Macrolide Compounds

Abstract

Disclosed herein are macrolide compounds with ATP synthase inhibitory activity. The macrolides can be used to treat cancer and other proliferative disorders. The macrolides can be used to treat leukemia, including acute myeloid leukemia. The macrolides can be used to treat cancer, including AML, in patients who have developed multidrug resistance. In some embodiments, the macrolide is a derivative of ammocidin A.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Patent Application No. 63 / 193,959, filed May 27, 2021, the contents of which are incorporated herein in their entirety.

[0002] Acknowledgement of Government Support This invention was made with Government support under Grant No. CA226833 awarded by the National Institutes of Health. The Government has certain rights in this invention.

[0003] The present invention is directed to novel macrolide compounds that have ATP synthase / ATPase inhibitory activity. The compounds are useful in a variety of therapeutic settings, including as anticancer agents, antibiotics, and immunosuppressants. [Background technology]

[0004] Cancer is a disease that may be characterized by abnormal clonal cell proliferation, poor cell differentiation, and infiltration into other tissues and organs, bone marrow, and peripheral blood. Acute myeloid leukemia (AML) is a malignant clonal disease derived from hematopoietic stem and progenitor cells, with a 5-year survival rate of less than 30%. AML is often treated with cytotoxic chemotherapy, which can result in 30-75% remission, but relapse is common. Resistance of leukemic cells to standard cytotoxic chemotherapeutic agents remains the main obstacle in the treatment of AML. Tumor drug resistance is mainly divided into primary drug resistance and acquired drug resistance. Primary drug resistance refers to the natural lack of sensitivity to drug therapy prior to the use of antitumor drugs (e.g., cytotoxic chemotherapy, which cannot affect non-cycling cells in the non-proliferating G0 phase). Acquired resistance refers to the loss of sensitivity to chemotherapy over time due to tumor-derived molecular resistance mechanisms that render previously effective drugs ineffective.

[0005] Macrolides comprise a structurally and pharmacologically diverse class of natural products that selectively address equally diverse cellular targets such as immunosuppressive signaling (FK-506, FKB12 ​​calcineurin), splicing factors (SF3b, pladienolides), ribosomes (azithromycin, erythromycin), and ion channels (ivermectin, glutamate-gated chloride channels). Furthermore, mutations within a structural family can have completely different targeting properties, which has motivated significant activity towards the generation of modified macrolides, both via chemical synthesis and biosynthetic pathway engineering. Correspondingly, the impact of macrolides is being realized with both clinical and chemical biological tools to reveal new insights into cell biology.

[0006] Apoptolidin A (Figure 1) was originally discovered in a screen for selective inducers of apoptosis in E1A oncogene-transformed cell lines and was isolated from Nocardiopsis sp. FU40. ‡ Apoptolidin was isolated from a strain designated F. Remarkably, apoptolidin was 1,000 times more potent against transformed cells (10 nM) than against a non-transformed cell line (10 μM). Further evaluation of the compound against the NCI-60 harvest revealed that activity was maximal (low nanomolar) in cell lines that did not display the Warburg effect, but instead were dependent on oxidative phosphorylation. A series of indirect cellular and biochemical studies demonstrated that F O This is consistent with the molecular target of apoptolysin A being F ATP synthase. However, the cell line differential selectivity and cellular response profile of apoptolysin are not consistent with the validated F OThe differences in the binding of apoptolidin family macrolides compared to the subunit inhibitor oligomycin suggest a different mode of action for this family. Recent studies have demonstrated that apoptolidin family macrolides bind to the F1 subcomplex of ATP synthase at a different allosteric site compared to the previously mentioned ATP synthase inhibitors. A structurally related natural product, ammocidin A (Figure 2), was isolated from Saccharothrix sp. AJ9571 in a screen against Ras oncogene transformed cell lines. Another structurally related macrolide, amycolatopsin (Figure 3), was isolated from Amycolatopsis sp. MST-108494.

[0007] Although naturally occurring macrolides have interesting physiological properties, they are not always ideal as drug substances due to limitations related to solubility, bioavailability, stability, or selectivity. There remains a need for ATP synthase inhibitors with increased potency, improved pharmacokinetic profile, chemical stability, and / or reduced toxicity. There remains a need for improved therapies for the treatment of proliferative disorders, including cancer, particularly leukemia. There remains a need for improved therapeutic agents for the treatment of AML and other cancers that do not lose effectiveness over time. There remains a need for improved cancer therapies that do not activate or exacerbate multidrug resistance in cancer in general, and leukemia in particular. There remains a need for improved cancer therapies that are effective against cancers that have developed multidrug resistance. Summary of the Invention

[0008] In accordance with the objectives of the disclosed materials and methods, as embodied and broadly described herein, the disclosed subject matter relates in one aspect to compounds, compositions, and methods of making and using the compounds and compositions.

[0009] Additional advantages will be set forth in part in the description which follows and in part will be obvious from the description or may be learned by practice of the aspects described hereinafter. The advantages described below will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing summary and the following detailed description are exemplary and explanatory only and are not restrictive.

[0010] The details of one or more embodiments are set forth in the description below. Other features, objects, and advantages will become apparent from the description and the claims. [Brief description of the drawings]

[0011] [Figure 1] 1 shows the chemical structure of apoptolidin A. [Diagram 2] The chemical structure of ammocidin A is shown. [Diagram 3] The chemical structure of amycolatopsin A is shown. [Figure 4-1] A: ApoJ / K analysis and biochemical bypass. ApoJ / K is required for methoxymalonyl-ACP loading but not for biochemical bypass using starter unit surrogates. B: Starter unit surrogates synthesized and incubated with an apoJK null strain. [Figure 4-2] C: HPLC / MS analysis of targeted deletion and chemical complementation of apoJK with natural loading units, higher diketide, and starter unit analogs. [Figure 5A] The structure of KK-32-011 / VU936203A is shown. [Figure 5B] The structure of KK-32-011 / VU936203B is shown. [Figure 5C] The structure of KK-32-012 / VU936195A is shown. [Figure 5D] The structure of KK-32-012 / VU936195B is shown. [Figure 6]FIG. 1 shows viability in the MV-4-11 human leukemia cell line as a function of ammocidin derivatives in an MTT viability assay. [Figure 7] 1 shows comparable efficacy in an MV-4-11 cell line-derived xenograft model between ammocidin A (square data points), venetoclax + azacitidine (triangle data points), and venetoclax + azacitidine + ammocidin (star data points). [Figure 8] Pharmacokinetic data in mice administered ammocidin A and ammocidin derivatives at 0.25 mg / kg. In the case of analogs VU936195B (195B) and VU936203B (203B), only free ammocidin A could be detected. [Figure 9A] 1 shows comparative data for venetoclax and ammocidin A on the AML cell line Molm-13 over a 72 hour period. [Figure 9B] 1 shows comparative data for venetoclax and ammocidin A on the AML cell line MV-4-11 over a 72 hour period. [Figure 9C] 1 shows comparative data for venetoclax and ammocidin A on the AML cell line OCI-AML-3 over a 72 hour period. [Figure 9D] 1 shows comparative data for venetoclax and ammocidin A on the AML cell line KMS-12 over a 72 hour period. [Figure 10A] 4 shows Molm-13 cell viability after 48 hours of exposure to a combination of ammocidin A and venetoclax. [Figure 10B] 1 shows the synergy score of the combination of ammocidin A and venetoclax on Molm-13 cells. [Figure 11A] Shows MV-4-11 cell viability 48 hours after exposure to a combination of ammocidin A and venetoclax. [Figure 11B] Shows synergy scores for the combination of ammocidin A and venetoclax on MV-4-11 cells. [Figure 12A]1 shows primary AML cell viability 48 hours after exposure to a combination of ammocidin A and venetoclax. [Figure 12B] 1 shows synergy scores for the combination of ammocidin A and venetoclax against primary AML cells from patients. [Figure 13A] 4 shows KMS-12 cell viability after 72 hours of exposure to a combination of ammocidin A and venetoclax. [Figure 13B] 4 shows the synergy score of the combination of ammocidin A and venetoclax on KMS-12 cells. [Figure 14A] Ammocidin A activity against three primary AML patient samples using survival GI50 data from Prism (GI50 from double logarithmic regression). 18-12-001=0.0302μM (0.0312μM); 17-03-006=0.0103μM (0.0321μM); 29-09-002=0.0158μM (0.0217μM). Data collected on July 8, 2021. [Figure 14B] Ammocidin A activity against two primary AML patient samples using survival GI50 data from Prism (GI50 from double logarithmic regression). 18-12-001 = 0.0284 μM (0.07536 μM); 17-03-006 = 0.0095 μM (0.0308 μM). Data collected on March 8, 2022. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Before the present methods and systems are disclosed and described, it is to be understood that the methods and systems are not limited to specific synthetic methods, specific components, or particular compositions. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting.

[0013] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value, and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximations, by using the antecedent "about," it will be understood that the particular value forms another embodiment. It will be further understood that each of the endpoints of a range is significant both in relation to the other endpoint, and independently of the other endpoint.

[0014] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes instances in which the event or circumstance occurs and instances in which the event or circumstance does not occur.

[0015] Throughout the description and claims of this specification, the word "comprise" and variations of words such as "comprising" and "comprises" mean "including, but not limited to," and are not intended to exclude, for example, other additives, components, elements, or steps. "Exemplary" means "an example of" and is not intended to convey an indication of a preferred or ideal embodiment. "Etc." is used for descriptive purposes, not limiting.

[0016] Components that can be used to carry out the disclosed methods and systems are disclosed. These and other components are disclosed herein, and when combinations, subsets, interactions, groups, etc. of these components are disclosed, it is understood that each is specifically contemplated and described herein for all methods and systems, although specific references to each of the various individual and collective combinations and permutations of these may not be expressly disclosed. This applies to all aspects of this application, including, but not limited to, steps in the disclosed methods. Thus, when there are various additional steps that can be carried out, it is understood that each of these additional steps can be carried out in any particular embodiment or combination of embodiments of the disclosed methods.

[0017] Unless stated to the contrary, formulas having chemical bonds shown only as solid lines and not as wedges or dashed lines contemplate each possible isomer, e.g., each enantiomer, diastereomer, and meso compound, as well as mixtures of isomers, e.g., racemic or scalemic mixtures. Unless stated to the contrary, a formula showing one or more stereochemical features does not exclude the presence of other isomers.

[0018] Throughout the definition, "C n -C m " denotes a range inclusive of the endpoints, where n and m are integers and indicate the number of carbons. Examples include, but are not limited to, C1-C4, C1-C6, etc.

[0019] The term "alkyl" as used herein refers to a branched or unbranched hydrocarbon group, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, dodecyl, etc. In various embodiments, an alkyl group is a group having 1 to 24 carbon atoms (C1-C 24 ), 1 to 12 carbon atoms (C1-C 12 ), 1 to 10 carbon atoms (C1-C 10), 1 to 8 carbon atoms (C1-C8), 1 to 6 carbon atoms (C1-C6), 1 to 4 carbon atoms (C1-C4), 1 to 3 carbon atoms (C1-C3), or 1 to 2 carbon atoms (C1-C2). Alkyl groups can also be substituted or unsubstituted. Unless otherwise indicated, the term "alkyl" contemplates both substituted and unsubstituted alkyl groups. Alkyl groups include, but are not limited to, C1-C 10 Alkoxy, C1-C 10 Alkenyl, C1-C 10 Alkynyl, C3-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C6-C 18 Aryl, C1-C 10 It may be substituted with one or more groups including heteroaryl, aldehyde, amino, carboxylic acid, oxo, halide, hydroxy, cyano, nitro, silyl, sulfo-oxo, or thiol. Alkyl groups that do not contain double or triple carbon-carbon bonds are called saturated alkyl groups, while alkyl groups with one or more such bonds are called unsaturated alkyl groups. Unsaturated alkyl groups with double bonds can be called alkenyl groups, and unsaturated alkyl groups with triple bonds can be called alkynyl groups.

[0020] As used herein, the term "cycloalkyl" refers to a non-aromatic carbon-based ring composed of at least three carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like. The term "heterocycloalkyl" refers to a cycloalkyl group as defined above in which at least one of the carbon atoms of the ring is replaced with a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur, selenium, or phosphorus. Cycloalkyl and heterocycloalkyl groups can be substituted or unsubstituted. Unless otherwise indicated, the terms "cycloalkyl" and "heterocycloalkyl" contemplate both substituted and unsubstituted cycloalkyl and heterocycloalkyl groups. Cycloalkyl and heterocycloalkyl groups include, but are not limited to, C1-C2-C3-C4-C5-C6-C7-C8-C9-C10-C11-C12-C13-C14-C15-C16-C17-C18-C19-C21-C22-C33-C24-C34-C25-C35-C36-C37-C38-C39-C47-C48-C49 ... 10 Alkyl, C1-C10 Alkoxy, C1-C 10 Alkenyl, C1-C 10 Alkynyl, C3-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C6-C 18 Aryl, C1-C 10 It may be substituted with one or more groups including heteroaryl, aldehyde, amino, carboxylic acid, halide, hydroxy, cyano, oxo, nitro, silyl, sulfo-oxo, or thiol. Cycloalkyl groups that do not contain double or triple carbon-carbon bonds are referred to as saturated cycloalkyl groups, and cycloalkyl groups that have one or more such bonds (that are not still aromatic) are referred to as unsaturated cycloalkyl groups.

[0021] As used herein, the term "aryl" is an aromatic ring composed of carbon atoms. Examples of aryl groups include, but are not limited to, phenyl and naphthyl. The term "heteroaryl" is an aryl group as defined above in which at least one of the carbon atoms of the ring is replaced with a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur, selenium, or phosphorus. The aryl and heteroaryl groups may be substituted or unsubstituted. Unless otherwise stated, the terms "aryl" and "heteroaryl" contemplate both substituted and unsubstituted aryl and heteroaryl groups. The aryl and heteroaryl groups include, but are not limited to, C1-C2 alkyl groups, C2-C3 alkyl groups, C3-C4 alkyl groups, C4-C5 alkyl groups, C5-C6 alkyl groups, C6-C7 alkyl groups, C7-C8 alkyl groups, C8-C9 alkyl groups, C9-C10 alkyl groups, C11-C12 alkyl groups, C12-C14 alkyl groups, C13-C15 alkyl groups, C14-C16 alkyl groups, C15-C17 alkyl groups, C16-C18 alkyl groups, C17-C19 alkyl groups, C18-C21 alkyl groups, C19-C22 alkyl groups, C12-C23 alkyl groups, C13-C24 alkyl groups, C14-C25 alkyl groups, C15-C26 alkyl groups, C16-C27 alkyl groups, C17-C28 alkyl groups, C18-C29 alkyl groups, C19-C31 alkyl groups, C19-C21 alkyl groups, C19-C22 alkyl groups, C19-C23 alkyl groups, C19-C24 alkyl groups, C19-C25 alkyl groups, C21-C25 alkyl groups, C22-C26 alkyl groups, C23-C27 alkyl groups, C24-C28 alkyl groups, C25-C29 alkyl groups, C25-C29 alkyl 10 Alkyl, C1-C 10 Alkoxy, C1-C 10 Alkenyl, C1-C 10 Alkynyl, C3-C 10 Cycloalkyl, C1-C 10 Heterocycloalkyl, C6-C 18 Aryl, C1-C 10 It may be substituted with one or more groups including heteroaryl, aldehyde, amino, carboxylic acid, halide, hydroxy, cyano, oxo, nitro, silyl, sulfo-oxo, or thiol.

[0022] Exemplary heteroaryl and heterocyclyl rings include benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzoxazolinyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, 4aH carbazolyl, carbolinyl, chromanyl, chromenylsilonolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3b]tetrahydrofuran, furanyl, Furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isatinoyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, methylenedioxyphenyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl, 1,2,5-oxadiazolyl , 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxindolyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, piperidonyl, 4-piperidonyl, piperonyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl nyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinoxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, tetrazolyl, 6H-1,2,5-thiadiazolyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, and xanthenyl.

[0023] The terms "alkoxy", "cycloalkoxy", "heterocycloalkoxy", "cycloalkoxy", "aryloxy" and "heteroaryloxy" have the above meanings of alkyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl, further provided that such group is linked through an oxygen atom.

[0024] As used herein, the term "substituted" is intended to include all permissible substituents of organic compounds. In a broad aspect, the permissible substituents include acyclic and cyclic, branched and unbranched, carbocyclic and heterocyclic, and aromatic and nonaromatic substituents of organic compounds. Exemplary substituents include, for example, those described below. The permissible substituents can be one or more and can be the same or different for appropriate organic compounds. For purposes of this disclosure, heteroatoms such as nitrogen can have hydrogen substituents and / or any permissible substituents of organic compounds described herein that satisfy the valence of the heteroatom. This disclosure is not intended to be limited in any manner by the permissible substituents of organic compounds. The terms "substituted" or "substituted with" also include the implicit proviso that such substitution is in accordance with the permissible valences of the substituted atom and substituent, and that the substitution results in a stable compound, e.g., a compound that does not spontaneously transform by rearrangement, cyclization, elimination, etc. Unless specifically stated otherwise, a substituent referred to as "substituted" means that the substituent can be substituted with one or more of alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aldehyde, amino, carboxylic acid, ester, ether, halide, hydroxy, ketone, nitro, silyl, sulfo-oxo, or thiol.

[0025] Unless otherwise specified, the term "patient" refers to any mammal, including, but not limited to, a human.

[0026] As used herein, a "nucleoside analog" is a compound that has the ability to mimic natural purine or pyrimidine nucleosides, which can disrupt metabolic and regulatory pathways.

[0027] As used herein, azanucleoside is a modified nucleoside in which one or more atoms in the furanosyl or aromatic ring are replaced by nitrogen atoms.In some embodiments, azanucleoside is a modified nucleoside with a modified cytosine, adenine, guanine, thymine or uracil ring, and one or more carbon atoms in the ring are replaced by nitrogen atoms.Modified nucleosides and azanucleosides, unless otherwise specified, include prodrugs, such as esters, phosphate esters, and phosphoramidates at 5' and / or 4' and 3' carbons.

[0028] Pharmaceutically acceptable salts are salts that retain the desired biological activity of the parent compound and do not impart undesired toxicological effects. Examples of such salts are acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, and nitric acid; 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, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, and polygalacturonic acid; salts formed from elemental anions such as chloride, bromide chloride, and iodide; salts formed from metal hydroxides such as sodium hydroxide, potassium hydroxide, calcium hydroxide, lithium hydroxide, and magnesium hydroxide; salts formed from metal carbonates such as sodium carbonate, potassium carbonate, calcium carbonate, and magnesium carbonate; metal bicarbonates such as sodium bicarbonate and potassium bicarbonate; metal sulfates such as sodium sulfate and potassium sulfate; and metal nitrates such as sodium nitrate and sodium nitrate. Pharmaceutically acceptable and non-pharmacologically acceptable salts can be prepared using procedures well known in the art, for example, by reacting a sufficiently basic compound, such as an amine, with a suitable acid that contains a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts of carboxylic acids can also be made.

[0029] As used herein, a compound of formula (1): [ka] and pharma- ceutically acceptable salts thereof are disclosed, During the ceremony, R 1 is H, OH, C 1-8 Alkyl and OC 1-8 alkyl, preferably CH3. R 2 is H, OH, C 1-8 Alkyl and OC 1-8alkyl, preferably OCH3. R 3 is H, OH, C 1-8 Alkyl and OC 1-8 It is selected from alkyl, preferably H or OH. R 4 is H, OH, C 1-8 Alkyl and OC 1-8 It is selected from alkyl, preferably H or OH. Q 1 teeth, [ka] is a group having the formula During the ceremony, R 2a -R 2a* , -OR 2a* , OP(O)(OR 2a* )2, OP(O)(OR 2a* )(N(R 2a* )2, -N(R 2a* )2, -N(R 2a* )3, -C(O)R 2a* , -C(O)OR 2a* , -OC(O)R 2a* , -OC(O)OR 2a* , -NR 2a* C(O)R 2a* , -C(O)N(R 2a* )2, NR 2a* C(O)OR 2a* , -OC(O)N(R 2a* )2, -NR 2a* C(O)N(R 2a* )2;-Cl, -F, -Br, -I, -NO2, -CN, -N3, -(OCH2CH2) m -OR p is selected from R 2a* is, in each occurrence independently, H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, Aryl, C 3-10 Cyclorkil, C 3-10 Cycloalkenyl, C 7-10Cycloalkynyl, aryl, C 1-10 Heterocysil, C 1-10 heteroaryl, and each R 2a* are -OH, -COOH, -NH2, -Cl, -F, -Br, -I, -NO2, -CN, -N3, PO(OH)2, -(OCH2CH2) m -OR p ;C 1-8 Heterocycyl, aryl, -OC 1-8 Heterocysil, or C 1-8 may be substituted one or more times by alkoxy; R 2a* any two or more of may together form a ring, R 3a* -R 3a* , -OR 3a* , OP(O)(OR 3a* )2, OP(O)(OR 3a* )(N(R 3a* )2, -N(R 3a* )2, -N(R 3a* )3, -C(O)R 3a* , -C(O)OR 3a* , -OC(O)R 3a* , -OC(O)OR 3a* , -NR 3a* C(O)R 3a* , -C(O)N(R 3a* )2, NR 3a* C(O)OR 3a* , -OC(O)N(R 3a* )2, -NR 3a* C(O)N(R 3a* )2;-Cl, -F, -Br, -I, -NO2, -CN, -N3, -(OCH2CH2) m -OR p is selected from R 3a* is, in each occurrence independently, H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, Aryl, C 3-10 Cyclorkil, C 3-10 Cycloalkenyl, C 7-10 Cycloalkynyl, aryl, C 1-10 Heterocysil, C 1-10heteroaryl, and each R 3a* are -OH, -COOH, -NH2, -Cl, -F, -Br, -I, -NO2, -CN, -N3, PO(OH)2, -(OCH2CH2) m -OR p ;C 1-8 Heterocycyl, aryl, -OC 1-8 Heterocysil, or C 1-8 may be substituted one or more times by alkoxy; R 3a* any two or more of may together form a ring, R 4a -R 4a* , -OR 4a* , OP(O)(OR 4a* )2, OP(O)(OR 4a* )(N(R 4a* )2, -N(R 4a* )2, -N(R 4a* )3, -C(O)R 4a* , -C(O)OR 4a* , -OC(O)R 4a* , -OC(O)OR 4a* , -NR 4a* C(O)R 4a* , -C(O)N(R 4a* )2, NR 4a* C(O)OR 4a* , -OC(O)N(R 4a* )2, -NR 4a* C(O)N(R 4a* )2;-Cl, -F, -Br, -I, -NO2, -CN, -N3, -(OCH2CH2) m -OR p is selected from R 4a* is, in each occurrence independently, H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, Aryl, C 3-10 Cyclorkil, C 3-10 Cycloalkenyl, C 7-10 Cycloalkynyl, aryl, C 1-10 Heterocysil, C 1-10 heteroaryl, and each R 4a*are -OH, -COOH, -NH2, -Cl, -F, -Br, -I, -NO2, -CN, -N3, PO(OH)2, -(OCH2CH2) m -OR p ;C 1-8 Heterocycyl, aryl, -OC 1-8 Heterocysil, or C 1-8 may be substituted one or more times by alkoxy; R 4a* any two or more of may together form a ring, R 5a -R 5a* , -OR 5a* , OP(O)(OR 5a* )2, OP(O)(OR 5a* )(N(R 5a* )2, -N(R 5a* )2, -N(R 5a* )3, -C(O)R 5a* , -C(O)OR 5a* , -OC(O)R 5a* , -OC(O)OR 5a* , -NR 5a* C(O)R 5a* , -C(O)N(R 5a* )2, NR 5a* C(O)OR 5a* , -OC(O)N(R 5a* )2, -NR 5a* C(O)N(R 5a* )2;-Cl, -F, -Br, -I, -NO2, -CN, -N3, -(OCH2CH2) m -OR p is selected from R 5a* is, in each occurrence independently, H, C 1-10 Alkyl, C 2-10 Alkenyl, C 2-10 Alkynyl, Aryl, C 3-10 Cyclorkil, C 3-10 Cycloalkenyl, C 7-10 Cycloalkynyl, aryl, C 1-10 Heterocysil, C 1-10 heteroaryl, and each R 5a*are -OH, -COOH, -NH2, -Cl, -F, -Br, -I, -NO2, -CN, -N3, PO(OH)2, -(OCH2CH2) m -OR p ;C 1-8 Heterocycyl, aryl, -OC 1-8 Heterocysil, or C 1-8 may be substituted one or more times by alkoxy; R p In each case, H, C 1-10 selected from alkyl, and aryl; R 5a* any two or more of may together form a ring, R 2a , R 3a , R 4a , and R 5a any two or more of may together form a ring, Q 2 teeth, [ka] is a group having the formula During the ceremony, R 1b is H, OH, C 1-8 Alkyl and OC 1-8 alkyl, R 2b -R 2b* , -OR 2b* , OP(O)(OR 2b* )2, OP(O)(OR 2b* )(N(R 2b* )2, -N(R 2b* )2, -N(R 2b* )3, -C(O)R 2b* , -C(O)OR 2b* , -OC(O)R 2b* , -OC(O)OR 2b* , -NR 2b* C(O)R 2b* , -C(O)N(R 2b* )2, NR 2b* C(O)OR 2b* , -OC(O)N(R 2b* )2, -NR 2b*C(O)N(R 2b* )2;-Cl, -F, -Br, -I, -NO2, -CN, -N3, -(OCH2CH2) m -OR p is selected from R 2b* is, in each occurrence independently, H, C 1-8 Alkyl, C 1-8 Alkenyl, C 1-10 Alkynyl, Aryl, C 3-8 Cyclorkil, C 3-8 Cycloalkenyl, C 7-10 Cycloalkynyl, aryl, C 1-8 Heterocysil, C 3-8 heteroaryl, and each R 2b* are -OH, -COOH, -NH2, -Cl, -F, -Br, -I, -NO2, -CN, -N3, PO(OH)2, -(OCH2CH2) m -OR p ;C 1-8 Heterocycyl, aryl, -OC 1-8 Heterocysil, or C 1-8 may be substituted one or more times by alkoxy; R 2b* any two or more of may together form a ring, R 3b is selected from H or a group having the formula: [ka] During the ceremony, R 3d is selected from H and CH3; R 4d is selected from H and the group having the formula: [ka]

[0030] In some embodiments, the compound has the following formula (2a), formula (2b), or formula (2c): [ka] The compound may be:

[0031] In some embodiments, Q 1 teeth, [ka] It is an (L) sugar with the conformation

[0032] Q 1 Exemplary stereoisomers of include: [ka] Examples include:

[0033] Q 2 teeth, [ka] The formula may be:

[0034] Q 2 but, [ka] 2. A compound according to any one of the preceding claims, which may have the formula:

[0035] R 3b teeth, [ka] It may be a group having the formula:

[0036] R 4d teeth, [ka] It can be a group having the formula:

[0037] The compounds disclosed herein preferably comprise R 5a is selected from CH3 and CH2OH, most preferably CH3.

[0038] In some embodiments, the compound is 3a Derivatized at position R 4a and R 2a Each is OH. In another embodiment, the compound is 4a Derivatized at position R 3a and R 2a Each is OH. In a further embodiment, the compound is 2a Derivatized at position R 3a and R 4a Each of R is OH. 4a and R 2a Either both are derivatized or R 3a and R 2a are both derivatized, or R 4a and R 3a Other embodiments are also contemplated in which both R 2a , R 3a , and R 4a Also within the scope of this disclosure are further embodiments in which all of R are derivatized. 2b may be as naturally found in ammocidin (-CHCHCHOCH), although it is contemplated that the disclosed compounds may also be derivatized at this position. 2a , R 3a , and R 4a None of the R 2b is not -CH2CH2CH2OCH3.

[0039] The macrolide derivatives disclosed herein include R 2a , R 3a , and R 4a At least one of is not OH or R 2bis not -CH2CH2CH2OCH3.

[0040] In certain embodiments, the macrolide derivatives may contain an azide, tetrazine, cyclooctyne, or trans-cyclooctene group. Such groups are useful in "click" cycloaddition reactions that can be used to further refine the macrolide analogs. Suitable cyclooctynes ​​include bicyclo[6.1.0]nonyne ("BCN"), dibenzocyclooctyne, dibenzocyclooctyne-amine, and substituted derivatives of each of these groups. Exemplary groups include: [ka] Examples include:

[0041] In certain embodiments, one or more hydroxyl groups on the naturally occurring (L)-fucose moiety may be modified. As used herein, modification includes oxidation, stereochemical inversion, and / or functionalization via chemical reaction. These compounds may be accessed by exploiting the different chemical reactivity of each hydroxyl in the fucose ring. In general, R 3a The position is the most reactive towards electrophiles and this position can be selectively modified. To modify other positions, first R 3a It may be necessary to protect the hydroxyl (and other positions) and such techniques are known to those skilled in the art.

[0042] In some embodiments, the macrolide derivatives may include a mitochondrial targeting moiety. Exemplary moieties include quaternary phosphonium and ammonium ions, such as triphenylphosphonium, trialkylammonium (e.g., trimethylammonium, triethylammonium), guanidinium, (including both cyclic and acyclic guanidinium), pyridinium, rhodamine, dequalinium, (E)-4-(1H-indol-3-ylvinyl)-N-methylpyridinium iodide, and the like.

[0043] In some embodiments, R 3a -OR 3a* , -OC(O)R 3a* , OP(O)(OR 3a* )2, OP(O)(OR 3a* )(N(R 3a* )2, -OC(O)OR 3a , -OC(O)N(R 3a* )2, or -(OCH2CH2) m -OR 3a* (m can be 1 to 100). In each case, R 3a* are independently 3a OR 3a* (except when 1-8 Alkyl, C 1-8 Alkenyl, or C 1-10 Alkynyl, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-8 Heterocyclyl, C 1-8 Alkyl C 7-10 Cyclorynyl, C 1-8 Alkyl C 7-10 Cycloalkenyl, or C 1-8 Alkyl C 1-8 Heterocyclyl. Such R 3a* The groups may be further substituted one or more times, exemplary substituents include COOR, F, Cl, Br, I, SO3R, OSO3R, -PPh3, -CH2NEt3, -CH2NMe3, P(O)(OR)2, OP(O)(OR)2, N3, heterocyclyl, heteroaryl, aryl, cycloalkyl, where R, at each occurrence, is independently selected from H, alkyl, and cycloalkyl.

[0044] In certain embodiments, R 3a is an ether or ester, e.g., -OR 3a* Or -OC(O)R 3a* and others are derivatized as phosphonate or phosphoramidate products, e.g., -OP(O)(OR 3a* )2 or OP(O)(OR 3a* )(N(R 3a*)2 and derivatized as R 3a is preferably H, aryl, and C 1-8 alkyl or an amino acid, e.g. [ka] Derived from, R p is as defined above.

[0045] In some embodiments, R 3a* teeth, [ka] may be a moiety having the formula: In the formula, the sum of z and z' is 7 or less, and Q 2a and Q 3a are each hydrogen or both C 1-8 Forming heterocysyl, Q 1a COOH, aryl, -C≡CH, -CF3, N3, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-10 Heteroaryl, or C 1-10 Particularly preferred Q is selected from heterocysyl. 1a Groups include COOH, N, and CF. In other embodiments, Q 1a is aryl and C 1-10 In another embodiment, Q is selected from heteroaryl, optionally substituted one or more times as described above. 1a is dibenzocyclooctyne, dibenzocyclooctyne-amine, or trans-cyclooctene.

[0046] In certain embodiments, Q 1a teeth, [ka] may be a moiety having the formula: In the formula, Xdq is an alkylene chain having 2-12 CH2 units, preferably 5-12 CH2 units, more preferably 7-11 CH2 units, and especially preferably 8-10 CH2 units. Those skilled in the art will recognize that although some of the above moieties are shown in electronically neutral form, these moieties may be protonated and paired with an appropriate counterion. Similarly, the quinolinium cations shown above are accompanied by a charge balancing ion or ions, such as, for example, dichloride, dibromide, diiodide, diacetate, etc.

[0047] In some embodiments, R 2a -OR 2a* , -OC(O)R 2a* , OP(O)(OR 2a* )2, OP(O)(OR 2a* )(N(R 2a* )2, -OC(O)OR 2a , -OC(O)N(R 2a* )2, or -(OCH2CH2) m -OR 2a* (m can be 1 to 100). In each case, R 2a* are independently 2a OR 2a* (except when 1-8 Alkyl, C 1-8 Alkenyl, or C 1-10 Alkynyl, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-8 Heterocyclyl, C 1-8 Alkyl C 7-10 Cyclorynyl, C 1-8 Alkyl C 7-10 Cycloalkenyl, or C 1-8 Alkyl C 1-8 Heterocyclyl. Such R 2a*The groups may be further substituted one or more times, exemplary substituents include COOR, F, Cl, Br, I, SO3R, OSO3R, -PPh3, -CH2NEt3, -CH2NMe3, P(O)(OR)2, OP(O)(OR)2, N3, heterocyclyl, heteroaryl, aryl, cycloalkyl, where R, at each occurrence, is independently selected from H, alkyl, and cycloalkyl.

[0048] In certain embodiments, R 2a is an ether or ester, e.g., -OR 2a* Or -OC(O)R 2a* and others are derivatized as phosphonate or phosphoramidate products, e.g., -OP(O)(OR 2a* )2 or OP(O)(OR 2a* )(N(R 2a* )2 and derivatized as R 2a is preferably H, aryl, and C 1-8 alkyl or an amino acid, e.g. [ka] Derived from, R p is as defined above.

[0049] In some embodiments, R 2a* teeth, [ka] may be a moiety having the formula: In the formula, the sum of y and y' is 7 or less, and Q 2b and Q 3b are each hydrogen or both C 1-8 Forming heterocysyl, Q 1b COOH, aryl, -C≡CH, -CF3, N3, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-10 Heteroaryl, or C 1-10Particularly preferred Q is selected from heterocysyl. 1b Groups include COOH, N, and CF. In other embodiments, Q 1b is aryl and C 1-10 In another embodiment, Q is selected from heteroaryl, optionally substituted one or more times as described above. 1b is dibenzocyclooctyne, dibenzocyclooctyne-amine, or trans-cyclooctene.

[0050] In certain embodiments, Q 1b teeth, [ka] may be a moiety having the formula: In the formula, X dq is an alkylene chain having 2-12 CH2 units, preferably 5-12 CH2 units, more preferably 7-11 CH2 units, and especially preferably 8-10 CH2 units. Those skilled in the art will recognize that although some of the above moieties are shown in electronically neutral form, these moieties may be protonated and paired with an appropriate counterion. Similarly, the quinolinium cations shown above are accompanied by a charge balancing ion or ions, such as, for example, dichloride, dibromide, diiodide, diacetate, etc.

[0051] In some embodiments, R 4a -OR 4a* , -OC(O)R 4a* , OP(O)(OR 4a* )2, OP(O)(OR 4a* )(N(R 4a* )2, -OC(O)OR 4a , -OC(O)N(R 4a* )2, or -(OCH2CH2) m -OR 4a* (m can be 1 to 100). In each case, R 4a* are independently 4a OR 4a* (except when 1-8 Alkyl, C1-8 Alkenyl, or C 1-10 Alkynyl, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-8 Heterocyclyl, C 1-8 Alkyl C 7-10 Cyclorynyl, C 1-8 Alkyl C 7-10 Cycloalkenyl, or C 1-8 Alkyl C 1-8 Heterocyclyl. Such R 4a* The groups may be further substituted one or more times, exemplary substituents include COOR, F, Cl, Br, I, SO3R, OSO3R, -PPh3, -CH2NEt3, -CH2NMe3, P(O)(OR)2, OP(O)(OR)2, N3, heterocyclyl, heteroaryl, aryl, cycloalkyl, where R, at each occurrence, is independently selected from H, alkyl, and cycloalkyl.

[0052] In certain embodiments, R 4a is an ether or ester, e.g., -OR 4a* Or -OC(O)R 4a* and others are derivatized as phosphonate or phosphoramidate products, e.g., -OP(O)(OR 4a* )2 or OP(O)(OR 4a* )(N(R 4a* )2 and derivatized as R 4a is preferably H, aryl, and C 1-8 alkyl or an amino acid, e.g. [ka] Derived from, R p is as defined above.

[0053] In some embodiments, R 4a* teeth, [ka] may be a moiety having the formula: In the formula, the sum of x and x' is 7 or less, and Q 2c and Q 3c are each hydrogen or both C 1-8 Forming heterocysyl, Q 1c COOH, aryl, -C≡CH, -CF3, N3, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-10 Heteroaryl, or C 1-10 Particularly preferred Q is selected from heterocysyl. 1c Groups include COOH, N, and CF. In other embodiments, Q 1c is aryl and C 1-10 In another embodiment, Q is selected from heteroaryl, optionally substituted one or more times as described above. 1c is dibenzocyclooctyne, dibenzocyclooctyne-amine, or trans-cyclooctene.

[0054] In certain embodiments, Q 1c teeth, [ka] may be a moiety having the formula: In the formula, X dq is an alkylene chain having 2-12 CH2 units, preferably 5-12 CH2 units, more preferably 7-11 CH2 units, and especially preferably 8-10 CH2 units. Those skilled in the art will recognize that although some of the above moieties are shown in electronically neutral form, these moieties may be protonated and paired with an appropriate counterion. Similarly, the quinolinium cations shown above are accompanied by a charge balancing ion or ions, such as, for example, dichloride, dibromide, diiodide, diacetate, etc.

[0055] In some embodiments, macrolide derivatives can be produced, for example, by modification of fermentation conditions as described herein. 2b It can be modified at position R.2b -OR 2b* , -OC(O)R 2b* , -PPh3, -CH2NEt3, -CH2NMe3, OP(O)(OR 2b* )2, OP(O)(OR 2b* )(N(R 2b* )2, -OC(O)OR 2b , -OC(O)N(R 2b* )2, or -(OCH2CH2) m -OR 2b* (m can be 1 to 100). In each case, R 2b* are independently 2b OR 2b* (except when 1-8 Alkyl, C 1-8 Alkenyl, or C 1-10 Alkynyl, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-8 Heterocyclyl, C 1-8 Alkyl C 7-10 Cyclorynyl, C 1-8 Alkyl C 7-10 Cycloalkenyl, or C 1-8 Alkyl C 1-8 Heterocyclyl. Such R 2b* The groups may be further substituted one or more times, exemplary substituents include COOR, F, Cl, Br, I, SO3R, OSO3R, P(O)(OR)2, OP(O)(OR)2, N3, C 1-10 Heterocyclyl, C 1-10 Heteroaryl, C 6-18 Aryl, C 3-10 cycloalkyl, where R, at each occurrence, is independently selected from H, alkyl, and cycloalkyl.

[0056] In certain embodiments, R 2b is an ether or ester, e.g., -OR 2b* Or -OC(O)R 2b* and others are derivatized as phosphonate or phosphoramidate products, e.g., -OP(O)(OR 2b* )2 or OP(O)(OR2b* )(N(R 2b* )2 and derivatized as R 2b is preferably H, aryl, and C 1-8 alkyl or an amino acid, e.g. [ka] Derived from, In the formula, R p is as defined above.

[0057] In some embodiments, R 2b* teeth, [ka] may be a moiety having the formula: In the formula, the sum of w and w' is 7 or less, and Q 2d and Q 3d are each hydrogen or both C 1-8 Forming heterocysyl, Q 1d COOH, aryl, -C≡CH, -CF3, N3, C 7-10 Cyclorynyl, C 7-10 Cycloalkenyl, C 1-10 Heteroaryl, or C 1-10 Particularly preferred Q is selected from heterocysyl. 1d Groups include COOH, N, and CF. In other embodiments, Q 1d is aryl and C 1-10 In another embodiment, Q is selected from heteroaryl, optionally substituted one or more times as described above. 1d is dibenzocyclooctyne, dibenzocyclooctyne-amine, or trans-cyclooctene.

[0058] In certain embodiments, Q 1d teeth, [ka] may be a moiety having the formula: In the formula, X dq is an alkylene chain having 2-12 CH2 units, preferably 5-12 CH2 units, more preferably 7-11 CH2 units, and especially preferably 8-10 CH2 units. Those skilled in the art will recognize that although some of the above moieties are shown in electronically neutral form, these moieties may be protonated and paired with an appropriate counterion. Similarly, the quinolinium cations shown above are accompanied by a charge balancing ion or ions, such as, for example, dichloride, dibromide, diiodide, diacetate, etc.

[0059] In addition, the following formulas (3a), (3b), and (3c) [ka] Also disclosed is a compound of the formula: In the formula, R 2f -R f* , -C(O)R f* , -C(O)OR f* , -OC(O)R 2b* , -OC(O)OR f* , C(O)N(R f* )2, or -(OCH2CH2) m -OR f* and In the formula, R 3f -R f* , -C(O)R f* , -C(O)OR f* , -OC(O)R 2b* , -OC(O)OR f* , C(O)N(R f* )2, or -(OCH2CH2) m -OR f* and In the formula, R 4f -R f* , -C(O)R f* , -C(O)OR f* , -OC(O)R 2b* , -OC(O)OR f* , C(O)N(R f* )2, or -(OCH2CH2) m -OR f*and R f* is, in each occurrence, independently H, substituted or unsubstituted C 1-8 Alkyl, substituted or unsubstituted C 1-8 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-8 Cycloalkenyl, substituted or unsubstituted C 7-10 Cycloalkynyl, aryl, substituted or unsubstituted C 1-8 Heterocyclic, substituted or unsubstituted C 3-8 heteroaryl; R s is H, substituted or unsubstituted C 1-8 Alkyl, substituted or unsubstituted C 1-8 Alkenyl, substituted or unsubstituted C 1-10 Alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C 3-8 Cycloalkyl, substituted or unsubstituted C 3-8 Cycloalkenyl, substituted or unsubstituted C 7-10 Cycloalkynyl, substituted or unsubstituted aryl, C 1-8 Heterocyclic, substituted or unsubstituted C 3-8 heteroaryl; However, R s If is CH2CH2CH2OCH3, R 2f , R 3f , and R 4f At the same time, it is not H.

[0060] As used herein, a substituent is a group in which one or more hydrogen atoms bonded to a carbon atom are replaced with a non-hydrogen group. Exemplary non-hydrogen groups include halo (F, Cl, Br, I), hydroxyl, sulfhydryl, C1-C 24 Alkoxy, C5-C 24 Aryloxy, acyl (C2-C 24 Alkylcarbonyl (-CO-alkyl) and C6-C 24 Arylcarbonyl (-CO-C5-C 24 aryl), haloacyl (C2-C 24Haloalkylcarbonyl (-C)-haloalkyl)C6-C 24 Haloarylcarbonyl (-CO-aryl) (including C2-C 24 Thioacyloxy (including -O-(CS)-alkyl and -O-(CS)-aryl), C2-C 24 Thiohaloacyloxy (including -O-(CS)-haloalkyl and -O-(CS)-haloaryl), acyloxy (-O-acyl), C2-C 24 Alkoxycarbonyl (-(CO)-O-alkyl), C6-C 24 Aryloxycarbonyl (-(CO)-O-aryl), C7-C 24 Alkaryloxycarbonyl (-(CO)-O-alkaryl), C7-C 24 Aralkyloxycarbonyl (-(CO)-O-aralkyl), C2-C 24 Haloalkylcarbonate (-O-(CO)-O-haloalkyl), C6-C 24 Haloarylcarbonate (-O-(CO)-O-haloaryl), C2-C 24 Alkylthiocarbonate (-O-(CS)-O-alkyl), C6-C 24 Arylthiocarbonate (-O-(CS)-O-aryl), C2-C 24 Haloalkylthiocarbonato (-O-(CS)-O-haloalkyl), and C6-C 24 Haloarylthiocarbonates (-O-(CS)-O-haloaryl), C6-C 24 Aryloxycarbonyl (-(CO)-O-aryl), halocarbonyl (-CO)-X where X is halo), C2-C 24 Alkylcarbonate (-O-(CO)-O-alkyl), C6-C 24 Arylcarbonato (-O-(CO)-O-aryl), carboxy (-COOH), carboxylato (-COO - ), carbamoyl (-(CO)-NH2), mono-(C1-C 24 Alkyl)-substituted carbamoyl (-(CO)-NH(C1-C 24 Alkyl)), di-(C1-C 24 Alkyl)-substituted carbamoyl (-(CO)-N(C1-C24 Alkyl)2), mono-(C6-C 24 aryl)-substituted carbamoyl (-(CO)-NH-aryl), di-(C6-C 24 aryl)-substituted carbamoyl (-(CO)-N(aryl)2), di-N-(C1-C 24 Alkyl), N-(C6-C 24 aryl)-substituted carbamoyl, thiocarbamoyl (-(CS)-NH2), carbamido (-NH-(CO)-NH2), formyl (-(CO)-H), thioformyl (-(CS)-H), amino (-NH2), mono-(C1-C 24 Alkyl)-substituted amino, di-(C1-C 24 Alkyl)-substituted amino, mono-(C5-C 24 aryl)-substituted amino, di-(C5-C 24 Aryl)-substituted amino, C2-C 24 Alkylamide (-NH-(CO)-alkyl), C6-C 24 Arylamide (-NH-(CO)-aryl), imino (-CR=NH, where R=hydrogen, C1-C 24 Alkyl, C5-C 24 Aryl, C6-C 24 Alkaryl, C6-C 24 aralkyl, alkylimino (R=hydrogen-CR=N(alkyl), C1-C 24 Alkyl, C5-C 24 Aryl, C6-C 24 Alkaryl, C6-C 24 aralkyl, arylimino (R=hydrogen, -CR=N(aryl), C1-C 24 Alkyl, C5-C 24 Aryl, C6-C 24 Alkaryl, C6-C 24 aralkyl, etc.), nitro (-NO2), nitroso (-NO), sulfo (-SO2-OH), sulfonamide (-SO2-NH2), sulfonato (-SO2-O - ), C1-C 24 Alkylsulfanyl (-S-alkyl, also known as "alkylthio"), arylsulfanyl (-S-aryl, also known as "arylthio"), C1-C24 Alkylsulfinyl (-(SO)-alkyl), C5-C 24 Arylsulfinyl (-(SO)-aryl), C1-C 24 Alkylsulfonyl (-SO2-alkyl), C5-C 24、 Arylsulfonyl (-SO2-aryl), phosphono (-P(O)(OH)2), phosphonato (-P(O)(O - )2), phosphinato (-P(O)(O - )), phospho (-PO2), and phosphino (-PH2); and hydrocarbyl moieties C1-C 24 Alkyl (preferably C1-C 18 Alkyl, more preferably C1-C 12 alkyl, most preferably C1-C6 alkyl), C2-C 24 Alkenyl (preferably C2-C 18 Alkenyl, more preferably C2-C 12 alkenyl, most preferably C2-C6 alkenyl), C2-C 24 Alkynyl (preferably C2-C 18 Alkynyl, more preferably C2-C 12 alkynyl, most preferably C2-C6 alkynyl), C5-C 24 Aryl (preferably C5-C 14 Aryl), C6-C 24 Alkaryl (preferably C6-C 18 Alkaryl), and C6-C 24 Aralkyl (preferably C6-C 18 aralkyl).

[0061] In some embodiments, R 2f is -C(O)R 2f* and R 3f and R 4f are H and R s is CH2CH2CH2OCH3. R 2f* Exemplary groups include substituted or unsubstituted C 1-8 Alkyl or substituted or unsubstituted C 3-8 Exemplary C 3-8Heteroaryl includes pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl. 1-8 Alkyl includes methyl, ethyl, isobutyl, and isopentyl.Preferred substituents include amino, carboxy, and sulfhydryl.

[0062] In some embodiments, R 2f teeth, [ka] The structure is In the formula, R aa is H, CH3, CH(CH3)2, CH2-phenyl, CH2-4-(hydroxyphenyl), CH2-(1H-imidazol-4-yl), CH2-(1H-indol-3-yl), CH2CH(CH3)2, CH2OH, CH(OH)CH3, CH2SH, CH2COOH, CH2CH2COOH, CH2CONH2, CH2CH2CONH2, (CH2)4NH2, CH2CH2CH2NHC(=NH)NH2, or CH2CH2SCH3.

[0063] In other embodiments, R 3f is -C(O)R f* and R 2f and R 4f are H and R s is CH2CH2CH2OCH3. R 2f* Exemplary groups include substituted or unsubstituted C 1-8 Alkyl or substituted or unsubstituted C 3-8 Exemplary C 3-8 Heteroaryl includes pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl. 1-8 Alkyl includes methyl, ethyl, isobutyl, and isopentyl.Preferred substituents include amino, carboxy, and sulfhydryl.

[0064] In some embodiments, R3f teeth, [ka] The structure is In the formula, R aa is H, CH3, CH(CH3)2, CH2-phenyl, CH2-4-(hydroxyphenyl), CH2-(1H-imidazol-4-yl), CH2-(1H-indol-3-yl), CH2CH(CH3)2, CH2OH, CH(OH)CH3, CH2SH, CH2COOH, CH2CH2COOH, CH2CONH2, CH2CH2CONH2, (CH2)4NH2, CH2CH2CH2NHC(=NH)NH2, or CH2CH2SCH3.

[0065] In a further embodiment, R 4f is -C(O)R f* and R 2f and R 3f are H and R s is CH2CH2CH2OCH3. R 2f* Exemplary groups include substituted or unsubstituted C 1-8 Alkyl or substituted or unsubstituted C 3-8 Exemplary C 3-8 Heteroaryl includes pyridin-2-yl, pyridin-3-yl, and pyridin-4-yl. 1-8 Alkyl includes methyl, ethyl, isobutyl, and isopentyl.Preferred substituents include amino, carboxy, and sulfhydryl.

[0066] In some embodiments, R 4f teeth, [ka] The structure is In the formula, R aais H, CH3, CH(CH3)2, CH2-phenyl, CH2-4-(hydroxyphenyl), CH2-(1H-imidazol-4-yl), CH2-(1H-indol-3-yl), CH2CH(CH3)2, CH2OH, CH(OH)CH3, CH2SH, CH2COOH, CH2CH2COOH, CH2CONH2, CH2CH2CONH2, (CH2)4NH2, CH2CH2CH2NHC(=NH)NH2, or CH2CH2SCH3.

[0067] In certain embodiments, R f* teeth, [ka] is selected from In the formula, n is 0, 1, 2, 3, 4, 6, 7, 8, 9, or 10, preferably 0, 1, 2, 3, or 4; Q f is CF3, -C≡CH, N3, -C≡N, COOH, -(CO)-O-alkyl, aryl (e.g., 4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl), adamantyl, C 1-10 Heterocyclyl, substituted or unsubstituted C 7-10 In some embodiments, n is 0, while in other embodiments, n is 3. In certain embodiments, n is 2, 3, or 4, and Q is cycloalkynyl. ... f C has the following formula: 1-10 is heterocyclyl, [ka] In the formula, X f is null, CH2, O, S, or NR xf and R xf is H or C 1-3 It is alkyl, preferably methyl.

[0068] In certain embodiments, R f* is -(CO)-O-alkyl, where alkyl is methyl or ethyl.

[0069] In some embodiments, R s teeth, [ka] is selected from In the formula, n is 0, 1, 2, 3, 4, 6, 7, 8, 9, or 10; Q s is CF3, -PPh3, -CH2NEt3, -CH2NMe3, -C≡CH, N3, -C≡N, COOH, -(CO)-O-alkyl, aryl (e.g., 4-fluorophenyl, 3-fluorophenyl, 2-fluorophenyl), adamantyl, C 1-8 Heterocyclyl, substituted or unsubstituted C 7-10 It is cycloalkynyl.

[0070] In certain embodiments, Q f teeth, [ka] may be a moiety having the formula: In the formula, X dqis an alkylene chain having 2-12 CH2 units, preferably 5-12 CH2 units, more preferably 7-11 CH2 units, and especially preferably 8-10 CH2 units. One skilled in the art will recognize that while some of the above moieties are shown in electronically neutral form, these moieties may be protonated and paired with appropriate counterions. Similarly, the quinolinium cations shown above are accompanied by a charge-balancing ion or ions, such as, for example, dichloride, dibromide, diiodide, diacetate, etc. The biosynthesis of naturally occurring apoptolidine and related macrolides by Nocardiopsis sp. FU40 begins with (R)-methoxymalonate, a substrate for (R)-2-methoxymalonyl-acyl carrier protein ("MeOM-ACP"). This protein is encoded by a five gene contiguous gene cassette (apoK-M2) and can be disrupted by double genetic replacement of apoJK to completely abolish the production of all apoptolidine macrolides. Fermentation with mutant Nocardiopsis sp. FU40 apoJK::aac(3)IV carrying N-acetylcysteamine, the (NAC) thioester of (R)-2-methoxymalonate, restores apoptolidin biosynthesis. 2b -C(O)SR st or R s -C(O)SR st (In the formula, R st Fermentation of Nocardiopsis sp. FU40 apoJK::aac(3)IV with thioesters of the corresponding R 2b or R s The resulting macrolide has a hydroxyl group.

[0071] As used herein, the phrase "therapeutic agent" refers to any agent that has a therapeutic and / or diagnostic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject.

[0072] Ammocidin compounds (e.g., ammocidin A, ammocidin B, ammocidin C, ammocidin D, ammocidin E, especially ammocidin A) and compounds of formula (1) (and subformulas thereof) may be effectively used for the treatment of proliferative disorders, including cancer and similar diseases. The compounds may be used to treat cancers characterized by one or more solid tumors. In other embodiments, the proliferative disorder is a hematological cancer. In some embodiments, the proliferative disorder is a myelodysplastic syndrome. In some embodiments, the proliferative disorder is a leukemia.

[0073] In a preferred embodiment, a naturally occurring ammocidin (e.g., ammocidin A) or a compound of formula (1), (2a), (2b), (2c), (3a), (3b), or (3c) may be used to treat a proliferative disorder such as acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, multiple myeloma, mixed lineage leukemia, brain tumor, glioblastoma, or lymphoma. Preferably, the agent used is ammocidin A or a compound of formula (3c).

[0074] The compounds disclosed herein are intended to be used in the treatment of acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), cancer in adrenocortical carcinoma, adrenal cortical carcinoma, AIDS-related cancer, Kaposi's sarcoma, AIDS-related lymphoma, primary CNS lymphoma, anal cancer, appendix cancer, carcinoid tumors, astrocytoma, atypical teratoid / rhabdoid tumors, basal cell carcinoma, skin cancer (non-melanoma), bile duct cancer, extrahepatic bladder cancer, bladder cancer, bone cancer (including Ewing's sarcoma and osteosarcoma and malignant fibrous histiocytoma), brain tumors, breast cancer, bronchial tumors, Burkitt's lymphoma (non-Hodgkin's), carcinoid tumors, cardiac tumors, heart tumors, tumors), atypical teratoid / rhabdoid tumor, germinal tumor, germ cell tumor, lymphoma, primary-cervical cancer, cholangiocarcinoma, chordoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), chronic myeloproliferative neoplasm, colorectal cancer, colorectal cancer, craniopharyngioma, cutaneous T-cell lymphoma, ductal carcinoma in situ (DCIS), germinal tumor, central nervous system, endometrial cancer, ependymoma, esophageal, nasal neuroblastoma, extracranial germ cell tumor, extragonadal germ cell tumor, eye cancer, intraocular melanoma, retinoblastoma, fallopian tube cancer, fibrous histiocytoma of bone, malignant and osteosarcoma, gallbladder cancer, stomach cancer, gastriccancer), gastric cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor (GIST), germ cell tumor, central nervous system, extracranial, extragonadal, ovarian testicular, gestational trophoblastic disease, glioma, hairy cell leukemia, head and neck cancer, cardiac tumor, hepatocellular (liver) cancer, histiocytosis, Langerhans cell, Hodgkin's lymphoma, hypopharyngeal cancer, intraocular melanoma, pancreatic islet cell tumor, pancreatic neuroendocrine tumor, Kaposi's sarcoma, renal-Langerhans cell histiocytosis, laryngeal cancer, laryngeal cancer and papillomatosis, leukemia, lip and oral cavity cancer, liver cancer (primary), lung cancer, lung cancer, lymphoma -Macroglobulinemia, Waldenström's non-Hodgkin's lymphoma, Male breast cancer, Malignant fibrous histiocytoma and osteosarcoma of bone, Melanoma, Intraocular (eye), Merkel cell carcinoma, Mesothelioma, Malignant, Mesothelioma, Metastatic squamous cell carcinoma of the neck with unknown primary site, Midline adenocarcinoma with NUT gene, Oral cancer, Multiple endocrine neoplasia syndrome, Multiple myeloma / plasma cell neoplasm, Mycosis fungoides, Myelodysplastic syndrome, Myelodysplastic / myeloproliferative neoplasms and chronic myeloproliferative neoplasms, Myeloid leukemia, chronic (CML), Myeloid leukemia, acute (AML), Nasal cavity and paranasal sinus cancer, Nasopharyngeal carcinoma, Nasopharyngeal carcinoma, Neuroblastoma Cell tumors, non-Hodgkin's lymphoma, non-small cell lung cancer, oral cavity cancer, lip and oral cavity cancer and oropharyngeal cancer, osteosarcoma and malignant fibrous histiocytoma of bone, ovarian cancer, pancreatic cancer and pancreatic neuroendocrine tumors (islet cell tumors), papillomatosis, paraganglioma, paraganglioma, paranasal sinus and nasal cancer, parathyroid cancer, penile cancer, pharyngeal cancer, pheochromocytoma, pheochromocytoma, pituitary tumor, plasma cell neoplasm / multiple myeloma, pleuropulmonary blastoma, pregnancy and breast cancer, primary central nervous system (CNS) lymphoma, primary peritoneal cancer, prostate cancer, rectal cancer, renal cell (kidney) cancer, retinoblastoma, rhabdomyosarcoma, salivary gland It may be used to treat cancer, salivary gland tumors, Ewing's sarcoma, Kaposi's sarcoma, osteosarcoma, rhabdomyosarcoma, uterine sarcoma, vascular tumors, Sezary syndrome, skin cancer, small cell lung cancer, small intestine cancer, soft tissue sarcoma, squamous cell carcinoma, cervical squamous cell carcinoma of unknown primary, metastatic, stomach cancer, gastric cancer, T-cell lymphoma, skin, testicular cancer, throat cancer, thymoma and thymic carcinoma, thyroid cancer, ureter and renal pelvis, transitional cell carcinoma, urethral cancer, uterine cancer, endometrial and uterine sarcoma, vaginal cancer, vaginal cancer, vascular tumors, vulvar cancer, Waldenstrom's macroglobulinemia, Wilms' tumor.

[0075] In the field of medical oncology, it is common practice to use a combination of different forms of treatment to treat each patient with cancer.In medical oncology, in addition to the composition of the present invention, the other component(s) of such combination treatment can be, for example, surgery, radiation therapy, chemotherapy, signal transduction inhibitors, and / or monoclonal antibodies.Therefore, the compounds disclosed herein can be administered as part of a combination treatment regime, for example, before or after surgery, or before or after radiation therapy.

[0076] In some embodiments, the compounds disclosed herein may be administered in combination with one or more anti-cancer agents, such as mitotic inhibitors, alkylating agents, antimetabolites, antisense DNA or RNA, intercalating antibiotics, growth factor inhibitors, signal transduction inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators, proteasome inhibitors, topoisomerase inhibitors, epigenetic inhibitors, biological response modifiers, antimetabolites, glycolysis inhibitors, glutamine metabolism inhibitors, antihormones, angiogenesis inhibitors, cytostatics antiandrogens, targeted antibodies, HMG-CoA reductase inhibitors, and prenyl-protein transferase inhibitors.

[0077] Exemplary anti-cancer agents include nucleoside analogs, antifolates, antimetabolites, topoisomerase I inhibitors, anthracyclines, podophyllotoxins, taxanes, vinca alkaloids, alkylating agents, platinum compounds, proteasome inhibitors, nitrogen mustards and estrogen analogs, monoclonal antibodies, tyrosine kinase inhibitors, mTOR inhibitors, retinoids, immunomodulators, histone deacetylase inhibitors, DNA methyl-transferase inhibitors, BCL-2 family protein inhibitors, and combinations thereof.

[0078] In some embodiments, the additional anticancer agent may be one or more nucleoside analogs, such as one or more azanucleosides.

[0079] In certain embodiments, the anticancer drug is abiraterone acetate, methotrexate, paclitaxel albumin stabilized nanoparticles, brentuximab vedotin, ado-trastuzumab emtansine, doxorubicin hydrochloride, afatinibdi maleate, everolimus, netupitant, palonosetron hydrochloride, imiquimod, aldesleukin, alectinib, alemtuzumab, melphalan hydrochloride, melphalan, pemetrexed disodium, chlorambucil, aminolevulinic acid, anastrozole, aprepitant, pamidronate disodium, exemceme. Stan, nelarabine, arsenic trioxide, ofatumumab, asparaginase erwinia chrysanthemi, atezolizumab, bevacizumab, axitinib, azacitidine, carmustine, belinostat, bendamustine hydrochloride, bevacizumab, bexarotene, tositumomab, bicalutamide, bleomycin, blinatumomab, bortezomib, bosutinib, busulfan, cabazitaxel, cabozantinib, alemtuzumab, irinotecan hydrochloride, capecitabine, fluorouracil, carboplatin, carfilzomib, bicalutamide, lomustine , ceritinib, daunorubicin hydrochloride, cetuximab, chlorambucil, cyclophosphamide, clofarabine, cobimetinib, dactinomycin, cobimetinib, crizotinib, ifosfamide, ramucirumab, cytarabine, dabrafenib, carbazine, decitabine, daratumumab, dasatinib, daunorubicin hydrochloride, decitabine, efibrotide sodium, defibrotide sodium, degarelix, denileukin diftitox, denosumab, dexamethasone, dexrazoxane hydrochloride, dinutuximab, docetaxel, doxorubicin hydrochloride, carba gin, rasburicase, epirubicin hydrochloride, elotuzumab, oxaliplatin, eltrombopag olamine, aprepitant, elotuzumab, enzalutamide, epirubicin hydrochloride, cetuximab, eribulin mesylate, vismodegib, erlotinib hydrochloride, etoposide, raloxifene hydrochloride, melphalan hydrochloride, toremifene, panobinostat, fulvestrant, letrozole, filgrastim, fludarabine phosphate, flutamide, methotrexate, pralatrexate, recombinant HPV quadrivalent vaccine, recombinant HPV nonavalent vaccine, obinutuzumab,Gefitinib, gemcitabine hydrochloride, gemtuzumab ozogamicin, afatinib dimaleate, imatinib mesylate, glucarpidase, goserelin acetate, eribulin mesylate, trastuzumab, topotecan hydrochloride, palbociclib, ibritumomab tiuxetan, ibrutinib, ponatinib hydrochloride, idarubicin hydrochloride, idelalisib, imiquimod, axitinib, recombinant interferon alpha-2b, tositumomab, ipilimumab, gefitinib, romidepsin, ixabepilone, ixazomib citrate, ruxolitinib phosphate, cabazita Xel, ado-trastuzumab emtansine, palifermin, pembrolizumab, lanreotide acetate, lapatinib ditosylate, lenalidomide mesylate, leuprolide acetate, olaparib, vincristine sulfate, procarbazine hydrochloride, mechlorethamine hydrochloride, megestrol acetate, trametinib, mercaptopurine, temozolomide, mitoxantrone hydrochloride, plerixafor, busulfan, azacitidine, gemtuzumab ozogamicin, vinorelbine tartrate, necitumumab, nelarabine, sorafenib tosylate, nilotinib, que Ixazomib nitrate, nivolumab, romiplostim, obinutuzumab, ofatumumab, olaparib, omacetaxine mepesuccinate, pegaspargase, ondansetron hydrochloride, osimertinib, panitumumab, panobinostat, peginterferon alfa-2b, pembrolizumab, pertuzumab, plerixafor, pomalidomide, ponatinib hydrochloride, necitumumab, pralatrexate, procarbazine hydrochloride, aldesleukin, denosumab, ramucirumab, rasburicase, regorafenib, lenalidomide, rituximab, salt Rolapitant acid, romidepsin, ruxolitinib phosphate, siltuximab, dasatinib, sunitinib malate, thalidomide, dabrafenib, osimertinib, talimozine, atezolizumab, temsirolimus, thalidomide, dexrazoxane hydrochloride, trabectedin, trametinib, trastuzumab, lapatinib ditosylate, dinutuximab, vandetanib, rolapitant hydrochloride, bortezomib, venetoclax, crizotinib, enzalutamide, ipilimumab, trabectedin, ziv-aflibercept, idelalisib, ceritinib,and pharma- ceutically acceptable salts thereof.

[0080] The additional agents may be combined with the disclosed compounds in a single pharmaceutical formulation (as defined herein) or may be administered separately.

[0081] In a preferred embodiment, a naturally occurring ammocidin (e.g., ammocidin A) or a compound of formula (1), (2a), (2b), (2c), (3a), (3b), or (3c) may be combined with one or more agents for the treatment of leukemia, such as acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, or mixed lineage leukemia. Exemplary agents include vincristine, azacitidine, decitabine, cytarabine, daunomycin, venetoclax, ibrutinib, idelalisib, doxorubicin, idarubicin (or another anthracycline), L-asparaginase, PEG-L-asparaginase, cyclophosphamide, nelarabine, cladribine, fludarabine, mitoxantrone, etoposide, hydroxyurea, methotrexate, 6-mercaptopurine, azacitidine, decitabine, prednisone, dexamethasone, or other corticosteroids. Preferably, one or more agents for leukemia are combined with either ammocidin A or a compound of formula (3c).

[0082] In some preferred embodiments, leukemia may be treated using a naturally occurring ammocidin (e.g., ammocidin A) or a compound of formula (1), (2a), (2b), (2c), (3a), (3b), or (3c) in combination with venetoclax, one or more nucleoside analogs or azanucleosides (e.g., azacitidine, decitabine, cedazuridine) or both, and venetoclax and one or more nucleoside analogs or azanucleosides. Preferably, azacitidine, venetoclax, or both azacitidine and venetoclax are combined with either ammocidin A or a compound of formula (3c). In other embodiments, ammocidin A or a compound of formula (3c) is administered in combination with decitabine and cedazuridine.

[0083] In some embodiments, the compounds disclosed herein may be administered to a patient in need of the compound without a corresponding increase in multidrug resistance. In such embodiments, the disclosed compounds may be administered alone or in combination with one or more additional anticancer agents as defined herein. In some embodiments, the disclosed compounds may be used to treat cancers in patients who have developed multidrug resistance, such as leukemias, such as AML.

[0084] Multidrug resistance may be identified using methods known in the art. In some embodiments, the patient may have multidrug resistance characterized by abnormal levels of P-glycoprotein, including elevated levels of P-glycoprotein (P-gp). In some cases, the patient may have multidrug resistance characterized by upregulation of P-gp, downregulation of anti-apoptotic protein B-cell lymphoma (Bcl-2), or both upregulation of P-gp and downregulation of Bcl-2. The patient may have multidrug resistance characterized by overexpression of multidrug resistance associated protein (MRP1) or overexpression of the ABCC1 gene. The patient may have multidrug resistance characterized by overexpression of lung resistance protein (LRP). The patient may have multidrug resistance characterized by overexpression of glutathione S-transferase (GST), including GSTα, GSTμ, or GSTπ. The patient may have multidrug resistance characterized by overexpression of PKCα, PKC e, and PKC q The patient may have multidrug resistance characterized by upregulation of protein kinase C (PKC), including FMS-like tyrosine kinase 3 (FLT3). The patient may have multidrug resistance characterized by expression of Wilms' tumor (WT1). The patient may have multidrug resistance characterized by RAS mutations, e.g., KRAS mutations, HRAS mutations, or NRAS mutations. The patient may have multidrug resistance characterized by mutations in one or more of IDH1, TP53, ASXL1, DNMT3A, CEBPA, IDH2, PTPN11. The patient may have multidrug resistance characterized by differentiation status, such as FAB classification, including primitive (M0) or monocytic (M5) differentiation.

[0085] The pharmaceutical compositions described herein may be prepared by any method known or hereafter developed in the art of pharmacy. In general, such methods of preparation include the step of bringing into association the active ingredient with one or more excipients and / or one or more other accessory ingredients, and then, if necessary and / or desired, shaping and / or packaging the product into the desired single or multiple dosage unit.

[0086] Dosage forms for topical or transdermal administration of the scaffold include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, and / or patches. In general, the active component is mixed under sterile conditions with a pharma- ceutically acceptable excipient and any necessary preservatives or buffers as necessary. Additionally, the present invention contemplates the use of transdermal patches, which often have the added advantage of providing controlled delivery of the active ingredient to the body. Such dosage forms can be prepared, for example, by dissolving and / or dispensing the active ingredient in a suitable medium. Alternatively or additionally, the rate can be controlled by either providing a rate-controlling membrane and / or dispersing the active ingredient in a polymer matrix and / or gel.

[0087] Pharmaceutically acceptable excipients used in the manufacture of pharmaceutical compositions include, but are not limited to, inert diluents, dispersing and / or granulating agents, surface active agents and / or emulsifying agents, disintegrating agents, binders, preservatives, buffers, lubricants, and / or oils. Such excipients may be optionally included in the formulation of the present invention. Excipients such as cocoa butter and suppository waxes, coloring agents, coating agents, sweetening agents, flavoring agents, and perfumes may be present in the composition according to the judgment of the formulator.

[0088] Exemplary diluents include, but are not limited to, calcium carbonate, sodium carbonate, calcium phosphate, dicalcium phosphate, calcium sulfate, monobasic calcium phosphate, sodium phosphate, lactose, sucrose, cellulose, microcrystalline cellulose, kaolin, mannitol, sorbitol, inositol, sodium chloride, dry starch, corn starch, powdered sugar, and the like, and combinations thereof.

[0089] Exemplary granulating and / or dispersing agents include, but are not limited to, potato starch, corn starch, tapioca starch, sodium starch glycolate, clays, alginic acid, guar gum, citrus pulp, agar, bentonite, cellulose, and wood products, natural sponge, cation exchange resins, calcium carbonate, silicates, sodium carbonate, cross-linked polyvinylpyrrolidone) (crospovidone), sodium carboxymethyl starch (sodium starch glycolate), carboxymethylcellulose, cross-linked sodium carboxymethylcellulose (croscarmellose), methylcellulose, partially pregelatinized starch (starch 1500), microcrystalline starch, water insoluble starch, calcium carboxymethylcellulose, magnesium aluminum silicate (Veegum), sodium lauryl sulfate, quaternary ammonium compounds, and the like, and combinations thereof.

[0090] Exemplary surfactants and / or emulsifiers include, but are not limited to, natural emulsifiers (e.g., acacia, agar, alginic acid, sodium alginate, tragacanth, chondrux, cholesterol, xanthan, pectin, gelatin, egg yolk, casein, wool fat, cholesterol, wax, and lecithin), colloidal clays (e.g., bentonite [aluminum silicate] and Veegum [magnesium aluminum silicate]), long chain amino acid derivatives, high molecular weight alcohols (e.g., stearyl alcohol, cetyl alcohol, oleyl alcohol, triacetin monostearate, distearate, glyceryl s ... Examples of suitable cellulose derivatives include polyoxyethylene sorbitan monolaurate, polyoxyethylene sorbitan monolaurate, polyvinyl alcohol, polyacrylic acid, acrylic acid polymers, and carboxyvinyl polymers, carrageenan, cellulose derivatives (e.g., sodium carboxymethylcellulose, powdered cellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, methylcellulose), sorbitan fatty acid esters (e.g., polyoxyethylene sorbitan monolaurate [Tween 20], polyoxyethylene sorbitan [Tween 60], polyoxyethylene sorbitan monooleate [Tween 80], sorbitan monopaltimate [Span 40], sorbitan monooleate [Span 60], sorbitan tristearate [Span 65], glyceryl monooleate, sorbitan monooleate [Span 80]), polyoxyethylene esters (e.g., polyoxyethylene monostearate [Myrj 45], polyoxyethylene hydrogenated castor oil, polyethoxylated castor oil, polyoxymethylene stearate, and Solutol), sucrose fatty acid esters, polyethylene glycol fatty acid esters (e.g., Cremophor), polyoxyethylene ethers, (e.g., polyoxyethylene lauryl ether [Brij30]), poly(vinyl-pyrrolidone), diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium lauryl sulfate, Pluronic F 68, Poloxamer 188, cetrimonium bromide, cetylpyridinium chloride, benzalkonium chloride, docusate sodium, and the like, and / or combinations thereof.

[0091] Exemplary binders include, but are not limited to, starches (e.g., corn starch and starch paste), gelatin, sugars (e.g., sucrose, glucose, dextrose, dextrin, molasses, lactose, lactitol, mannitol), natural and synthetic gums (e.g., acacia, sodium alginate, extract of Irish moss, panwar gum, ghatti gum, mucilage of isapol husk, carboxymethylcellulose, methylcellulose, ethylcellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, microcrystalline cellulose, cellulose acetate, polyvinylpyrrolidone), magnesium aluminum silicate (Veegum), and larch arabogalactan), alginates, polyethylene oxide, polyethylene glycol, inorganic calcium salts, silicic acid, polymethacrylates, waxes, water, alcohol, and combinations thereof.

[0092] Exemplary preservatives may include antioxidants, chelating agents, antibacterial preservatives, antifungal preservatives, alcohol preservatives, acidic preservatives, and other preservatives. Exemplary antioxidants include, but are not limited to, alpha tocopherol, ascorbic acid, acorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, monothioglycerol, potassium metabisulfite, propionic acid, propyl formate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, and sodium sulfite. Exemplary chelating agents include ethylenediaminetetraacetic acid (EDTA), citric acid monohydrate, disodium edetate, dipotassium edetate, edetic acid, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, and trisodium edetate. Exemplary antibacterial preservatives include, but are not limited to, benzalkonium chloride, benzethonium chloride, benzyl alcohol, bronopol, cetrimide, cetylpyridinium chloride, chlorhexidine, chlorobutanol, chlorocresol, chloroxylenol, cresol, ethyl alcohol, glycerin, hexetidine, imidurea, phenol, phenoxyethanol, phenylethyl alcohol, phenylmercuric nitrate, propylene glycol, and thimerosal. Exemplary antifungal preservatives include, but are not limited to, butylparaben, methylparaben, ethylparaben, propylparaben, benzoic acid, hydroxybenzoic acid, potassium benzoate, potassium sorbate, sodium benzoate, sodium propionate, and sorbic acid. Exemplary alcohol preservatives include, but are not limited to, ethanol, polyethylene glycol, phenol, phenolic compounds, bisphenol, chlorobutanol, hydroxybenzoate, and phenylethyl alcohol. Exemplary acidic preservatives include, but are not limited to, vitamin A, vitamin C, vitamin E, beta-carotene, citric acid, acetic acid, dehydroacetic acid, ascorbic acid, sorbic acid, and phytic acid.Other preservatives include, but are not limited to, tocopherol, tocopherol acetate, dethoxime mesylate, cetrimide, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), ethylenediamine, sodium lauryl sulfate (SLS), sodium lauryl ether sulfate (SLES), sodium sulfite, sodium metabisulfite, potassium sulfite, potassium metabisulfite, Glydant Plus, Phenonip, methylparaben, Germall 115, Germaben II, Neolone, Kathon, and Euxyl. In certain embodiments, the preservative is an antioxidant. In other embodiments, the preservative is a chelating agent.

[0093] Exemplary buffering agents include, but are not limited to, citrate buffer, acetate buffer, phosphate buffer, ammonium chloride, calcium carbonate, calcium chloride, calcium citrate, calcium glubionate, calcium gluceptate, calcium gluconate, D-gluconic acid, calcium glycerophosphate, calcium lactate, propanoic acid, calcium levulinate, pentanoic acid, dicalcium phosphate, phosphoric acid, tricalcium phosphate, calcium hydroxide phosphate, potassium acetate, potassium chloride, potassium gluconate, potassium mixtures, dipotassium phosphate, monopotassium phosphate, potassium phosphate mixtures, sodium acetate, sodium bicarbonate, sodium chloride, sodium citrate, sodium lactate, disodium phosphate, monosodium phosphate, sodium phosphate mixtures, tromethamine, magnesium hydroxide, aluminum hydroxide, alginic acid, pyrogen-free water, isotonic saline, Ringer's solution, ethyl alcohol, and the like, and combinations thereof.

[0094] Exemplary lubricants include, but are not limited to, magnesium stearate, calcium stearate, stearic acid, silica, talc, malt, glyceryl behanate, hydrogenated vegetable oils, polyethylene glycol, sodium benzoate, sodium acetate, sodium chloride, leucine, magnesium lauryl sulfate, sodium lauryl sulfate, and the like, and combinations thereof.

[0095] Exemplary oils include, but are not limited to, almond, apricot kernel, avocado, babassu, bergamot, black currant seed, borage, cade, chamomile, canola, caraway, carnauba, castor, cinnamon, cocoa butter, coconut, cod liver, coffee, corn, cottonseed, emu, eucalyptus, evening primrose, fish, linseed, geraniol, gourd, grape seed, hazelnut, hyssop, isopropyl myristate, jojoba, kukui nut, and lavandin. , lavender, lemon, litsea cubeba, macadamia nut, mallow, mango seed, meadowfoam seed, mink, nutmeg, olive, orange, orange roughy, palm, palm kernel, peach kernel, peanut, poppy seed, pumpkin seed, rapeseed, rice bran, rosemary, safflower, sandalwood, camellia, peppermint, sea buckthorn, sesame, shea butter, silicone, soybean, sunflower, tea tree, thistle, camellia, vetiver, walnut, and wheat germ oil. Exemplary oils include, but are not limited to, butyl stearate, caprylic triglyceride, capric triglyceride, cyclomethicone, diethyl sebacate, dimethicone 360, isopropyl myristate, mineral oil, octyldodecanol, oleyl alcohol, silicone oil, and combinations thereof.

[0096] Liquid dosage forms for oral and parenteral administration include, but are not limited to, pharma- ceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups, and elixirs.In addition to the active ingredient, liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (e.g., cottonseed, peanut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof.In addition to inert diluents, oral compositions may contain adjuvants, such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents, and perfuming agents. In certain embodiments for parenteral administration, the conjugate is mixed with a solubilizing agent such as Cremophor, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers, and combinations thereof.

[0097] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, can be formulated according to known techniques using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations can be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, for example, solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that can be used are water, Ringer's solution, USP, and isotonic sodium chloride solution. In addition, sterile fixed oils are conventionally used as solvents or suspending media. For this purpose, any non-irritating fixed oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid can be used in the preparation of injectables.

[0098] In order to prolong the effect of active ingredient, it is often desirable to delay the absorption of active ingredient from subcutaneous or intramuscular injection.This can be achieved by using a liquid suspension of crystalline or amorphous substance that is poorly water-soluble.Then, the absorption rate of active ingredient depends on its dissolution rate, which in turn depends on crystal size and crystalline form.In some embodiments, the absorption delay of parenterally administered active ingredient is achieved by dissolving or suspending the drug in oil vehicle.

[0099] Compositions for rectal or vaginal administration can typically be prepared by mixing the conjugate with a suitable non-irritating excipient, such as cocoa butter, polyethylene glycol, or a suppository wax, which are solid at ambient temperature, but liquid at body temperature, and will melt in the rectum or vaginal cavity and release the active ingredient.

[0100] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active ingredient is mixed with at least one inert, pharma- ceutically acceptable excipient such as sodium citrate or dicalcium phosphate, and / or (a) fillers or extenders such as starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; (c) humectants such as glycerol; (d) disintegrating agents such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (e) solution retarding agents such as paraffin; (f) absorption accelerators such as quaternary ammonium compounds; (g) wetting agents such as, for example, cetyl alcohol and glycerol monostearate; (h) absorbents such as kaolin and bentonite clay; and (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may comprise buffering agents.

[0101] Solid compositions of a similar type may be used as fillers for soft and hard filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols. Solid dosage forms of tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical art. They may optionally contain opacifying agents, and may optionally be of a composition that releases the active ingredient(s) only, or preferentially, in a certain part of the gastrointestinal tract in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may be used as fillers for soft and hard filled gelatin capsules using excipients such as lactose or milk sugar as well as high molecular weight polyethylene glycols.

[0102] The compounds disclosed herein are useful for treating various conditions in which inhibition of ATP synthase is beneficial.Specifically, the compounds are useful for treating blood cancers, including myeloid leukemia (both chronic and acute), lymphocytic leukemia (including chronic lymphocytic leukemia, non-Hodgkin's lymphoma, and mantle cell lymphoma), and myeloma.The compounds can also be used to treat glioblastoma, including ENO1-deficient glioblastoma, prostate cancer, and lung cancer. EXAMPLES

[0103] The following examples are intended only to illustrate the invention and are not intended to limit the scope of the invention in any manner.

[0104] Example 1: Ammocidin A fermentation Ammocidin A was obtained by culturing Saccharothrix sp. AJ9571, provided by Ajinomoto Co., Inc. (Kawasaki, Japan). The microorganism was plated on Bennett agar (0.1% yeast extract, 0.1% beef extract, 0.2% NZ amine type A, 1.0% dextrose, 2.0% agar, pH 7.0) and incubated at 30°C for 3-7 days until sporulation. Spores scraped from solid cultures were used to start seed cultures in 250-mL Erlenmeyer flasks containing 50 mL of seed medium (1.0% soluble starch, 1.0% molasses (Plantation Blackstrap, non-sulfurized), 1.0% peptone, 1.0% beef extract, pH 7.0) and incubated at 30°C for 7 days with shaking at 220 RPM. Production cultures were carried out in multiple 250 mL Erlenmeyer flasks containing 50 mL of production medium (2.0% glycerol, 1.0% molasses, 0.5% casamino acids, 0.1% peptone, 0.4% calcium carbonate, pH 7.2) and incubated at 30° C. for 7 days with shaking at 220 RPM.

[0105] After 7 days of fermentation, mycelia were separated from the culture broth by centrifugation at 3000g × 30 min. The culture broth was extracted three times with one volume of ethyl acetate, and the combined organic layers were washed with brine, dried over Na2SO4, and concentrated in vacuum. The crude extract was then subjected to chromatography with LH-20 resin using methanol as the mobile phase, and glycomacrolide-containing fractions were identified by thin-layer chromatography and pooled. The LH-20 fraction was then subjected to reverse-phase HPLC using a Waters XBridge Prep C18 19 × 150 mm column with a 20 min gradient of 70%A / 30%B to 20%A / 80%B (Buffer A: 95% water, 5% acetonitrile, 10 mM ammonium acetate; Buffer B: 5% water, 95% acetonitrile, 10 mM ammonium acetate). Ammocidin A-RT 9.0 min. Fractions containing the pure compound were lyophilized using a Genevac HT-6 to give a white solid.

[0106] Example 2: Ammocidin A Derivatization Derivatization with 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propanoic acid To a solution of 3-(3-(but-3-yn-1-yl)-3H-diazirin-3-yl)propanoic acid (4.5 mg, 0.027 mmol, Enamine, Kyiv, Ukraine) in dichloromethane (4.0 mL) on ice was added bromo-tris-pyrrolidinophosphonium hexafluorophosphate (PyBrop, 13.3 mg, 0.28 mmol) and diisopropylethylamine (DIPEA, 31 μL, 0.177 mmol). The resulting solution was stirred at 0° C. for 10 min. Ammocidin A (20 mg 0.018 mmol) was added, followed by a crystal of 4-dimethylaminopyridine (DMAP). The resulting solution was allowed to warm to room temperature overnight (16 h). The reaction was monitored by TLC (90:10 CHCl3:MeOH), quenched with 100 μL MeOH, and then concentrated. The resulting residue was diluted in EtOAc (20 mL) and washed with 1M HCl (5 mL). The aqueous layer was extracted twice with EtOAc (2×10 mL). The organic extracts were combined, washed with NaHCO3 (5 mL) and brine (5 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The resulting residue was dissolved in 800 μL of MeOH and purified by reverse-phase HPLC using a Waters XBridge Prep C18 19×150 mm column with a 20 min gradient of 32% to 77% acetonitrile in water containing 25 mM ammonium bicarbonate. Fractions containing the desired product (rt=16 min) were combined and lyophilized to give the product as a white solid (0.6 mg, 2.5% isolated yield). HRMS (ESI-TOF MS) m / z 1287.6854 (M+Na) + Calculated value 1287.6866, observed value (0.9 ppm). [ka] The regiochemistry of the addition was confirmed by multidimensional NMR. [ka]

[0107] Derivatization with 4-morpholinobutanoic acid To a solution of 4-morpholinobutanoic acid (2.2 mg, 0.013 mmol) in dichloromethane (2.0 mL) on ice was added bromo-tris-pyrrolidinophosphonium hexafluorophosphate (PyBrop, 7 mg, 0.14 mmol) and diisopropylethylamine (DIPEA, 16 μL, 0.09 mmol). The resulting solution was stirred at 0° C. for 10 min. Ammocidin A (10 mg 0.009 mmol) was added, followed by a crystal of 4-dimethylaminopyridine (DMAP). The resulting solution was allowed to warm to room temperature overnight (16 h). The reaction was monitored by TLC (90:10 CHCl3:MeOH), quenched with 100 μL MeOH, and then concentrated. The resulting residue was diluted in EtOAc (20 mL) and washed with 1 M HCl (5 mL). The aqueous layer was extracted twice with EtOAC (2 x 10 mL). The organic extracts were combined, washed with NaHCO3 (5 mL) and brine (5 mL), dried over anhydrous sodium sulfate, and concentrated in vacuo. The resulting residue was dissolved in 800 μL of MeOH and purified by reverse-phase HPLC using a Waters XBridge Prep C18 19 x 150 mm column with a 20 min gradient of 32% to 77% acetonitrile in water containing 25 mM ammonium bicarbonate. Two major products were seen as distinct peaks and collected in separate fractions. HRMS (ESI-TOF MS) confirmed the addition of the ester (M+H). + Calculated 1312.7412 m / z, observed 1312.7400 m / z (0.9 ppm). 2D NMR (HSQC, HMBC) confirmed the two products as the 2' and 3' esters.

[0108] 3-(4-Methylpiperazin-1-yl)butanoic acid ester was prepared using a similar method and the addition was confirmed by HRMS (ESI-TOF MS) (M+H). + 1325.7729 m / z calculated, 1325.7704 observed (1.9 ppm).

[0109] Example 3: Apoptolidin biosynthetic modification The translated sequence of the five gene (R)-2-methoxymalonyl-acyl carrier protein (MeOM-ACP) contiguous gene cassette (apoK-M2) has extensive sequence identity to fkbG-K, which encodes the biosynthesis of this extender unit from the FK520 gene cluster of Streptomyces hygroscopicus, and ApoJ shows 53% identity to the acyl carrier protein of the extender unit. However, although MeOMal-ACP has been reported to function as an extender unit by intercepting polyketide synthases in trans, it has never been reported as a chain initiator.

[0110] To disrupt apoJ in this cluster, a two-step PCR targeted replacement was used, where the gene was first replaced by an antibiotic resistance marker in a fosmid containing the apo gene cluster, which was then transferred to Nocardiopsis to select for a double crossover event. Attempts to replace the 291 bp apoJ did not yield any recombinant clones, but double gene replacement of apoJK was successful, resulting in the mutant strain Nocardiopsis sp.FU40 apoJK::aac(3)IV. The translated apoK gene shares 65% identity with FkbK, the oxidase responsible for the 3-OH dehydrogenation of glyceryl-ACP en route to hydroxymalonate, and its deletion is predicted to have no effect on downstream apoptolysin biosynthesis. LC-MS analysis of extracts of fermentation cultures of this strain demonstrated complete abolition of the production of all apoptolysins, supporting the hypothesis of (R)-2-methoxymalonyl-ACP biosynthesis initiation (Figure 4).

[0111] Synthetic thioesters of chain initiation and elongation building blocks and intermediates have been shown to load the KS active site cysteine ​​thiol domain in vitro and have also been used successfully in chemical complementation studies of blocked polyketide biosynthetic pathways. Chemical rescue of apoJK knockout strains was performed with the N-acetylcysteamine, (NAC) thioester of (R)-2-methoxymalonate. Initial studies of synthetic MeOMe-SNAC added to early stage Nocardiopsis growth cultures substantially restored apoptolidin A biosynthesis. Optimal incorporation efficiency was determined by evaluating pulse dosing schedules where 60ug / mL was fed in equal portions over a 7 day fermentation. It was determined that pulse supplementation of MeOMe-SNAC in 50μL aliquots of 8mg / mL in DMSO starting on day 2 of the seed culture yielded the best results, where apoptolidin A production was restored to near wild type levels. These results are consistent with the acylation site of the first polyketide synthase protein, ApoS1, being the active cysteine ​​in the second KS domain.

[0112] Example 4: Bypass fermentation with synthetic starter units Thiophenyl esters of methoxyacetate 9, 2-azidoacetate 10, 2-hydroxyacetate 11, and 2-bromoacetate 12 were prepared and supplemented to Nocardiopsis sp. FU40 apoJK::aac(3)IV cultures. LC / MS analysis of the extracts revealed that the phenyl thioester of methoxyacetate successfully complemented the apoJK deletion (Figure 4C, trace v), and that the 2-azidoacetate bypass yielded a new metabolite of m / z = 1139 with UV max of 290 and 330 nm, consistent with the properties of other apoptolysins, suggesting successful incorporation of the azide at C28 (Figure 4C, trace vi). To confirm the identity of this newly observed compound, collision-induced dissociation studies were performed on the putative azide-containing analog. The three sugars of apoptolysin provide diagnostic fragments for identification. Loss of the C27 sugar by dehydration yielded fragments of m / z 306 and 835. Subsequent loss of the C9 sugar resulted in fragments at m / z 163 and 675. Finally, the observation of fragmentation across the C22-C23 bond and dehydration (m / z of 457) confirmed the incorporation of an azide at the terminus of apoptolysin.

[0113] Example 5: Cell proliferation assay (CellTitre-Glo) Compounds were diluted in DMSO (less than 0.05% DMSO) and dispensed into 384-well plates using an Echo 555 liquid handler (Labcyte). After compound addition, cells were pipetted into 384-well plates at a concentration of 2,000-8,000 cells per well in IMDM or RPMI medium, as described above, supplemented with 10% FBS, and incubated at 37°C, 5% CO2 in a tissue culture incubator. Plates were incubated for 48 hours, and cell viability was measured using CellTiter-Glo reagent (Promega). Percent viability was defined as the relative luminescence units (RLU) of each well divided by the RLU of cells in the DMSO control. Dose-response curves and GI50 values ​​were determined using linear regression of double log-transformed data (GraphPad Prism version 6.0h). Control bone marrow-derived CD34+ cells were purchased from STEMCELL Technologies.

[0114] 100,000 cells ml -1 100 μl of suspended cells were added to wells of microtiter plates precoated with 0.5 μl of test compound in dimethyl sulfoxide at 200x and incubated for 48-72 h. MTT reagent was dissolved in fresh medium at 1 mg ml-1 and 100 μl was added to each well to achieve a final concentration of 0.5 mg ml-1 and incubated at 37 °C for 2 h. Cells were centrifuged at 800 g for 5 min and decanted. MTT crystals were redissolved in 100 μl of dimethyl sulfoxide, allowed to incubate at room temperature for 5 min and read at 560 nm using a SpectraMax plus 384 plate reader (Molecular Devices). Absorbance values ​​were normalized by background subtraction (wells without cells) such that vehicle-treated cells had a viability of 1.0. Concentration-response curves were fitted using the DRC R package with a four-parameter log-logistic function. Statistical tests of differences between concentration-response curves were performed using the EDcomp function with default parameters for comparison of half-maximal inhibitory concentration values ​​and the paramcomp function for comparison of other parameters.

[0115] The compounds listed in the table below were prepared using the techniques described herein. [Table 1]

[0116] Example 6: Pharmacokinetic studies All animal studies were performed in accordance with guidelines approved by the IACUC at Vanderbilt University Medical Center. The pharmacokinetics of ammocidin in NSGS male mice in biological triplicates was evaluated in whole blood after intraperitoneal administration of ammocidin alone at 0.5 mg kg-1. Whole blood samples were collected into EDTA tubes for analysis of plasma. Plasma was mixed 1:1 with an internal standard solution consisting of 1 μM Apop A in PBS. Metabolites were extracted with 200 μl ethyl acetate, evaporated to dryness, and resuspended in 50 μl MeOH. Ammo A concentrations were determined using liquid chromatography-MS (Thermo TSQ Quantum Access Max) with isocratec 60 / 40 HO / acetonitrile + 10 mM ammonium acetate at 250 μl min−1 in technical duplicate on a 50 × 1.8 mm C18 column in electrospray ionization-positive mode monitoring to be Ammo A (1139.7→208.8, collision energy (CE) 19 V, retention time (RT) = 1.00 min) and Apop A (1146.68→805.46, CE 19 V, RT = 1.66 min).

[0117] The compositions and methods of the appended claims are not limited in scope by the specific compositions and methods described herein, which are intended as illustrations of some aspects of the claims, and any compositions and methods that are functionally equivalent are intended to fall within the scope of the claims. Various modifications of the compositions and methods in addition to those shown and described herein are intended to fall within the scope of the appended claims. Furthermore, while only certain representative compositions and method steps disclosed herein have been specifically described, other combinations of compositions and method steps are also intended to fall within the scope of the appended claims even if not specifically recited. Thus, although a step, element, component, or combination of components may be explicitly recited or less herein, other combinations of steps, elements, components, and components are included even if not explicitly recited. As used herein, the term "comprising" and variations thereof are used synonymously with the term "including" and variations thereof, and are open-ended, non-limiting terms. Although the terms "comprising" and "including" have been used herein to describe various embodiments, the terms "consisting essentially of" and "consisting of" can be used in place of "comprising" and "including" to provide more specific embodiments of the present invention, and are also disclosed. All numbers expressing amounts of ingredients, reaction conditions, and the like used in the specification and claims should be understood, at the very least, as not an attempt to limit the application of the doctrine of equivalents to the scope of the specification and claims, and should be construed in light of the number of significant digits and ordinary rounding approaches.

Claims

1. A compound having the following formula, 【Chemical Formula 1】 or a pharmaceutically acceptable salt thereof, wherein, R １ is selected from H, OH, C １－８ alkyl, and OC １－８ alkyl, R ２ is selected from H, OH, C １－８ alkyl, and OC １－８ alkyl, and is R ３ is selected from H, OH, C １－８ alkyl, and OC １－８ alkyl, and R ４ is selected from H, OH, C １－８ alkyl, and OC １－８ alkyl, and is Q １ is 【Chemical Formula 2】 is a group having the formula of, wherein, R ２ａ is -R ２ａ＊ 、 -OR ２ａ＊ 、 OP(O)(OR ２ａ＊ ) ２ 、 OP(O)(OR ２ａ＊ )(N(R ２ａ＊ )) ２ 、 -N(R ２ａ＊ )) ２ 、 -N(R ２ａ＊ )) ３ 、 -C(O)R ２ａ＊ 、 -C(O)OR ２ａ＊ 、 -OC(O)R ２ａ＊ 、 -OC(O)OR ２ａ＊ 、 -NR ２ａ＊ C(O)R ２ａ＊ 、 -C(O)N(R ２ａ＊ )) ２ 、 NR ２ａ＊ C(O)OR ２ａ＊ 、 -OC(O)N(R ２ａ＊ )) ２ 、 -NR ２ａ＊ C(O)N(R ２ａ＊ )) ２ ; -Cl, -F, -Br, -I, -NO ２ 、 -CN, -N ３ 、 -(OCH ２ CH ２ )) ｍ -OH, selected from R ２ａ＊ is, in each case, independently, H, C １－１０ alkyl, C ２－１０ alkenyl, C ２－１０ alkynyl, aryl, C ３－１０ cycloalkyl, C ３－１０ cycloalkenyl, C ７－１０ cycloalkynyl, aryl, C １－１０ heterocyclyl, C １－１０ heteroaryl, and each R ２ａ＊ is -OH, -COOH, -, -PPh ３ , -CH ２ N Et ３ , -CH ２ N Me ３ , -NHC(=NH)NH ２ , rhodamine dye, bisquinolinium, -NH ２ , -Cl, -F, -Br, -I, -NO ２ , -CN, -N ３ , PO(OH) ２ , -(OCH ２ CH ２ ) ｍ -OH; C １－８ heterocyclyl, aryl, -OC １－８ heterocyclyl, or C １－８ may be substituted one or more times by alkoxy, R ２ａ＊ Any two or more of them may together form a ring, R ３ａ＊ is -R ３ａ＊ 、-OR ３ａ＊ 、OP(O)(OR ３ａ＊ ) ２ 、OP(O)(OR ３ａ＊ )(N(R ３ａ＊ )) ２ 、-N(R ３ａ＊ )) ２ 、-N(R ３ａ＊ )) ３ 、-C(O)R ３ａ＊ 、-C(O)OR ３ａ＊ 、-OC(O)R ３ａ＊ 、-OC(O)OR ３ａ＊ 、-NR ３ａ＊ C(O)R ３ａ＊ 、-C(O)N(R ３ａ＊ )) ２ 、NR ３ａ＊ C(O)OR ３ａ＊ 、-OC(O)N(R ３ａ＊ )) ２ 、-NR ３ａ＊ C(O)N(R ３ａ＊ )) ２ ; -Cl, -F, -Br, -I, -NO ２ 、-CN, -N ３ 、-(OCH ２ CH ２ )) ｍ -OH, selected from R ３ａ＊ is, in each case, independently, H, C １－１０ alkyl, C ２－１０ alkenyl, C ２－１０ alkynyl, aryl, C ３－１０ cycloalkyl, C ３－１０ cycloalkenyl, C ７－１０ cycloalkynyl, aryl, C １－１０ heterocyclyl, C １－１０ heteroaryl, and each R ３ａ＊ is selected from -OH, -COOH, -PPh ３ , -CH ２ N Et ３ , -CH ２ N Me ３ , -NH C(=NH)NH ２ , rhodamine dye, bisquinolinium, -NH ２ , -Cl, -F, -Br, -I, -NO ２ , -CN, -N ３ , PO(OH) ２ , -(OCH ２ CH ２ ) ｍ -OH; C １－８ heterocyclyl, aryl, -OC １－８ heterocyclyl, or C １－８ may be substituted one or more times by alkoxy, R ３ａ＊ Any two or more of them may together form a ring, R ４ａ is -R ４ａ＊ 、 -OR ４ａ＊ 、 OP(O)(OR ４ａ＊ ) ２ 、 OP(O)(OR ４ａ＊ )(N(R ４ａ＊ )) ２ 、 -N(R ４ａ＊ )) ２ 、 -N(R ４ａ＊ )) ３ 、 -C(O)R ４ａ＊ 、 -C(O)OR ４ａ＊ 、 -OC(O)R ４ａ＊ 、 -OC(O)OR ４ａ＊ 、 -NR ４ａ＊ C(O)R ４ａ＊ 、 -C(O)N(R ４ａ＊ )) ２ 、 NR ４ａ＊ C(O)OR ４ａ＊ 、 -OC(O)N(R ４ａ＊ )) ２ 、 -NR ４ａ＊ C(O)N(R ４ａ＊ )) ２ ; -Cl, -F, -Br, -I, -NO ２ 、 -CN, -N ３ 、 -(OCH ２ CH ２ )) ｍ -OH, selected from R ４ａ＊ is, in each case, independently, H, C １－１０ alkyl, C ２－１０ alkenyl, C ２－１０ alkynyl, aryl, C ３－１０ cycloalkyl, C ３－１０ cycloalkenyl, C ７－１０ cycloalkynyl, aryl, C １－１０ heterocyclyl, C １－１０ heteroaryl, and each R ４ａ＊ is -OH, -COOH, -PPh ３ , -CH ２ Net ３ , -CH ２ NMe ３ , -NHC(=NH)NH ２ , rhodamine dye, bisquinolinium, -NH ２ , -Cl, -F, -Br, -I, -NO ２ , -CN, -N ３ , PO(OH) ２ , -(OCH ２ CH ２ ) ｍ -OH; C １－８ heterocyclyl, aryl, -OC １－８ heterocyclyl, or C １－８ may be substituted one or more times by alkoxy, R ４ａ＊ Any two or more of them may together form a ring, R ５ａ is -R ５ａ＊ 、 -OR ５ａ＊ 、 OP(O)(OR ５ａ＊ ) ２ 、 OP(O)(OR ５ａ＊ )(N(R ５ａ＊ )) ２ 、 -N(R ５ａ＊ )) ２ 、 -N(R ５ａ＊ )) ３ 、 -C(O)R ５ａ＊ 、 -C(O)OR ５ａ＊ 、 -OC(O)R ５ａ＊ 、 -OC(O)OR ５ａ＊ 、 -NR ５ａ＊ C(O)R ５ａ＊ 、 -C(O)N(R ５ａ＊ )) ２ 、 NR ５ａ＊ C(O)OR ５ａ＊ 、 -OC(O)N(R ５ａ＊ )) ２ 、 -NR ５ａ＊ C(O)N(R ５ａ＊ )) ２ ; -Cl, -F, -Br, -I, -NO ２ 、 -CN, -N ３ 、 -(OCH ２ CH ２ )) ｍ -OH selected from, R ５ａ＊ is, in each case, independently, H, C １－１０ alkyl, C ２－１０ alkenyl, C ２－１０ alkynyl, aryl, C ３－１０ cycloalkyl, C ３－１０ cycloalkenyl, C ７－１０ cycloalkynyl, aryl, C １－１０ heterocyclyl, C １－１０ heteroaryl, and each R ５ａ＊ is selected from -OH, -COOH, -PPh ３ , -CH ２ N Et ３ , -CH ２ N Me ３ , -NHC(=NH)NH ２ , rhodamine dye, bisquinolinium, -NH ２ , -Cl, -F, -Br, -I, -NO ２ , -CN, -N ３ , PO(OH) ２ , -(OCH ２ CH ２ ) ｍ -OH; C １－８ heterocyclyl, aryl, -OC １－８ heterocyclyl, or C １－８ may be substituted one or more times by alkoxy, R ｐ which, in each case, is selected from H, C １－１０ alkyl, and aryl R ５ａ＊ Any two or more of them may together form a ring, R ２ａ 、R ３ａ 、R ４ａ 、and R ５ａ any two or more of which may together form a ring, Q ２ is [Chemical Formula 3] is a group having the formula of, wherein, R １ｂ is selected from H, OH, C １－８ alkyl, and OC １－８ alkyl, and is R ２ｂ is -R ２ｂ＊ 、-OR ２ｂ＊ 、OP(O)(OR ２ｂ＊ ) ２ 、OP(O)(OR ２ｂ＊ )(N(R ２ｂ＊ )) ２ 、-N(R ２ｂ＊ )) ２ 、-N(R ２ｂ＊ )) ３ 、-C(O)R ２ｂ＊ 、-C(O)OR ２ｂ＊ 、-OC(O)R ２ｂ＊ 、-OC(O)OR ２ｂ＊ 、-NR ２ｂ＊ C(O)R ２ｂ＊ 、-C(O)N(R ２ｂ＊ )) ２ 、NR ２ｂ＊ C(O)OR ２ｂ＊ 、-OC(O)N(R ２ｂ＊ )) ２ 、-NR ２ｂ＊ C(O)N(R ２ｂ＊ )) ２ ; -Cl, -F, -Br, -I, -NO ２ 、-CN, -N ３ 、-(OCH ２ CH ２ )) ｍ -OH, selected from R ２ｂ＊ which, in each case, independently, is H, C １－８ alkyl, C １－８ alkenyl, C １－１０ alkynyl, aryl, C ３－８ cycloalkyl, C ３－８ cycloalkenyl, C ７－１０ cycloalkynyl, aryl, C １－８ heterocyclyl, C ３－８ heteroaryl, and each R ２ｂ＊ is -OH, -COOH, -PPh ３ , -CH ２ NEt ３ , -CH ２ NMe ３ , -NHC(=NH)NH ２ , rhodamine dye, bisquinolinium, -NH ２ , -Cl, -F, -Br, -I, -NO ２ , -CN, -N ３ , PO(OH) ２ , -(OCH ２ CH ２ ) ｍ -OH; C １－８ heterocyclyl, aryl, -OC １－８ heterocyclyl, or C １－８ may be substituted one or more times by alkoxy, R ２ｂ＊ Any two or more of them may together form a ring, R ３ｂ is selected from H or a group having the following formula, [Chemical Formula 4] wherein, R ３ｄ is selected from H and CH ３ and R ４ｄ is selected from H and a group having the following formula: 【Chemical Formula 5】 in the formula, m is selected from 1 to 100 in each case, However, R ２ａ , R ３ａ , and R ４ａ are all OH, R ２ｂ is not CH ２ CH ２ CH ２ OCH ３ , a compound, or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, wherein the compound is a compound of the following formula (2a), (2b), or (2c) [Chemical Formula 6]

3. The compound according to claim 1, which is an (L)-sugar having the following three-dimensional structure Q １ is 【Chemical Formula 7】

4. The compound according to claim 1, which has the following structure Q １ is 【Chemical 8】

5. The compound according to claim 1, which has the following formula Q ２ is 【Chemical Formula 9】

6. The compound according to claim 1, which has the following formula Q ２ is

7. The compound according to claim 1, which has the following formula R ４ｄ is 【Chemical 11】

8.

9. R ５ａ is CH ３ and CH ２ OH, the compound according to claim 2, selected from

10. R ４ａ and R ２ａ are each OH, the compound according to claim 2.

11. R ３ａ and R ２ａ are each OH, the compound according to claim 2.

12. R ３ａ and R ４ａ are each OH, the compound according to claim 2.

13. The compound according to claim 1, which has the following formula (3a), (3b), or (3c) 【Chemical 20】 or has a pharmaceutically acceptable salt thereof, wherein, R ２ｆ is -C(O)R ｆ＊ -C(O)OR ｆ＊ -OC(O)R ２ｂ＊ -OC(O)OR ｆ＊ or C(O)N(R ｆ＊ ) ２ , and wherein, R ３ｆ is -C(O)R ｆ＊ , -C(O)OR ｆ＊ , -OC(O)R ２ｂ＊ , -OC(O)OR ｆ＊ , or C(O)N(R ｆ＊ )2, and In the formula, R ４ｆ is -C(O)R ｆ＊ -C(O)OR ｆ＊ -OC(O)R ２ｂ＊ -OC(O)OR ｆ＊ or C(O)N(R ｆ＊ )2, and R ｆ＊ is, in each case, independently, H, substituted or unsubstituted C １－８ alkyl, substituted or unsubstituted C １－８ alkenyl, substituted or unsubstituted C １－１０ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C ３－８ cycloalkyl, substituted or unsubstituted C ３－８ cycloalkenyl, substituted or unsubstituted C ７－１０ cycloalkynyl, aryl, substituted or unsubstituted C １－８ heterocyclyl, substituted or unsubstituted C ３－８ heteroaryl, and is selected from R ｓ is H, substituted or unsubstituted C １－８ alkyl, substituted or unsubstituted C １－８ alkenyl, substituted or unsubstituted C １－１０ alkynyl, substituted or unsubstituted aryl, substituted or unsubstituted C ３－８ cycloalkyl, substituted or unsubstituted C ３－８ cycloalkenyl, substituted or unsubstituted C ７－１０ cycloalkynyl, substituted or unsubstituted aryl, C １－８ heterocyclyl, substituted or unsubstituted C ３－８ is selected from heteroaryl, However, when R ｓ is CH ２ CH ２ CH ２ OCH ３ the compound according to claim 1, wherein R ２ｆ , R ３ｆ , and R ４ｆ are not simultaneously H.

14. R ２ｆ is -C(O)R ２ｆ＊ wherein R ３ｆ and R ４ｆ are each H, and R ｓ is CH ２ CH ２ CH ２ OCH ３ The compound according to claim 12

15. The compound according to claim 14, wherein Rf* is C1-8 alkyl substituted with C1-8 heterocyclyl or aryl. R ３ｆ is -C(O)R ｆ＊ wherein R ２ｆ and R ４ｆ are each H, and R ｓ is CH ２ CH ２ CH ２ OCH ３ The compound according to claim 12

16.

17. The compound according to claim 15, wherein Rf* is C1-8 alkyl substituted with morpholinyl or substituted piperazinyl. R ４ｆ is -C(O)R ｆ＊ wherein R ２ｆ and R ３ｆ are each H, and R ｓ is CH ２ CH ２ CH ２ OCH ３ The compound according to claim 12.

18. A pharmaceutical composition for treating cancer, comprising the compound according to claim 1.

19. The pharmaceutical composition according to claim 17, wherein the cancer includes chronic myeloid leukemia, acute myeloid leukemia, lymphocytic leukemia, chronic lymphocytic leukemia, non-Hodgkin lymphoma, mantle cell lymphoma, or myeloma.

20. A pharmaceutical composition for treating leukemia, comprising amocidin. ​

Images