group, and a C1-C6 alkyl group; and Y is selected from phenyl, thiophenyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl, where Y may be unsubstituted or may be selected from hydroxyl, oxo group, C1-C6 alkyl group, C3-C5 cycloalkyl group, hydroxy C1-C6 alkyl group, C1-C6 alkoxy group, methoxy C1-C6 alkyl group, -NR 11 R 12 basis, [ka] and wherein R 11 and R 12 are each independently selected from hydrogen and a C1 to C6 alkyl group. DETAILED DESCRIPTION OF THE INVENTION

[0010] Also provided herein are compounds of formula II: [ka] and pharmaceutically acceptable salts thereof [In formula: X is selected from O, an NR′ group, and CH2, where R′ is selected from hydrogen and a C1-C6 alkyl group; R1 is methyl, an NR11R12 group, [ka] Groups, and [ka] is selected from R10 is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a haloC1-C6 alkyl group, wherein the C3-C8 cycloalkyl group can be unsubstituted or substituted 1 to 3 times with groups independently selected from a C1-C6 alkyl group, a hydroxyl group, a halogen group, and a C1-C6 alkoxy group; R11 and R12 are each independently selected from a C1 to C6 alkyl group; Either R2 or R3 is hydrogen or a C1-C6 alkyl group, and the other is selected from hydrogen, hydroxy, and a C1-C6 alkyl group; Either R4 or R5 is hydrogen, and the other is hydrogen, hydroxy, and [ka] Selected from; R6 and R7 are each independently selected from a C1 to C6 alkyl group; R8 and R9 are each independently selected from hydrogen and a C1-C6 alkyl group; or R8 and R9 together form a cyclopropyl ring; and Y is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, methoxy, and a -NR13R14 group, where R13 and R14 are each independently selected from hydrogen, a C1-C6 alkyl group, and a methoxyC1-C6 alkyl group; or R13 and R14 together with N are [ka] forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; wherein Y may be unsubstituted or a C1-C6 alkyl group, hydroxy, a hydroxy C1-C6 alkyl group, methoxy, a methoxy C1-C6 alkyl group, halo, a halo C1-C6 alkyl group, -C(O)NH2, -NHCOO-C1-C6 alkyl group, -COOH, [ka] and -NR15R16 groups, wherein R15 and R16 are each independently selected from hydrogen and a C1-C6 alkyl group.

[0011] Also provided herein are compounds of formula III: [ka] and pharmaceutically acceptable salts thereof [In formula: n is selected from 0, 1, 2 and 3; m is selected from 1, 2, and 3; R1 is a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a -NR11R12 group; [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] selected from the group R11 is selected from hydrogen, -NR16R17 group, C1-C6 alkyl group, C3-C8 cycloalkyl group, -(C1-C6 alkyl)-CO2H group, -(C1-C6 alkyl)-CO2R12 group, -(C1-C6 alkyl)-NR16R17 group, and C3-C8 heterocyclyl group, wherein -NR11R12 group, C1-C6 alkyl group, C3-C8 cycloalkyl group, and C3-C8 heterocyclyl group may be unsubstituted or substituted 1 to 3 times with groups independently selected from C1-C6 alkyl group, -(C1-C6 alkyl)-CO2H group, hydroxy, halogen group, and C1-C6 alkoxy group; R12 is selected from hydrogen and a C1-C6 alkyl group; Either R2 or R3 is selected from hydrogen and a C1-C6 alkyl group, and the other is selected from hydrogen, —OR10, —OC(O)R10, —OC(O)R1 and a C1-C6 alkyl group; R4 is selected from hydrogen and hydroxy; R5 and R6 are each independently selected from a C1 to C6 alkyl group; R7 and R8 are each independently selected from hydrogen, hydroxy, a C1-C6 alkoxy group, and a C1-C6 alkyl group; and R9 and R10 are each independently selected from hydrogen, a C1-C6 alkyl group, hydroxy, and a C1-C6 alkoxy group; or one of R9 or R10 is oxo, and the other is absent; Z is selected from a C1-C6 alkyl group, a —C(O)—C1-C6 alkyl group, a —OR13 group, and a —NR14R15 group; wherein R13 is selected from hydrogen, a C1-C6 alkyl group, and a —C(O)—C1-C6 alkyl group; wherein R14 and R15 are each independently selected from hydrogen, a C1-C6 alkyl group, and a methoxy C1-C6 alkyl group; or R14 and R15 together with N represent [ka] forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; In the formula, Z may be unsubstituted or may be a C1 to C6 alkyl group, a C3 to C5 cycloalkyl group, a hydroxy C1 to C6 alkyl group, a C1 to C6 alkoxy group, a methoxy C1 to C6 alkyl group, a -NR16R17 group, [ka] wherein R16 and R17 are each independently selected from hydrogen and a C1-C6 alkyl group.

[0012] Also disclosed herein are pharmaceutical compositions comprising at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, the pharmaceutical compositions further comprise at least one pharmaceutically acceptable carrier.

[0013] Also disclosed herein are methods of treating a subject with cancer, comprising administering to the subject a therapeutically acceptable amount of at least one compound of Formula I, at least one compound of Formula II, at least one compound of Formula III, and / or a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the cancer may be selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, cholangiocarcinoma, and / or lung cancer. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in the splicing factor 3B subunit 1 (SF3B1) gene or protein. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in spliceosome genes or proteins, such as those listed in Table 1. In some embodiments, administration of at least one compound of formula I, at least one compound of formula II, at least one compound of formula III, and / or a pharmaceutically acceptable salt of any of the foregoing induces at least one neo-antigen and / or T cell response.

[0014] Also disclosed herein is the use of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing in methods of therapeutic treatment, e.g., methods of treating cancer. In some embodiments, the cancer may be selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, cholangiocarcinoma, and / or lung cancer. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in the splicing factor 3B subunit 1 (SF3B1) gene or protein. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in spliceosome genes or proteins, such as those listed in Table 1. In some embodiments, administration of at least one compound of formula I, at least one compound of formula II, at least one compound of formula III, and / or a pharmaceutically acceptable salt of any of the foregoing induces at least one neo-antigen and / or T cell response.

[0015] Also disclosed herein is the use of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing in the preparation of a medicament. In some embodiments, the medicament is useful for treating cancer. In some embodiments, the cancer may be selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, cholangiocarcinoma, and / or lung cancer. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in the splicing factor 3B subunit 1 (SF3B1) gene or protein. In some embodiments, the cancer is selected from cancers that test positive for one or more mutations in spliceosome genes or proteins, such as those listed in Table 1. In some embodiments, administration of at least one compound of formula I, at least one compound of formula II, at least one compound of formula III, and / or a pharmaceutically acceptable salt of any of the foregoing induces at least one neo-antigen and / or T cell response.

[0016] Also disclosed herein is the use of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing, to target the spliceosome, e.g., subunit 1 of the SF3B spliceosome. As used herein, the following definitions shall apply unless otherwise indicated.

[0017] Also disclosed herein is a method for inducing at least one neoantigen, comprising contacting neoplastic cells with a therapeutically effective amount of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, such contacting can induce the production of at least one neoantigen.

[0018] Also disclosed herein is a method for inducing at least one neoantigen and / or T cell response in a subject having or suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0019] Also disclosed herein are methods for treating a subject having or suspected of having a neoplastic disorder. In some embodiments, the methods include administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, wherein administration can result in the induction of at least one neoantigen and / or T-cell response. In some embodiments, the methods can also include detecting one or more neoantigens and / or T-cell responses in the subject after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the methods can also include continuing administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing if one or more neoantigens and / or T-cell responses are detected.

[0020] Also provided herein is a method for treating a subject having or suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of at least one compound selected from at least one compound selected from compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing.

[0021] Also provided herein are neoantigen vaccines comprising at least one neoantigen peptide, in some embodiments, the at least one neoantigen peptide comprises a modified or novel neoantigen sequence derived by contacting neoplastic cells with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0022] The methods and uses provided herein may, in some embodiments, further comprise administering at least one additional therapy, hi some embodiments, the methods and uses provided herein may result in reduced systemic toxicity and / or improved tolerability.

[0023] Also disclosed herein is a method for treating cancer in a subject in need thereof, comprising administering at least one compound selected from compounds of Formula I, II, III, and pharmaceutically acceptable salts of any of the foregoing, and at least one additional therapy. Also disclosed herein is a method for treating a subject having or suspected of having a neoplastic disorder, comprising administering at least one compound selected from compounds of Formula I, II, III, and pharmaceutically acceptable salts of any of the foregoing, and at least one additional therapy.

[0024] As described herein, the disclosed compounds may be substituted with one or more substituents, such as those generally described herein or as exemplified by the specific classes, subclasses, and species of the present disclosure. In general, the term "substituted" refers to the replacement of a hydrogen radical in a given structure with a group of the specified substituent. Unless otherwise indicated, a substituted group may have a substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents may be the same or different at every position. Combinations of substituents envisioned by the present disclosure are those that result in the formation of stable or chemically feasible compounds.

[0025] "Stable" refers to a compound that does not change substantially chemically and / or physically when subjected to conditions that allow for its production, detection, and its recovery, purification, and use for one or more of the purposes disclosed herein. In some embodiments, a stable compound or chemically feasible compound is one that does not change substantially when placed at a temperature of 40° C. or less in the absence of moisture or other chemically reactive conditions for at least one week.

[0026] "Isomers" refer to compounds that have the same number and kind of atoms, and therefore the same molecular weight, but differ in the arrangement or configuration of the atoms. "Stereoisomers" refer to compounds that have the same sequence of bonding of the atoms, but differ in the arrangement of those atoms in space. "Diastereoisomers" or "diastereomers" refer to stereoisomers that are not enantiomers. "Enantiomers" refer to stereoisomers that are non-superimposable mirror images of each other.

[0027] Enantiomers as taught herein may include "enantiopure" isomers that contain substantially a single enantiomer at a particular asymmetric center or centers, e.g., greater than 90%, 92%, 95%, 98%, or 99%, or even equal to 100%, of a single enantiomer. An "asymmetric center" or "chiral center" refers to a tetrahedral carbon atom containing four different substituents.

[0028] As used herein, "stereoisomerically pure" refers to a compound or composition thereof that comprises one stereoisomer of the compound and is substantially free of other stereoisomers of the compound. For example, a stereoisomerically pure composition of a compound having one chiral center will be substantially free of the opposite enantiomer of the compound. In some embodiments, a stereoisomerically pure composition of a compound having two chiral centers will be substantially free of diastereomers of the compound and substantially free of the opposite enantiomer. In some embodiments, a stereoisomerically pure compound will be greater than about 80% by weight of one stereoisomer of the compound and less than about 20% by weight of the other stereoisomer of the compound, e.g., greater than about 90% by weight of one stereoisomer of the compound and less than about 10% by weight of the other stereoisomer of the compound, further e.g., greater than about 95% by weight of one stereoisomer of the compound and less than about 5% by weight of the other stereoisomer of the compound, and further e.g., greater than about 97% by weight of one stereoisomer of the compound and less than about 3% by weight of the other stereoisomer of the compound. See, for example, U.S. Patent No. 7,189,715.

[0029] The terms "R" and "S" as descriptive of isomers describe the stereochemical configuration at an asymmetrically substituted carbon atom. The designation of an "R" or "S" at an asymmetrically substituted carbon atom is made by application of the Cahn-Ingold-Prelog precedence rules, as is well known to those skilled in the art, and is set forth in the International Union of Pure and Applied Chemistry (IUPAC) Rules for the Nomenclature of Organic Chemistry, Section E, Stereochemistry.

[0030] An "amine oxide" or "amine-N-oxide" or "N-oxide" is a chemical compound containing the functional group R3N+-O-, which is an N-O bond with three additional hydrogens and / or hydrocarbon side chains attached to the N. It is sometimes written as R3N→O.

[0031] "Ar" or "aryl" refers to an aromatic carbocyclic moiety having one or more closed rings. Examples include, without limitation, phenyl, naphthyl, anthracenyl, phenanthracenyl, biphenyl, and pyrenyl.

[0032] "Heteroaryl" refers to a cyclic moiety having one or more closed rings, at least one of which has one or more heteroatoms (oxygen, nitrogen, or sulfur), where at least one of the rings is aromatic, and one or more of the rings can be independently fused and / or bridged. Examples include, without limitation, phenyl, thiophenyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl.

[0033] "Alkyl" or "alkyl group," as used herein, means a completely saturated straight-chain (i.e., unbranched), branched, or cyclic hydrocarbon chain. In some embodiments, an alkyl group contains 1 to 8 carbon atoms. In some embodiments, an alkyl group contains 1 to 6 carbon atoms ("C1-C6 alkyl group"). In some embodiments, an alkyl group contains 1 to 3 carbon atoms. In still other embodiments, an alkyl group contains 2 to 3 carbon atoms, and in some embodiments, an alkyl group contains 1 to 2 carbon atoms. In some embodiments, the term "alkyl" or "alkyl group" refers to a cycloalkyl group, also known as a carbocycle. Non-limiting examples of alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, cyclopropyl, and cyclohexyl.

[0034] "Alkoxy," as used herein, refers to an alkyl group, as defined above, attached to the main carbon chain through an oxygen ("alkoxy") atom.

[0035] "Haloalkyl" refers to an alkyl group substituted with one or more halo atoms (F, Cl, Br, I). For example, "fluoromethyl" refers to a methyl group substituted with one or more fluoro atoms (e.g., monofluoromethyl, difluoromethyl, or trifluoromethyl).

[0036] "Heteroatom" refers to O, S or N.

[0037] "Heterocyclyl" or "heterocyclic," as used herein, means a monocyclic heterocycle, a bicyclic heterocycle, or a tricyclic heterocycle containing at least one heteroatom in the ring.

[0038] Monocyclic heterocycles are 3-, 4-, 5-, 6-, 7-, or 8-membered rings containing at least one heteroatom independently selected from O, N, and S. In some embodiments, heterocycles are 3- or 4-membered rings containing one heteroatom selected from O, N, and S. In some embodiments, heterocycles are 5-membered rings containing zero or one double bond and one, two, or three heteroatoms selected from O, N, and S. In some embodiments, heterocycles are 6-, 7-, or 8-membered rings containing zero, one, or two double bonds and one, two, or three heteroatoms selected from O, N, and S. Representative examples of monocyclic heterocycles include, but are not limited to, azetidinyl, azepanyl, aziridinyl, diazepanyl, 1,3-dioxanyl, 1,3-dioxolanyl, dihydropyranyl (including 3,4-dihydro-2H-pyran-6-yl), 1,3-dithiolanyl, 1,3-dithianyl, imidazolinyl, imidazolidinyl, isothiazolinyl, isothiazolidinyl, isoxazolinyl, isoxazolidinyl, morpholinyl, oxadiazolinyl, oxadiazolidinyl , oxazolinyl, oxazolidinyl, piperazinyl, piperidinyl, pyranyl, pyrazolinyl, pyrazolidinyl, pyrrolinyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydropyranyl (including tetrahydro-2H-pyran-4-yl), tetrahydrothienyl, thiadiazolinyl, thiadiazolidinyl, thiazolinyl, thiazolidinyl, thiomorpholinyl, 1,1-dioxidethiomorpholinyl (thiomorpholinesulfone), thiopyranyl, and trithianyl.

[0039] Bicyclic heterocycles of the present disclosure include monocyclic heterocycles fused to an aryl group, or monocyclic heterocycles fused to a monocyclic cycloalkyl, or monocyclic heterocycles fused to a monocyclic cycloalkenyl, or monocyclic heterocycles fused to a monocyclic heterocycle. Examples of bicyclic heterocycles include, but are not limited to, 3,4-dihydro-2H-pyranyl, 1,3-benzodioxolyl, 1,3-benzodithiolyl, 2,3-dihydro-1,4-benzodioxinyl, 2,3-dihydro-1-benzofuranyl, 2,3-dihydro-1-benzothienyl, 2,3-dihydro-1H-indolyl, and 1,2,3,4-tetrahydroquinolinyl.

[0040] In some embodiments, the bicyclic heterocycle is a spiro heterocycle. As known in the art, a "spiro" heterocycle is a bicyclic moiety in which the rings are connected by only one atom. This connecting atom, also referred to as the spiro atom, is most often a quaternary atom such as carbon or nitrogen. Spiro compounds may be designated by the infix spiro, followed by the number of atoms in the small ring, excluding the spiro atom itself, and the number of atoms in the large ring, separated by a dot, in square brackets. A non-limiting example of such a compound is 2,6-diazaspiro[3.3]heptane.

[0041] A tricyclic heterocycle is a bicyclic heterocycle fused to an aryl group, a bicyclic heterocycle fused to a monocyclic cycloalkyl group, a bicyclic heterocycle fused to a monocyclic cycloalkenyl group, or a bicyclic heterocycle fused to a monocyclic heterocycle. Representative examples of tricyclic heterocycles include, but are not limited to, 2,3,4,4a,9,9a-hexahydro-1H-carbazolyl, 5a,6,7,8,9,9a-hexahydrodibenzo[b,d]furanyl, and 5a,6,7,8,9,9a-hexahydrodibenzo[b,d]thienyl.

[0042] The heterocyclic groups of the present disclosure are connected to the parent molecular moiety through any substitutable carbon atom or any substitutable nitrogen, oxygen, or sulfur atom contained within the group and may contain one or two alkylene bridges of 1, 2, 3, or 4 carbon atoms, each linking two non-adjacent carbon atoms of the group. Examples of such "bridged" heterocyclic groups include, but are not limited to, oxatricyclo[3.3.1.13,7]decyl (including 2-oxatricyclo[3.3.1.13,7]decyl), 2,4-dioxabicyclo[4.2.1]nonyl, oxabicyclo[2.2.1]heptyl (including 2-oxabicyclo[2.2.1]heptyl), and 2,5-diazabicyclo[2.2.1]heptane.

[0043] In the above heteroaryl and heterocycles, the nitrogen or sulfur atom may be optionally oxidized to various oxidation states. In specific examples, the group S(O)0-2 refers to -S- (sulfide), -S(O)- (sulfoxide), and -SO2- (sulfone), respectively. For convenience, nitrogen, particularly but not limited to, defined as a cyclic aromatic nitrogen, is intended to include its corresponding N-oxide form. Thus, for example, for compounds of the present disclosure having a pyridyl ring, the corresponding pyridyl-N-oxide is intended to be included as another compound of the present disclosure.

[0044] "Treatment", "treating" or "treating" cancer refers to reversing (e.g., resolving a cell differentiation blockage), ameliorating (e.g., improving one or more symptoms, such as fatigue due to anemia, low blood counts, etc.), and / or slowing its progression (e.g., slowing the progression of the disease state, such as transformation to AML) of cancer as described herein.

[0045] "Subject," as used herein, means an animal subject, such as a mammalian subject, particularly a human.

[0046] The term "antibody" is used in its broadest sense to refer to an immunoglobulin molecule that recognizes and specifically binds to a target, such as a protein, polypeptide, carbohydrate, polynucleotide, lipid, or a combination thereof, via at least one antigen recognition site within the variable region of the immunoglobulin molecule. The heavy chain of an antibody is composed of a heavy chain variable domain (VH) and a heavy chain constant domain (CH). The light chain is composed of a light chain variable domain (VL) and a light chain constant domain (CL). For purposes of this application, mature heavy and light chain variable domains each contain three complementarity-determining regions (CDR1, CDR2, and CDR3) within four framework regions (FR1, FR2, FR3, and FR4) aligned from N-terminus to C-terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. An "antibody" may be naturally occurring or artificial, such as a monoclonal antibody produced by conventional hybridoma technology. The term "antibody" includes full-length monoclonal and polyclonal antibodies, as well as antibody fragments such as Fab, Fab', F(ab')2, Fv, and single-chain antibodies. Antibodies may belong to any one of the five major immunoglobulin classes: IgA, IgD, IgE, IgG, and IgM, or their subclasses (e.g., isotypes IgG1, IgG2, IgG3, IgG4). The term also encompasses human antibodies, chimeric antibodies, humanized antibodies, and any modified immunoglobulin molecule containing an antigen recognition site, so long as it exhibits the desired biological activity (e.g., binding to a target antigen, internalizing into a target antigen-expressing cell).

[0047] As used herein, the term "pharmaceutically acceptable carrier" refers to a non-toxic carrier, adjuvant, or vehicle that does not destroy the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions of the present disclosure include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts, or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, cyclodextrins, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat.

[0048] A "pharmaceutically acceptable salt" is a salt that retains the desired biological activity of the parent compound and does not impart undesired toxicological effects. Examples of such salts are (a) acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, nitric acid, and the like; and salts formed with organic acids such as acetic acid, oxalic acid, tartaric acid, succinic acid, maleic acid, fumaric acid, gluconic acid, citric acid, malic acid, ascorbic acid, benzoic acid, tannic acid, palmitic acid, alginic acid, polyglutamic acid, naphthalenesulfonic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, polygalacturonic acid, and the like; and (b) salts formed from elemental anions such as chlorine, bromine, and iodine. See, e.g., Haynes et al., "Commentary: Occurrence of Pharmaceutically Acceptable Anions and Cations in the Cambridge Structural Database," J. Pharmaceutical Sciences, vol. 94, no. 10 (2005), and Berge et al., "Pharmaceutical Salts," J. Pharmaceutical Sciences, vol. 66, no. 1 (1977), which are incorporated herein by reference.

[0049] Unless otherwise indicated, the nomenclature used to describe chemical groups or moieties as used herein follows the convention that the name is read from left to right and the point of attachment to the remainder of the molecule is to the right of the name. For example, the group "(C1-3 alkoxy)C1-3 alkyl" is attached to the remainder of the molecule at the alkyl terminus. Further examples include methoxyethyl, where the point of attachment is at the ethyl terminus, and methylamino, where the point of attachment is at the amine terminus.

[0050] Unless otherwise indicated, when a chemical group is depicted by a chemical formula or structure having a terminal attachment point designated by "-", it will be understood that the "-" represents the point of attachment.

[0051] Unless otherwise specified, compounds depicted herein include all enantiomeric, diastereomeric, and geometric (or conformational) forms of the structure; for example, the R and S conformations for each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the present disclosure. Unless otherwise specified, all tautomeric forms of the disclosed compounds are within the scope of the present disclosure. Additionally, unless otherwise specified, structures depicted herein include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the formula disclosed herein except for the replacement of hydrogen with deuterium or tritium, or the replacement of a carbon with a C- or C-enriched carbon, are within the scope of the present disclosure. Such compounds may be useful, for example, as analytical tools or probes in biological assays.

[0052] According to some embodiments, provided herein is a compound of formula I: [ka] and pharmaceutically acceptable salts thereof [In formula: n is selected from 0, 1, 2 or 3; R1 is a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a -NR9R10 group; [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] selected from the group R9 is selected from hydrogen, -NR11R12 group, C1-C6 alkyl group, -(C1-C6 alkyl)-CO2H group, C3-C8 cycloalkyl group, and C3-C8 heterocyclyl group, wherein -NR11R12 group, C1-C6 alkyl group, C3-C8 cycloalkyl group, and C3-C8 heterocyclyl group may be unsubstituted or substituted 1 to 3 times with groups independently selected from C1-C6 alkyl group, -(C1-C6 alkyl)-CO2H group, hydroxy, halogen group, and C1-C6 alkoxy group; R10 is selected from hydrogen and a C1-C6 alkyl group; Either R2 or R3 is selected from hydrogen and a C1-C6 alkyl group, and the other is selected from hydrogen, —OR10, —OC(O)R10, —OC(O)R1, and a C1-C6 alkyl group; R4 is selected from hydrogen and hydroxy; R5 and R6 are each independently selected from a C1 to C6 alkyl group; R7 and R8 are each independently selected from hydrogen, hydroxy, a C1-C6 alkoxy group, and a C1-C6 alkyl group; and Y is selected from phenyl, thiophenyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl, where Y is unsubstituted or is an oxo group, a C1-C6 alkyl group, a C3-C5 cycloalkyl group, a hydroxy C1-C6 alkyl group, a C1-C6 alkoxy group, a methoxy C1-C6 alkyl group, a -NR11R12 group, [ka] wherein R11 and R12 are each independently selected from hydrogen and a C1 to C6 alkyl group.

[0053] In some embodiments, in Formula I, Y is [ka] is.

[0054] In some embodiments, in Formula I, Y is selected from optionally substituted phenyl groups.

[0055] In some embodiments, in Formula I, R1 is methyl, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] groups, wherein R9, R10, R11, and R12 are defined as above.

[0056] Also provided herein are compounds of formula II: [ka] and pharmaceutically acceptable salts thereof [In formula: X is selected from O, an NR′ group, and CH2, where R′ is selected from hydrogen and a C1-C6 alkyl group; R1 is methyl, an NR11R12 group, [ka] Groups, and [ka] selected from the group R10 is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, and a haloC1-C6 alkyl group, wherein the C3-C8 cycloalkyl group can be unsubstituted or substituted 1 to 3 times with groups independently selected from a C1-C6 alkyl group, a hydroxyl group, a halogen group, and a C1-C6 alkoxy group; R11 and R12 are each independently selected from a C1 to C6 alkyl group; Either R2 or R3 is selected from hydrogen and a C1-C6 alkyl group, and the other is selected from hydrogen, hydroxy, and a C1-C6 alkyl group; Either R4 or R5 is hydrogen, and the other is hydrogen, hydroxy, and [ka] Selected from; R6 and R7 are each independently selected from a C1 to C6 alkyl group; R8 and R9 are each independently selected from hydrogen and a C1-C6 alkyl group; or R8 and R9 together form a cyclopropyl ring; and Y is selected from a C1-C6 alkyl group, a C3-C8 cycloalkyl group, methoxy, and a -NR13R14 group, where R13 and R14 are each independently selected from hydrogen, a C1-C6 alkyl group, and a methoxyC1-C6 alkyl group; or R13 and R14 together with N are [ka] forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; wherein Y may be unsubstituted or a C1-C6 alkyl group, hydroxy, a hydroxy C1-C6 alkyl group, methoxy, a methoxy C1-C6 alkyl group, halo, a halo C1-C6 alkyl group, -C(O)NH2, -NHCOO-C1-C6 alkyl group, -COOH, [ka] and -NR15R16 groups, wherein R15 and R16 are each independently selected from hydrogen and a C1-C6 alkyl group.

[0057] Also provided herein are compounds of formula III: [ka] and pharmaceutically acceptable salts thereof [In formula: n is selected from 0, 1 and 2; m is selected from 1, 2, and 3; R1 is a C1-C6 alkyl group, a C3-C8 cycloalkyl group, or a -NR11R12 group; [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] is selected from the group R11 is selected from hydrogen, -NR16R17 group, C1-C6 alkyl group, -(C1-C6 alkyl)-CO2H group, -(C1-C6 alkyl)-CO2R12 group, -(C1-C6 alkyl)-NR16R17 group, C3-C8 cycloalkyl group, and C3-C8 heterocyclyl group, wherein -NR16R17 group, C1-C6 alkyl group, C3-C8 cycloalkyl group, and C3-C8 heterocyclyl group may be unsubstituted or substituted 1 to 3 times with groups independently selected from C1-C6 alkyl group, -(C1-C6 alkyl)-CO2H group, hydroxy, halogen group, and C1-C6 alkoxy group; R12 is selected from hydrogen and a C1-C6 alkyl group; Either R2 or R3 is selected from hydrogen and a C1-C6 alkyl group, and the other is selected from hydrogen, —OR10, —OC(O)R10, —OC(O)R1, and a C1-C6 alkyl group; R4 is hydrogen or hydroxy; R5 and R6 are each independently selected from a C1 to C6 alkyl group; R7 and R8 are each independently selected from hydrogen, hydroxy, a C1-C6 alkoxy group, and a C1-C6 alkyl group; and R9 and R10 are each independently selected from hydrogen, a C1-C6 alkyl group, hydroxy, and a C1-C6 alkoxy group; or one of R9 or R10 is oxo, and the other is absent; Z is selected from a C1-C6 alkyl group, a —C(O)—C1-C6 alkyl group, a —OR13 group, and a —NR14R15 group; wherein R13 is selected from hydrogen, a C1-C6 alkyl group, and a —C(O)—C1-C6 alkyl group; wherein R14 and R15 are each independently selected from hydrogen, a C1-C6 alkyl group, and a methoxy C1-C6 alkyl group; or R14 and R15 together with N represent [ka] forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; In the formula, Z may be unsubstituted or may be a C1 to C6 alkyl group, a C3 to C5 cycloalkyl group, a hydroxy C1 to C6 alkyl group, a C1 to C6 alkoxy group, a methoxy C1 to C6 alkyl group, a -NR16R17 group, [ka] wherein R16 and R17 are each independently selected from hydrogen and a C1-C6 alkyl group.

[0058] In some embodiments, in Formula III, R1 is methyl, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] basis, [ka] Groups, and [ka] groups, wherein R11, R12, R16, and R17 are defined as above.

[0059] Further, in this specification, [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] and pharmaceutically acceptable salts thereof.

[0060] Further, in this specification, [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-9-oxo-9-pyrrolidin-1-ylnona-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[[(2R,3R)-3-hydroxypentan-2-yl]carbamoyloxy]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-(propylcarbamoyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-4-oxidepiperazin-4-ium-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(dimethylcarbamoyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-(diethylcarbamoyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propan-2-yl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[butyl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[butan-2-yl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-carbamoyloxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R)-2-(methoxymethyl)pyrrolidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[2-methoxyethyl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]azetidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2S)-2-methylpyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2S)-2-methylpyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]piperidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R)-2-(hydroxymethyl)pyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](3R)-3-hydroxypyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]morpholine-4-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]4-methylpiperazine-1-carboxylate; 3-Thiazolidinecarboxylic acid [(2R,3E,5E)-6-[(2R,3S,4E,6R,7R,10R)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl] ester; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]1,3-dihydroisoindole-2-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]indole-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[2-(1-hydroxyethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,2-dimethylpyrrolidine-1-carbonyl)oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2S,5S)-2,5-dimethylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2,3-dihydroindole-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2-oxa-5-azaspiro[3.4]octane-5-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-6-[6-[(2R)-1-hydroxypropan-2-yl]pyridin-2-yl]hepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridazin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3R,4E,6S,7R,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6R)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-propan-2-ylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-tert-butylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclopentylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(oxan-4-yl)piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]6-cycloheptyl-2,6-diazaspiro[3.3]heptane-2-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-3-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclobutylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N-methyl-N-(1-methylpiperidin-4-yl)carbamate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]morpholine-4-carboxylate; [(2R,3R,4E,6S,7R,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6R)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl](1S,4R)-5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]8-cycloheptyl-3,8-diazabicyclo[3.2.1]octane-3-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methyl-1,4-diazepane-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclohexylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-1,4-diazepane-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-hydroxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(azepan-1-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(8,8-difluoro-3-azabicyclo[3.2.1]octan-3-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-9-oxo-9-pyrrolidin-1-ylnona-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10R)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-10-(pyrrolidine-1-carbonyloxy)-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(2S)-2-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3R)-3-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3R)-3-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-carbamoylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[(2R)-2-(methoxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2S,5S)-2,5-dimethylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,2-dimethylpyrrolidine-1-carbonyl)oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-2-[(2E,4E)-6,6-dimethyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E)-6,6-dimethyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-7-hydroxy-3,7-dimethyl-12-oxo-10-(pyrrolidine-1-carbonyloxy)-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; (2R)-1-[(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienoxy]carbonylpyrrolidine-2-carboxylic acid; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(3-oxopyrrolidine-1-carbonyl)oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2-oxa-7-azaspiro[3.4]octane-7-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-5-[1-(pyrrolidine-1-carbonyloxymethyl)cyclopropyl]penta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3S,4R)-3,4-dihydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; (3S)-1-[(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienoxy]carbonylpyrrolidine-3-carboxylic acid; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3S)-3-(dimethylamino)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,5-dihydropyrrole-1-carbonyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3S)-3-[(2-methylpropan-2-yl)oxycarbonylamino]pyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]3-azabicyclo[3.1.0]hexane-3-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-2-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-(2-pyrrolidin-1-ylpyrimidin-4-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrazin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(3-methylpyridin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(4-methylpyridin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridazin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-4-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(4-methylpyrimidin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-(6-pyrrolidin-1-ylpyridin-2-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-10-hydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-10-hydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N,N-dimethylcarbamate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6R)-6-(dimethylcarbamoyloxy)-3-methyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6R)-6-(dimethylcarbamoyloxy)-3-methyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](3R)-3-hydroxypyrrolidine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-[(2S)-2-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-(2,2,2-trifluoroethyl)piperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N,N-dimethylcarbamate; [(2S,3S,4E,6R)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-(2-pyrrolidin-1-ylpyrimidin-4-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-[2-[(3S)-3-triethylsilyloxypyrrolidin-1-yl]pyrimidin-4-yl]hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E)-6-[2-[(3R)-3-hydroxypyrrolidin-1-yl]pyrimidin-4-yl]hepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3S)-3-(1-phenyltetrazol-5-yl)oxypyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonylamino)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]amino]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonylamino)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[methyl(pyrrolidine-1-carbonyl)amino]hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(4-cyclopropyltriazol-1-yl)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-methoxycarbonyloxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-9-methoxy-6-methyl-9-oxonona-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(cyclopentanecarbonylamino)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(cyclopentanecarbonylamino)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; 4-Cycloheptyl-1-piperazinecarboxylic acid [(2R,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-7-[oxo(1-pyrrolidinyl)methoxy]hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl] ester; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-azacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-2-methyl-6-[(2S,3S,4E,6R)-3-methyl-6-[(4-methylpiperazine-1-carbonyl)amino]-12-oxo-1-oxacyclododec-4-en-2-yl]hepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2S,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R,3R)-3-hydroxy-2-methylpentanoate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-hydroxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-4-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-7-methyl-6-pyridin-2-ylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[(2R,3R)-3-[(2R,3R)-3-acetyloxypentan-2-yl]oxiran-2-yl]-6-hydroxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-methyl-8-phenylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-phenylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-thiophen-2-ylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-phenylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-6-(6-methoxypyridin-2-yl)hepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-[6-(2-methylpropoxy)pyridin-2-yl]hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-8-pyridin-2-ylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-7-pyridin-2-ylhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E,6R)-6-hydroxy-6-methyl-8-phenylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-2-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-4-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-8-(4-hydroxyphenyl)-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-8-phenylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[2-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[4-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[3-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6S)-6-hydroxy-6-methyl-7-[(2R,3R)-3-[(2S)-3-oxopentan-2-yl]oxiran-2-yl]hepta-2,4-dien-2-yl]-3,7-dimethyl-10,12-dioxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6E,8S)-8-pyridin-2-ylnona-2,4,6-trien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methyl-4-oxidepiperazin-4-ium-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(4-fluoropiperidin-1-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(4,4-difluoropiperidin-1-yl)piperidine-1-carboxylate; (4S,7S,8S,9E,11S,12S)-4,7,8-trihydroxy-7,11-dimethyl-12-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-9-en-2-one; [(2S,3S,4E,6S,7S,10S)-7-acetyloxy-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((S,2E,4E)-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-6-acetyloxy-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-7-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7-acetyloxy-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7R,10R)-7-ethoxy-10-hydroxy-2-[(2E,4E,6R)-6-hydroxy-7-[(2R,3R)-3-[(2S,3S)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-6-acetyloxy-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-7-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]N-methyl-N-[2-(methylamino)ethyl]carbamate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]N-methyl-N-[2-(dimethylamino)ethyl]carbamate; 3-[4-[[(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]oxycarbonyl]piperazin-2-yl]propanoic acid; 4-[4-[[(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]oxycarbonyl]piperazin-1-yl]butanoic acid; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; (2S,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-propylpiperazine-1-carboxylate; (2R,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((2S,6R,E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhept-4-en-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 4-(2-hydroxyethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 4-methylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-(2-aminoethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7,10-dihydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-methylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7,10-dihydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; and pharmaceutically acceptable salts thereof.

[0061] Disclosed herein are compositions comprising at least one compound of the present disclosure (e.g., compounds of Formula I, Formula II, and Formula III) and / or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier. The at least one pharmaceutically acceptable carrier can be selected depending on the particular route of administration for which the composition is intended.

[0062] The pharmaceutical compositions of the present disclosure may be formulated for parenteral, oral, inhalation spray, topical, rectal, nasal, buccal, vaginal, or implanted reservoir administration, etc. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the compositions are administered intravenously, orally, subcutaneously, or intramuscularly. The compositions of the present disclosure in sterile injectable form may be aqueous or oily suspensions. Such suspensions may be formulated using suitable dispersing or wetting agents and suspending agents according to techniques known in the art. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic saline solution. In addition, sterile fixed oils are conventionally employed as solvents or suspending media.

[0063] Any non-irritating fixed oil can be used, including synthetic mono- or diglycerides.Fatty acids such as oleic acid and its glyceride derivatives are useful in preparing injections, as are natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated versions.These oil solutions or suspensions can also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions.Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability promoters commonly used in the manufacture of pharmaceutically acceptable solid, liquid, or other dosage forms, can also be used for formulation purposes.

[0064] For oral administration, the compounds of the present disclosure (e.g., Formula I, Formula II, or Formula III) and / or their pharmaceutically acceptable salts may be provided in acceptable oral dosage forms, including, but not limited to, capsules, tablets, aqueous suspensions, or solutions. For tablets intended for oral use, commonly used carriers include lactose and cornstarch. Lubricants such as magnesium stearate may also be added. For oral administration in capsule form, useful diluents include lactose and dry cornstarch. When aqueous suspensions are required for oral use, the active ingredient may be combined with emulsifying and / or suspending agents. If necessary, certain sweetening, flavoring, or coloring agents may also be added.

[0065] The compounds and compositions of the present disclosure may be used to treat various cancers, including those that respond to drugs targeting the spliceosome, including SF3B1. As mentioned above, the antitumor activity of pladienolide B has been reported to be related to its targeting of the SF3b complex, inhibition of splicing, and alteration of gene expression patterns (Kotake et al., "Splicing factor SF3b as a target of the antitumor natural product pladienolide," Nature Chemical Biology 2007, 3, 570-575). Mutations in the splicing factor 3B subunit 1 (SF3B1) protein are known to be involved in several cancers, including hematological malignancies and solid tumors. Scott et al., “Acquired mutations that affect pre-mRNA splicing in hematologic malignancies and solid tumors,” JNCI 105, 20, 1540-1549.

[0066] Thus, the compounds of the present disclosure (e.g., compounds of Formula I, Formula II, and Formula III and pharmaceutically acceptable salts thereof) and compositions may be used to treat hematological malignancies, such as cancers of the blood (leukemia) and cancers of the lymph nodes (lymphoma). Leukemias include acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myelocytic leukemia (CML), chronic myelomonocytic leukemia (CMML), acute monocytic leukemia (AMoL), and the like. Lymphomas include Hodgkin's lymphoma and non-Hodgkin's lymphoma. Other hematological malignancies may include myelodysplastic syndromes (MDS).

[0067] Solid tumors include carcinomas such as adenocarcinomas, for example, breast cancer, pancreatic cancer, prostate cancer, colon or colorectal cancer, lung cancer, gastric cancer, cervical cancer, endometrial cancer, ovarian cancer, bile duct cancer, glioma, melanoma, and the like.

[0068] The compounds of the present disclosure (e.g., compounds of Formula I, Formula II, and Formula III) and pharmaceutically acceptable salts and compositions thereof may also be used to treat cancers that may respond to drugs that target spliceosomal genes or proteins other than SF3B1. The following are non-limiting examples of cancers that may respond to drugs that target the spliceosomal. Thus, the compounds of the present disclosure may be administered to subjects for the treatment of various such cancers or conditions, particularly patients or subjects suffering from:

[0069] a) Myelodysplastic syndromes (MDS): see, for example, "SF3B1 mutations in myelodysplastic syndromes: clinical associations and prognostic implications," Damm F. et al. Leukemia, 2011, 1-4; "Frequent pathway mutations in splicing machinery in myelodysplasia," Yoshida K. et al. Nature, 2011, 478, 64-69; "Clinical significance of SF3B1 mutations in myelodysplastic syndromes and myelodysplastic / myeloproliferative neoplasms," Malcovati L. et al. al., Blood, 2011, 118, 24, 6239-6246; “Mutations in the spliceosome machinery, a novel and ubiquitous pathway in leukemogenesis,” Makishima et al, Blood, 2012, 119, 3203-3210; “Somatic SF3B1 mutation in myelodysplasia with ring sideroblasts,” Pappaemannuil, E. et al, New England J. Med. 2011, DOI 10.1056 / NEJMoa1103283.

[0070] b) Chronic lymphocytic leukemia (CLL): e.g., “Defects in the spliceosomal machinery: a new pathway of leukaemogenesis,” Maciejewski, JP, Padgett, RA, Br. J. Hematology, 2012, 1-9; “Mutations in the SF3B1 splicing factor in chronic lymphocytic leukemia: associations with progression and fludarabine-refractility,” Rossi et al., Blood, 2011, 118, 6904-6908; “Exome sequencing identifies recurrent mutations of the splicing factor SF3B1 gene in chronic lymphocytic See “Exome sequencing identifies recurrent mutations in the splicing factor SF3B1 gene in chronic lymphocytic leukemia,” Quesada et al, Nature Genetics, 2011, 44, 47-52.

[0071] c) Chronic myelomonocytic leukemia (CMML): See, e.g., Yoshida et al, Nature 2011; “Spliceosomal gene mutations are frequent events in the diverse mutational spectrum of chronic myelomonocytic leukemia but largely absent in juvenile myelomonocytic leukemia,” Kar SA et al, Haematologia, 2012, DOI: 10.3324 / haematol.2012.064048.

[0072] d) Acute myeloid leukemia (AML): See, for example, Malcovati et al., Blood 2011; Yoshida et al., Nature 2011.

[0073] e) Breast cancer: See, e.g., “Whole genome analysis informs breast cancer response to aromatase inhibition,” Ellis et al, Nature, 2012, 486, 353-360.

[0074] f) Uveal melanoma: See, e.g., “SF3B1 mutations are associated with alternative splicing in uveal melanoma”, Furney et al, Cancer Disc. 2013, 10, 1122-1129.

[0075] g) Endometrial cancer: See, e.g., Tefferi et al., "Myelodysplastic syndromes." N Engl J Med. 2009;361:1872-85.

[0076] h) Gastric cancer: See, e.g., Int J Cancer. 2013 Jul;133(1):260-5, “Mutational analysis of splicing machinery genes SF3B1, U2AF1 and SRSF2 in myelodysplasia and other common tumors.” Je et al.

[0077] i) Ovarian cancer: See, e.g., Int J Cancer. 2013 Jul;133(1):260-5, “Mutational analysis of splicing machinery genes SF3B1, U2AF1 and SRSF2 in myelodysplasia and other common tumors.” Je et al.

[0078] j) Biliary tract cancers, such as cholangiocarcinoma, and pancreatic cancer: See, e.g., Biankin et al., “Pancreatic cancer genomes reveal aberrations in axon guidance pathway genes,” Nature 2012, 491, 399-405.

[0079] k) Lung cancer: See, e.g., “Exome sequencing identifies recurrent mutations of the splicing factor SF3B1 gene in chronic lymphocytic leukemia,” Quesada et al., Nature Genetics 44, 47-52 (2012); Scott et al., “Acquired mutations that affect pre-mRNA splicing in hematologic malignancies and solid tumors,” JNCI 105, 20, 1540-1549.

[0080] In addition, the Catalogue of somatic mutations in cancer (COSMIC) (Wellcome Trust Sanger Institute, Genome Research Limited, UK) reports SF3B1 mutations found in various cancer samples.

[0081] A compound of the present disclosure (e.g., a compound of Formula I, Formula II, or Formula III) may be administered to a subject in a therapeutically effective amount. The amount of a compound of the present disclosure that may be combined with a carrier material to produce a single dosage form of the composition will vary depending on the subject being treated and the particular route of administration. In some embodiments, a dose of 0.01 mg / kg to 100 mg / kg of body weight / day of at least one compound disclosed herein is administered. In some embodiments, the dose is 0.01 mg to 50 mg of at least one compound disclosed herein. In some embodiments, 0.1 mg to 25 mg of at least one compound disclosed herein is provided. In some embodiments, 5 mg to 40 mg of at least one compound disclosed herein is provided.

[0082] Those skilled in the art will understand that the specific dosage and treatment regimen for a particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combinations, the judgment of the treating physician, and the severity of the particular disease being treated. The amount of at least one compound disclosed herein will also depend on the specific compound / salt used.

[0083] In some embodiments, the cancer is tested for and / or positive for one or more mutations in the splicing factor 3B subunit 1 (SF3B1) gene or protein, where the presence of one or more mutations ("positive") indicates that the subject's cancer will respond to a treatment method comprising administering at least one compound disclosed herein that targets this protein and / or the spliceosome. Examples of such spliceosomal genes include, but are not limited to, those provided in Table 1.

[0084] [Table 1]

[0085] In some embodiments, the subject's cancer is one that may respond to a treatment method involving administration of a compound that targets a spliceosomal gene or protein, even in the absence of such a mutation in the protein and / or spliceosome.

[0086] Screening or testing for mutations may be performed by any known means, e.g., genotyping, phenotyping, etc., using nucleic acid amplification, electrophoresis, microarray, blot, functional assay, immunoassay, etc. Screening methods may, for example, include obtaining a biological sample comprising cancerous cells / tissues from said subject.

[0087] In some embodiments, a subject having cancer as described herein can be treated with at least one compound selected from a compound of formula I, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable salt of any of the foregoing, and at least one additional therapy.

[0088] In some embodiments, the at least one additional therapy comprises a cytokine or cytokine analog therapy, such as any cytokine or cytokine analog therapy disclosed herein. Cytokines are a broad category of small proteins that have been shown to be involved in autocrine, paracrine, and / or endocrine signaling as immunomodulators. Exemplary cytokines are disclosed herein and include chemokines, interferons, interleukins, lymphokines, and tumor necrosis factors. As used herein, the term "cytokine" refers to a polypeptide secreted by a cell that mediates an immune response by affecting the function of other cells, and the term "cytokine therapy" refers to the administration and / or induction of secretion of such peptides. In some embodiments, the cytokine is a recombinant cytokine or an analog thereof. In some embodiments, the cytokine is a cytokine analog. The terms "cytokine analog" and "cytokine analog therapy" refer to modified cytokines, in which one or more amino acid residues of a naturally occurring cytokine are substituted with other naturally occurring or non-naturally occurring amino acid residues and / or one or more naturally occurring or non-naturally occurring amino acid residues are added to the naturally occurring cytokine. In some embodiments, cytokine or cytokine analog therapy comprises administering at least one cytokine or cytokine analog to a patient in need of such treatment.

[0089] In some embodiments, the at least one additional therapy comprises one or more engineered tumor-targeting T cells (e.g., CAR-T or other cell-based therapy), such as any CAR-T therapy disclosed herein. The terms "CAR-T" and "CAR-T therapy" are used interchangeably to refer to a cell or cell population (e.g., a T cell or T cell population) modified with a CAR. In some embodiments, a chimeric T cell receptor (CAR) can be engineered with an antigen recognition sequence so that, when the CAR is expressed on a cell (e.g., a T cell), the CAR and / or the cell is reactive to a target antigen. For example, in some embodiments, a CAR can be engineered by first identifying an antibody that recognizes an antigen protein domain expressed on the cell surface. The antigen recognition sequence of such an antibody can then be fused to a T cell receptor domain for selective targeting and activation. In some embodiments, this CAR sequence is cloned and expanded into a patient-derived T cell population using currently available protocols. In some embodiments, the engineered T cells are then infused back into the patient's circulation before, concurrently with, or after treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. After treatment with the at least one compound and / or pharmaceutically acceptable salt, in some embodiments, tumor cells may begin to present antigens, e.g., antigens targeted by the engineered T cell population. In some embodiments, the engineered T cell population can associate with and kill antigen-presenting tumor cells.

[0090] In some embodiments, the at least one additional therapy comprises a checkpoint inhibitor therapy, e.g., any checkpoint inhibitor therapy disclosed herein. Immune checkpoints are inhibitory pathways that slow or halt the immune response, preventing excessive tissue damage due to uncontrolled immune cell activity. As used herein, the terms "checkpoint inhibitor" and "checkpoint inhibitor therapy" are used interchangeably to refer to any therapeutic agent that inhibits one or more inhibitory pathways, thereby enabling broad immune activity, including any small molecule chemical compound, antibody, nucleic acid molecule, or polypeptide, or any fragment thereof. In some embodiments, checkpoint inhibitor therapy comprises administering at least one checkpoint inhibitor to a patient in need of such treatment.

[0091] In some embodiments, at least one additional therapy comprises a neoantigen vaccine. In some embodiments, the treatment comprises administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and administering a neoantigen vaccine. In some embodiments, the neoantigen vaccine comprises a tumor neoantigen and / or a neoantigen induced by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the treatment further comprises administering a checkpoint inhibitor therapy. In some embodiments, the checkpoint inhibitor therapy targets PD1 / PDL1, CTLA4, OX40, CD40, LAG3, TIM3, GITR, and / or KIR. In some embodiments, the checkpoint inhibitor therapy targets PD1 / PDL1 (e.g., an anti-PD1 antibody or an anti-PDL1 antibody). In some embodiments, the checkpoint inhibitor therapy targets CTLA4 (e.g., an anti-CTLA4 antibody). In some embodiments, the treatment includes initially (i) administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, followed by administering a combination therapy including a neoantigen vaccine; and (ii) detecting the presence of a neoantigen (e.g., a neoantigen from a neoantigen vaccine). In some embodiments, neoantigen expression is monitored during the course of treatment. In some embodiments, if a neoantigen is not detected, the treatment is discontinued.

[0092] Also disclosed herein, in some embodiments, are methods of treating a patient by inducing neoantigens in tumor cells that can be targeted for clearance by the patient's immune system. Without being bound by theory, in some embodiments, administration of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing can result in the production of neoantigens that induce an immune response, such as the induction of a double-stranded RNA immune response as a result of the re-expression of an intron-resident endogenous retrovirus, and / or the production of neoantigens that induce immunogenic cell death.

[0093] As used herein, the term "neoantigen" refers to any antigen to which the immune system has not previously been exposed, resulting from one or more tumor-specific mutations and / or from exposure of the tumor to at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Tumor-specific mutations may include missense mutations, frameshifts, translocations, and mRNA splicing variants, as well as mutations that affect post-translational processing, such as phosphorylation and glycosylation. These exemplary mutations may, in some embodiments, result from nonsynonymous coding changes and / or mutations that alter mRNA processing (e.g., splicing). Any of these exemplary mutations may, in some embodiments, result in a molecular change that can be recognized by an appropriate T cell receptor. In some embodiments, the exemplary neoantigen is a neoantigen induced by delivery of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, delivery of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may induce de novo mRNA splicing, resulting in the translation of a protein containing one or more novel peptide domains to which the immune system has not previously been exposed. In some embodiments, the tumor-specific mutation may be an mRNA splice variant resulting from delivery or administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0094] Without being bound by theory, in some embodiments, delivery of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing may induce de novo mRNA splicing (e.g., exon skipping, intron retention), resulting in alterations in the open reading frame and / or coding sequence of various genes. In some embodiments, these altered genes, when translated, become proteins containing one or more novel peptide domains that the immune system recognizes as foreign. In some embodiments, the one or more novel peptide domains are not present in the protein in the absence of compound treatment, or in any other part of the human proteome. In some embodiments, proteins containing the one or more novel peptide domains may be degraded by the proteasome to generate novel peptide fragments that serve as substrates for the immune peptide presentation machinery, e.g., by MHC presentation. In some embodiments, the novel peptide fragments, corresponding to neoantigens, may be presented in the MHC1-bound peptidome, e.g., on tumor cells.

[0095] In some embodiments, delivery of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing can lead to one or more tumor cell-specific events (e.g., cell growth arrest). In some embodiments, the one or more tumor cell-specific events can include (1) enhanced engagement by phagocytes (Bracci et al. (2014) Cell Death Differ. 21(1):15-25); (2) trafficking of the novel peptide fragments to tumor-draining lymph nodes resulting in engagement with antigen-presenting cells; (3) processing of the novel peptide fragments from phagocytosed tumor cells by antigen-presenting cells and presentation of the fragments as neo-antigens to circulating naive T cell populations; (4) interaction of the novel peptide fragments with T cells expressing receptors that recognize the fragments as neo-antigens; (5) maturation and activation of effector T cell responses (e.g., CD4+ and / or CD8+ T cells); and / or (6) T cell engagement with additional tumor cells that have been exposed to compound treatment and that present the novel peptide fragments corresponding to the neo-antigens on their surface MHC1 complexes. In some embodiments, one or more tumor cell-intrinsic events can directly or indirectly result in effector T cell engagement and / or killing of the neo-antigen-presenting tumor cells.

[0096] Also, without being bound by theory, in some embodiments, delivery of at least one compound selected from a compound of formula I, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable salt of any of the foregoing may result in the re-expression of intron-resident endogenous retroviruses, leading to a double-stranded RNA immune response.

[0097] Furthermore, without being bound by theory, in some embodiments, delivery of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can lead to immunogenic cell death caused by the compound-induced release of mutation-derived neoantigens. In some embodiments, delivery of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can induce a double-stranded RNA immune response. In some embodiments, the double-stranded RNA immune response can be caused by the re-expression of intron-resident endogenous retroviruses. In some embodiments, the double-stranded RNA immune response can result in tumor cell death. In some embodiments, delivery of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can induce immunogenic cell death. In some embodiments, the immunogenic cell death can be caused by the release of mutation-derived neoantigens and / or a host immune response against tumor cells.

[0098] Thus, in some embodiments, a therapeutic method is disclosed that includes inducing neoantigens by administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the method includes administering a reduced dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, relative to what would be required absent the induction of neoantigens. In some embodiments, the method includes administering one or more initial loading doses to produce neoantigens and induce an immune response (e.g., converting naive T cells to memory cells), followed by a reduced dosage or frequency of administration (i.e., due to the combined effect of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and immune targeting of the neoantigens). In some embodiments, the treatment may involve administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce a neoantigen-based immune response in combination with at least one additional therapy (e.g., a second anticancer therapy). For example, in some embodiments, the treatment may involve administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce a neoantigen-based immune response in combination with one or more checkpoint inhibitors. In some embodiments, the treatment may involve administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce a neoantigen-based immune response in combination with one or more cytokines or cytokine analogs. In some embodiments, treatment may involve administering at least one compound selected from a compound of formula I, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce a neoantigen-based immune response, in combination with one or more neoantigen vaccines.In some other embodiments, treatment may involve administering at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce a neo-antigen-based immune response in combination with one or more engineered tumor-targeting T cells (e.g., CAR-T).

[0099] In some embodiments, neoantigens can be used to monitor the effectiveness of treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. For example, a patient sample (e.g., a tumor biopsy) can be obtained after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and screened for neoantigens or for identifiers of an immune or inflammatory response. If neoantigens and / or an immune response are detected, further treatment can be provided, for example, at a reduced dosage.

[0100] In some embodiments, a method of treatment is disclosed that includes inducing a double-stranded RNA immune response by administering at least one compound selected from a compound of formula I, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0101] In some embodiments, a method of treatment is disclosed that includes inducing immunogenic cell death by administering at least one compound selected from a compound of formula I, a compound of formula II, a compound of formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0102] In some embodiments, administration of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing can be combined with any known anticancer therapy. Examples of immune activation strategies currently available for oncology treatment include, but are not limited to, treatment with immune checkpoint inhibitor (ICI) molecules, treatment with cytokines or cytokine analogs, tumor-associated vaccination, and modification of tumor-targeting T cells (e.g., expansion of tumor-infiltrating lymphocytes or CAR-T cells). These techniques primarily focus on enhancing or inducing an immune response against pre-existing tumor antigens (either mutations or aberrant expression of cell surface proteins). One or more of these strategies may involve one or more mutations capable of inducing an antigenic T cell response. For example, patient response to checkpoint inhibition may be correlated with nonsynonymous mutation burden. In addition, cancer vaccine approaches that rely on pre-existing mutations and the antigenicity of those mutations may be used.

[0103] Compounds of Formula I, II, III, and pharmaceutically acceptable salts of any of the foregoing can induce widespread transcriptome changes across multiple lineages. These mRNA changes, when translated, can result in robust and reproducible protein changes that produce MHC1-binding neopeptides with high affinity across multiple HLA isotypes. Without being bound by theory, due to numerous transcriptome and proteome changes, treatment with at least one compound selected from compounds of Formula I, II, III, and pharmaceutically acceptable salts of any of the foregoing can enhance the number of potentially responsive neoantigens and promote the engagement of adaptive immune responses.

[0104] In some embodiments, the present disclosure provides methods for inducing at least one neoantigen by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the present disclosure provides methods for inducing a double-stranded RNA immune response by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the present disclosure provides methods for inducing immunogenic cell death by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0105] In some embodiments, the neoplastic cells are present in an in vitro cell culture. In some embodiments, the neoplastic cells are obtained from a subject. In some embodiments, the neoplastic cells are present in a subject. In some embodiments, the neoplastic cells are derived from a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from a B-cell malignancy, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the solid tumor is selected from breast cancer (e.g., HER2-positive breast cancer), gastric cancer (e.g., gastric adenocarcinoma), prostate cancer, ovarian cancer, lung cancer (e.g., lung adenocarcinoma), uterine cancer (e.g., serous endometrial carcinoma), salivary duct cancer, melanoma, colon cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, and prostate cancer.

[0106] In some embodiments, the present disclosure further provides a method of inducing at least one neoantigen and / or T cell response in a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Also provided herein, in some embodiments, is a method of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, wherein administration of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces at least one neoantigen and / or T cell response.

[0107] In various other embodiments, the present disclosure provides methods of inducing a double-stranded RNA immune response in a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Also provided herein, in some embodiments, are methods of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, wherein administration of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces a double-stranded RNA immune response.

[0108] In still other embodiments, the present disclosure provides methods of inducing immunogenic cell death in a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Also provided herein, in some embodiments, are methods of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, including at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, wherein administration of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces immunogenic cell death.

[0109] In some embodiments, the present disclosure further provides a method of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in combination with one or more additional therapies, including a second agent, wherein administration of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces immunogenic cell death.

[0110] In some embodiments of the therapeutic methods described herein, the administered amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or a second agent, is reduced due to induction of at least one neo-antigen and / or T cell response when compared to a standard dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or a second agent. In some embodiments, the dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or a second agent is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to the standard dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or a second agent. In some embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or the second agent is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently than a standard administration regimen of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or the second agent. In some embodiments, the dose and / or dosage of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or the second agent results in reduced systemic toxicity and / or improved tolerability.

[0111] As used herein, the term "standard dosage" or "standard administration regimen" refers to any usual or routine administration regimen for a therapeutic agent, e.g., a regimen suggested by a manufacturer, approved by a regulatory agency, or otherwise tested in human subjects to meet the needs of the average patient. In some embodiments, the therapeutic agent is at least one compound selected from compounds of Formula I, II, III, and pharmaceutically acceptable salts of any of the foregoing, which have anticancer activity.

[0112] For example, a standard dosing regimen for trastuzumab, an exemplary anti-HER2 antibody, may be 8 mg / kg administered intravenously over 90 minutes (week 1), followed by 6 mg / kg administered intravenously over 30-90 minutes every three weeks (week 4 through the end of the therapy cycle) (Herceptin® (trastuzumab) FDA Labeling Supplement, 2017).

[0113] As another example, a standard dosing regimen for ipilimumab, an exemplary anti-CTLA4 checkpoint inhibitor antibody, may be 3 mg / kg administered intravenously over 90 minutes every three weeks for four doses (Yervoy® (ipilimumab) FDA Labeling Supplement, 2018). Another standard dosing regimen for ipilimumab may be 10 mg / kg administered intravenously over 90 minutes every three weeks for four doses, followed by 10 mg / kg every 12 weeks for up to three years (Yervoy® (ipilimumab) FDA Labeling Supplement, 2018).

[0114] As another example, the standard dosing regimen for nivolumab, an exemplary anti-PD1 checkpoint inhibitor antibody, may be 3 mg / kg administered intravenously over 60 minutes every two weeks (Opdivo® (nivolumab) FDA Labeling, 2015).

[0115] As another example, the standard dosing regimen for atezolizumab, an exemplary anti-PDL1 checkpoint inhibitor antibody, may be 1200 mg administered intravenously over 60 minutes every three weeks (Tecentriq® (atezolizumab) FDA Labeling Supplement, 2018).

[0116] As yet another example, the standard dosing regimen for T-DM1, an exemplary anti-HER2 antibody drug conjugate, may be 3.6 mg / kg administered intravenously over 90 minutes every three weeks (Kadcyla® (T-DM1) FDA Labeling Supplement, 2016).

[0117] In some embodiments, the methods described herein may further include administering at least one additional therapy (e.g., a checkpoint inhibitor, a neoantigen vaccine, a cytokine or cytokine analog, CAR-T, etc.). In some embodiments, the administered amount of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or the at least one additional therapy is reduced due to induction of at least one neoantigen and / or T cell response when compared to a standard dosage of the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or the at least one additional therapy. In some embodiments, the administered amount of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or at least one additional therapy is reduced due to induction of a double-stranded RNA immune response when compared to a standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or at least one additional therapy. In some embodiments, the administered amount of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or at least one additional therapy is reduced due to induction of immunogenic cell death when compared to a standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or at least one additional therapy.In some embodiments, the dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to the standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or at least one additional therapy. In some embodiments, the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently when compared to a standard administration regimen of the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy. In some embodiments, the dose and / or dosage of the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy results in reduced systemic toxicity and / or improved tolerability.

[0118] In some embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated before administration of at least one additional therapy. In other embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated after administration of at least one additional therapy. In yet other embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated simultaneously with administration of at least one additional therapy.

[0119] In some embodiments, the administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is repeated at least once after the initial administration. In some embodiments, the amount used for the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the amount used for the initial administration. In some embodiments, the amount used for the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the standard dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the amount used for repeat administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to a standard dosage or initial dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0120] In some embodiments, the administration of the at least one additional therapy is repeated at least once after the initial administration. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced compared to the amount used for the initial administration. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced compared to the standard dosage of the at least one additional therapy. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage or the initial dosage of the at least one additional therapy.

[0121] In some embodiments, the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is concurrent with the repeated administration of at least one additional therapy. In some embodiments, the administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is sequential or staggered with the repeated administration of at least one additional therapy.

[0122] In some embodiments, the at least one additional therapy comprises administering a checkpoint inhibitor, e.g., any checkpoint inhibitor disclosed herein. In some embodiments, the subject is intolerant, refractory, or non-responsive to a checkpoint inhibitor when administered alone. In some embodiments, the checkpoint inhibitor targets PD1 / PDL1, CTLA4, OX40, CD40, LAG3, TIM3, GITR, and / or KIR. In some embodiments, the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR. In some embodiments, the checkpoint inhibitor is an antibody with inhibitory or agonistic activity against its target. In some embodiments, the checkpoint inhibitor is targeted with an inhibitory antibody or other similar inhibitory molecule. In other embodiments, the checkpoint inhibitor is targeted with an agonist antibody or other similar agonist molecule.

[0123] In some other embodiments, the at least one additional therapy comprises administering a neoantigen vaccine, such as any neoantigen vaccine disclosed herein. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered before administration of the neoantigen vaccine. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered after administration of the neoantigen vaccine. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered simultaneously with administration of the neoantigen vaccine. In some embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is repeated at least once after the initial administration. In some embodiments, the amount used for repeat administration of at least one compound selected from the compound of formula I, the compound of formula II, the compound of formula III, and pharmaceutically acceptable salts of any of the foregoing is reduced when compared to the amount used for the initial administration.

[0124] In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide. In some embodiments, the at least one neo-antigen peptide is in the range of about 10 to about 50 amino acids in length. In some embodiments, the at least one neo-antigen peptide is in the range of about 10 to about 35 amino acids in length. In some embodiments, the at least one neo-antigen peptide is in the range of about 15 to about 25 amino acids in length. In some embodiments, the at least one neo-antigen peptide comprises one or more neo-antigen sequences.

[0125] In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50 amino acids in length. In some embodiments, at least one neo-antigenic peptide ranges from about 10 to about 35 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 15 to about 25 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 20 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion does not exclusively overlap with or consist of a reference peptide sequence (e.g., any of the exemplary reference peptide sequences underlined in Table 13).

[0126] The term "antigenic portion" or "antigenic fragment" of a neo-antigen sequence, as used herein, refers to one or more fragments of a neo-antigen sequence that retain the ability to induce a T cell response (e.g., antigen-specific expansion and / or maturation of one or more effector T cell populations). The antigenic portion may also, in some embodiments, retain the ability to be internalized, processed, and / or presented by antigen-presenting cells (e.g., dendritic cells). In some embodiments, the antigenic portion also retains T cell priming function. In some embodiments, the antigenic portion of the neo-antigen sequence ranges from about 10 to about 50 amino acids in length. In some embodiments, the antigenic portion of the neo-antigen sequence ranges from about 10 to about 35 amino acids in length. In some embodiments, the antigenic portion of the neo-antigen sequence ranges from about 15 to about 25 amino acids in length. In some embodiments, the antigenic portion of the neo-antigen sequence ranges from about 10 to about 20 amino acids in length. In some embodiments, an antigenic portion of a neo-antigen sequence (eg, an antigenic portion of any one of SEQ ID NOs: 30-57), or its encoding mRNA, is formulated as a neo-antigen vaccine.

[0127] An exemplary embodiment of an antigenic portion is one or more regions flanking amino acids 45-53 of SEQ ID NO: 30. Another exemplary embodiment of an antigenic portion is one or more regions flanking amino acids 82-90 of SEQ ID NO: 30. In some embodiments, the antigenic portion is capable of binding to at least one HLA allele (e.g., HLA-A*02:01) expressed in a subject. In some other embodiments, the antigenic portion is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder. In some embodiments, the antigenic portion is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of a population of subjects suffering from a neoplastic disorder.

[0128] In some embodiments, the antigenic portion does not exclusively overlap with or consist of a reference peptide sequence. The term "reference peptide sequence," as used herein, refers to any contiguous peptide sequence present in the human proteome in the absence of contact with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing (e.g., in the absence of contact with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing), and / or in the absence of contact with a compound to which the immune system has previously been exposed. In some embodiments, the reference peptide sequence is derived from and / or encoded by a reference transcript open reading frame. Exemplary reference peptide sequences are underlined in Table 13.

[0129] In some embodiments, when at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered, the reference peptide sequence can be derived from and / or encoded by the in-frame 24 nucleotides immediately 5' preceding the aberrant splicing event induced by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Thus, in some embodiments, the reference peptide sequence comprises or consists of the 8 amino acids immediately N-terminal to the neo-antigen sequence induced by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, when a 5' exon sequence terminates at the termination nucleotide of a codon, the reference peptide sequence terminates at the end of that exon. In some other embodiments, when the 5' exon sequence terminates at one or two of the three nucleotides of a codon, the reference peptide sequence is derived from and / or encoded by the 24 nucleotides preceding the incomplete codon. In some embodiments, the mRNA sequence 3' to the aberrant splicing event can be translated within the same open reading frame derived from the 5' exon until it reaches a stop codon at which translation can terminate. In some embodiments, when the reference transcript open reading frame is preserved as a result of an aberrant splicing event (e.g., exon skipping), the C-terminal sequence can be translated, with an additional 24 nucleotides encoding the C-terminal 8 amino acids. In this regard, in some embodiments, only the region spanning the aberrant exon junction can encode the neo-antigen sequence. In some embodiments, when the open reading frame is shifted (e.g., intron retention), the complete C-terminal sequence (encoded by the 3' mRNA) can encode the neo-antigen sequence.

[0130] In some embodiments, antigenic portions of neo-antigen sequences are selected by comparing the neo-antigen sequence to a reference peptide sequence and selecting portions of the neo-antigen sequence that do not exclusively overlap, consist of, and / or align with the reference peptide sequence. Antigenic portions of neo-antigen sequences can, in some embodiments, be screened for antigenicity and / or T cell priming function in the same manner as full-length neo-antigen sequences (e.g., the neo-antigen sequence from which the antigenic portion is derived). In some embodiments, antigenic portions of neo-antigen sequences are assessed for antigenicity and / or T cell priming function using a T cell priming assay, such as the exemplary T cell priming experiments described herein.

[0131] In some embodiments, the neoantigen sequence is a neoantigen sequence specific to the subject. In some embodiments, the neoantigen sequence is a neoantigen vaccine personalized for the subject. In some embodiments, the neoantigen sequence used to create a neoantigen vaccine personalized for the subject has the ability to bind to at least one HLA allele expressed in the subject. In some embodiments, the personalized neoantigen vaccine is selected by, for example, identifying neoantigens expressed in the subject's tumor after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and selecting a vaccine comprising a neoantigen sequence found in the patient's tumor.

[0132] The term "individualized," when used in describing a neoantigen vaccine, refers to a vaccine created by identifying one or more neoantigens produced in a patient, preferably those identified in the patient after exposure to at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and then using one or more of those neoantigens as the basis for a vaccine for the same patient. Thus, in some embodiments, at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered to a patient and screened for neoantigens produced by the treatment. In some embodiments, the selected neoantigen vaccine comprises a neoantigen peptide or mRNA disclosed herein and identified to be present in the patient after exposure to at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, at least one compound and / or peptide or mRNA vaccine selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be administered to a patient once or repeatedly. Then, in some embodiments, one or more of these neoantigens are used to create a personalized vaccine that is administered to the patient. In some embodiments, one or more neoantigens used to create the personalized vaccine have binding affinity for one or more patient-specific HLA alleles. In some embodiments, the patient expresses one or more MHC1 alleles that bind to one or more neoantigens. Prediction of whether a given neoantigen binds to a particular MHC1 allele can be determined using any computational prediction method known in the art. Exemplary computational prediction methods are disclosed, for example, in Meydan et al. (2013) BMC Bioinformatics 14(Suppl. 2):S13, which is incorporated herein by reference for such methods.

[0133] In some other embodiments, the neo-antigen sequence is a universal neo-antigen sequence. In some embodiments, the neo-antigen sequence is a universal neo-antigen vaccine.

[0134] The term "universal," when used to describe a neoantigen vaccine, preferably refers to a vaccine having peptide or mRNA sequences based on one or more common or known neoantigens recognized by sequencing neoantigens produced in multiple patients and / or patient tissue samples after exposure to at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing. The peptide or mRNA sequence used in the vaccine need not be present in every patient; rather, it is sufficient that it be observed in at least some patients or patient tissue samples. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing and / or a peptide or mRNA vaccine may be administered to a patient once or repeatedly. Thereafter, in some embodiments, the peptide or mRNA sequence is used to vaccinate additional patients. In some embodiments, a patient is administered at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, followed by administration of a peptide or mRNA vaccine of a known neoantigen to enhance the immune response to the neoantigen produced by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, a patient is administered a universal peptide or mRNA vaccine followed by administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the neoantigen sequence (or sequences) used to create a universal neoantigen vaccine are selected based on the overall MHC1 allele frequency in a given patient population (Maiers et al. (2007) Hum. Immunol. 68(9):779-88).

[0135] In some embodiments, the neoantigen (e.g., universal neoantigen) sequence is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder. In some embodiments, the neoantigen sequence is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of subjects suffering from a neoplastic disorder.

[0136] In some embodiments, the neo-antigenic sequence is identified by sequencing at least one neo-antigenic peptide induced in a subject by administering a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, or by sequencing its encoding mRNA. In some embodiments, the at least one neo-antigenic peptide comprises a neo-antigenic sequence induced by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the neoplastic cell is present in an in vitro cell culture. In some embodiments, the neoplastic cell is obtained from a subject. In some embodiments, the neoplastic cell is present in a subject.

[0137] In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable carrier (e.g., any of the exemplary carriers described herein). In some embodiments, the at least one neo-antigen peptide is linked to a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine. In some embodiments, the neo-antigen peptide and the pharmaceutically acceptable carrier are covalently linked via a linker. In some embodiments, the neo-antigen peptide and the pharmaceutically acceptable carrier are expressed as a fusion protein. In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable diluent. In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide and a pharmaceutically acceptable adjuvant.

[0138] In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen mRNA, hi some embodiments, the at least one neo-antigen mRNA encodes one or more neo-antigen sequences.

[0139] In some embodiments, the neo-antigen sequence is a subject-specific neo-antigen sequence. In some embodiments, the neo-antigen sequence is a neo-antigen vaccine personalized for the subject. In some embodiments, the neo-antigen sequence has the ability to bind to at least one HLA allele expressed in the subject.

[0140] In some other embodiments, the neo-antigen sequence is a universal neo-antigen sequence. In some embodiments, the neo-antigen sequence is a universal neo-antigen vaccine. In some embodiments, the neo-antigen sequence is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder. In some embodiments, the neo-antigen sequence is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of subjects suffering from a neoplastic disorder.

[0141] In some embodiments, the neoantigen sequence is identified by sequencing the protein sequence of at least one neoantigen. In some embodiments, the neoantigen sequence is identified by sequencing at least one mRNA encoding a neoantigen induced in a subject by administering a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the at least one neoantigen mRNA encodes a neoantigen sequence induced by contacting neoplastic cells with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the neoplastic cells are present in an in vitro cell culture. In some embodiments, the neoplastic cells are obtained from a subject. In some embodiments, the neoplastic cells are present in a subject.

[0142] In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable carrier (e.g., any of the exemplary carriers described herein). In some embodiments, the at least one neoantigen mRNA is linked to a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine. In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent. In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant. In some embodiments, the neoantigen mRNA is encapsulated in an encapsulating agent. In some embodiments, the encapsulating agent is a liposome. In some embodiments, the encapsulating agent is a nanoparticle.

[0143] In some embodiments, the at least one additional therapy comprises administering a cytokine or cytokine analog, such as any cytokine or cytokine analog disclosed herein. In some embodiments, the subject is intolerant, refractory, or poorly responsive to a cytokine or cytokine analog when administered alone. In some embodiments, the cytokine or cytokine analog comprises a T cell enhancer. In some embodiments, the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα. In some embodiments, the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, and / or IL-15. In some embodiments, administration of the cytokine or cytokine analog enhances T cell priming following administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing due to the induction and presentation of neoantigens.

[0144] In some embodiments, the at least one additional therapy comprises administering engineered tumor-targeting T cells (i.e., CAR-T), such as any CAR-T therapy disclosed herein.

[0145] In some embodiments, the methods described herein may further include detecting one or more neoantigens and / or T-cell responses in the subject after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and, optionally, continuing administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing if one or more neoantigens and / or T-cell responses are detected. In some embodiments, detection of one or more neoantigens and / or T-cell responses in the subject indicates the effectiveness of treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if one or more neoantigens and / or T-cell responses are detected, treatment with an additional therapy is continued in conjunction with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if one or more neo-antigen and / or T cell responses are detected, treatment is continued at a reduced dosage and / or frequency.

[0146] In some embodiments, the methods described herein may further include detecting a double-stranded RNA immune response in the subject after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and optionally continuing administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing if a double-stranded RNA immune response is detected. In some embodiments, detection of a double-stranded RNA immune response in the subject indicates the effectiveness of treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if a double-stranded RNA immune response is detected, treatment with an additional therapy is continued in conjunction with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if a double-stranded RNA immune response is detected, treatment is continued at a reduced dosage and / or frequency.

[0147] In some embodiments, the methods described herein may further include detecting immunogenic cell death in the subject after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and optionally, continuing administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing if immunogenic cell death is detected. In some embodiments, detection of immunogenic cell death in the subject indicates the effectiveness of treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if immunogenic cell death is detected, treatment with additional therapy is continued in conjunction with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, if immunogenic cell death is detected, treatment is continued at a reduced dosage and / or frequency.

[0148] In some embodiments, the subject has a nonsynonymous mutation load of about 150 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 100 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 50 mutations or less. In some embodiments, the subject has or is suspected of having a neoplastic disorder, such as a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, and prostate cancer.

[0149] In some embodiments, the present disclosure further provides a method of treating a subject having or suspected of having a neoplastic disorder, the method comprising: (a) administering to the subject a therapeutically effective amount of at least one compound selected from the group consisting of compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing, wherein administration of the at least one compound selected from the group consisting of compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing induces at least one neoantigen and / or T-cell response; (b) detecting one or more neoantigen and / or T-cell responses in the subject after administration of the at least one compound selected from the group consisting of compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing; and (c) if the one or more neoantigen and / or T-cell responses are detected, continuing the administration of the at least one compound selected from the group consisting of compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing. In some embodiments, detection of one or more neo-antigens and / or T cell responses in a subject indicates the efficacy of treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the one or more neo-antigens comprise the amino acid sequence of any one of SEQ ID NOs: 1-29. In some embodiments, the one or more neo-antigens comprise the amino acid sequence of SEQ ID NO: 1. In some embodiments, the one or more neo-antigens comprise the amino acid sequence of SEQ ID NO: 3. In some embodiments, the one or more neo-antigens comprise the amino acid sequence of any one of SEQ ID NOs: 10-13.

[0150] In some embodiments, a patient having a cancer as described herein can be treated with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in combination with checkpoint inhibitor therapy.

[0151] Treatment of patients with immune checkpoint blockade has shown robust efficacy in certain clinical indications. Recently, the FDA approved the use of checkpoint inhibitors in patients with tumors exhibiting high microsatellite instability, regardless of histology. This approval was based, in part, on the observation that response rates are positively correlated with mutational burden (Rizvi et al. (2015) Science 348(6230):124-8; Hellmann et al. (2018) Cancer Cell 33(5):853-861). Estimates from the literature vary in absolute numbers and across lineages, but generally support an increased probability of response above a threshold of approximately 150-250 mutations. Analysis of TCGA data indicates that the majority of adult-onset tumor lineages have a relatively low nonsynonymous mutation burden (Vogelstein et al. (2013) Science 339:1549-58). For most lineages, the median nonsynonymous mutation rate is approximately 30–80 per patient, well below the threshold at which checkpoint inhibitors improve the likelihood of response.

[0152] For example, HER2-positive breast cancers have been shown to have a median of approximately 60 nonsynonymous mutations per patient sample. However, as discussed above, the therapeutic efficacy threshold for checkpoint inhibitors is estimated to be in the range of approximately 150-250 nonsynonymous mutations; patients above this threshold are likely to experience complete remission, partial remission, and / or stable disease, while patients below this threshold are likely to experience disease progression. Therefore, strategies to increase the apparent number of nonsynonymous mutations and / or neoantigens displayed on tumor cells are desirable and may, for example, increase the overall likelihood of success with checkpoint inhibitor therapy. Because cytokines (and their analogs) operate via a similar mechanism of action, such strategies may also increase the overall likelihood of success with cytokine-based therapies.

[0153] Current response rates in HER2-positive breast cancer are approximately 15-25% (CTI NCT02129556). In some embodiments disclosed herein, treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in combination with checkpoint inhibitor and / or cytokine therapy may improve such response rates. In some embodiments, treatment with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in combination with checkpoint inhibitor and / or cytokine therapy, may be applied to any adult-onset tumor, particularly those with an estimated median nonsynonymous mutation rate below a threshold of approximately 150 mutations. In some embodiments, exemplary cancer types suitable for treatment with a therapeutically effective amount of at least one compound selected from the group consisting of the compound of Formula I, the compound of Formula II, the compound of Formula III, and any of the pharmaceutically acceptable salts thereof, alone or in combination with additional therapy (e.g., checkpoint inhibitor therapy, cytokine therapy) include, but are not limited to, esophageal cancer, non-Hodgkin's lymphoma, colorectal cancer, head and neck cancer, gastric cancer, endometrial cancer, pancreatic adenocarcinoma, ovarian cancer, prostate cancer, hepatocellular carcinoma, glioblastoma, breast cancer (e.g., HER2-positive breast cancer), lung cancer (e.g., non-small cell lung cancer), chronic lymphocytic leukemia, and acute myeloid leukemia.Other exemplary suitable cancer types are identified, for example, in Vogelstein et al. (2013) Science 339:1549-58 (which is incorporated herein by reference in its entirety).

[0154] Because many checkpoint inhibitor therapies are based on chronic tumor-associated antigen expression, periodic therapeutic boosts are necessary for efficacy and to "re-boost" responding T cell populations. The inducible nature of neoantigens generated by at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, as described herein, provides therapeutic administration regimens that can be designed to enhance neoantigen-responsive T cell immune responses while suppressing T cell exhaustion often caused by chronic antigen stimulation. For example, in some embodiments, a subject is administered an initial dose of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, to trigger aberrant splicing and the production of neoantigenic peptides. After sufficient time for protein production and antigen presentation has passed, in some embodiments, the subject is then administered an initial dose of a checkpoint inhibitor to boost and / or enhance effector T cell priming and expansion. In some embodiments, the waiting period between doses of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and a checkpoint inhibitor is about 2, about 3, about 4, about 5, about 6, or about 7 days, In some embodiments, the waiting period is about 3 to about 5 days.

[0155] In some embodiments, the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR. In some embodiments, the combination therapeutic benefit of a checkpoint inhibitor with a therapeutically effective amount of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing may be additive or supra-additive.

[0156] In some embodiments, administration of a therapeutically effective amount of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing is initiated prior to administration of the checkpoint inhibitor.

[0157] In some embodiments, administration of a therapeutically effective amount of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing is initiated after administration of a checkpoint inhibitor.

[0158] In some embodiments, administration of a therapeutically effective amount of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing begins simultaneously with administration of the checkpoint inhibitor, e.g., in a single formulated product or in separate formulated products administered in a single procedure.

[0159] In some embodiments, after a period sufficient for T cell priming and expansion has elapsed, the subject is then administered a second or subsequent dose of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to elicit re-presentation of neoantigen peptides. In some embodiments, the waiting period between the first dose of checkpoint inhibitor and the second or subsequent dose of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is about 2, about 3, about 4, or about 5 weeks. In some embodiments, the waiting period is about 3 weeks. Upon receiving the second or subsequent dose of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in some embodiments, the immune system may engage neoantigen-presenting tumor cells and / or induce tumor cell death. In some embodiments, after allowing for secondary T cell priming and expansion, the subject is then administered a second or subsequent dose of a checkpoint inhibitor to further expand the memory effector T cell population.

[0160] In some embodiments, the waiting period between the first dose of a therapeutically effective amount of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and the second or subsequent dose of a checkpoint inhibitor is about 2, about 3, about 4, or about 5 weeks, hi some embodiments, the waiting period is about 3 weeks.

[0161] In some embodiments, the administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can be pulsed after this exemplary initial treatment regimen, i.e., a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can be administered at intervals long enough to allow for antigen presentation, T cell engagement and / or tumor cell killing, and / or recovery of memory T cell populations (e.g., about every 4 weeks, about every 5 weeks, about every 6 weeks). At a later time point, in some embodiments, treatment with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can be combined with one or more checkpoint inhibitors targeted to restore effector function to depleted T cell populations. For example, in some embodiments, at a later time, treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be combined with one or more checkpoint inhibitors targeting PD1 / PDL1, LAG3, and / or TIM3. In some embodiments, the pulsatile nature of the neoantigen presentation and priming allows for less frequent and / or lower doses of the checkpoint inhibitor and / or at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the pulsatile nature of neo-antigen presentation may provide one or more therapeutic benefits to a checkpoint inhibitor (e.g., an anti-CTLA4 antibody such as ipilimumab) compared to the checkpoint inhibitor when administered without the concomitant administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, e.g., by potentially reducing the risk of adverse reactions often observed with standard dosing regimens of checkpoint inhibitors.

[0162] In certain embodiments, the checkpoint inhibitor is an inhibitor of the cytotoxic T-lymphocyte-associated antigen (CTLA4) pathway. CTLA4, also known as CD152, is a protein receptor that downregulates immune responses. CTLA4 is constitutively expressed on regulatory T cells, but is upregulated on conventional T cells only after activation. As used herein, the term "CTLA4 inhibitor" is intended to refer to any inhibitor of CTLA4 and / or the CTLA4 pathway. Exemplary CTLA4 inhibitors include, but are not limited to, anti-CTLA4 antibodies. CTLA4-blocking antibodies for human use have been developed based on preclinical activity seen in mouse models of anti-tumor immunity. Exemplary anti-CTLA4 antibodies include, but are not limited to, ipilimumab (MDX-010) and tremelimumab (CP-675,206), both of which are fully human. Ipilimumab is an IgG1 with a plasma half-life of approximately 12-14 days; tremelimumab is an IgG2 with a plasma half-life of approximately 22 days. See, e.g., Phan et al. (2003) Proc Natl Acad Sci USA. 100:8372-7; Ribas et al. (2005) J Clin Oncol. 23:8968-77; Weber et al. (2008) J Clin Oncol. 26:5950-6. In some embodiments, the anti-CTLA4 antibody is ipilimumab.

[0163] In certain embodiments, the checkpoint inhibitor is an inhibitor of the programmed death-1 (PD1) pathway. The programmed cell death 1 (PD1) pathway represents a major immune regulatory switch that tumor cells can engage to evade activated T cell immune surveillance. PD1 ligands (PDL1 and PDL2) are constitutively expressed or can be induced in various tumors. High expression of PDL1 (and, to a lesser extent, PDL2) on tumor cells has been shown to correlate with poor prognosis and survival in various other solid tumor types. Furthermore, PD1 has been suggested to control tumor-specific T cell expansion in melanoma patients. These observations suggest that the PD1 / PDL1 pathway plays a critical role in tumor immune evasion and may be considered an attractive target for therapeutic intervention. As used herein, the term "PD1 inhibitor" is intended to refer to any inhibitor of PD1 and / or the PD1 pathway. Exemplary PD1 inhibitors include, but are not limited to, anti-PD1 and anti-PDL1 antibodies. In certain embodiments, the checkpoint inhibitor is an anti-PD1 antibody. Exemplary anti-PD1 antibodies include, but are not limited to, nivolumab and pembrolizumab (MK-3475). For example, nivolumab is a fully human immunoglobulin G4 (IgG4) PD1 immune checkpoint inhibitor antibody that disrupts the interaction of the PD1 receptor with its ligands PDL1 and PDL2, thereby suppressing cellular immune responses (Guo et al. (2017) J Cancer 8(3):410-6). In some embodiments, the anti-PD1 antibody is nivolumab. For example, pembrolizumab is a potent, highly selective humanized mAb of the IgG4 / κ isotype designed to directly block the interaction between PD1 and its ligands PDL1 and PDL2. Pembrolizumab potently enhances T lymphocyte immune responses in cultured blood cells from healthy human donors, cancer patients, and primates. Pembrolizumab has also been reported to modulate levels of interleukin-2 (IL-2), tumor necrosis factor alpha (TNFα), interferon gamma (IFNγ), and other cytokines. Exemplary anti-PDL1 antibodies include, but are not limited to, atezolizumab, avelumab, and durvalumab.For example, atezolizumab is an IgG1 humanized mAb that has been reported to block PD1 / PDL1 interaction by targeting PDL1, which is expressed on a wide variety of malignant cells. This blockade of the PD1 / PDL1 pathway can stimulate anti-tumor immune defense mechanisms (Abdin et al. (2018) Cancers (Basel) 10(2):32). In some embodiments, the anti-PDL1 antibody is atezolizumab.

[0164] In certain embodiments, checkpoint inhibitors target PD1 / PDL1, CTLA4, OX40, CD40, LAG3, TIM3, GITR, and / or KIR. In certain embodiments, checkpoint inhibitors target CTLA4, OX40, CD40, and / or GITR. In certain embodiments, checkpoint inhibitors target with inhibitory antibodies or other similar inhibitory molecules (e.g., inhibitory anti-CTLA4 or anti-PD1 / PDL1 antibodies). In certain other embodiments, checkpoint inhibitors target with agonists of the target; examples of this class include the stimulatory targets OX40, CD40, and / or GITR. In some embodiments, checkpoint inhibitors targeting OX40, CD40, and / or GITR are agonistic antibodies. Agonistic antibodies directed against OX40 can have the dual role of inhibiting the suppression of regulatory T cells while enhancing effector T cell function. Agonistic anti-GITR antibodies have also been shown to enhance the resistance of effector T cells to suppression induced by regulatory T cells (Karaki et al. (2016) Vaccines (Basel) 4(4):37). Similarly, agonistic CD40 antibodies have demonstrated T cell-dependent antitumor activity. Activation of CD40 on dendritic cells increases cross-presentation of tumor antigens, resulting in increased numbers of activated tumor-targeting effector T cells (Ellmark et al. (2015) Oncoimmunol. 4(7):e1011484).

[0165] In certain embodiments, the checkpoint inhibitor targets CTLA4 (e.g., an anti-CTLA4 antibody). In certain embodiments, targeting CTLA4 promotes the priming and activation of naive T cells. In certain embodiments, the checkpoint inhibitor targets OX40 (e.g., an anti-OX40 antibody). In certain embodiments, targeting OX40 enhances the expansion of effector T cells. In certain embodiments, the checkpoint inhibitor targets CD40 (e.g., an anti-CD40 antibody). In certain embodiments, targeting CD40 inhibits "tolerogenic" priming of T cells and / or the formation of regulatory T cells. In certain embodiments, the checkpoint inhibitor targets GITR (e.g., an anti-GITR antibody). In certain embodiments, targeting GITR inhibits the activity of regulatory T cells. In certain embodiments, the benefit (e.g., effect on at least one symptom or risk / rate of disease progression) of combination therapy with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a CTLA4-targeting, OX40-targeting, CD40-targeting, and / or GITR-targeting agent is additive. In some embodiments, the benefit of combination therapy with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a CTLA4-targeting, OX40-targeting, CD40-targeting, and / or GITR-targeting agent is supra-additive (i.e., synergistic).

[0166] Checkpoint inhibitor therapeutic strategies are based on the hypothesis that treatment can promote and / or enhance the priming of T cell responses to weakly or poorly antigenic tumors (e.g., CTLA4) or restore and / or reactivate T cells that respond to tumor antigens but become "exhausted" due to the chronic nature of antigen presentation (e.g., PD1, PDL1) (Chen and Mellman (2013) Immunity 39(1):1-10). Examples of suitable checkpoint inhibitor therapies and agents, such as anti-PD1, anti-PDL1, or anti-CTLA4 antibodies, are known in the art. See, e.g., WO 2001 / 014424, WO 2013 / 173223, and WO 2016 / 007235.

[0167] Combining these primed T cell responses following checkpoint inhibitor therapy with a treatment that induces neoantigens in tumor cells to which the immune system can respond (e.g., by administering a therapeutically effective amount of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing) can result in beneficial synergy. Because neoantigens resulting from a compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing have not yet been presented for T cell priming, combination with a CTLA4 inhibitor can be particularly beneficial. In some embodiments, the treatment involves administering a therapeutically effective amount of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce neoantigen production, followed by, prior to, concurrently with, or following, an initial administration of a CTLA4 inhibitor to stimulate CD8 T cell priming. In some embodiments, an additional dose of a CTLA4 inhibitor is provided to the patient, e.g., to further stimulate priming and / or activation of neoantigen-reactive CD8 populations. In some embodiments, neoantigen presentation by the tumor can be increased by providing the patient with an additional therapeutically effective dose of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. The therapeutically effective dose of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and the repeated administration of the checkpoint inhibitor therapy can be administered simultaneously or at staggered intervals. In some embodiments, the treatment further includes administering a PD1 / PDL1 inhibitor co-therapy, e.g., to restore effector function of neoantigen-targeting T cells that have been depleted within the tumor microenvironment.

[0168] The terms "combination" or "combination therapy," as used herein, refer to the administration of a therapeutically effective amount of at least one compound selected from the group consisting of compounds of Formula I, Formula II, Formula III, and pharmaceutically acceptable salts of any of the foregoing, together with an additional agent or therapy (e.g., a checkpoint inhibitor, a cytokine or cytokine analog, a neoantigen vaccine, or a CAR-T) as part of a treatment regimen intended to provide a beneficial (i.e., additive or synergistic) effect due to the co-action of one or more of the administered agents. In some embodiments, the combination may also include one or more additional agents, including, but not limited to, chemotherapeutic agents, antiangiogenic agents, and agents that reduce immunosuppression (e.g., a second checkpoint inhibitor). Beneficial effects of the combination include, but are not limited to, pharmacokinetic or pharmacodynamic co-actions resulting from the combination of therapeutic agents. The administration of these therapeutic agents in combination typically occurs over a defined period of time (e.g., minutes, hours, days, or weeks, depending on the combination selected).

[0169] As used herein, "administered in combination" or "co-administration" refers to the delivery, in any order, of two or more different therapies to a subject while the subject is suffering from a medical condition (e.g., cancer or a neoplastic disorder). For example, in some embodiments, two or more therapies are delivered after a subject is diagnosed with a disease or disorder and before the disease or disorder is cured or resolved, or when the subject is identified as being at risk for the disease but before the subject develops symptoms. In some embodiments, the delivery of one treatment is still occurring when the delivery of a second treatment begins, thereby resulting in overlap. In some embodiments, the first and second treatments are initiated simultaneously. This type of delivery is sometimes referred to herein as "simultaneous," "concurrent," or "combined" delivery. In other embodiments, the delivery of one treatment ends before the delivery of the second treatment begins. This type of delivery is sometimes referred to herein as "continuous" or "sequential" delivery.

[0170] In some embodiments, the two treatments (e.g., at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a checkpoint inhibitor) are contained in the same composition. Such a composition may be administered in any suitable form and by any suitable route. In other embodiments, the two treatments (e.g., at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a checkpoint inhibitor) are administered in separate compositions, in any suitable form, and by any suitable route. For example, in some embodiments, a composition comprising a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a composition comprising a checkpoint inhibitor may be administered concurrently or sequentially at different times in any order; in either case, they must be administered sufficiently closely in time to produce the desired therapeutic or prophylactic effect.

[0171] In either simultaneous or sequential delivery embodiments, the combined administration can result in more effective treatment. In some embodiments, the first treatment is more effective than would be observed if the first treatment were administered in the absence of the second treatment, e.g., an equivalent effect is observed with less of the first treatment (e.g., a lower dose). In some embodiments, the first treatment is more effective, such that the reduction in symptoms or other parameters associated with the disease or disorder is greater than would be observed if the first treatment were administered in the absence of the second treatment. In other embodiments, a similar situation is observed with the second treatment. In some embodiments, the benefit of the combination therapy (e.g., effect on at least one symptom or risk / rate of disease progression) is additive. In some embodiments, the benefit of the combination therapy is supra-additive.

[0172] In some embodiments, the present disclosure provides a method of treating cancer in a subject in need thereof and / or having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and at least one additional therapy (e.g., checkpoint inhibitor therapy, cytokine or cytokine analog, neoantigen vaccine, CAR-T). In some embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces at least one neoantigen and / or T cell response. In some embodiments, administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces a double-stranded RNA immune response. In some embodiments, administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing induces immunogenic cell death. In some embodiments, the at least one additional therapy can include at least one, at least two, at least three, at least four, or at least five additional therapies. For example, in some embodiments, a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be administered in combination with two checkpoint therapies, i.e., using two different checkpoint inhibitors. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be administered in combination with checkpoint inhibitor therapy and a neoantigen vaccine.

[0173] In some embodiments of the combination therapy, the dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or at least one additional therapy. In some embodiments, the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently than a standard dosing regimen of the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy. In some embodiments, the dose and / or dosage of the at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and / or the at least one additional therapy results in reduced systemic toxicity and / or improved tolerability.

[0174] In some embodiments, administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated before administration of at least one additional therapy. In some embodiments, administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated after administration of at least one additional therapy. In some embodiments, administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated simultaneously with administration of at least one additional therapy.

[0175] In some embodiments, the administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is repeated at least once after the initial administration. In some embodiments, the amount used for the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the amount used for the initial administration. In some embodiments, the amount used for the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the standard dosage of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the amount used for repeat administration of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of at least one compound selected from the group consisting of compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing.

[0176] In some embodiments, the administration of the at least one additional therapy is repeated at least once after the initial administration. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced compared to the amount used for the initial administration. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced compared to the standard dosage of the at least one additional therapy. In some embodiments, the amount used for the repeated administration of the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of the at least one additional therapy.

[0177] In some embodiments, the repeated administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is concurrent with the repeated administration of at least one additional therapy. In some embodiments, the repeated administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is sequential or staggered with the repeated administration of at least one additional therapy.

[0178] In some embodiments, the present disclosure provides methods of treating cancer in a subject in need thereof and / or a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and checkpoint inhibitor therapy. In some embodiments, the checkpoint inhibitor therapy comprises administering at least one checkpoint inhibitor. In some embodiments, the subject is intolerant, refractory, or refractory to at least one checkpoint inhibitor administered alone. In some embodiments, the subject may be considered refractory or refractory to at least one checkpoint inhibitor, for example, as determined using immune-related Response Criteria (irRC) and / or Response Evaluation Criteria in Solid Tumors (irRECIST). See, e.g., Wolchok et al. (2009) Clin Cancer Res. 15(23):7412-20; Bohnsack et al. "Adaptation of the Immune-Related Response Criteria: irRECIST" (Abstract 4958) ESMO 2014. Exemplary criteria may include those used in the art to define when a cancer patient's tumor improves ("responds"), stays the same ("stabilizes"), or worsens ("progresses") during treatment, when the treatment being evaluated is an immuno-oncology drug (e.g., a checkpoint inhibitor). In some embodiments, a subject may be considered intolerant to at least one checkpoint inhibitor if the subject experiences one or more adverse (grade 2 or higher) events identified for each checkpoint inhibitor (e.g., ipilimumab).In some embodiments, a subject may be considered intolerant to ipilimumab treatment if the subject experiences one or more adverse events selected from, for example, enterocolitis, hepatitis, dermatitis (including toxic epidermal necrolysis), neuropathy, and endocrinopathy (Yervoy® (ipilimumab) FDA Labeling Supplement, 2018).

[0179] In some embodiments, checkpoint inhibitors target PD1 / PDL1, CTLA4, OX40, CD40, LAG3, TIM3, GITR, and / or KIR. In some embodiments, checkpoint inhibitors target CTLA4, OX40, CD40, and / or GITR. In some embodiments, checkpoint inhibitors are targeted with inhibitory antibodies or other similar inhibitory molecules. In some other embodiments, checkpoint inhibitors are targeted with agonist antibodies or other similar agonist molecules. In some embodiments, checkpoint inhibitors comprise cytotoxic T-lymphocyte-associated antigen 4 pathway (CTLA4) inhibitors. In some embodiments, the CTLA4 inhibitor is an anti-CTLA4 antibody. In some embodiments, the anti-CTLA4 antibody is ipilimumab. In some embodiments, checkpoint inhibitors comprise programmed death-1 pathway (PD1) inhibitors. In some embodiments, the PD1 inhibitor is an anti-PD1 antibody. In some embodiments, the anti-PD1 antibody is nivolumab. In some embodiments, the PD1 inhibitor is an anti-PDL1 antibody. In some embodiments, the anti-PDL1 antibody is atezolizumab. In some embodiments, the checkpoint inhibitor comprises a CTLA4 inhibitor and a PD1 inhibitor. In some embodiments, the checkpoint inhibitor targets OX40. In some embodiments, the checkpoint inhibitor targets CD40. In some embodiments, the checkpoint inhibitor targets GITR. In some embodiments, the benefit (e.g., effect on at least one symptom or risk / rate of disease progression) of combination therapy with a therapeutically effective amount of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and a checkpoint inhibitor (e.g., a CTLA4-targeting, PD1 / PDL1-targeting, OX40-targeting, CD40-targeting, and / or GITR-targeting antibody or molecule) is additive.In some embodiments, the benefit of combination therapy with a therapeutically effective amount of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing, and a checkpoint inhibitor (e.g., a CTLA4-targeting, PD1 / PDL1, OX40-targeting, CD40-targeting, and / or GITR-targeting antibody or molecule) is supra-additive (i.e., synergistic).

[0180] In some embodiments, the present disclosure provides a method of treating cancer in a subject in need thereof and / or having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from compounds of Formula I, compounds of Formula II, compounds of Formula III, and pharmaceutically acceptable salts of any of the foregoing; and cytokine or cytokine analog therapy. In some embodiments, the cytokine or cytokine analog therapy comprises administering at least one cytokine or cytokine analog. In some embodiments, the subject is intolerant, refractory, or non-responsive to at least one cytokine or cytokine analog when administered alone.

[0181] In some embodiments, the cytokine or cytokine analog comprises a T cell enhancer. In some embodiments, the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα. In some embodiments, the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, and / or IL-15. In some embodiments, administration of the cytokine or cytokine analog enhances T cell priming following administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, due to the induction and presentation of neoantigens.

[0182] In some embodiments, the cytokine or cytokine analog comprises IL-2. In some embodiments, IL-2 boosts signaling to effector cells, promoting their expansion (Rosenberg (2014) J Immunol. 192(12):5451-8). In some embodiments, the cytokine or cytokine analog comprises IL-10. In some embodiments, IL-10 boosts CD8+ T cell priming and activation (Mumm et al. (2011) Cancer Cell 20(6):781-96). In some embodiments, the cytokine or cytokine analog comprises IL-12. In some embodiments, IL-12 links innate and adaptive immune responses, boosting antigen-specific priming and targeting (Tugues et al. (2015) Cell Death Differ. 22(2):237-46). In some embodiments, the cytokine or cytokine analog comprises IL-15. In some embodiments, IL-15 boosts T effector (CD8) cell priming and / or activation. In some embodiments, the cytokine or cytokine analog comprises IFNγ. In some embodiments, IFNγ complements T effector cell secretion of IFNγ. In some embodiments, the cytokine or cytokine analog comprises TNFα. In some embodiments, TNFα complements T effector cell secretion of TNFα.

[0183] In some embodiments, a subject is administered an initial therapeutically effective dose of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to induce aberrant splicing and production of neo-antigenic peptides. After sufficient time for protein production and antigen presentation has elapsed, in some embodiments, the subject is then administered an initial dose of a cytokine or cytokine analog to boost and / or enhance effector T cell priming and expansion. In some embodiments, the waiting period between doses of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and the cytokine or cytokine analog is about 2, about 3, about 4, about 5, about 6, or about 7 days. In some embodiments, the waiting period is about 3 to about 5 days. In some embodiments, the cytokine or cytokine analog is IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα. In some embodiments, the combination therapy benefit of at least one compound selected from compounds of formula I, compounds of formula II, compounds of formula III, and pharmaceutically acceptable salts of any of the foregoing with a cytokine or cytokine analog may be additive or superadditive.

[0184] In some embodiments, after a period of time sufficient for T cell priming and expansion has elapsed, the subject is then administered a second or subsequent dose of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing to elicit re-presentation of neo-antigenic peptides. In some embodiments, the waiting period between the first dose of cytokine or cytokine analog and the second or subsequent dose of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is about 2, about 3, about 4, or about 5 weeks. In some embodiments, the waiting period is about 3 weeks. In some embodiments, the subsequent dose of cytokine or cytokine analog may be administered intermittently, for example, between subsequent doses of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. Upon receiving a second or subsequent dose of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, the immune system may, in some embodiments, engage neoantigen-presenting tumor cells and / or induce tumor cell killing. In some embodiments, the administration of a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, after this exemplary initial treatment regimen, may be pulsed, i.e., the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, may be administered at intervals long enough to allow for antigen presentation, T cell engagement and / or tumor cell killing, and / or recovery of memory T cell populations (e.g., about every 4 weeks, about every 5 weeks, about every 6 weeks).

[0185] In some embodiments, the subject has a nonsynonymous mutation load of about 150 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 100 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 50 mutations or less. In some embodiments, the subject has or is suspected of having a neoplastic disorder, such as a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, and prostate cancer.

[0186] In some embodiments, the subject is in need of a method for treating cancer. In some embodiments, the cancer is a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, and prostate cancer.

[0187] In some embodiments, patients with cancer as described herein can be treated with a neoantigen vaccine in combination with at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and pharmaceutically acceptable salts of any of the foregoing. Without being bound by theory, vaccines used alone or in combination with immune checkpoint inhibitor (ICI) molecules have shown promise in early trials (Ott et al. (2017) Nature 547(7662):217-21; Sahin et al. (2017) Nature 547(7662):222-6), but generally require sequencing of patient tumor mutations (Ott et al. (2017) Nature 547(7662):217-21; Aldous and Dong (2018) Bioorg. Med. Chem. 26(10):2842-9). Thus, vaccines often rely on a sufficient number of antigenic nonsynonymous mutations. Generally, tumors with very low mutational burden offer few antigen candidates, and rapidly growing tumors limit the time available for identifying and producing patient-specific vaccines.

[0188] To date, attempts to develop vaccines that can be broadly immunogenic in most patients have focused on proteins that are either frequently mutated, ectopically overexpressed, or amplified, and / or that exist as "self" proteins in the organism. In addition, these proteins are often expressed in immunologically restricted tissues (e.g., neuronal markers expressed in neuroendocrine tumor types), while others may be expressed normally during embryonic development (e.g., carcinoembryonic antigen). Therefore, the usefulness of vaccines that use such proteins as antigens is often limited to specific tumor lineages or subsets in which one or more of the antigens are presented. The usefulness of the vaccine may also need to be confirmed by sequencing patient tumor samples, which can be time-consuming.

[0189] Furthermore, if these antigens are present as "self" proteins, the immune system may be primed to recognize them as "self" and thus not respond. Alternatively, if the immune system is able to mount an effector response against such antigens, this could lead to on-target side effects in tissues where the antigens may be expressed. In both of these cases, one of the key challenges is that many antigenic peptides are derived from "passenger" genes (i.e., genes that mutate or amplify during tumor development but do not play a critical role in the continued survival or growth of the tumor itself). In this way, these genes can be silenced without significantly affecting tumor progression, thereby allowing tumors to "escape" the immune response against their antigens. Without wishing to be bound by theory, this mechanism may play a role in tumor evolution, where random mutations with strong antigenicity are often "counterselected" by tumors early in tumor development (Dunn et al. (2004) Annu. Rev. Immunol. 22:329-60).

[0190] Additionally, evidence suggests that chronic antigen presentation and immune stimulation can lead to immune cell anergy and exhaustion (Pardoll (2012) Nat. Rev. Cancer 12(4):252-64). These phenotypes form the basis for the therapeutic rationale behind current ICI treatments, as ICIs have been shown to either suppress immune cell exhaustion phenotypes (α-PD1 / PD-L1) or promote additional immune cell responses (α-CTLA4). Of note, some patients with α-CTLA4 therapy have been reported to experience severe immune-related adverse events, which may be attributed to the promotion of T cell activation and the disruption of immune tolerance mechanisms that suppress autoreactive immune responses.

[0191] Both of these approaches (i.e., eliciting or enhancing a de novo immune response to a neoantigen or reversing anergy or exhaustion of a pre-existing immune response) involve chronic immune activation, and as such, are susceptible to anergy, editing, and other tumor-mediated mechanisms designed to suppress immune engagement.

[0192] In contrast, treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can induce an immune response against novel sequences corresponding to neoantigens. In some embodiments, presentation of neoantigens allows the adaptive immune system to engage and activate a greater variety of targets. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing can acutely induce alternative splicing and the resulting neoantigens, thereby reducing the risk of immune system exhaustion due to chronic exposure to mutation-derived neoantigens and / or limiting the ability of tumor cells to adapt to evade therapy. In some embodiments, administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing in combination with a neoantigen vaccine enhances the immune response against neoantigens generated by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered before, during, or after vaccination. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and / or the vaccine may be administered once or more than once during a course of treatment. In some embodiments, the vaccine is administered once during a course of treatment, and at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered more than once. In some embodiments, the vaccine is administered once during a course of treatment, followed by one or more booster doses.

[0193] As used herein, the term "neo-antigen vaccine" refers to a pooled preparation of one or more immunogenic neo-antigen peptides or mRNAs, e.g., at least two, at least three, at least four, at least five, or more neo-antigen peptides. The term "vaccine" refers to a composition that generates immunity for the prevention and / or treatment of disease (e.g., neoplastic disorders, e.g., hematological malignancies or solid tumors). Thus, a vaccine is a pharmaceutical product containing an immunogenic agent and intended for use in humans or animals to generate specific immune defenses and protection following vaccination. Neo-antigen vaccines can further comprise pharmaceutically acceptable carriers, diluents, excipients, and / or adjuvants.

[0194] As used herein, the term "immunogenic" refers to any agent or composition that is capable of eliciting an immune response, such as a T cell response. The immune response can be antibody-mediated or cell-mediated, or both.

[0195] In some embodiments, a patient is administered at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, followed by administration of a peptide or mRNA vaccine of a known neoantigen to enhance the immune response to the neoantigen produced by the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some other embodiments, a patient is administered at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and screened for neoantigens produced by the treatment. One or more of these neoantigens are then used to create a personalized vaccine to be administered to the patient. In any of these embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and / or the peptide or mRNA vaccine may be administered to the patient once or repeatedly.

[0196] In some embodiments, neoantigens suitable for vaccines can be identified by screening a panel of transcripts with splicing alterations and robust expression from one or more patient tissue samples (e.g., from tumor biopsies). In some embodiments, mutant protein sequences are identified in the screened samples based on translation spanning aberrantly spliced ​​mRNA junctions while retaining a portion of the protein sequence (up to 12 amino acids) adjacent to the amino acid changes spanning the junction. In some embodiments, peptide fragments spanning these junctions are scanned for high-affinity binding to MHC1 alleles using tools such as NetMHC1 (Nielsen et al. (2003) Protein Sci 12(5):1007-17; Andreatta and Neilsen (2016) Bioinformatics 32(4):511-7). These results allow for filtering neopeptides for predicted high-affinity binders to unique patient HLA allele configurations, as well as assembling pools of neopeptides predicted to bind broadly to HLA alleles occurring at high frequency in various populations (Maiers et al. (2007) Hum Immunol 68(9):779-88). In some embodiments, the identified neopeptides are then formulated as vaccines, e.g., by conjugation with a suitable carrier or adjuvant (Ott et al. (2017) Nature 547(7662):217-21), or formulated for delivery as mRNA (Sahin et al. (2017) Nature 547(7662):222-6).

[0197] In some embodiments, the neoantigens selected are based on screening individual patent tumor responses to at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, to identify one or more neoantigens resulting from the treatment for use in subsequent vaccination. In other embodiments, neoantigens are selected based on, for example, screening a panel of samples from different patients to identify common neoantigens produced by at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, which are then used as a universal vaccine in future patients.

[0198] Without being bound by theory, in some embodiments, the use of a universal neoantigen vaccine may eliminate the need to sequence and analyze the unique mutational status of each patient's tumor because the selected neoantigens are not dependent on tumor mutations but rather mimic neoantigens produced by at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and which are generally recognized as foreign by the body. Additionally, in some embodiments, the use of a neoantigen vaccine may be particularly effective because a patient's tumor cells may be more likely to mutate away from producing one or more neoantigens dependent on tumor mutations, compared to those that mimic neoantigens produced by at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. This may enable the formulation of a bulk vaccine that may have broad immunogenicity across a large proportion of patients, accelerating the initiation of a treatment regime. Patients may be vaccinated according to the schedule outlined herein and may be further treated with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, prior to completion of subsequent vaccinations, e.g., to induce expression of neoantigen peptides. In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be administered to the patient before, simultaneously with, or after vaccination. In some embodiments, the patient is administered at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, screened for one or more neoantigens found in a panel of universal neoantigens, and vaccinated with a universal neoantigen vaccine comprising at least one universal neoantigen identified in the subject.In some embodiments, at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing may be administered to a patient one or more times after vaccination. At least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing and / or a vaccine may be administered one or more times during the course of treatment.

[0199] In some embodiments, vaccines may include one or more neo-antigenic peptides or mRNAs. In some embodiments, vaccines may include one or more long neo-antigenic peptides. Such "long" neo-antigenic peptides, in some embodiments, undergo efficient internalization, processing, and cross-presentation in professional antigen-presenting cells, such as dendritic cells. Similarly, long vaccine peptides have been shown in other contexts to induce cytotoxic T cells in humans (Melief and van der Burg (2008) Nat Rev Cancer 8(5):351-60). In some embodiments, neo-antigenic peptides are extended to include the neo-antigenic peptide sequence itself plus adjacent amino acid sequences. In some embodiments, the extended peptide sequence facilitates protein uptake by antigen-presenting cells, such as dendritic cells. In some embodiments, the extended peptide sequence enables efficient antigen presentation and T cell priming in various HLA isotype models. In some embodiments, longer neo-antigenic peptides and / or extended peptide sequences exhibit increased uptake by antigen-presenting cells (e.g., dendritic cells), increased antigen presentation, and / or increased T cell priming compared to shorter neo-antigenic peptides and / or shorter peptide sequences (e.g., peptide sequences less than about 10 amino acids in length or less than about 5 amino acids in length). In some embodiments, long neo-antigenic peptides range from about 5 to about 50 amino acids in length. In some embodiments, long neo-antigenic peptides range from about 10 to about 50 amino acids in length. In some embodiments, at least one neo-antigenic peptide ranges from about 10 to about 35 amino acids in length. In some embodiments, long neo-antigenic peptides range from about 15 to about 25 amino acids in length.

[0200] In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 35 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 15 to about 25 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 20 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion does not exclusively overlap with or consist of a reference peptide sequence (e.g., any of the exemplary reference peptide sequences underlined in Table 13).

[0201] The amino acid sequences of exemplary long neo-antigenic peptides are shown in Table 13.

[0202] These exemplary neo-antigenic peptides are generated following administration of an ADC containing a pladienolide splicing modulator; however, given a similar mechanism of action (i.e., a similar splicing modulation mechanism), similar neo-antigenic peptides can be produced by a compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0203] [Table 2]

[0204] The protein sequences of the 29 neopeptides listed in Table 12 can be extended. The extended protein sequences incorporate both the neopeptide sequence itself and additional flanking amino acid sequences. The extended protein sequences better facilitate protein uptake by dendritic cells and enable antigen presentation and T cell priming in various HLA isotype models. The amino acid sequences of the 29 extended neopeptides are shown in Table 13.

[0205] [Table 3]

[0206] [Table 4]

[0207] As used herein, a neo-antigenic peptide or mRNA vaccine encompasses the use of a fragment of a neo-antigenic peptide or its encoding mRNA, so long as the fragment retains its immunogenic potential.

[0208] In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide. In some embodiments, the neo-antigen vaccine comprises at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 12, at least 15, or at least 20 neo-antigen peptides. In some embodiments, one or more neo-antigen peptides range from about 5 to about 50 amino acids in length. In some embodiments, one or more neo-antigen peptides range from about 10 to about 50 amino acids in length. In some embodiments, at least one neo-antigen peptide ranges from about 10 to about 35 amino acids in length. In some embodiments, one or more neo-antigen peptides ranges from about 15 to about 25 amino acids in length.

[0209] In some embodiments, the present disclosure provides a method of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and a neoantigen vaccine. The neoantigen vaccine may be, for example, a peptide or mRNA neoantigen vaccine. In some embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered before the administration of the neoantigen vaccine. In some embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered after the administration of the neoantigen vaccine. In some embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is administered simultaneously with the administration of the neoantigen vaccine. In some embodiments, the administration of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is repeated at least once after the initial administration. In some embodiments, the amount used in the repeated administration of at least one compound selected from the group consisting of a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the amount used in the initial administration.

[0210] In some embodiments, the present disclosure further provides a combination comprising at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and a neoantigen vaccine (e.g., a universal neoantigen vaccine) for use in treating a subject having or suspected of having a neoplastic disorder. In some embodiments, the neoantigen vaccine is a peptide or mRNA neoantigen vaccine. In some embodiments, the combination further comprises at least one additional therapy. In some embodiments, the at least one additional therapy comprises at least 1, at least 2, at least 3, at least 4, or at least 5 additional therapies.

[0211] In some embodiments, the present disclosure further provides methods of treating a subject having or suspected of having a neoplastic disorder by: (a) administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; (b) detecting one or more neoantigens in the subject after administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; (c) comparing the one or more neoantigens to a panel of universal neoantigens; and (d) administering to the subject a universal neoantigen vaccine comprising at least one universal neoantigen present in the subject. In some embodiments, the universal neoantigen vaccine is administered alone or in combination with at least one additional therapy. In some embodiments, the at least one additional therapy comprises at least 1, at least 2, at least 3, at least 4, or at least 5 additional therapies.

[0212] In some embodiments, the at least one additional therapy comprises repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated before administration of the universal neo-antigen vaccine. In some embodiments, the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated after administration of the universal neo-antigen vaccine. In some embodiments, the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is initiated simultaneously with administration of the universal neo-antigen vaccine. In some embodiments, the amount used for the repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced compared to the amount used for the initial administration. In some embodiments, the amount used for the initial administration and / or repeat administration of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced when compared to the standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing when used without vaccine therapy. In some embodiments, the amount used for the initial administration and / or repeat administration of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to the standard dosage of at least one compound selected from the compound of Formula I, the compound of Formula II, the compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing.

[0213] In some embodiments, the at least one additional therapy comprises administering a checkpoint inhibitor (e.g., any of the exemplary checkpoint inhibitors described herein). In some embodiments, administration of the checkpoint inhibitor is initiated before administration of the universal neo-antigen vaccine and / or repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, administration of the checkpoint inhibitor is initiated after administration of the universal neo-antigen vaccine and / or repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, administration of the checkpoint inhibitor is initiated simultaneously with administration of the universal neo-antigen vaccine and / or repeated administration of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, administration of the checkpoint inhibitor is repeated at least once after the initial administration. In some embodiments, the amount used in the repeated administration of the checkpoint inhibitor is reduced compared to the amount used in the initial administration. In some embodiments, the amount used for repeated administration of the checkpoint inhibitor is reduced when compared to the standard dosage of the checkpoint inhibitor. In some embodiments, the amount used for repeated administration of the checkpoint inhibitor is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% when compared to the standard dosage of the checkpoint inhibitor. In some embodiments, the subject is intolerant, refractory, or non-responsive to a checkpoint inhibitor when administered alone.

[0214] Also provided herein, in some embodiments, is a neo-antigen vaccine comprising at least one neo-antigen peptide or at least one neo-antigen mRNA. In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide. In some other embodiments, the neo-antigen vaccine comprises at least one neo-antigen mRNA.

[0215] Also provided herein, in some embodiments, are kits comprising at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and a neo-antigen vaccine (e.g., a universal neo-antigen vaccine). In some embodiments, the neo-antigen vaccine is a peptide or mRNA neo-antigen vaccine. In some embodiments, the kits further comprise one or more additional components, including, but not limited to, instructions for use; other medicinal agents, such as one or more additional therapeutic agents; a device, container, or other material for preparing the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or the neo-antigen vaccine for therapeutic administration; a pharmaceutically acceptable carrier; and a device, container, or other material for administering the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or the neo-antigen vaccine to a patient. The instructions for use may include guidance regarding therapeutic application, including recommended dosages and / or administration methods, for example, in patients with or suspected of having a neoplastic disorder. In some embodiments, the kit further includes instructions for therapeutic use, e.g., at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a neoantigen vaccine, to treat or prevent a neoplastic disorder in a patient. In some embodiments, the kit further includes at least one additional therapeutic agent (e.g., for administration together with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and a neoantigen vaccine, e.g., a checkpoint inhibitor). In some embodiments, the at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, and / or the neoantigen vaccine is formulated as a pharmaceutical composition.

[0216] In some embodiments of the methods and compositions disclosed herein, the neo-antigen vaccine comprises at least one neo-antigen peptide. In some embodiments, the at least one neo-antigen peptide is in the range of about 10 to about 50 amino acids in length. In some embodiments, the at least one neo-antigen peptide is in the range of about 10 to about 35 amino acids in length. In some embodiments, the at least one neo-antigen peptide is in the range of about 15 to about 25 amino acids in length.

[0217] In some embodiments, at least one neo-antigen peptide comprises one or more neo-antigen sequences disclosed herein.

[0218] In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 35 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 15 to about 25 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 20 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion does not exclusively overlap with or consist of a reference peptide sequence (e.g., any of the exemplary reference peptide sequences underlined in Table 13).

[0219] In some embodiments, the neo-antigen sequence is a subject-specific neo-antigen sequence. In some embodiments, the neo-antigen sequence is a neo-antigen vaccine personalized for the subject. In some embodiments, the neo-antigen sequence has the ability to bind to at least one HLA allele expressed in the subject.

[0220] In some other embodiments, the neo-antigen sequence is a universal neo-antigen sequence. In some embodiments, the neo-antigen sequence is a universal neo-antigen vaccine. In some embodiments, the neo-antigen sequence is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder. In some embodiments, the neo-antigen sequence is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of subjects suffering from a neoplastic disorder.

[0221] In some embodiments, the neo-antigenic sequence is identified by sequencing at least one neo-antigenic peptide induced in a subject by administering a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the at least one neo-antigenic peptide comprises a neo-antigenic sequence induced by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the neoplastic cell is present in an in vitro cell culture. In some embodiments, the neoplastic cell is obtained from a subject. In some embodiments, the neoplastic cell is present in a subject.

[0222] In some embodiments, a neo-antigen vaccine comprises at least one neo-antigen peptide or mRNA and a pharmaceutically acceptable carrier. In some embodiments, the neo-antigen peptide or mRNA can be linked to a suitable carrier to aid in eliciting an immune response. Exemplary carriers for linking an immunogenic agent (e.g., a neo-antigen peptide or mRNA) include serum albumin, keyhole limpet hemocyanin, immunoglobulin molecules, thyroglobulin, ovalbumin, tetanus toxoid, or toxoids or attenuated toxin derivatives derived from other pathogenic bacteria, such as diphtheria, E. coli, cholera, or H. pylori. Other carriers that stimulate or enhance an immune response include cytokines, such as IL-1, IL-1α and β peptides, IL-2, γ-INF, IL-10, GM-CSF, and chemokines, such as M1P1α and β and RANTES. The immunogenic agent can also be linked to a peptide that enhances tissue transport, for example, as described in WO 97 / 17613 and WO 97 / 17614. In some embodiments, the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

[0223] In some embodiments, the neoantigenic peptide or mRNA may be linked to a pharmaceutically acceptable carrier. The immunogenic agent can be linked to the carrier by chemical crosslinking. Techniques for linking the immunogenic peptide to the carrier include disulfide bond formation using N-succinimidyl-3-(2-pyridyl-thio)propionate (SPDP) and succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) (if the peptide lacks a sulfhydryl group, this may be provided by the addition of a cysteine ​​residue). These reagents create a disulfide bond between themselves and the peptide cysteine ​​residue on one protein and an amide bond through the ε-amino on a lysine or other free amino group on another amino acid. Various such disulfide / amide-forming agents are described in Jansen et al. ((1982) Immun Rev. 62:185). Other bifunctional coupling agents form thioethers rather than disulfide bonds. Many of these thioether-forming agents are commercially available and include reactive esters of 6-maleimidocaproic acid, 2-bromoacetic acid, and 2-iodoacetic acid, 4-(N-maleimido-methyl)cyclohexane-1-carboxylic acid. The carboxyl group can be activated by combining it with succinimide or 1-hydroxyl-2-nitro-4-sulfonic acid, sodium salt. In some embodiments, the neoantigenic peptide and the pharmaceutically acceptable carrier are covalently linked via a linker.

[0224] Neoantigens and other such immunogenic peptides can also be expressed as fusion proteins with a carrier. The immunogenic peptide can be linked to the carrier at the amino terminus, carboxyl terminus, or anywhere within the peptide (internally). In some embodiments, multiple repeats of the immunogenic peptide can be present in the fusion protein. In some embodiments, the neoantigen peptide and a pharmaceutically acceptable carrier are expressed as a fusion protein.

[0225] In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide or its encoding mRNA and a pharmaceutically acceptable diluent. In some embodiments, the neo-antigen vaccine comprises at least one neo-antigen peptide or its encoding mRNA and a pharmaceutically acceptable adjuvant (e.g., an adjuvant as described herein).

[0226] In some embodiments of the methods and compositions disclosed herein, the neo-antigen vaccine comprises at least one neo-antigen mRNA, hi some embodiments, the at least one neo-antigen mRNA encodes one or more neo-antigen sequences.

[0227] In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 50 amino acids in length. In some embodiments, at least one neo-antigenic peptide ranges from about 10 to about 35 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 15 to about 25 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion ranges from about 10 to about 20 amino acids in length. In some embodiments, the neo-antigenic sequence and / or antigenic portion does not exclusively overlap with or consist of a reference peptide sequence (e.g., any of the exemplary reference peptide sequences underlined in Table 13).

[0228] In some embodiments, the neo-antigen sequence is a subject-specific neo-antigen sequence. In some embodiments, the neo-antigen sequence is a neo-antigen vaccine personalized for the subject. In some embodiments, the neo-antigen sequence has the ability to bind to at least one HLA allele expressed in the subject.

[0229] In some other embodiments, the neo-antigen sequence is a universal neo-antigen sequence. In some embodiments, the neo-antigen sequence is a universal neo-antigen vaccine. In some embodiments, the neo-antigen sequence is capable of binding to at least one HLA allele expressed in at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, or at least 45% of subjects in a population of subjects suffering from a neoplastic disorder. In some embodiments, the neo-antigen sequence is capable of eliciting a T cell response against tumors present in at least 1%, at least 5%, or at least 10% of subjects suffering from a neoplastic disorder.

[0230] In some embodiments, the neoantigen sequence is identified by sequencing at least one neoantigen mRNA induced in a subject by administering a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the at least one neoantigen mRNA encodes a neoantigen sequence induced by contacting a neoplastic cell with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. In some embodiments, the neoplastic cell is present in an in vitro cell culture. In some embodiments, the neoplastic cell is obtained from a subject. In some embodiments, the neoplastic cell is present in a subject.

[0231] In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable carrier. In some embodiments, the at least one neoantigen mRNA is linked to a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutically acceptable carrier is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

[0232] In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable diluent. In some embodiments, the neoantigen vaccine comprises at least one neoantigen mRNA and a pharmaceutically acceptable adjuvant (e.g., an adjuvant as described herein).

[0233] In some embodiments, the neo-antigen mRNA is encapsulated in an encapsulating agent. In some embodiments, the encapsulating agent protects the neo-antigen mRNA from degradation and improves vaccine delivery (McNamara et al. (2015) J Immunol Res. 2015:794528). In some embodiments, the encapsulating agent is a liposome. In some embodiments, the liposome is a cationic liposome such as N-[1-(2,3-dioleoloxy)propyl]-N,N,N-trimethylammonium chloride 1 (DOTAP). In some embodiments, the encapsulating agent is a nanoparticle. In some embodiments, the nanoparticle protects the neo-antigen mRNA from nuclease degradation and / or enhances cellular uptake and / or delivery efficiency. In some embodiments, the nanoparticle may be modified to be fully degradable. In some embodiments, the nanoparticles are biodegradable core-shell nanoparticles with a pH-responsive poly-(b-amino ester) (PBAE) core surrounded by a phospholipid shell (Su et al. (2011) Mol Pharm. 8(3):774-87). In some embodiments, such nanoparticles are particularly efficient for delivering mRNA and eliciting anti-tumor immune responses in vivo.

[0234] In some embodiments, the subject has a nonsynonymous mutation load of about 150 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 100 mutations or less. In some embodiments, the subject has a nonsynonymous mutation load of about 50 mutations or less. In some embodiments, the subject has or is suspected of having a neoplastic disorder, such as a hematological malignancy or a solid tumor. In some embodiments, the hematological malignancy is selected from B-cell malignancies, leukemia, lymphoma, and myeloma. In some embodiments, the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma. In some embodiments, the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer. In some embodiments, the solid tumor is selected from HER2-positive breast cancer, gastric adenocarcinoma, and prostate cancer.

[0235] As used herein, "adjuvant" refers to a substance capable of increasing, amplifying, or modulating the immune response to an associated immunogenic agent, e.g., a neoantigen peptide or mRNA. In certain embodiments, the neoantigens of the present disclosure can be administered in combination with an adjuvant, i.e., a substance that does not itself elicit an adaptive immune response but amplifies or modulates the response to the associated neoantigen. Various adjuvants can be used in combination with the disclosed neoantigens to elicit an immune response. In some embodiments, one or more adjuvants are selected to enhance the intrinsic response to the neoantigen without causing conformational changes in the neoantigen that could affect the qualitative form of the response. In some embodiments, one or more adjuvants are selected to enhance T effector (e.g., CD8) cell priming and / or activation.

[0236] In certain embodiments, the adjuvant is an aluminum salt (alum), such as aluminum hydroxide, aluminum phosphate, and aluminum sulfate. Such adjuvants can be used with or without other specific immunostimulants, such as 3-O-deacylated monophosphoryl lipid A (MPL) or 3-DMP, polymers such as polyglutamic acid or polylysine, or monomeric amino acids. Such adjuvants can be used with or without other specific immunostimulants, such as muramyl peptides (e.g., N-acetylmuramyl-L-threonyl-D-isoglutamine (thr-MDP), N-acetyl-normuramyl-L-alanyl-D-isoglutamine (nor-MDP), N-acetylmuramyl-L-alanyl-D-isoglutaminyl-L-alanine-2-(1'-2'dipalmitoyl-sn-glycero-3-hydroxyphosphoryloxy)-ethylamine (MTP-PE), N-acetylglucsaminyl-N-acetylmuramyl-L-Al-D-isoglu-L-Ala-dipalmitoxypropylamide (DTP-DPP)), or other bacterial cell wall components.Other adjuvants are oil-in-water emulsions such as (a) MF59 (WO 90 / 14837) containing 5% squalene, 0.5% Tween 80, and 0.5% Span 85 (optionally containing various amounts of MTP-PE) formulated into submicron particles using a microfluidizer such as a Model 110Y Microfluidics; (b) SAF containing 10% squalene, 0.4% Tween 80, 5% Pluronic blocked polymer L121, and thr-MDP, either microfluidized into a submicron emulsion or vortexed to produce a larger particle size emulsion; and (c) 2% squalene, 0.2% Tween 85. and the Ribi™ Adjuvant System (RAS) (Ribi ImmunoChem), which contains one or more bacterial cell wall components from the group consisting of monophosphoryl lipid A (MPL), trehalose dimycolate (TDM), and cell wall skeleton (CWS), e.g., MPL-FCWS (Detox™). In some embodiments, the adjuvant is a saponin, such as Stimulon™ (QS21), or particles produced therefrom, such as ISCOMs (immunostimulating complexes) and ISCOMATRIX. Other adjuvants include complete Freund's adjuvant (CFA) and incomplete Freund's adjuvant (IFA), cytokines, such as interleukins (IL-1, IL-2, and IL-12), macrophage colony-stimulating factor (M-CSF), and tumor necrosis factor (TNF).

[0237] The adjuvant can be administered with the immunogenic agent (e.g., neo-antigenic peptide or mRNA) as a single composition, or can be administered before, simultaneously with, or after administration of the immunogenic agent. In some embodiments, the immunogenic agent and adjuvant can be packaged and supplied in the same vial, or packaged in separate vials and mixed before use. In some embodiments, the immunogenic agent and adjuvant can be packaged with labeling indicating the intended therapeutic application. In some embodiments, if the immunogenic agent and adjuvant are packaged separately, the packaging can include instructions for mixing before use. The choice of adjuvant and / or carrier depends on the stability of the immunogenic formulation containing the adjuvant, the route of administration, the dosing schedule, and the efficacy of the adjuvant for the species being vaccinated. For humans, a pharmaceutically acceptable adjuvant is one that has been approved or will be approved for human administration by the relevant regulatory agency. For example, complete Freund's adjuvant is not suitable for human administration. However, alum, MPL, or incomplete Freund's adjuvant (Chang et al. (1998) Adv Drug Deliv Rev. 32:173-186), alone or optionally in combination with any of alum, QS21, and MPL, and any combination thereof, are suitable for human administration.

[0238] In some embodiments, the present disclosure further provides a method for screening and identifying at least one neoantigen. More specifically, in some embodiments, the present disclosure provides a method for identifying at least one neoantigen by: (a) contacting neoplastic cells with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; (b) detecting at least one alternatively spliced ​​mRNA transcript after contacting the neoplastic cells with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; (c) predicting translation of the at least one alternatively spliced ​​mRNA transcript into at least one peptide; and (d) comparing the at least one peptide to a reference proteome, wherein the at least one neoantigen is identified if the at least one peptide does not match any peptide in the reference proteome. In some embodiments, the method further includes contacting one or more additional neoplastic cells to identify at least one universal neoantigen. In some embodiments, the method is repeated with one or more additional neoplastic cells or samples (e.g., tissue biopsies) to confirm suitable neoantigens (e.g., for use in a neoantigen vaccine) and / or to identify one or more universal neoantigens.

[0239] In various other embodiments, the present disclosure provides methods for identifying at least one neoantigen by: (a) contacting neoplastic cells with a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; (b) detecting at least one peptide comprising a potential neoantigen sequence after contacting the neoplastic cells with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and (c) comparing the at least one peptide to a reference proteome, wherein at least one neoantigen is identified if the at least one peptide does not match any peptide in the reference proteome. In some embodiments, the method further includes contacting one or more additional neoplastic cells to identify at least one universal neoantigen. In some embodiments, the method is repeated with one or more additional neoplastic cells or samples (e.g., tissue biopsies) to confirm suitable neoantigens (e.g., for use in a neoantigen vaccine) and / or identify one or more universal neoantigens.

[0240] In some embodiments of the neoantigen identification methods described herein, detecting at least one alternatively spliced ​​mRNA transcript comprises RNAseq. In some embodiments, predicting translation of at least one alternatively spliced ​​mRNA transcript comprises quantifying a change in percent spliced-in (dPSI) value for the at least one transcript. In some embodiments, predicting translation of at least one alternatively spliced ​​mRNA transcript comprises RiboSeq and / or ribosome profiling.

[0241] In some embodiments of the neoantigen identification methods described herein, the method further comprises evaluating at least one peptide for predicted major histocompatibility complex (MHC) binding. In some embodiments, the predicted MHC binding is determined by measuring an uncorrected affinity predicted binding strength of at least one peptide. In some embodiments, an uncorrected affinity predicted binding strength of about 500 nM or greater indicates MHC binding. In some embodiments, the predicted MHC binding is determined by identifying a distribution of predicted binding strengths for a set of random peptides; and comparing the predicted binding strength of at least one peptide to the distribution. In some embodiments, a predicted binding strength in the top 2% of the distribution indicates weak MHC binding. In some embodiments, a predicted binding strength in the top 0.5% of the distribution indicates strong MHC binding.

[0242] In some embodiments of the neoantigen identification methods described herein, the neoplastic cells are present in an in vitro cell culture. In some embodiments, the neoplastic cells are obtained from a subject. In some embodiments, the neoplastic cells are present in a subject.

[0243] Also provided herein, in some embodiments, are methods of making a neoantigen vaccine by (a) identifying at least one neoantigen (e.g., at least one neoantigen peptide or its encoding mRNA) using any of the exemplary identification methods disclosed herein; and (b) formulating the at least one neoantigen with a pharmaceutically acceptable carrier, diluent, or adjuvant (e.g., any of the pharmaceutically acceptable carriers, diluents, or adjuvants described herein).

[0244] In some embodiments, at least one neo-antigen and / or antigenic moiety ranges from about 10 to about 50 amino acids in length. In some embodiments, at least one neo-antigen peptide ranges from about 10 to about 35 amino acids in length. In some embodiments, at least one neo-antigen and / or antigenic moiety ranges from about 15 to about 25 amino acids in length. In some embodiments, at least one neo-antigen and / or antigenic moiety ranges from about 10 to about 20 amino acids in length. In some embodiments, at least one neo-antigen and / or antigenic moiety does not exclusively overlap with or consist of a reference peptide sequence (e.g., any of the exemplary reference peptide sequences underlined in Table 13).

[0245] In some embodiments, at least one neo-antigen used in the vaccine is linked to a pharmaceutically acceptable carrier, which in some embodiments is selected from a peptide, serum albumin, keyhole limpet hemocyanin, immunoglobulin, thyroglobulin, ovalbumin, a toxoid or attenuated toxoid derivative, a cytokine, and a chemokine.

[0246] In some embodiments, patients having cancer as described herein can be treated with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in combination with one or more engineered tumor-targeting T cells (i.e., CAR-T). Accordingly, in some embodiments, the present disclosure provides a method of treating a subject having or suspected of having a neoplastic disorder by administering to the subject a therapeutically effective amount of at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing; and engineered tumor-targeting T cells (i.e., CAR-T). In some embodiments, a chimeric T cell receptor can be engineered with an antigen recognition sequence reactive to an identified neoantigen.

[0247] For example, in some embodiments, to target changes in the extracellular domain of a cell surface protein induced by at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, a chimeric antigen-reactive T cell receptor (CAR) can be engineered by first identifying an antibody that recognizes a cell surface-expressed neo-antigen protein domain. .

[0248] In various other embodiments, a strategy is used to integrate neoantigens generated from at least one compound selected from the group consisting of compounds of Formula I, Formula II, Formula III, and pharmaceutically acceptable salts of any of the foregoing with the antigen presentation machinery of tumor cells. In some embodiments, cells containing a known frequently occurring HLA allele (e.g., HLA-A*02:01) can be treated with at least one compound selected from the group consisting of compounds of Formula I, Formula II, Formula III, and pharmaceutically acceptable salts of any of the foregoing, and MHC1-binding neoantigens are identified by ligand mics. In some embodiments, these peptides can be used to prime and / or expand T cells from healthy donors expressing the same HLA allele. In some embodiments, such T cells can be isolated and their T cell receptor (TCR) alpha and beta chains can be sequenced to identify the cognate antigen recognition / variable regions. In some embodiments, the cognate CAR can then be engineered.

[0249] In some embodiments, the CAR sequence is cloned and expanded into a patient-derived T cell population using currently available protocols. In some embodiments, the engineered T cells are then infused back into the patient's circulation before, simultaneously with, or after treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing. After treatment with at least one compound selected from a compound of Formula I, a compound of Formula II, a compound of Formula III, and a pharmaceutically acceptable salt of any of the foregoing, in some embodiments, tumor cells may begin to present antigens, e.g., antigens targeted by the engineered T cell population. In some embodiments, the engineered T cell population can engage and kill antigen-presenting tumor cells.

[0250] In order that the disclosure set forth herein may be more fully understood, the following examples are set forth, it being understood that these examples are for illustrative purposes only and are not to be construed as limiting the disclosure in any manner whatsoever. [Example]

[0251] Examples 1 to 205 Summary: Microwave heating was performed using a Biotage Emrys Liberator or Initiator microwave. Column chromatography was performed using an Isco Rf200d. Solvent removal was performed using a Büchi rotary evaporator or a Genevac centrifugal evaporator. Preparative LC / MS was performed under acidic mobile phase conditions using a Waters autopurifier and a 19 x 100 mm XTerra 5 micron MS C18 column. NMR spectra were recorded using a Varian 400 MHz spectrometer.

[0252] When the term "inert" is used to describe a reactor (e.g., a reactor tank, flask, glass reactor, etc.), it means that the air within the reactor has been replaced with an essentially anhydrous or dry inert gas (nitrogen, argon, etc.).

[0253] General methods and experimental procedures for the preparation of compounds of the present disclosure are set forth below. In some cases, specific compounds are described by way of example. However, it will be understood that in each case, a series of compounds of the present disclosure were prepared according to the schemes and experimental procedures described below.

[0254] The following abbreviations are used herein: COMU: (1-cyano-2-ethoxy-2-oxoethylideneaminooxy)dimethylamino-morpholino-carbenium hexafluorophosphate DMAP: 4-(dimethylamino)pyridine DMP: Dess-Martin periodinane EDC: N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide KHMDS: potassium bis(trimethylsilyl)amide LCMS: Liquid Chromatography-Mass Spectrometry Pd2(dba)3: Tris(dibenzylideneacetone)dipalladium(0) TBAF: Tetrabutylammonium fluoride TBSCl: tert-butyldimethylsilyl chloride TBSOTf: tert-butyldimethylsilyl trifluoromethanesulfonate TESCl: chlorotriethylsilane THF: tetrahydrofuran TLC: Thin Layer Chromatography pTsOH: p-toluenesulfonic acid PPTS: Pyridinium p-toluenesulfonate

[0255] Materials: The following compounds are commercially available and / or can be prepared by several methods well known to those skilled in the art of organic synthesis. More specifically, the disclosed compounds can be prepared using the reactions and techniques described herein. In the description of synthetic methods set forth below, it should be understood that all proposed reaction conditions, including the choice of solvent, reaction atmosphere, reaction temperature, experimental duration, and workup procedures, can be selected to be standard conditions for the reaction in question unless otherwise indicated. It is understood by those skilled in the art of organic synthesis that the functional groups present on the various portions of the molecule must be compatible with the proposed reagents and reactions. Substituents incompatible with the reaction conditions will be apparent to those skilled in the art, and alternative methods are indicated accordingly. The starting materials in these examples are either commercially available or readily prepared by standard methods from known materials.

[0256] LCMS information: Mobile phase: A (0.1% formic acid in H2O) and B (0.1% formic acid in acetonitrile). Gradient: 5% to 95% B in 1.8 min. Column: Acquity BEH C18 column (1.7 um, 2.1 x 50 mm).

[0257] U.S. Patent Nos. 7,884,128 and 7,816,401, both entitled "Process for Total Synthesis of Pladienolide B and Pladienolide D," describe methods for synthesizing pladienolides B and D known in the art. The synthesis of pladienolides B and D may also be carried out using methods known in the art and described in Kanada et al., "Total Synthesis of the Potent Antitumor Macrolides Pladienolide B and D," Angew. Chem. Int. Ed. 46:4350-4355 (2007). Kanada et al. and WO 2003 / 099813, entitled "Novel Physiologically Active Substances," describe methods known in the art for synthesizing E7107 (compound 45 of WO '813) from pladienolide D (compound 11107D of WO '813). The corresponding U.S. patent is U.S. Pat. No. 7,550,503 to Kotake et al.

[0258] Exemplary Synthesis of Compounds Compounds 1-60 (Table I) were prepared by the method of Scheme 1. [ka]

[0259] General synthetic protocol for compounds 1-60: Step 1: A solution of pladienolide D (A, 5.3 g, 9.7 mmol, 1.0 equiv.) in DMF (80 mL, 0.1 M) at 0° C. under nitrogen was treated with imidazole (4.6 g, 67.8 mmol, 7.0 equiv.) and TBSCl (7.3 g, 48.4 mmol, 5.0 equiv.). The reaction was allowed to warm to room temperature and stirred for 20 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was extracted with ethyl acetate, and the organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (B, 7.5 g, 9.6 mmol, 99%).

[0260] Step 2: To a solution of olefin B (7.6 g, 9.7 mmol, 1.0 equiv) in degassed THF:HO (210 mL:21 mL, 0.01 M) under nitrogen at 0 °C, osmium tetroxide (24.4 mL, 1.9 mmol, 0.2 equiv, 2.5% solution in tert-butanol) was added, followed by N-methylmorpholine N-oxide (2.3 g, 19.5 mmol, 2.0 equiv). The reaction was allowed to warm to room temperature and stirred for 13 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium sulfite, diluted with ethyl acetate, and the organic layer was washed with water, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (dichloromethane / methanol as eluent) to give the desired product (C, 6.8 g, 8.3 mmol, 86%).

[0261] Step 3: To a solution of diol C (7.9 g, 9.7 mmol, 1.0 equiv) in benzene (350 mL, 0.03 M) at room temperature under nitrogen was added lead tetraacetate (8.6 g, 19.4 mmol, 2.0 equiv). The reaction was stirred for 30 minutes or until the reaction was complete as determined by LCMS or TLC. The reaction was concentrated and purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (D, 2.5 g, 5.26 mmol, 54%).

[0262] Step 4: To a solution of aldehyde D (1.4 g, 2.9 mmol, 1.0 equiv) in THF (9.5 mL, 0.5 M) at room temperature was added ethoxyethene (11.1 mL, 40.0 equiv) and pyridinium p-toluenesulfonate (0.07 g, 0.3 mmol, 0.1 equiv). The reaction was stirred for 24 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium bicarbonate and diluted with ethyl acetate. The ethyl acetate was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (E, 1.2 g, 2.2 mmol, 75%).

[0263] Step 5: To a solution of the corresponding sulfone (1.5 equiv.) in THF (0.02 M) at −78° C. under nitrogen, KHMDS (1.5 equiv.) was added dropwise, and the reaction was stirred for 20 minutes. Aldehyde E (1.0 equiv.) in THF was then added dropwise. The reaction was stirred at −78° C. for 90 minutes and then allowed to warm to −20° C. for 1 hour. The reaction was quenched with ammonium chloride, diluted with ethyl acetate, and warmed to room temperature. The organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (F).

[0264] Step 6: To a solution of acetate salt F (1.0 equiv.) in methanol (0.1 M) at room temperature was added potassium carbonate (1.1 equiv.). The reaction was run for 24 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with water, diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil (G) was carried on to the next step without further purification.

[0265] Step 7: To a solution of alcohol (G) (1.0 equiv.) in dichloromethane (0.1 M) at room temperature, N,N-dimethylaminopyridine (0.5 equiv.) was added, followed by 4-nitrophenyl chloroformate (2.0 equiv.). The reaction was stirred at room temperature for 3 hours. The corresponding amine (3.0 equiv.) was then added at room temperature. After stirring for 1 hour, the reaction was quenched with water and diluted with dichloromethane. The organic layer was washed with 1N sodium hydroxide solution, and the organic layer was concentrated. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (H).

[0266] Step 8: To a solution of the silyl ether (H, 1.0 equiv.) in methanol (0.1 M) at room temperature was added p-methoxytoluenesulfonic acid (3.0 equiv.). The reaction was stirred for 3 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium bicarbonate, diluted with ethyl acetate, washed with water and brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired products (1-59).

[0267] Exemplary synthesis protocol for compound 46 Steps 1 to 4 are as above.

[0268] Step 5: To a solution of (S)-2-(1-((1-phenyl-1H-tetrazol-5-yl)sulfonyl)propan-2-yl)pyridine (233.0 mg, 0.7 mmol, 1.4 equiv) in THF (2.5 mL, 0.2 M) at −78° C. under nitrogen, KHMDS (1.5 mL, 0.75 mmol, 1.5 equiv) was added dropwise and the reaction was stirred for 20 minutes. Aldehyde E (280.0 mg, 0.5 mmol, 1.0 equiv) in THF (0.5 mL) was then added dropwise. The reaction was stirred at −78° C. for 90 minutes and then allowed to warm to −20° C. over 1 hour. The reaction was quenched with ammonium chloride, diluted with ethyl acetate, and warmed to room temperature. The organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluent: hexane / ethyl acetate) to give the desired Julia product (F, 180 mg, 0.3 mmol, 54%).

[0269] Step 6: To a solution of acetate salt F (250.0 mg, 0.4 mmol, 1.0 equiv) in methanol (3 mL, 0.1 M) at room temperature was added potassium carbonate (58.0 mg, 0.4 mmol, 1.1 equiv). The reaction was run for 24 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with water, diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting foamy solid (G, 235 mg, 0.4 mmol, 100%) was carried on to the next step without further purification.

[0270] Step 7: To a solution of alcohol G (22.0 mg, 0.04 mmol, 1.0 equiv) in dichloromethane (0.5 mL, 0.1 M) at room temperature, N,N-dimethylaminopyridine (2.1 mg, 0.02 mmol, 0.5 equiv) was added, followed by 4-nitrophenyl chloroformate (14.4 mg, 0.08 mmol, 2.0 equiv). The reaction was stirred at room temperature for 3 hours. Next, 1-(tetrahydro-2H-pyran-4-yl)piperazine (20.4 mg, 0.12 mmol, 3.0 equiv) was added at room temperature. After stirring for 1 hour, the reaction was quenched with water and diluted with dichloromethane. The organic layer was washed with 1N sodium hydroxide solution, and the organic layer was concentrated. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (H, 26.0 mg, 0.03 mmol, 80%).

[0271] Step 8: To a solution of the silyl ether (H, 26.0 mg, 0.03 mmol, 1.0 equiv) in methanol (0.3 mL, 0.1 M) at room temperature was added p-methoxytoluenesulfonic acid (17.0 mg, 0.09 mmol, 3.0 equiv). The reaction was stirred for 3 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium bicarbonate, diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (compound 46, 16.3 mg, 0.025 mmol, 85%). 1H NMR(400MHz,chloroform-d)δ:8.48(ddd,J=4.9,1.9,1.0Hz,1H),7.54(td,J=7.7,1.9Hz,1H),7.09(d,J=8.0Hz,1H),7.05( t,J=6.1Hz,1H),6.15-6.34(m,1H),6.04(d,J=10.8Hz,1H),5.93(dd,J=15.1,7.5Hz,1H),5.48-5.67(m,2H),5.08(d,J =10.5Hz,1H),4.94(d,J=9.5Hz,1H),3.95(dd,J=11.3,3.8Hz,2H),3.53-3.76(m,2H),3.37-3.49(m,5H),3.22-3.37(m ,2H),2.35-2.57(m,7H),1.88(s,1H),1.44-1.70(m,11H),1.14-1.39(m,8H),0.72-0.89(m,3H),MS(ES+)=626.6[M+H].

[0272] [Table 5]

[0273] [Table 6]

[0274] [Table 7]

[0275] Table 8

[0276] Table 9

[0277] Table 10

[0278] Table 11

[0279] Table 12

[0280] Table 13

[0281] Table 14

[0282] Table 15

[0283] Table 16

[0284] Table 17

[0285] [Table 18]

[0286] [Table 19]

[0287] [Table 20]

[0288] [Table 21]

[0289] [Table 22]

[0290] [Table 23]

[0291] [Table 24]

[0292] [Table 25]

[0293] [Table 26]

[0294] Compounds 61-104 (Table 2) were prepared by the method of Scheme 2. [ka]

[0295] General synthetic protocol for compounds 61-104: Step 1: A solution of E7107 (I, 3.7 g, 5.1 mmol, 1.0 equiv) in DMF (100 mL, 0.05 M) at 0 °C under nitrogen was treated with imidazole (2.5 g, 36.1 mmol, 7.0 equiv) and TBSCl (3.9 g, 25.7 mmol, 5.0 equiv) was added. The reaction was allowed to warm to room temperature and stirred for 20 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was diluted with ethyl acetate, and the organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (J, 4.7 g, 5.0 mmol, 96%).

[0296] Step 2: To a solution of olefin J (4.7 g, 5.0 mmol, 1.0 equiv) in THF:HO (10:1, 133 mL:13 mL, 0.03 M) under nitrogen at 0 °C was added osmium tetroxide (12.4 mL, 1.0 mmol, 0.2 equiv, 2.5% solution), followed by N-methylmorpholine N-oxide (1.16 g, 9.9 mmol, 2.0 equiv). The reaction was allowed to warm to room temperature and stirred for 13 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium sulfite, diluted with ethyl acetate, and the organic layer was washed with water, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (dichloromethane / methanol as eluent) to give the desired product (K, 4.8 g, 4.9 mmol, 99%).

[0297] Step 3: To a solution of diol K (4.4 g, 4.5 mmol, 1.0 equiv) in benzene (100 mL, 0.05 M) at room temperature under nitrogen was added lead tetraacetate (4.0 g, 9.0 mmol, 2.0 equiv). The reaction was stirred for 30 minutes or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium sulfite and diluted with dichloromethane. The organic layer was washed with water, dried over sodium sulfate, filtered, and concentrated in vacuo. The desired product (L, 1.5 g, 2.3 mmol, 52%) was carried forward crude.

[0298] Step 4: To a solution of the corresponding sulfone (2.5 equiv.) in THF (0.02 M) at −78° C. under nitrogen, KHMDS (2.5 equiv.) was added dropwise, and the reaction was stirred for 10 minutes. Aldehyde L (1.0 equiv.) in THF (0.5 M) was then added dropwise. The reaction was stirred at −78° C. for 5 hours and then allowed to warm to room temperature overnight. The reaction was quenched with water and diluted with ethyl acetate. The organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (M).

[0299] Step 5: A solution of silyl ether M (1.0 equiv.) in MeOH (0.02 M) at room temperature under nitrogen was treated with pTsOH (2.0 equiv.). The reaction was stirred for 2 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was then diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by preparative TLC (dichloromethane / methanol as eluent) to give the desired product (61-104).

[0300] Exemplary synthesis protocol for compound 63 Steps 1 to 3 are as above.

[0301] Step 4: To a solution of (S)-2-methyl-3-((1-phenyl-1H-tetrazol-5-yl)sulfonyl)propylpyrrolidine-1-carboxylate (45.0 mg, 0.12 mmol, 2.5 equiv) in THF (2.0 mL, 0.02 M) at −78° C. under nitrogen, KHMDS (0.23 mL, 0.12 mmol, 2.5 equiv) was added dropwise and the reaction was stirred for 10 minutes. Aldehyde L (30.0 mg, 0.05 mmol, 1.0 equiv) in THF (0.2 mL) was then added dropwise. The reaction was stirred at −78° C. for 5 hours and then allowed to warm to room temperature overnight. The reaction was quenched with water and diluted with ethyl acetate. The organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluent: hexane / ethyl acetate) to give the desired product (M, 35 mg, 0.04 mmol, 76%).

[0302] Step 5: A solution of silyl ether M (35.0 mg, 0.04 mmol, 1.0 equiv) in MeOH (2.0 mL, 0.02 M) at room temperature under nitrogen was treated with pTsOH (15.0 mg, 0.08 mmol, 2.0 equiv). The reaction was stirred for 2 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was then diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by preparative TLC (dichloromethane / methanol as eluent) to give the desired product (Compound 63, 22.2 mg, 32 mmol, 80%). 1H NMR(400MHz, methanol-d4)δ:0.90(d,J=6.65Hz,3H)1.09(d,J=6.78Hz,3H)1.24(s,3H)1.32-1.45(m,2H)1.47-1.85(m ,15H)1.85-1.94(m,4H)1.95-2.10(m,2H)2.50-2.68(m,4H)2.96-3.08(m,4H)3.09-3.21(m,1H)3.34-3.39(m,4H)3 .52-3.88(m,5H)3.92-4.06(m,2H)4.97(d,J=9.66Hz,1H)5.07(d,J=10.67Hz,1H)5.61(dd,J=15.18,9.79Hz,1H)5. 72(d,J=9.79Hz,2H)6.12(dd,J=10.79,1.00Hz,1H)6.37(ddd,J=15.12,10.85,0.88Hz,1H).MS(ES+)=688.5[M+H]+.

[0303] [Table 27]

[0304] [Table 28]

[0305] [Table 29]

[0306] Table 30

[0307] Table 31

[0308] Table 32

[0309] Table 33

[0310] Table 34

[0311] Table 35

[0312] Table 36

[0313] Table 37

[0314] Table 38

[0315] Table 39

[0316] Table 40

[0317] [Table 41]

[0318] Compounds 105-115 were prepared by the method in Scheme 3. [ka]

[0319] General synthetic protocol for compounds 105-115: Step 1: A solution of 6-deoxypladienolide D (N, 100.0 mg, 0.2 mmol, 1.0 equiv) in DMF (8 mL, 0.2 M) at 0 °C under nitrogen was treated with imidazole (89.2 mg, 1.3 mmol, 7.0 equiv) and TBSCl (140.3 mg, 0.9 mmol, 5.0 equiv). The reaction was allowed to warm to room temperature and stirred for 20 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was extracted with ethyl acetate, and the organic layer was washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (O, 143.0 mg, 0.19 mmol, 100%).

[0320] Step 2: To a solution of olefin O (30.0 mg, 0.04 mmol, 1.0 equiv) in degassed THF:HO (10:1, 1.0 mL:0.1 mL, 0.01 M) under nitrogen at 0 °C, osmium tetroxide (0.1 mL, 0.008 mmol, 0.2 equiv, 2.5% solution in tert-butanol) was added, followed by N-methylmorpholine N-oxide (9.2 mg, 0.08 mmol, 2.0 equiv). The reaction was allowed to warm to room temperature and stirred for 30 minutes or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium sulfite, diluted with ethyl acetate, and the organic layer was washed with water, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (dichloromethane / methanol as eluent) to give the desired product (P, 29.2 mg, 0.04 mmol, 93%).

[0321] Step 3: To a solution of triol P (498.2 mg, 0.6 mmol, 1.0 equiv) in benzene (25 mL, 0.03 M) at room temperature under nitrogen was added lead tetraacetate (553.4 mg, 1.2 mmol, 2.0 equiv). The reaction was stirred for 30 minutes or until the reaction was complete as determined by LCMS or TLC. The reaction was concentrated and purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (Q, 232 mg, 0.5 mmol, 80%).

[0322] Step 4: To a solution of the corresponding sulfone (2.5 equiv.) in THF (0.02 M) at −78° C. under nitrogen, KHMDS (2.5 equiv.) was added dropwise, and the reaction was stirred for 20 minutes. Aldehyde Q (1.0 equiv.) in THF (0.5 M) was then added dropwise. The reaction was stirred at −78° C. for 90 minutes and then allowed to warm to −20° C. over 1 hour. The reaction was quenched with aqueous ammonium chloride, diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (R).

[0323] Step 5: To a solution of acetate salt R (1.0 equiv.) in methanol (0.1 M) at room temperature was added potassium carbonate (2.5 equiv.). The reaction was run for 24 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with water, diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil (S) was carried on crude to the next step.

[0324] Step 6: To a solution of alcohol (S) (1.0 equiv.) in dichloroethane (0.1 M) at room temperature, N,N-dimethylaminopyridine (0.3 equiv.) was added, followed by 4-nitrophenyl chloroformate (4.0 equiv.). The reaction was stirred at room temperature for 24 hours. The corresponding amine (10.0 equiv.) was then added at room temperature. After stirring for 1 hour, the reaction was concentrated, and the resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (T).

[0325] Step 7: To a solution of silyl ether T in methanol (0.1 M) at room temperature was added p-methoxytoluenesulfonic acid (2.5 equiv.). The reaction was stirred for 3 h or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium bicarbonate, diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired products (105–115). (Table 3)

[0326] Exemplary Synthesis Protocol for Compound 114 Steps 1 to 3 are as above.

[0327] Step 4: To a solution containing (S)-2-(1-((1-phenyl-1H-tetrazol-5-yl)sulfonyl)propan-2-yl)pyridine (44.0 mg, 0.1 mmol, 2.5 equiv) and THF (2.0 mL, 0.02 M) at −78° C. under nitrogen, KHMDS (0.27 mL, 0.1 mmol, 2.5 equiv) was added dropwise and the reaction was stirred for 20 minutes. Aldehyde Q (25 mg, 0.05 mmol, 1.0 equiv) in THF (0.1 mL) was then added dropwise. The reaction was stirred at −78° C. for 90 minutes and then allowed to warm to −20° C. over 1 hour. The reaction was quenched with aqueous ammonium chloride, diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluent: hexane / ethyl acetate) to give the desired product (R, 21.0 mg, 0.04 mmol, 69%).

[0328] Step 5: To a solution of acetate salt R (15.2 mg, 0.03 mmol, 1.0 equiv) in methanol (2 mL, 0.1 M) at room temperature was added potassium carbonate (9.1 mg, 0.07 mmol, 2.5 equiv). The reaction was run for 24 h or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with water, diluted with ethyl acetate, washed with brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil (S, 14 mg, 0.03 mmol, 100%) was carried on crude to the next step.

[0329] Step 6: To a solution of the alcohol (S, 4.2 mg, 0.008 mmol, 1.0 equiv) in dichloromethane (1 mL, 0.1 M) at room temperature, N,N-dimethylaminopyridine (0.3 mg, 0.002 mmol, 0.3 equiv) was added, followed by 4-nitrophenyl chloroformate (6.4 mg, 0.03 mmol, 4.0 equiv). The reaction was stirred at room temperature for 24 hours. N-methylpiperazine (0.009 mL, 0.08 mmol, 10.0 equiv) was then added at room temperature. After stirring for 1 hour, the reaction was concentrated, and the resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (T, 4.9 mg, 0.007 mmol, 94%).

[0330] Step 7: To a solution of silyl ether T (4.9 mg, 0.007 mmol, 1.0 equiv) in methanol (0.7 mL, 0.1 M) at room temperature was added p-methoxytoluenesulfonic acid (3.6 mg, 0.02 mmol, 2.5 equiv). The reaction was stirred for 3 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was quenched with sodium bicarbonate, diluted with ethyl acetate, washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (Compound 114, 3.6 mg, 0.007 mmol, 89%). 1H NMR(400MHz,chloroform-d)δ:0.88(d,J=6.78Hz,3H)0.99(d,J=6.90Hz,3H)1.13- 1.33(m,2H)1.44(d,J=6.90Hz,3H)1.47-1.51(m,1H)1.73(d,J=0.75Hz,3H)1. 74-1.81(m,1H)1.84-1.97(m,1H)2.30(s,3H)2.36(br.s.,4H)2.39-2.61(m,3 H)3.41(m,1H)3.49(br.s.,4H)3.67-3.74(m,2H)4.86(t,J=10.04Hz,1H)5.13( d,J=10.67Hz,1H)5.35(dd,J=14.93,9.66Hz,1H)5.54(dd,J=15.06,9.91Hz,1 H)6.00(dd,J=15.12,7.47Hz,1H)6.12(d,J=10.92Hz,1H)6.32(ddd,J=15.09, 10.82,1.07Hz,1H)7.11(ddd,J=7.53,4.89,1.13Hz,1H)7.16(d,J=7.91Hz,1H )7.61(td,J=7.65,1.88Hz,1H)8.55(d,J=4.96Hz,1H),MS(ES+):540.3[M+H]+.

[0331] Compound 116 was prepared by the method in Scheme 4. [ka]

[0332] Step 1: To a solution of Intermediate Q (35 mg, 0.062 mmol, 1 equiv.) and SPE-11 (23.20 mg, 0.068 mmol, 1.1 equiv.) in tetrahydrofuran (3 mL) at room temperature under a nitrogen atmosphere, triphenylarsine (18.98 mg, 0.062 mmol, 1 equiv.), silver(I) oxide (71.8 mg, 0.31 mmol, 5 equiv.), and tris(dibenzylideneacetone)dipalladium(0) (11.35 mg, 0.012 mmol, 0.2 equiv.) were added sequentially and stirred at the same temperature for 16 h in the dark. The solid was removed by filtration through Celite, and the pad was washed with EtOAc. Excess solvent was removed under reduced pressure, and the resulting residue was purified by silica gel chromatography (0-50% EtOAc / hexanes) to give the desired product (SPE-12, 17.6 mg, 0.027 mmol, 43.6%).

[0333] Step 2: To a solution of SPE-12 (17.6 mg, 0.027 mmol, 1 equiv.) in THF (2 mL) at 0 °C, TBAF (0.216 mL, 0.216 mmol, 8 equiv.) was added, and the reaction mixture was then gradually warmed to room temperature and stirred for 2 h. The reaction mixture was directly applied to silica gel and purified by silica gel chromatography (0-30% EtOAc / hexanes) to give the desired product (compound 116, 5.2 mg, 9.69 μmol, 35.8%). H NMR (400 MHz, methanol-d4) δ: ppm 0.86-1.02(m,15H)1.37(d,J=3.51Hz,8H)1.47-1.55(m,2H)1.62-1.71(m,3H)1.79(s, 3H)1.85-1.96(m,2H)2.02(s,3H)2.42-2.48(m,1H)2.55-2.63(m,2H)2.65-2.76(m,1H) 2.83-2.94(m,1H)3.50-3.62(m,1H)3.80(s,2H)4.89-4.97(m,1H)5.01-5.09(m,1H)5. 39-5.56(m,2H)5.82-5.96(m,1H)6.11-6.20(m,1H)6.49-6.62(m,1H).MS(ES+):535.56 [MH]-.

[0334] [Table 42]

[0335] [Table 43]

[0336] [Table 44]

[0337] [Table 45]

[0338] [Table 46]

[0339] Compounds 117-134 were prepared by the method in Scheme 5. [ka]

[0340] General synthetic protocol for compounds 117-134: Step 1: To a solution of NaH (8.3 g, 207 mmol, 1.2 equiv) in diethyl ether (400 mL, 0.1 M) at 0 °C was added diethyl 2-methylmalonate (U, 30 g, 172 mmol, 1.0 equiv) dropwise. The reaction was gradually warmed to reflux and stirred at reflux for 3 h. The reaction was then cooled to room temperature and iodoform (67.8 g, 172 mmol, 1.0 equiv) was added dropwise. The reaction was again heated at reflux for 24 h or until the reaction was complete as determined by LCMS or TLC. The reaction was cooled to 0 °C, quenched with 10% aqueous hydrochloric acid, diluted with ether, and washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was then dissolved in ethanol / water / methanol (400 mL, 3:1:1) and KOH (48.3 g, 861 mmol, 5.0 equiv.) was added at room temperature. The solution was then heated to 75° C. for 24 hours. The reaction was cooled to room temperature and concentrated in vacuo. The resulting oil was diluted with ethyl acetate and water, extracted with ethyl acetate, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (V, 26 g, 123 mmol, 71%).

[0341] Step 2: To a solution of acid V (25.0 g, 118 mmol, 1.0 equiv) in THF (400 mL, 0.3 M) at 0 °C was added lithium aluminum hydride (4.9 g, 130 mmol, 1.1 equiv). The reaction was gradually warmed to room temperature and stirred for 4 hours or until the reaction was complete as determined by LCMS or TLC. The reaction was cooled to 0 °C and quenched with water. The resulting suspension was charged with Rochelle's salt solution (20% by volume) and stirred at room temperature for 3 hours. The mixture was filtered, washing with ethyl acetate, and the volume of the filtrate was reduced in vacuo. Ethyl acetate was added, and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (W, 15 g, 76 mmol, 64%).

[0342] Step 3: To a solution of alcohol W (60 mg, 0.3 mmol, 1.0 equiv) in diethyl ether (2 mL, 0.1 M) at room temperature was added manganese dioxide (395 mg, 4.5 mmol, 15.0 equiv). The reaction was stirred for 2 h or until the reaction was complete as determined by LCMS or TLC. The reaction was filtered through Celite® and the filtrate was concentrated in vacuo. The crude product (X, 59 mg, 0.30 mmol, 99%) was carried forward without purification.

[0343] Step 4: To a solution of (1R,2S)-2-(N-benzyl-2,4,6-trimethylphenylsulfonamido)-1-phenylpropylpropionate (1.9 g, 4.4 mmol, 1.0 equiv.), prepared as previously described (Masamune et al. J. Am. Chem. Soc. 1997, 119, 2586-2587), in dichloromethane (40 mL, 0.1 M) at −78° C., triethylamine (1.7 mL, 12.3 mmol, 3.0 equiv.) was added, followed by the dropwise addition of dicyclohexyl(((trifluoromethyl)sulfonyl)oxy)borane (2.67 g, 8.0 mmol, 2.0 equiv.). The reaction was stirred at −78° C. for 2 hours. A solution of (£)-3-iodo-2-methylacrylaldehyde (X, 1.2 g, 6.2 mmol, 1.5 equiv) in dichloromethane (3 mL) was then added dropwise over 30 minutes. The reaction was stirred at -78 °C for 2 hours and then allowed to warm to 0 °C. The reaction was quenched by the addition of aqueous hydrogen peroxide (16 mL, 20.5 mmol), and the reaction was allowed to warm gradually to room temperature. The solvent volume was reduced in vacuo, and the solution was diluted with dichloromethane and water. The organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (eluting with hexane / ethyl acetate) to give the desired product (Y, 1.9 g, 2.8 mmol, 69%).

[0344] Step 5: To a solution of alcohol Y (2.8 g, 4.1 mmol, 1.0 equiv) in dichloromethane (50 mL, 0.1 M) at −78 °C, 2,6-lutidine (1.0 mL, 8.3 mmol, 2.0 equiv) was added, followed by tert-butyldimethylsilyl trifluoromethanesulfonate (1.1 mL, 4.9 mL, 1.2 equiv). The reaction was gradually warmed to room temperature and quenched with aqueous ammonium chloride. Ethyl acetate was added, and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (Z, 2.8 g, 3.5 mmol, 85%).

[0345] Step 6: To a solution of ester Z (2.8 g, 3.5 mmol, 1.0 equiv) in dichloromethane (40 mL, 0.1 M) at 0 °C was added DIBAL (8.9 mL, 8.9 mmol, 2.5 equiv). The reaction was stirred for 1 h, then quenched with Rochelle's salt solution (20% by volume) and stirred at room temperature for 3 h. The mixture was filtered through Celite®, washing with ethyl acetate, and the volume of the filtrate was reduced in vacuo. Ethyl acetate was added, and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (AA, 1.1 g, 2.8 mmol, 80%).

[0346] Step 7: To a solution of alcohol AA (2.97 g, 8.0 mmol, 1.0 equiv) in dichloromethane (80 mL, 0.1 M) at 0 °C was added Dess-Martin periodinane (4.4 g, 10.4 mmol, 1.3 equiv). The reaction was stirred for 2 h or until the reaction was complete as determined by LCMS or TLC. The reaction was concentrated in vacuo, and the resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (BB, 2.6 g, 7.1 mmol, 88%).

[0347] Step 8: To a solution of methyltriphenylphosphonium bromide (11.8 g, 33.0 mmol, 3.0 equiv) in THF (110 mL, 0.1 M) at 0 °C was added n-butyllithium (13.2 mL, 33.0 mmol, 3.0 equiv). The reaction was stirred for 30 min and then cooled to -78 °C. Aldehyde BB (4.1, 11.0 mmol, 1.0 equiv) in THF (0.5 M) was added dropwise and the reaction was stirred for 1 h. The reaction was quenched with ammonium chloride and warmed to room temperature. Ethyl acetate was added, and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (CC, 3.8 g, 10.4 mmol, 94%).

[0348] Step 9: To a solution of olefin CC (0.1 g, 0.4 mmol, 1.0 equiv) in THF (4 mL, 0.1 M) at 0 °C was added TBAF (0.45 mL, 0.4 mmol, 1.1 equiv). The reaction was stirred for 30 minutes or until the reaction was complete as determined by LCMS or TLC. Diethyl ether was added and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The crude product (DD, 0.1 g, 0.4 mmol, 99%) was carried forward without purification.

[0349] Step 10: To a solution of alcohol DD (0.15 g, 0.4 mmol, 1.0 equiv) in dichloromethane (4 mL, 0.1 M) at 0 °C, EDC (0.10 g, 0.5 mmol, 1.3 equiv) was added, followed by nonenoic acid (0.08 g, 0.4 mmol, 1.1 equiv) and DMAP (catalytic). The reaction was gradually warmed to room temperature and stirred overnight. Ethyl acetate was added, and the organic layer was washed with water, brine, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (EE, 0.13 g, 0.33 mmol, 81%).

[0350] Step 11: To a solution of ester EE (0.5 g, 1.3 mmol, 1.0 equiv) in degassed toluene (65 mL, 0.05 M) at room temperature was added benzoquinone (0.007 g, 0.06 mmol, 0.05 equiv), followed by Hoyveda-Grubbs catalyst (0.08 g, 0.13 mmol, 0.1 equiv). The reaction was gradually warmed to 60 °C and stirred overnight. Upon completion as determined by TLC or LCMS, the reaction was concentrated. This crude material (FF) was used in the following step without further purification.

[0351] Step 12: To a solution of macrocycle FF (1.0 equiv.) in dioxane (65 mL, 0.05 M) at room temperature was added selenium dioxide (0.4 g, 3.8 mmol, 3.0 equiv.). The reaction was heated to 80 °C for 3 h. Ethyl acetate was added, and the organic layer was washed with water and saturated sodium bicarbonate, dried over magnesium sulfate, filtered, and concentrated in vacuo. The resulting oil was purified by silica gel column chromatography (hexane / ethyl acetate as eluent) to give the desired product (GG, 0.3 g, 0.8 mmol, 64%).

[0352] St...

Claims

1. Compounds of Formula I: 【Chemistry 1】 and a pharmaceutically acceptable salt thereof. [In the formula: n is selected from 0, 1, 2 or 3; R １ is C １ ~C ６ Alkyl group, C ３ ~C ８ cycloalkyl group, —NR ９ R １０ , 【Chemistry 2】 basis, 【Transformation 3】 basis, 【Chemistry 4】 basis, 【Transformation 5】 basis, 【Transformation 6】 basis, 【Transformation 7】 basis, 【Transformation 8】 Groups, and 【Chemistry 9】 is selected from the group R ９ is hydrogen, -NR １１ R １２ Group, C １ ~C ６ alkyl group, -(C １ ~C ６ alkyl)-CO ２ H group, C ３ ~C ８ Cycloalkyl groups, and C ３ ~C ８ heterocyclyl groups, wherein said —NR １１ R １２ Group, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, and C ３ ~C ８ The heterocyclyl group may be unsubstituted or C １ ~C ６ alkyl group, -(C １ ~C ６ alkyl)-CO ２ H group, hydroxy, halogen group, and C １ ~C ６ optionally substituted 1 to 3 times with groups independently selected from alkoxy groups; R １０ is hydrogen and C １ ~C ６ alkyl groups; R ２ or R ３ One of the two is hydrogen and C １ ~C ６ alkyl group, and the other is selected from hydrogen, —OR １０ , -OC(O)R １０ , -OC(O)R １ , and C １ ~C ６ alkyl groups; R ４ is selected from hydrogen and hydroxy; R ５ and R ６ are each independently C １ ~C ６ alkyl groups; R ７ and R ８ are each independently hydrogen, hydroxy, C １ ~C ６ Alkoxy groups, and C １ ~C ６ alkyl groups; and Y is selected from phenyl, thiophenyl, triazolyl, pyridinyl, pyrimidinyl, pyridazinyl, and pyrazinyl, where Y may be unsubstituted or may be an oxo group, C １ ~C ６ Alkyl group, C ３ ~C ５ Cycloalkyl group, hydroxy C １ ~C ６ Alkyl group, C １ ~C ６ Alkoxy group, methoxy C １ ~C ６ alkyl group, -NR １１ R １２ basis, 【Chemistry 10】 and optionally substituted 1 to 3 times with groups independently selected from １１ and R １２ are each independently hydrogen and C １ ~C ６ alkyl groups].

2. Y is 【Chemistry 11】 2. The compound of claim 1, wherein:

3. 2. The compound of claim 1, wherein Y is optionally substituted phenyl.

4. R １ But methyl, 【Chemistry 12】 basis, 【Chemistry 13】 basis, 【Chemistry 14】 basis, 【Chemistry 15】 basis, 【Chemistry 16】 basis, 【Chemistry 17】 basis, [Chemistry 18] Groups, and 【Chemistry 19】 2. The compound of claim 1, wherein the compound is selected from the group:

5. Compound of Formula II: 【Chemistry 20】 and a pharmaceutically acceptable salt thereof. [In the formula: X is O, an NR′ group, and CH ２ wherein R′ is selected from hydrogen and C １ ~C ６ alkyl groups; R １ is methyl, -NR １１ R １２ basis, 【Chemistry 21】 Groups, and 【Chemistry 22】 is selected from the group R １０ is C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups and haloC １ ~C ６ alkyl group, wherein said C ３ ~C ８ The cycloalkyl group may be unsubstituted or may be C １ ~C ６ Alkyl groups, hydroxyl groups, halogen groups, and C １ ~C ６ optionally substituted 1 to 3 times with groups independently selected from alkoxy groups; R １１ and R １２ are each independently C １ ~C ６ alkyl groups; R ２ or R ３ One of the two is hydrogen and C １ ~C ６ alkyl group, and the other is selected from hydrogen, hydroxy and C １ ~C ６ alkyl groups; R ４ or R ５ is hydrogen, and the other is hydrogen, hydroxy, and 【Chemistry 23】 Selected from: R ６ and R ７ are each independently C １ ~C ６ alkyl groups; R ８ and R ９ are each independently hydrogen and C １ ~C ６ alkyl groups; or R ８ and R ９ together form a cyclopropyl ring; and Y is C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl groups, methoxy, and —NR １３ R １４ group, wherein R １３ and R １４ are each independently hydrogen, C １ ~C ６ Alkyl group, and methoxy C １ ~C ６ alkyl groups; or R １３ and R １４ together with N, 【Chemistry 24】 forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; where Y may be unsubstituted or C １ ~C ６ Alkyl group, hydroxyl group, hydroxyl C group １ ~C ６ Alkyl group, methoxy, methoxy C １ ~C ６ Alkyl group, halo group, halo C １ ~C ６ Alkyl group, —C(O)NH ２ , -NHCOO-C １ ~C ６ alkyl group, —COOH, 【Chemistry 25】 and -NR １５ R １６ groups, wherein R １５ and R １６ are each independently hydrogen and C １ ~C ６ alkyl groups].

6. Compound of Formula III: 【Chemistry 26】 and a pharmaceutically acceptable salt thereof. [In the formula: n is selected from 0, 1 and 2; m is selected from 1, 2, and 3; R １ is C １ ~C ６ Alkyl group, C ３ ~C ８ cycloalkyl group, —NR １１ R １２ basis, 【Chemistry 27】 basis, 【Chemistry 28】 basis, 【Chemistry 29】 basis, 【Transformation 30】 basis, 【Chemistry 31】 basis, 【Chemistry 32】 basis, 【Transformation 33】 basis, 【Transformation 34】 Groups, and 【Chemistry 35】 is selected from the group R １１ is hydrogen, -NR １６ R １７ Group, C １ ~C ６ alkyl group, -(C １ ~C ６ alkyl)-CO ２ H group, -(C １ ~C ６ alkyl)-CO ２ R １２ Group, -(C １ ~C ６ alkyl)-NR １６ R １７ Group, C ３ ~C ８ Cycloalkyl groups, and C ３ ~C ８ heterocyclyl groups, wherein said —NR １１ R １２ Group, C １ ~C ６ Alkyl group, C ３ ~C ８ Cycloalkyl group and C ３ ~C ８ The heterocyclyl group may be unsubstituted or C １ ~C ６ alkyl group, -(C １ ~C ６ alkyl)-CO ２ H group, hydroxy, halogen group, and C １ ~C ６ optionally substituted 1 to 3 times with groups independently selected from alkoxy groups; R １２ is hydrogen and C １ ~C ６ alkyl groups; R ２ or R ３ One of the two is hydrogen and C １ ~C ６ alkyl group, and the other is selected from hydrogen, —OR １０ , -OC(O)R １０ , -OC(O)R １ , and C １ ~C ６ alkyl groups; R ４ is hydrogen or hydroxy; R ５ and R ６ are each independently C １ ~C ６ alkyl groups; R ７ and R ８ are each independently hydrogen, hydroxy, C １ ~C ６ Alkoxy groups, and C １ ~C ６ alkyl groups; and R ９ and R １０ are each independently hydrogen, C １ ~C ６ Alkyl groups, hydroxyl groups, and C １ ~C ６ alkoxy groups; or R ９ or R １０ one of is oxo and the other is absent; Z is C １ ~C ６ alkyl group, —C(O)—C １ ~C ６ alkyl group, -OR １３ , and -NR １４ R １５ is selected from the group In the formula, R １３ is hydrogen, C １ ~C ６ Alkyl groups, and —C(O)—C １ ~C ６ alkyl groups, In the formula, R １４ and R １５ are each independently hydrogen, C １ ~C ６ Alkyl group, and methoxy C １ ~C ６ alkyl groups; or R １４ and R １５ together with N, 【Transformation 36】 forming a group selected from morpholine, piperidine, thiazolidine, indole, indoline, and isoindoline rings; wherein Z may be unsubstituted or C １ ~C ６ Alkyl group, C ３ ~C ５ Cycloalkyl group, hydroxy C １ ~C ６ Alkyl group, C １ ~C ６ Alkoxy group, methoxy C １ ~C ６ alkyl group, -NR １６ R １７ basis, 【Chemistry 37】 and optionally substituted 1 to 3 times with groups independently selected from １６ and R １７ are each independently hydrogen and C １ ~C ６ alkyl groups].

7. R １ But methyl, 【Transformation 38】 basis, 【Chemistry 39】 basis, 【Chemistry 40】 basis, 【Chemistry 41】 basis, 【Chemistry 42】 basis, 【Chemistry 43】 basis, 【Chemistry 44】 basis, 【Chemistry 45】 Groups, and 【Chemistry 46】 7. The compound of claim 6, wherein the compound is selected from the group:

8. [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-9-oxo-9-pyrrolidin-1-ylnona-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[[(2R,3R)-3-hydroxypentan-2-yl]carbamoyloxy]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-(propylcarbamoyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-4-oxidepiperazin-4-ium-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(dimethylcarbamoyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-(diethylcarbamoyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propan-2-yl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[butyl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[butan-2-yl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-carbamoyloxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R)-2-(methoxymethyl)pyrrolidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[2-methoxyethyl(methyl)carbamoyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]azetidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2S)-2-methylpyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2S)-2-methylpyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]piperidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R)-2-(hydroxymethyl)pyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](3R)-3-hydroxypyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]morpholine-4-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]4-methylpiperazine-1-carboxylate; 3-Thiazolidinecarboxylic acid [(2R,3E,5E)-6-[(2R,3S,4E,6R,7R,10R)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl] ester; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]1,3-dihydroisoindole-2-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]indole-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[2-(1-hydroxyethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,2-dimethylpyrrolidine-1-carbonyl)oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2S,5S)-2,5-dimethylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2,3-dihydroindole-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-6-(4-methylpiperazine-1-carbonyl)oxy-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2-oxa-5-azaspiro[3.4]octane-5-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-6-[6-[(2R)-1-hydroxypropan-2-yl]pyridin-2-yl]hepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridazin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3R,4E,6S,7R,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6R)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-propan-2-ylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-tert-butylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclopentylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(oxan-4-yl)piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]6-cycloheptyl-2,6-diazaspiro[3.3]heptane-2-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-3-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclobutylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N-methyl-N-(1-methylpiperidin-4-yl)carbamate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]morpholine-4-carboxylate; [(2R,3R,4E,6S,7R,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6R)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl](1S,4R)-5-methyl-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]8-cycloheptyl-3,8-diazabicyclo[3.2.1]octane-3-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methyl-1,4-diazepane-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cyclohexylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptyl-1,4-diazepane-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-hydroxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(azepan-1-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(8,8-difluoro-3-azabicyclo[3.2.1]octan-3-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-9-oxo-9-pyrrolidin-1-ylnona-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6S)-6-methyl-7-[methyl(propyl)carbamoyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10R)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-10-(pyrrolidine-1-carbonyloxy)-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(2S)-2-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3R)-3-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3R)-3-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-carbamoylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-[(2R)-2-(methoxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2S,5S)-2,5-dimethylpyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3R)-3-fluoropyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,2-dimethylpyrrolidine-1-carbonyl)oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-2-[(2E,4E)-6,6-dimethyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E)-6,6-dimethyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-7-hydroxy-3,7-dimethyl-12-oxo-10-(pyrrolidine-1-carbonyloxy)-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; (2R)-1-[(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienoxy]carbonylpyrrolidine-2-carboxylic acid; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(3-oxopyrrolidine-1-carbonyl)oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]2-oxa-7-azaspiro[3.4]octane-7-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-5-[1-(pyrrolidine-1-carbonyloxymethyl)cyclopropyl]penta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3S,4R)-3,4-dihydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; (3S)-1-[(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienoxy]carbonylpyrrolidine-3-carboxylic acid; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(3S)-3-(dimethylamino)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(2,5-dihydropyrrole-1-carbonyloxy)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3S)-3-[(2-methylpropan-2-yl)oxycarbonylamino]pyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-(4-cycloheptylpiperazine-1-carbonyl)oxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]3-azabicyclo[3.1.0]hexane-3-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-2-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-(2-pyrrolidin-1-ylpyrimidin-4-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrazin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(3-methylpyridin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(4-methylpyridin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridazin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyrimidin-4-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-(4-methylpyrimidin-2-yl)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-(6-pyrrolidin-1-ylpyridin-2-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-10-hydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-2-[(2E,4E,6R)-7-[(2R)-2-(fluoromethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-10-hydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N,N-dimethylcarbamate; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6R)-6-(dimethylcarbamoyloxy)-3-methyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2R,3E,5E)-6-[(2S,3S,4E,6R)-6-(dimethylcarbamoyloxy)-3-methyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](3R)-3-hydroxypyrrolidine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-2-[(2E,4E,6R)-6-methyl-7-[(2S)-2-methylpyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-(2,2,2-trifluoroethyl)piperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]N,N-dimethylcarbamate; [(2S,3S,4E,6R)-2-[(2E,4E)-6-[2-(dimethylamino)pyrimidin-4-yl]hepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-(2-pyrrolidin-1-ylpyrimidin-4-yl)hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-[2-[(3S)-3-triethylsilyloxypyrrolidin-1-yl]pyrimidin-4-yl]hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E)-6-[2-[(3R)-3-hydroxypyrrolidin-1-yl]pyrimidin-4-yl]hepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-3-methyl-12-oxo-2-[(2E,4E)-6-pyrimidin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6R)-7-[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[(3S)-3-(1-phenyltetrazol-5-yl)oxypyrrolidine-1-carbonyl]oxyhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S)-7-hydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonyloxy)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonylamino)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-[[(2R)-2-(hydroxymethyl)pyrrolidine-1-carbonyl]amino]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-(pyrrolidine-1-carbonylamino)hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E,6R)-6-methyl-7-[methyl(pyrrolidine-1-carbonyl)amino]hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(4-cyclopropyltriazol-1-yl)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6S)-7-methoxycarbonyloxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-9-methoxy-6-methyl-9-oxonona-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(cyclopentanecarbonylamino)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6R)-7-(cyclopentanecarbonylamino)-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; 4-cycloheptyl-1-piperazinecarboxylic acid [(2R,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-7-[oxo(1-pyrrolidinyl)methoxy]hepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl] ester; [(2S,3S,4E,6R,7R,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6R)-2-[(2E,4E,6R)-7-[(3R)-3-hydroxypyrrolidine-1-carbonyl]oxy-6-methylhepta-2,4-dien-2-yl]-3-methyl-12-oxo-1-azacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2R,3E,5E)-2-methyl-6-[(2S,3S,4E,6R)-3-methyl-6-[(4-methylpiperazine-1-carbonyl)amino]-12-oxo-1-oxacyclododec-4-en-2-yl]hepta-3,5-dienyl]pyrrolidine-1-carboxylate; [(2S,3E,5E)-6-[(2S,3S,4E,6S,7S,10S)-6-acetyloxy-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-2-yl]-2-methylhepta-3,5-dienyl](2R,3R)-3-hydroxy-2-methylpentanoate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E,6R)-7-hydroxy-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-4-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-7-methyl-6-pyridin-2-ylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-2-[(2E,4E,6S)-7-[(2R,3R)-3-[(2R,3R)-3-acetyloxypentan-2-yl]oxiran-2-yl]-6-hydroxy-6-methylhepta-2,4-dien-2-yl]-7,10-dihydroxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-methyl-8-phenylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-phenylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-6-thiophen-2-ylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-phenylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-6-(6-methoxypyridin-2-yl)hepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-[6-(2-methylpropoxy)pyridin-2-yl]hepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-8-pyridin-2-ylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-7-pyridin-2-ylhepta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E,6R)-6-hydroxy-6-methyl-8-phenylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-2-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-4-ylhexa-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6R,7R,10R)-7,10-dihydroxy-2-[(2E,4E)-6-hydroxy-8-(4-hydroxyphenyl)-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-2-[(2E,4E)-6-methyl-8-phenylocta-2,4-dien-2-yl]-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[2-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[4-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-2-[(2E,4E)-8-[3-(methoxymethyl)phenyl]-6-methylocta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S)-7-hydroxy-2-[(2E,4E,6S)-6-hydroxy-6-methyl-7-[(2R,3R)-3-[(2S)-3-oxopentan-2-yl]oxiran-2-yl]hepta-2,4-dien-2-yl]-3,7-dimethyl-10,12-dioxo-1-oxacyclododec-4-en-6-yl]acetate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6E,8S)-8-pyridin-2-ylnona-2,4,6-trien-2-yl]-1-oxacyclododec-4-en-6-yl]4-cycloheptylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-methyl-4-oxidopiperazin-4-ium-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(4-fluoropiperidin-1-yl)piperidine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7,10-dihydroxy-3,7-dimethyl-12-oxo-2-[(2E,4E)-6-pyridin-3-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-4-en-6-yl]4-(4,4-difluoropiperidin-1-yl)piperidine-1-carboxylate; (4S,7S,8S,9E,11S,12S)-4,7,8-trihydroxy-7,11-dimethyl-12-[(2E,4E,6S)-6-pyridin-2-ylhepta-2,4-dien-2-yl]-1-oxacyclododec-9-en-2-one; [(2S,3S,4E,6S,7S,10S)-7-acetyloxy-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((S,2E,4E)-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; [(2S,3S, 4E,6S,7S,10S)-6-acetyloxy-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-7-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-7-acetyloxy-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7R,10R)-7-ethoxy-10-hydroxy-2-[(2E,4E,6R)-6-hydroxy-7-[(2R,3R)-3-[(2S,3S)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-6-acetyloxy-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-7-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]piperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6R)-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]4-methylpiperazine-1-carboxylate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]N-methyl-N-[2-(methylamino)ethyl]carbamate; [(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]N-methyl-N-[2-(dimethylamino)ethyl]carbamate; 3-[4-[[(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]oxycarbonyl]piperazin-2-yl]propanoic acid; 4-[4-[[(2S,3S,4E,6S,7S,10S)-10-hydroxy-2-[(2E,4E,6S)-6-hydroxy-7-[(2R,3R)-3-[(2R,3R)-3-hydroxypentan-2-yl]oxiran-2-yl]-6-methylhepta-2,4-dien-2-yl]-7-methoxy-3,7-dimethyl-12-oxo-1-oxacyclododec-4-en-6-yl]oxycarbonyl]piperazin-1-yl]butanoic acid; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl(1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; (2S,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 2,5-diazabicyclo[2.2.1]heptane-2-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-propylpiperazine-1-carboxylate; (2R,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((2S,6R,E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhept-4-en-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 4-(2-hydroxyethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-6-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-7-yl 4-methylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-(2-aminoethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7-acetoxy-10-hydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-(2-ethoxy-2-oxoethyl)piperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7,10-dihydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-yl 4-methylpiperazine-1-carboxylate; (2S,3S,6S,7R,10R,E)-7,10-dihydroxy-2-((R,2E,4E)-6-hydroxy-7-((2R,3R)-3-((2R,3S)-3-hydroxypentan-2-yl)oxiran-2-yl)-6-methylhepta-2,4-dien-2-yl)-3,7-dimethyl-12-oxooxacyclododec-4-en-6-ylpiperazine-1-carboxylate; and pharmaceutically acceptable salts thereof.

9. The compound of any one of claims 1 to 8, wherein the compound is stereomerically pure.

10. A pharmaceutical composition comprising a compound and / or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9.

11. 11. The pharmaceutical composition of claim 10, formulated for intravenous, oral, subcutaneous, or intramuscular administration.

12. 12. A method for treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound and / or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11.

13. 13. The method of claim 12, wherein the cancer is selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, bile duct cancer, and lung cancer.

14. 14. The method of claim 13, wherein the cancer is selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, and acute myeloid leukemia.

15. 14. The method of claim 13, wherein the cancer is a myelodysplastic syndrome.

16. 14. The method of claim 13, wherein the cancer is chronic lymphocytic leukemia.

17. 14. The method of claim 13, wherein the cancer is chronic myelomonocytic leukemia.

18. 14. The method of claim 13, wherein the cancer is acute myeloid leukemia.

19. 14. The method of claim 13, wherein the cancer is colon cancer.

20. 14. The method of claim 13, wherein the cancer is pancreatic cancer.

21. 14. The method of claim 13, wherein the cancer is endometrial cancer.

22. 14. The method of claim 13, wherein the cancer is ovarian cancer.

23. 14. The method of claim 13, wherein the cancer is breast cancer.

24. 14. The method of claim 13, wherein the cancer is uveal melanoma.

25. 14. The method of claim 13, wherein the cancer is gastric cancer.

26. 14. The method of claim 13, wherein the cancer is cholangiocarcinoma.

27. 14. The method of claim 13, wherein the cancer is lung cancer.

28. 28. The method of any one of claims 12 to 27, wherein the cancer is positive for one or more mutations in spliceosomal genes or proteins.

29. The spliceosome gene or protein is selected from the group consisting of splicing factor 3B subunit 1 (SF3B1), U2 small nuclear RNA cofactor 1 (U2AF1), serine / arginine-rich splicing factor 2 (SRSF2), zinc finger (CCCH-type) RNA-binding motif and serine / arginine-rich 2 (ZRSR2), pre-mRNA processing-splicing factor 8 (PRPF8), U2 small nuclear RNA cofactor 2 (U2AF2), splicing factor 1 (SF1), splicing factor 3a subunit 29. The method of claim 28, wherein the protein is selected from the group consisting of PRP40 pre-mRNA processing factor 40 homolog B (PRPF40B), RNA-binding motif protein 10 (RBM10), poly(rC)-binding protein 1 (PCBP1), crooked-neck pre-mRNA splicing factor 1 (CRNKL1), DEAH (Asp-Glu-Ala-His) box helicase 9 (DHX9), peptidyl-prolyl cis-trans isomerase-like 2 (PPIL2), RNA-binding motif protein 22 (RBM22), small nuclear ribonucleoprotein Sm D3 (SNRPD3), putative ATP-dependent RNA helicase DDX5 (DDX5), pre-mRNA-splicing factor ATP-dependent RNA helicase DHX15 (DHX15), and polyadenylate-binding protein 1 (PABPC1).

30. 30. The method of claim 29, wherein the spliceosome gene or protein is splicing factor 3B subunit 1 (SF3B1).

31. Use of a compound and / or a pharmaceutically acceptable salt according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11, in the preparation of a medicament for the treatment of cancer.

32. 32. The use of claim 31 , wherein the cancer is selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, bile duct cancer, and lung cancer.

33. 33. The use of claim 32, wherein the cancer is selected from myelodysplastic syndrome, chronic lymphocytic leukemia, chronic myelomonocytic leukemia, and acute myeloid leukemia.

34. 33. The use of claim 32, wherein the cancer is myelodysplastic syndrome.

35. 33. The use of claim 32, wherein the cancer is chronic lymphocytic leukemia.

36. 33. The use of claim 32, wherein the cancer is chronic myelomonocytic leukemia.

37. 33. The use of claim 32, wherein the cancer is acute myeloid leukemia.

38. 33. The use of claim 32, wherein the cancer is colon cancer.

39. 33. The use of claim 32, wherein the cancer is pancreatic cancer.

40. 33. The use of claim 32, wherein the cancer is endometrial cancer.

41. 33. The use of claim 32, wherein the cancer is ovarian cancer.

42. 33. The use of claim 32, wherein the cancer is breast cancer.

43. 33. The use of claim 32, wherein the cancer is uveal melanoma.

44. 33. The use of claim 32, wherein the cancer is gastric cancer.

45. 33. The use of claim 32, wherein the cancer is cholangiocarcinoma.

46. 33. The use of claim 32, wherein the cancer is lung cancer.

47. 47. The use according to any one of claims 31 to 46, wherein the cancer is positive for one or more mutations in spliceosome genes or proteins.

48. The spliceosome gene or protein is selected from the group consisting of splicing factor 3B subunit 1 (SF3B1), U2 small nuclear RNA cofactor 1 (U2AF1), serine / arginine-rich splicing factor 2 (SRSF2), zinc finger (CCCH-type) RNA-binding motif and serine / arginine-rich 2 (ZRSR2), pre-mRNA processing-splicing factor 8 (PRPF8), U2 small nuclear RNA cofactor 2 (U2AF2), splicing factor 1 (SF1), splicing factor 3a subunit 48. The use of claim 47, wherein the protein is selected from the group consisting of PRP40 pre-mRNA processing factor 40 homolog B (PRPF40B), RNA-binding motif protein 10 (RBM10), poly(rC)-binding protein 1 (PCBP1), crooked-neck pre-mRNA splicing factor 1 (CRNKL1), DEAH (Asp-Glu-Ala-His) box helicase 9 (DHX9), peptidyl-prolyl cis-trans isomerase-like 2 (PPIL2), RNA-binding motif protein 22 (RBM22), small nuclear ribonucleoprotein Sm D3 (SNRPD3), putative ATP-dependent RNA helicase DDX5 (DDX5), pre-mRNA-splicing factor ATP-dependent RNA helicase DHX15 (DHX15), and polyadenylate-binding protein 1 (PABPC1).

49. 49. The use of claim 48, wherein the spliceosome gene or protein is splicing factor 3B subunit 1 (SF3B1).

50. A method for treating cancer in a subject in need thereof, comprising a therapeutically effective amount of a compound and / or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11; and At least one additional therapy to said subject.

51. 51. The method of claim 50, wherein the at least one additional therapy comprises at least 1, at least 2, at least 3, at least 4, or at least 5 additional therapies.

52. 51. The method of claim 50, wherein the dosage of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, and / or the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, and / or the at least one additional therapy.

53. 53. The method of any one of claims 50 to 52, wherein the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, and / or said at least one additional therapy is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently than a standard administration regimen of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, and / or said at least one additional therapy.

54. 54. The method of any one of claims 50 to 53, wherein the dose and / or dosage of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, and / or said at least one additional therapy results in reduced systemic toxicity and / or improved tolerability.

55. 51. The method of claim 50, wherein administration of the compound and / or pharmaceutically acceptable salt according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, is initiated before administration of the at least one additional therapy.

56. 51. The method of claim 50, wherein administration of the compound and / or pharmaceutically acceptable salt of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or 11, is initiated after administration of the at least one additional therapy.

57. 51. The method of claim 50, wherein administration of the compound and / or pharmaceutically acceptable salt of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or 11, is initiated simultaneously with administration of the at least one additional therapy.

58. 58. The method according to any one of claims 50 to 57, wherein the administration of the compound and / or pharmaceutically acceptable salt according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, is repeated at least once after the initial administration.

59. 59. The method according to claim 58, wherein the amount used for repeated administration of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, is reduced compared to the amount used for the initial administration.

60. 59. The method of claim 58, wherein the amount used for repeated administration of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, is reduced compared to the standard dosage of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11.

61. 59. The method of claim 58, wherein the amount used for repeated administration of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11, is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of the compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or the pharmaceutical composition according to claim 10 or 11.

62. 62. The method of any one of claims 50-61, wherein the administration of the at least one additional therapy is repeated at least once after the initial administration.

63. 63. The method of claim 62, wherein the amount used in repeat administrations of the at least one additional therapy is reduced compared to the amount used in the initial administration.

64. 63. The method of claim 62, wherein the amount used for the repeated administration of the at least one additional therapy is reduced compared to a standard dosage of the at least one additional therapy.

65. 63. The method of claim 62, wherein the amount used for repeat administration of the at least one additional therapy is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of the at least one additional therapy.

66. 66. The method of any one of claims 50 to 65, wherein the repeated administration of the compound and / or pharmaceutically acceptable salt of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or 11, is concurrent with the repeated administration of said at least one additional therapy.

67. 66. The method of any one of claims 50 to 65, wherein the repeated administration of the compound and / or pharmaceutically acceptable salt of any one of claims 1 to 9, or the pharmaceutical composition of claim 10 or 11, is sequential or staggered with the repeated administration of said at least one additional therapy.

68. 68. The method of any one of claims 50-67, wherein the at least one additional therapy comprises administering a checkpoint inhibitor.

69. 69. The method of claim 68, wherein the subject is intolerant, refractory, or non-responsive to the checkpoint inhibitor when administered alone.

70. 69. The method of claim 68, wherein the checkpoint inhibitor targets CTLA4, PD1, PDL1, OX40, CD40, GITR, LAG3, TIM3, and / or KIR.

71. 69. The method of claim 68, wherein the checkpoint inhibitor targets CTLA4, OX40, CD40, and / or GITR.

72. 72. The method of claim 70 or 71, wherein the checkpoint inhibitor comprises a cytotoxic T-lymphocyte-associated antigen 4 pathway (CTLA4) inhibitor.

73. 73. The method of claim 72, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

74. 74. The method of claim 73, wherein the anti-CTLA4 antibody is ipilimumab.

75. 72. The method of claim 70 or 71, wherein the checkpoint inhibitor comprises a programmed death-1 pathway (PD1) inhibitor.

76. 76. The method of claim 75, wherein the PD1 inhibitor is an anti-PD1 antibody.

77. 77. The method of claim 76, wherein the anti-PD1 antibody is nivolumab.

78. 76. The method of claim 75, wherein the PD1 inhibitor is an anti-PDL1 antibody.

79. 79. The method of claim 78, wherein the anti-PDL1 antibody is atezolizumab.

80. 72. The method of claim 70 or 71, wherein the checkpoint inhibitors comprise a CTLA4 inhibitor and a PD1 inhibitor.

81. 81. The method of claim 80, wherein the CTLA4 inhibitor is an anti-CTLA4 antibody.

82. 82. The method of claim 81, wherein the anti-CTLA4 antibody is ipilimumab.

83. 82. The method of claim 80 or 81, wherein the PD1 inhibitor is an anti-PD1 antibody.

84. 84. The method of claim 83, wherein the anti-PD1 antibody is nivolumab.

85. 82. The method of claim 80 or 81, wherein the PD1 inhibitor is an anti-PDL1 antibody.

86. 86. The method of claim 85, wherein the anti-PDL1 antibody is atezolizumab.

87. 68. The method of any one of claims 50 to 67, wherein the at least one additional therapy comprises administering a cytokine or cytokine analogue.

88. 88. The method of claim 87, wherein the subject is intolerant, refractory, or non-responsive to the cytokine or cytokine analog when administered alone.

89. 88. The method of claim 87, wherein the cytokine or cytokine analog comprises a T cell enhancer.

90. 88. The method of claim 87, wherein the cytokine or cytokine analog comprises IL-2, IL-10, IL-12, IL-15, IFNγ, and / or TNFα.

91. 68. The method of any one of claims 50 to 67, wherein the at least one additional therapy comprises administering engineered tumor-targeting T cells.

92. 92. The method of any one of claims 50-91, wherein the subject has a non-synonymous mutation load of about 150 mutations or less.

93. 93. The method of any one of claims 50-92, wherein the subject has a non-synonymous mutation load of about 100 mutations or less.

94. 94. The method of any one of claims 50-93, wherein the subject has a non-synonymous mutation load of about 50 mutations or less.

95. 95. The method of any one of claims 50 to 94, wherein the cancer is a hematological malignancy or a solid tumor.

96. 96. The method of claim 95, wherein the hematological malignancy is selected from a B-cell malignancy, leukemia, lymphoma, and myeloma.

97. 97. The method of claim 95 or 96, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

98. 96. The method of claim 95, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer.

99. 95. The method of any one of claims 50 to 94, wherein the cancer is selected from myelodysplastic syndrome, chronic lymphocytic leukemia, acute lymphoblastic leukemia, chronic myelomonocytic leukemia, acute myeloid leukemia, colon cancer, pancreatic cancer, endometrial cancer, ovarian cancer, breast cancer, uveal melanoma, gastric cancer, bile duct cancer, and lung cancer.

100. 12. A method for inducing at least one neoantigen, comprising contacting a therapeutically effective amount of a compound and / or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11, with a neoplastic cell, thereby inducing the production of at least one neoantigen.

101. 101. The method of claim 100, wherein the neoplastic cells are in an in vitro cell culture.

102. 102. The method of claim 100 or 101, wherein the neoplastic cells are obtained from a subject.

103. 101. The method of claim 100, wherein the neoplastic cells are present in a subject.

104. 104. The method of any one of claims 100 to 103, wherein the neoplastic cells are derived from a hematological malignancy or a solid tumor.

105. 105. The method of claim 104, wherein the hematological malignancy is selected from a B-cell malignancy, leukemia, lymphoma, and myeloma.

106. 106. The method of claim 104 or 105, wherein the hematological malignancy is selected from acute myeloid leukemia and multiple myeloma.

107. 105. The method of claim 104, wherein the solid tumor is selected from breast cancer, gastric cancer, prostate cancer, ovarian cancer, lung cancer, uterine cancer, salivary duct cancer, melanoma, colon cancer, and esophageal cancer.

108. 12. A method of inducing at least one neoantigen and / or T cell response in a subject having or suspected of having a neoplastic disorder, the method comprising administering to the subject a therapeutically effective amount of a compound and / or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11.

109. 12. A method of treating a subject having or suspected of having a neoplastic disorder, comprising administering to the subject a therapeutically effective amount of a compound and / or pharmaceutically acceptable salt thereof according to any one of claims 1 to 9, or a pharmaceutical composition according to claim 10 or 11, wherein administration of the compound and / or pharmaceutically acceptable salt thereof, or pharmaceutical composition induces at least one neoantigen and / or T cell response.

110. 110. The method of claim 109, wherein the amount of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition administered is reduced due to induction of at least one neo-antigen and / or T cell response compared to a standard dosage of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition.

111. 111. The method of claim 110, wherein the dosage of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition is reduced by 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% compared to the standard dosage of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition.

112. 112. The method of any one of claims 109 to 111, wherein the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition is administered at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 75%, or 90% less frequently than a standard administration regimen of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition.

113. 112. The method of any one of claims 109 to 111, wherein the amount and / or dosage of the compound and / or pharmaceutically acceptable salt, or pharmaceutical composition results in reduced systemic toxicity and / or improved tolerability.

114. 114. The method of any one of claims 108 to 113, further comprising administering at least one additional therapy.

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