Chemoenzymatic synthesis of polyketides

Abstract

Provided herein are synthetic methods and intermediates useful in the preparation of polyketides, including 12-membered ring macrolides. These synthetic methods and intermediates provide scalable access to polyketides, including polyketides with potent bioactive splicing modulator activity and therapeutic potential, including treating various types of cancer. The polyketides include 12-membered ring macrolides, including Compound A, Compound B, and pladienolides, including pladienolide A, pladienolide B, pladienolide C, pladienolide D, pladienolide E, pladienolide F, and pladienolide G.

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 432,011, filed December 12, 2022, and U.S. Provisional Patent Application No. 63 / 429,356, filed December 1, 2022, the entire contents of each of which are incorporated herein by reference. Summary of the Invention [Means for solving the problem]

[0002] overview A family of double-stranded RNA-specific adenosine deaminase enzymes is encoded by the ADAR family genes. ADAR (adenosine deaminase acting on RNA) proteins enzymatically perform base editing (e.g., converting adenosine to inosine), which subsequently disrupts otherwise orthogonal Watson-Crick pairings.

[0003] Unlike the therapeutic use of CRISPR-Cas9 (which acts at the gene editing level and is therefore a permanent or genetic intervention), ADAR enzymes act on double-stranded RNA (which is a temporary molecule). Therefore, ADAR modulation has the potential to be a temporary, tunable, non-genetic therapeutic intervention. ADAR enzymes were previously called double-stranded RNA adenosine deaminase (dsRAD) ("Toward the therapeutic editing of mutated RNA sequences," PNAS, 1995, 92, 8298-8302). Rather than using the CRISPR-Cas9 gene editing approach (which runs the risk of permanent off-target gene mutations), RNA editing allows for modifications to the code for regulated protein production without permanent gene editing. Therefore, there is a long-standing need for therapeutic intervention at the ribonucleic acid-to-protein stage of molecular biology's central dogma, including the need for modulators of the spliceosome and ADAR enzymes.

[0004] Therefore, provided herein are commercially scalable synthetic methods and intermediates useful for preparing polyketides.These polyketides include 12-membered ring macrolides (including Compound A, Compound B, and pladienolides (including pladienolide A, pladienolide B, pladienolide C, pladienolide D, pladienolide E, pladienolide F, and pladienolide G)).These compounds regulate spliceosome, and these compounds have shown the ability to down-regulate the level of ADAR enzymes. [Brief explanation of the drawings]

[0005] [Figure 1] FIG. 1 shows a synthetic scheme including chemoenzymatic transformations to prepare compound 14, which is a useful intermediate for preparing polyketides, including compound A.

[0006] [Figure 2]FIG. 2 shows a synthetic scheme involving chemoenzymatic transformations to prepare compound A, which involves combining compound 14 with compound 20 to prepare compound A. DETAILED DESCRIPTION OF THE INVENTION

[0007] Detailed Description definition Certain terms, whether used alone or as part of a phrase or another term, are defined below.

[0008] The articles "a" and "an" refer to one or to more than one of the grammatical object of the article.

[0009] Numerical values ​​relating to measurements are subject to measurement errors that impose limitations on their accuracy. For this reason, all numerical values ​​provided herein should be understood to be modified by the term "about" unless otherwise indicated. Accordingly, the last decimal place of a numerical value provided herein indicates its degree of accuracy. Unless other tolerances are given, the maximum permissible range is ascertained by applying rounding conventions to the last decimal place, or to the last significant digit if no decimals are present in the given numerical value.

[0010] The term "alkyl" refers to a saturated hydrocarbon, which may include a straight-chain, branched, or cyclic saturated hydrocarbon, or mixtures thereof.

[0011] The term "amelioration" refers to a decrease in the severity of at least one indicator of a condition or disease, e.g., a delay or slowing of the progression of one or more indicators of a condition or disease. The severity of an indicator can be determined by subjective or objective measures known to those skilled in the art.

[0012] The term "aryl" refers to an aromatic carbocyclic ring system containing one, two, three or more rings.

[0013] The term "composition" refers to a mixture of at least two or more components.

[0014] Modifier “C” x~y " refers to a moiety containing from x to y carbon atoms, where x and y are independently integers.

[0015] The terms "effective amount" and "therapeutically effective amount" refer to an effective amount of a therapeutic compound (e.g., a compound prepared as described herein, including Compound A, Compound B, pladienolide B, etc.) administered to a subject either in a single dose or as part of a series, which effective amount is effective to produce the desired therapeutic effect. Generally, a therapeutically effective amount can be initially estimated either in cell culture assays or in mammalian models (e.g., non-human primates, mice, rabbits, dogs, or pigs). The animal models can also be used to determine appropriate concentration ranges and routes of administration. Such information can then be used to determine useful dosages and routes of administration in non-human and human subjects.

[0016] The term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable substance, composition, or carrier (e.g., a liquid filler, solid filler, stabilizer, dispersant, suspending agent, diluent, excipient, thickener, solvent, or encapsulating substance) that is involved in carrying or transporting at least one compound described herein within or to a patient so that the compound can perform its intended function. A given carrier must be "acceptable" in the sense of being compatible with the other ingredients of a particular formulation (including the compounds described herein) and not harmful to the patient. Other ingredients that can be included in the pharmaceutical compositions described herein are known in the art and are described, for example, in "Remington's Pharmaceutical Sciences" (Genaro, ed., Mack Publishing Co., 1985), the entire contents of which are incorporated herein by reference.

[0017] The term " pharmaceutical composition " refers to the mixture of at least one compound described herein and pharmaceutically acceptable carrier.This pharmaceutical composition facilitates the administration of this compound to patient or subject.There are multiple techniques for administering compound, including but not limited to intravenous administration, oral administration, aerosol administration, parenteral administration, ophthalmic administration, pulmonary administration and topical administration.

[0018] The term "treatment" or "treating" refers to the application of one or more specific procedures used for the amelioration of a disease. "Prophylactic" treatment refers to reducing the rate of progression of the disease or condition being treated, delaying the onset of the disease or condition, or reducing the severity of its onset.

[0019] The recitation of ranges of values ​​herein is merely intended to serve as a shorthand method of individually referring to each separate value falling within the range. Unless otherwise indicated herein, each separate value is incorporated herein by reference as if it were individually set forth herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise expressly contradicted by context. The use of any and all examples or exemplary phrases (e.g., "such as") provided herein is merely intended to better illustrate the described subject matter and does not impose limitations on the scope of the otherwise claimed subject matter. No phrase herein should be construed as indicating any non-claimed element essential to practicing the described subject matter.

[0020] Groupings of alternative elements or embodiments of the present disclosure should not be construed as limitations. Each member of a group may be referred to and claimed individually or in any combination with other members of the group or other elements found herein. Furthermore, described members of a group may be included in or excluded from another described group for reasons of convenience or patentability. When any such inclusion or exclusion is made, the specification is deemed to include the modified group, and thus fulfills the description requirements of all Markush groups used in the appended claims.

[0021] References to patents and printed publications are made throughout this specification, each of which is individually incorporated herein by reference in its entirety.

[0022] It is to be understood that the embodiments of the present disclosure are illustrative, and therefore, the present disclosure is not limited to that precisely as shown and described.

[0023] Synthetic Methods and Intermediates Many synthetic schemes and intermediate compounds have been discovered that are useful for improving the commercial synthetic preparation of polyketides, including Compound A. Compound A has been described, including in WO2021 / 026273A1 and US10,675,267B2, which are incorporated herein by reference. Compound A has also been synthesized as described by Chan et al. (Cell Reports Physical Science, 2020, 1, 12, 100277). [ka]

[0024] In some embodiments, step 1, step 2, step 4, step 5, step 6, step 7, step 8, step 16, step 17, step 4a, step 5a, step 6a, step 7a, step 3b, step 4b, step 5b, or a combination thereof (as shown in Figures 1-4) have been discovered as novel and useful in the preparation of polyketides (e.g., Compound A). Additionally, in some embodiments, compound 2, compound 5, compound 6, compound 7, compound 8, compound 19-TPS, compound 19R-TPS, compound 19O, compound 20, compound 21, compound 22, compound 23, compound 24, compound 25, and compound 26 have been discovered as novel and useful in the preparation of polyketides (e.g., Compound A).

[0025] In some embodiments, the schemes and intermediates provided herein can be adapted by those skilled in the art to prepare polyketides other than Compound A, including Compound B or a stereoisomer thereof, pladienolide or a stereoisomer thereof (e.g., pladienolide B), and compounds of the same class of polyketides that are capable of binding to the SF3b complex of the spliceosome, inhibiting mRNA splicing activity, or downregulating ADAR levels. Pladienolide B has been previously synthesized, for example, as described by Rhoades et al. (Journal of the American Chemical Society, 2021, 143(13), 4915-4920, DOI: 10.1021 / jacs.1c01135). [ka] [ka]

[0026] The compounds described herein also include isotopically labeled compounds in which one or more atoms are replaced by an atom having the same atomic number but an atomic mass or mass number different from that predominantly found in nature. Examples of isotopes suitable for inclusion in the compounds described herein include: 2 H, 3 H, 11 C. 13 C. 14 C. 36 Cl, 18 F, 123 I, 125 I, 13 N, 15 N, 15 O. 17 O, or 18 In some embodiments, isotopically labeled compounds are useful in drug or substrate tissue distribution studies. In another embodiment, substitution with heavier isotopes (e.g., deuterium) provides for greater metabolic stability (e.g., increased in vivo half-life or reduced administration requirements). In yet another embodiment, positron-emitting isotopes (e.g., 11 C. 18 F, 15 O and 13 Substitution with N) is useful in positron emission tomography (PET) studies to examine substrate receptor occupancy. Isotopically labeled compounds are prepared by any suitable method or process that substitutes an appropriate isotopically labeled reagent for the unlabeled reagent used in other methods.

[0027] In some embodiments, the compounds described herein are labeled by other means, including but not limited to, the use of chromophores or fluorescent moieties, bioluminescent labels, or chemiluminescent labels.

[0028] The compounds described herein, and other related compounds having different substituents, can be synthesized using techniques and materials described herein, as well as in other publications, such as, for example, Fieser and Fieser's Reagents for Organic Synthesis, Vols. 1-17 (John Wiley and Sons, 1991); Rodd's Chemistry of Carbon Compounds, Vols. 1-5 and supplements (Elsevier Science Publishers, 1989); Organic Reactions, Vols. 1-40 (John Wiley and Sons, 1991); Larock's Comprehensive Organic Transformations (VCH Publishers Inc., 1989); March, Advanced Organic Chemistry, 4th ed. (Wiley, 1992); Carey and Sundberg, Advanced Organic Chemistry, 4th ed., Vols. A and B (Plenum, 2000, 2001), and Green and Wuts, Protective Groups in Organic Chemistry Synthesis, 3rd Edition (Wiley, 1999), all of which are incorporated by reference for their disclosure. The general methods for the preparation of the compounds described herein are modified by the use of reagents and conditions appropriate for the introduction of the various moieties found in the formulas provided herein.

[0029] The compounds described herein are synthesized using any suitable procedure starting from compounds available from commercial sources or prepared using the procedures described herein.

[0030] In some embodiments, the compounds described herein can be prepared by a synthetic method that includes any of the synthetic steps described in the Examples or in Figure 1, Figure 2, Figure 3, or Figure 4 associated with the compound being prepared.

[0031] In some embodiments, [ka] [ka] Provided herein is a compound selected from, or a salt thereof, wherein: R 1 is C 1~6 alkyl (e.g., ethyl), R 2 is R 2 Organotin moieties containing a tin (Sn) atom covalently bonded to a carbon to which is bonded (e.g., Sn(C 1~6 alkyl)3, for example, Sn(n-Bu3) or SnMe3).

[0032] In some embodiments, the compound is [ka] [ka] or salts thereof.

[0033] In some embodiments, the compound is [ka] is selected from.

[0034] In some embodiments, still further compounds useful in preparing the pladienolides herein are: [ka] (for example, [ka] ), wherein TPS is triphenylsilanyl.

[0035] In some embodiments, provided herein are compositions comprising the compounds provided herein.

[0036] In some embodiments, the compounds or compositions provided herein are for use in the preparation of a synthesized compound.

[0037] In some embodiments, provided herein are methods of preparing a synthesized compound, the methods comprising contacting a compound provided herein with one or more reagents to form the synthesized compound.

[0038] In some embodiments, provided herein are methods of preparing the compounds provided herein, the methods comprising contacting a precursor compound with one or more reagents to form a compound provided herein.

[0039] In some embodiments, provided herein is a method for preparing a compound to be synthesized, the method comprising at least one of the following steps: Step 1) Mixing at least one solvent, 10-camphorsulfonic acid, and Compound 1 [ka] and preparing a mixture containing Compound 2. [ka] forming a Step 2) Reacting at least one solvent, an acid, 1-(dimethoxymethyl)-4-methoxy-benzene, and Compound 2 [ka] and preparing a mixture containing Compound 3. [ka] forming a Step 4) Reacting at least one solvent with lithium diisopropylamide and Compound 4 [ka] and preparing a mixture containing Compound 5. [ka] forming a Step 5) Reacting at least one solvent, 2-iodoxybenzoic acid, and Compound 5 [ka] and preparing a mixture containing Compound 6. [ka] or At least one solvent, N,N'-dicyclohexylcarbodiimide, pyridine trifluoroacetic acid, and Compound 5 [ka] and preparing a mixture containing Compound 6. [ka] forming a Step 6) A method for producing a compound 6 by reacting at least one solvent, an enzyme having aldo-keto reductase activity, a reducing cofactor selected from NADPH or NADH, an auxiliary enzyme and a substrate for regenerating the cofactor, and [ka] and preparing a mixture containing Compound 7. [ka] forming a Step 7) Reacting Compound 7 with at least one solvent, tert-butyldimethylsilyl chloride, imidazole, tert-butyldimethylsilyl trifluoromethanesulfonate, a base, and [ka] and preparing a mixture containing Compound 8. [ka] forming a Step 8) Reacting at least one solvent, a base, and Compound 8 [ka] and preparing a mixture containing Compound 9. [ka] forming a Step 15b) Reacting at least one solvent, a silanyl chloride, and compound 19rac [ka] and forming a reaction product (e.g., triphenylsilanyl compound 19rac), which is purified by chromatography on silica gel to obtain compound 19rac having an enantiopurity at carbon 7 of greater than 99%. [ka] or compound 19R [ka] forming a Step 16) Reacting at least one solvent, tributyltin hydride, and compound 19 [ka] or compound 19R [ka] and preparing a mixture containing Compound 20. [ka] or compound 20R [ka] (or alternatively using trimethyltin hydride to form the corresponding vinyltrimethyltin compound); Step 17) Reacting at least one solvent, a Pd catalyst, compound 20 or compound 20R (or the corresponding vinyltrimethyltin compound), and compound 14 [ka] and preparing a mixture containing Compound A. [ka] or Compound B [ka] forming a Step 4a) Reacting Compound 4 with at least one solvent [ka] and Compound 21 [ka] and preparing a mixture containing Compound 22. [ka] forming a Step 5a) Reacting Compound 22 with at least one solvent [ka] and oxidizing compound 22 to form compound 23. [ka] forming a Step 6a) Reacting Compound 23 with at least one solvent [ka] and reducing compound 23 to form compound 24. [ka] forming a Step 7a) Reacting at least one solvent, tert-butyldimethylsilyl chloride, a base, and compound 24 [ka] and preparing a mixture containing Compound 11. [ka] forming a Step 3b) reacting at least one solvent with Compound 3 [ka] and oxidizing compound 3 to form compound 25. [ka] forming a Step 4b) Reacting Compound 25 with at least one solvent, a peptide coupling reagent, and [ka] and preparing a mixture containing Compound 26. [ka] or forming Step 5b) Reacting at least one solvent, dimethylhydroxylamine, a base, and Compound 26 [ka] and preparing a mixture containing Compound 6. [ka] A process of forming. In some embodiments, any of the compounds in the synthetic steps herein may be replaced with the respective general counterpart of that compound provided herein.

[0040] In some embodiments for step 6, the enzyme having aldo-keto reductase activity is a KRED or an enzymatically active variant thereof. KRED requires the reduced cofactor NAD(P)H. To reduce the cost of commercial synthesis procedures, a cofactor recycling system may be used in combination with the enzyme having aldo-keto reductase activity. Thus, in some embodiments, a cofactor-regenerating enzyme may be used. In some embodiments, the cofactor-regenerating enzyme used herein may include glucose-6-phosphate dehydrogenase, glucose dehydrogenase, and isocitrate dehydrogenase. In some embodiments, the substrate for the cofactor-regenerating enzyme may include glucose or isocitrate. Step 6 may further include an additional substrate isomerase, including aconitase, which isomerizes citrate to isocitrate. In some embodiments, the enzyme used in step 6 is bound to a solid support, is not bound to a solid support, or includes a combination of bound and unbound enzymes. In some embodiments, the enzymes include ketoreductase 1 (KRED1-Pglu) from Oogatea glycozyma, alcohol dehydrogenase (RADH) from Ralstonia sp., and alcohol dehydrogenase (LbADH) from Lactobacillus brevis. In some embodiments, KREDs include those classified under EC number 1.1.1. KREDs may include alcohol dehydrogenase, carbonyl reductase, lactate dehydrogenase, hydroxyacid dehydrogenase, hydroxyisocaproate dehydrogenase, β-hydroxybutyrate dehydrogenase, steroid dehydrogenase, sorbitol dehydrogenase, or aldoreductase, or enzymatically active variants thereof. NADPH-dependent KREDs include those classified under EC number 1.1.1.2. NADH-dependent KREDs include those classified under EC number 1.1.1.1.In some embodiments, the KRED enzyme is KRED-A6-P2D5, and the enzymatic reaction occurs in the presence of NADP and a solvent, optionally a buffer, and a nonionic detergent having a hydrophilic head and a hydrophobic / lipophilic tail with a hydrophilic-lipophilic balance (HLB) of about 10 to about 20 (e.g., 13-15) (e.g., octylphenoxypolyethoxyethanol (IGEPAL CA-630; HLB 13.4) or polyethylene glycol p-(1,1,3,3-tetramethylbutyl)-phenyl ether (e.g., Triton®-X100 (CAS No. 9002-93-1; HLB 13.4)). In some embodiments, the solvent includes isopropyl alcohol, water, or a mixture thereof. Similar to step 6, for the sequential steps 15c / d, aldo-keto reductase activity can be used to convert compound 19O to pure compound 19 or 19R.

[0041] In some embodiments, the compound synthesized is a polyketide.

[0042] In some embodiments, the polyketide is [ka] is.

[0043] In some embodiments, the compound to be synthesized is [ka] is.

[0044] In some embodiments, provided herein are compounds (whether intermediates or polyketides (e.g., Compound A, Compound B, or Pladienolide B)) prepared by one or more methods described herein.

[0045] In some embodiments, provided herein are compounds prepared by one or more of the methods described herein, wherein the compounds [ka] is.

[0046] In some embodiments, a composition is provided herein, the composition comprising: 1) Has greater than 99% enantiomeric purity at carbon 21 [ka] Contains or Has greater than 99% enantiomeric purity at carbon 21 [ka] Contains or Has enantiomeric purity greater than 99% at carbon 8 [ka] Contains or Has enantiomeric purity greater than 99% at carbon 8 [ka] Contains or Has enantiomeric purity greater than 99% at carbon 8 [ka] Contains or Has enantiomeric purity greater than 99% at carbon 8 [ka] Contains or Has greater than 99% enantiomeric purity at carbon 7 [ka] Contains, or Has greater than 99% enantiomeric purity at carbon 7 [ka] Includes; 2) Optionally, the composition contains less than 0.5 ppm Sn (eg, less than 0.5 ppm Sn other than Sn from the compounds (eg, organostannane compounds)).

[0047] In some embodiments, a composition is provided herein, the composition comprising: 1) have an enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains or Has enantiomeric purity greater than 99% [ka] Contains, or Has enantiomeric purity greater than 99% [ka] Includes; 2) Optionally, the composition contains less than 0.5 ppm Sn (eg, less than 0.5 ppm Sn other than Sn from the compounds (eg, organostannane compounds)).

[0048] Chemical abbreviations used herein include, but are not limited to: CSA for 10-camphorsulfonic acid; DCC for N,N'-dicyclohexylcarbodiimide; DMAP for dimethylaminopyridine; DMSO for dimethyl sulfoxide; GDH for glutamate dehydrogenase; HPLC for high performance liquid chromatography; IBX for 2-iodoxybenzoic acid; KRED for aldo-keto reductase; LDA for lithium diisopropylamide; MTBE for methyl tert-butyl ether; nicotinamide adenine dinucleotide instead of NAD or NAD+; nicotinamide adenine dinucleotide phosphate instead of NADP or NADP+; NADPH instead of the reduced form of NADP+; parts per million instead of ppm; tert-butyldimethylsilyl chloride instead of TBSCl; tert-butyldimethylsilyl trifluoromethanesulfonate instead of TBSOTf; (2,2,6,6-tetramethylpiperidin-1-yl)oxyl instead of TEMPO; trifluoroacetic acid instead of TFA; and tetrahydrofuran instead of THF.

[0049] Compositions and Uses As noted above, the compounds provided herein (including intermediates and final compounds, e.g., Compound A, Compound B, pladienolide B, etc.) can be prepared as described herein on a commercially relevant scale, making the compounds even more useful in preparative and therapeutic applications.

[0050] Therefore, in some embodiments, the present invention provides a composition comprising the compound provided herein.In some embodiments, the composition is a pharmaceutical composition comprising the compound described herein (such as, but not limited to, Compound A) prepared by a synthetic method comprising at least one synthetic step described herein.In some embodiments, the pharmaceutical composition referred to herein can comprise at least one pharmaceutically acceptable carrier.

[0051] In some embodiments, compounds described herein (such as, but not limited to, Compound A) prepared by a synthetic method comprising at least one synthetic step described herein are useful for treating neoplasms. In some embodiments, the neoplasm comprises cancer. In some embodiments, the neoplasm comprises a tumor. In some embodiments, the cancer is a malignant cancer. In some embodiments, the cancer is a benign cancer. In some embodiments, compounds described herein (such as, but not limited to, Compound A) prepared by a synthetic method comprising at least one synthetic step described herein are useful for modulating (e.g., inhibiting) spliceosome activity. In some embodiments, compounds described herein (such as, but not limited to, Compound A) prepared by a synthetic method comprising at least one synthetic step described herein are useful for modulating (e.g., inhibiting) ADAR activity. In some embodiments, compounds described herein (such as, but not limited to, Compound A) prepared by a synthetic method comprising at least one synthetic step described herein are useful for modulating (e.g., inhibiting) RNA editing activity. Such activity can occur in vivo or in vitro, including inside a subject (eg, a human subject).

[0052] In some embodiments, provided herein are methods of treating a neoplasm in a subject in need thereof, the methods comprising administering to the subject a therapeutically effective amount of an active agent or composition thereof prepared as described herein.

[0053] In some embodiments, provided herein are methods for modulating spliceosome activity in a cell, the methods comprising contacting the cell with an effective amount of an active agent or composition thereof prepared as described herein.

[0054] In some embodiments, provided herein are methods of modulating double-stranded RNA-specific adenosine deaminase (ADAR) activity in a cell, the method comprising contacting the cell with an effective amount of an active agent or composition thereof prepared as described herein.

[0055] In some embodiments, the cell is in vitro. In some embodiments, the cell is in vivo (e.g., in a subject, e.g., a mammalian subject, e.g., a human subject).

[0056] Thus, the actual dosage level of an active ingredient (e.g., Compound A, Compound B, pladienolide B, etc.) prepared as described herein, a composition provided herein, or a pharmaceutical composition may be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration without being toxic to the patient.

[0057] In particular, the selected dosage level depends on various factors, including the activity of the specific compound used, the timing of administration, the excretion rate of the compound, the duration of treatment, other drugs, compounds or substances used in combination with the compound, the age of the patient to be treated, the sex of the patient to be treated, the weight of the patient to be treated, the condition of the patient to be treated, the overall health status of the patient to be treated, and the past medical history of the patient to be treated, and similar factors well known in the medical field.A doctor (e.g., a physician or veterinarian) skilled in the art can easily determine and prescribe the effective amount of the pharmaceutical composition required.For example, a physician or veterinarian may start the administration of the compound used in the pharmaceutical composition at a level lower than that required to achieve the desired therapeutic effect, and gradually increase the dosage until the desired effect is achieved.

[0058] Administration route includes but is not limited to oral administration route, nasal administration route, rectal administration route, vaginal administration route, parenteral administration route, buccal administration route, sublingual administration route or topical administration route.In some embodiments, oral administration route or nasal administration route is oral inhalation administration route or nasal inhalation administration route.The compound for use described herein can be formulated for administration by any route suitable for achieving the specific method applied.

[0059] In some embodiments, provided herein are packaged compounds, packaged compositions, or packaged pharmaceutical compositions that include a container holding a therapeutically effective amount of a compound described herein (such as, but not limited to, Compound A) prepared by the synthetic methods described herein, and instructions for using the compound according to one or more of the methods provided herein.

[0060] The compounds and related materials can be completed into commercial products by conventional processes practiced in the field of the present invention (e.g., by appropriate sterilization and packaging). For example, the materials can be treated with UV / visible light irradiation (200 nm to 500 nm) using, for example, photoinitiators with various absorption wavelengths (e.g., Irgacure 184, 2959), preferably water-soluble initiators (e.g., Irgacure 2959). Such irradiation is typically carried out for irradiation times of 1 minute to 60 minutes, although longer irradiation times may be applied depending on the specific method. Materials according to the present disclosure can be terminally sterile wrapped and packaged in appropriate containers (such as boxes) (e.g., by adding a specific product information leaflet) to maintain sterility until use.

[0061] According to further embodiments, the compounds can also be provided in kit form in combination with other components necessary for administration of the substance to a patient. For example, the disclosed kits (e.g., for use in treating cancer) can further include, for example, administration substances.

[0062] The kit is designed in a variety of forms based on the particular defect it is designed to treat.

[0063] The compounds or compositions provided herein can be prepared and placed in a container for storage at ambient or elevated temperatures. When the compounds or compositions are stored in a polyolefin plastic container compared to a polyvinyl chloride plastic container, discoloration of the compounds or compositions, or adsorption of the compounds to the surface of the container, whether dissolved or suspended in a liquid composition (e.g., an aqueous or organic liquid solution) or as a solid, may be reduced. Without wishing to be bound by theory, the container may reduce exposure of the contents of the container to electromagnetic radiation, whether visible light (e.g., having a wavelength of about 380 nm to 780 nm) or ultraviolet (UV) light (e.g., having a wavelength of about 190 nm to 320 nm (UV B light) or about 320 nm to 380 nm (UV A light)). Some containers also include the ability to reduce exposure of the contents of the container to infrared light, or include a second component with such ability. The container that can be used includes the container made of polyolefin (for example, polyethylene, polypropylene, polyethylene terephthalate, polycarbonate, polymethylpentene, polybutene, or a combination thereof, particularly polyethylene, polypropylene, or a combination thereof).In some embodiments, the container is a glass container.The container can be further placed in a second container (for example, paper; cardboard; paperboard; metal film; or foil; or a combination thereof; a container that further reduces the exposure of the contents of the container to UV light, visible light, or infrared light).The compound or composition provided herein may require up to 3 months or more than 3 months of continuous storage; in some cases, up to 1 year or more than 1 year of continuous storage.The container can be any form (for example, bag, bottle, or box) that is suitable for containing its contents.

[0064] The following examples further illustrate aspects of the present disclosure, but in no way limit the teachings or disclosure set forth herein. [Example]

[0065] Example 1: Synthesis of intermediates and compound 14 as shown in Figure 1 Methods and materials are compared with Chan (2020) and WO2021 / 026273A1.

[0066] Step 1: 10-Camphorsulfonic acid (CSA), MeOH, 12 hours at 50°C, then buffer containing Ambersep 900(OH).

[0067] Advantages: Previous synthetic methods require large amounts of solvent to convert compound 1 to compound 3. The present method allows for a scalable preparation using 8 volumes of solvent, which is a key step in enabling access to kg-quantity production of compound 3.

[0068] Exemplary procedure: To a solution of compound 1 (10 g, 27.58 mmol, 1 equiv.) in MeOH (80 mL) was added CSA (961.03 mg, 4.14 mmol, 0.15 equiv.), and the mixture was stirred at 50 °C for 12 h. Ambersep 900(OH) (5 g) (strongly basic anion exchange resin; contains hydroxide bound to the resin via a quaternary ammonium) was added to adjust the pH between 7 and 8, and the solution was filtered and concentrated. The crude product (91% by NMR) was used directly in the next step.

[0069] Step 2: Buffer containing anisaldehyde dimethyl acetal, Amberlyst 15(H), CH2Cl2, then Ambersep 900(OH).

[0070] Advantages: As described above in step 1.

[0071] Exemplary procedure: 1-(Dimethoxymethyl)-4-methoxy-benzene (10.05 g, 55.18 mmol, 9.40 mL, 2 equiv.) and Amberlyst 15(H) (1 g, 27.59 mmol) (strongly acidic cation exchange resin; containing resin-bound sulfonic acid) were added sequentially to the crude product from step 1 (4.42 g, 27.59 mmol, 1 equiv.) in CHCl (120 mL) at 20 °C. The mixture was stirred at 20 °C for 2 h. The reaction mixture was filtered to remove the acid resin. Ambersep 900(OH) (5 g) was added to adjust the pH to 7-8, filtered, and concentrated under reduced pressure to give a residue. Purification by column chromatography using a gradient of heptane to 3:1 heptane / EtOAc afforded pure 2 (4.7 g, 61%).

[0072] Step 3: KBr, TEMPO, NaHCO3, NaClO, CH2Cl2.

[0073] Advantage: Addition of KBr as a co-catalyst.

[0074] Exemplary procedures: As described elsewhere in Chan (2020) or WO2021 / 026273A1.

[0075] Step 4: EtOAc, LDA, THF.

[0076] Advantages: High yield addition to aldehydes.

[0077] Exemplary procedure: This procedure was carried out under N2 atmosphere until quenched with water. LDA (0.69 M, 158 mL, 1.50 equiv) was slowly added to a solution of EtOAc (8.29 g, 94.1 mmol, 9.21 mL, 1.30 equiv) in THF (180 mL) at -70 °C. The mixture was stirred at -70 °C for 1 h. A solution of compound 4 (20.0 g, 72.4 mmol, 1.00 equiv) in THF (60 mL) was added at -60 °C. The mixture was stirred at -70 °C for 0.5 h. The reaction mixture was quenched with saturated NH4Cl (100 mL) at -40 °C. The resulting solution was extracted with EtOAc (2 x 70 mL). The combined organic layers were washed with HO and dried over Na2SO4. After filtration through filter paper, the organic layer was concentrated under reduced pressure to give a residue. Purification by column chromatography using a gradient of petroleum ether to 15:1 petroleum ether / EtOAc gave pure compound 5 (23.0 g, 84.9%).

[0078] Step 5: Two methods were found to be viable: IBX (3 equiv.), EtOAc, 70 °C, 24 h; or DCC (3 equiv.), pyridine·TFA (0.5 equiv.), DMSO, 20 °C, 12 h, 60% (Pfitzner-Moffatt).

[0079] Advantages: By allowing oxidation to an achiral species that allows chiral reduction in step 6, the IBX process has advantages at the gram scale, while the Pfitzner-Moffat process can be used to reach the kg scale.

[0080] Exemplary procedure (IBX): IBX (22.5 g, 80.3 mmol) was added to compound 5 (9.8 g, 26.7 mmol) in EtOAc (300 mL). The mixture was heated at 70° C. with rapid stirring for 24 hours. After this period, it was cooled to room temperature and filtered. Pure compound 6 (5.9 g, 61%) was obtained by flash plug chromatography eluting with aliquots of 4:1 heptane:EtOAc, 3:1 heptane:EtOAc, and 2:1 heptane:EtOAc.

[0081] Exemplary procedure (Pfitzner-Moffatt): Alcohol compound 5 (50 mg, 0.14 mmol) was dissolved in DMSO (5 mL) and DCC (84.9 mg, 0.41 mmol) was added. After stirring at room temperature for 12 h, HO (20 mL) was added. Extraction with EtOAc (4 × 50 mL), drying over NaSO, and concentration by rotary evaporation afforded the crude product. Pure compound 6 (0.30 g, 60%) was obtained by flash plug chromatography eluting with aliquots of 4:1 heptane:EtOAc, 3:1 heptane:EtOAc, and 2:1 heptane:EtOAc.

[0082] Step 6: GDH, ketoreductase, D-glucose, phosphate buffer pH 7.

[0083] Advantages: Highly stereoselective reduction process with chiral purity of 99.6%.

[0084] Exemplary procedure: To a mixture of D-glucose (10.8 g, 59.9 mmol, 6.04 equiv.), GDH from Bacillus megaterium (360 mg), NADP+ (360 mg), and KRED from Hansenula polymorpha (3.60 g) in phosphate buffer (pH 6.5-7) (180 mL) was added compound 6 (3.60 g, 9.93 mmol, 1.00 equiv.) in DMSO (10 mL) at 30 °C dropwise, and the mixture was stirred at 30 °C for 18 h. HPLC analysis indicated that 15.6% of compound 6 (retention time 3.033 min) remained, and 75.4% of the product, compound 7 (retention time 2.813 min), was detected. The reaction mixture was quenched with CH3CN (320 mL), filtered, and the filtrate was collected. The resulting solution was diluted with EtOAc (150 mL), and the organic phase was separated, washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated to give compound 7 (3.50 g, crude product, chiral purity: 99.6%) as a yellow oil.

[0085] Step 7: TBSCl, imidazole, CH2Cl2 or TBSOTf, 2,6-lutidine, CH2Cl2.

[0086] Advantages: The TBSCl procedure is more scalable than the TBSOTf procedure.

[0087] Exemplary procedure (TBSCl): Imidazole (2.00 g, 29.4 mmol, 3.06 equiv) and TBSCl (3.00 g, 19.90 mmol, 2.44 mL, 2.07 equiv) were added sequentially to a solution of compound 7 (3.50 g, 9.60 mmol, 1.00 equiv) in CHCl (3 mL) at 25 °C. The mixture was stirred at 25 °C for 12 h. HO (30 mL) was added to the reaction mixture, the resulting solution was extracted with CHCl (2 × 20 mL), and the combined organic layers were washed with HO and dried over NaSO. After filtration through paper, the organic layer was concentrated under reduced pressure to give a residue. Purification by column chromatography using a gradient of petroleum ether to 10:1 petroleum ether / EtOAc afforded pure compound 8 (4 g, crude product).

[0088] Step 8: KOH, MeOH.

[0089] Advantages: Provides easy conversion to compound 9.

[0090] Exemplary procedure: To a solution of compound 8 (4.00 g, 8.36 mmol, 1.00 equiv) in HO (10 mL) and MeOH (40 mL) at 20 °C was added KOH (1.20 g, 21.4 mmol, 2.56 equiv), and the mixture was then stirred at 20 °C for 1 h. The reaction mixture was adjusted to pH 5-6 with 5% citric acid (150 mL) below 0 °C, extracted with CHCl (200 mL), washed with HO (150 mL), dried over NaSO, filtered, and the filtrate was concentrated under reduced pressure to give a residue. Purification by column chromatography using a gradient of petroleum ether to 20:1 petroleum ether / EtOAc afforded pure compound 9 (2.5 g, 66.4%).

[0091] Step 9: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0092] Step 10: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0093] Step 11: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0094] Step 12: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0095] Example 2: Synthesis of intermediates and compound A as shown in Figure 2 Methods and materials are compared with Chan (2020) and WO2021 / 026273A1.

[0096] Step 13: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0097] Step 14: Exemplary procedure: As described in Chan (2020) or WO2021 / 026273A1.

[0098] Step 15: Exemplary procedure: Synthesis as described in Chan (2020) or WO2021 / 026273A1, but with improved purification methods (step 15b and / or step 15c / d).

[0099] Advantages: Chromatographic and chemoenzymatic methods were developed to ensure that compound 19 could be produced in >99% enantiomeric purity with <1 ppm of organostannane by-products.

[0100] Step 15b: Conversion of a mixture of compound 19 and compound 19R to pure compound 19 or pure compound 19R. [ka]

[0101] Chlorotriphenylsilane (3.91 g, 13.26 mmol, 0.3 equiv.) was added to a solution of crude compound 19 (10 g, 44.19 mmol, 1 equiv.; a mixture of compound 19 and compound 19R), DMAP (539.82 mg, 4.42 mmol, 0.1 equiv.), and pyridine (17.48 g, 220.93 mmol, 17.83 mL, 5 equiv.) in CHCl (70 mL) at 25 °C. The mixture was stirred at 25 °C for 12 h. TLC (5:1 petroleum ether / EtOAc) showed the formation of one new spot. The reaction mixture was quenched by the addition of HO (150 mL) at 0 °C and then extracted with MTBE (2 × 150 mL). The combined organic layers were washed with 10% citric acid until the pH of the aqueous phase reached 3–4. The organic layer was then dried over Na2SO4, filtered, and concentrated under reduced pressure to give a residue (22.5 g). The residue was dissolved in MTBE (150 mL) and 34 g of silica gel (100 mesh-200 mesh) was added. It was then concentrated under reduced pressure to remove the solvent and give a residue. 170 g of silica gel (200 mesh-300 mesh) was then loaded into a chromatography column. The residue was purified by column chromatography (170 g of silica gel, 200 mesh-300 mesh, 100:0 to 1:1 petroleum ether / EtOAc). Compound 19R (eluted with 50:1 petroleum ether / EtOAc) and compound 19 (eluted with 3:1 petroleum ether / EtOAc) were collected and then concentrated to give 5.5 g of enantiopure compound 19.

[0102] Step 15c / d: Conversion of a mixture of compound 19 and compound 19R to pure compound 19. [ka]

[0103] Exemplary procedure: 2-iodoxybenzoic acid (IBX) (8.4 g, 30 mmol) was added to a solution of crude compound 19 (5.01 g, 22.1 mmol; a mixture of compound 19 and compound 19R) in anhydrous EtOAc (500 mL) in a 1 L flask. The mixture was stirred at 23 °C for 4 h, at which point the resulting mixture was filtered through a pad of silica gel (200 g) and concentrated to give a crude colorless oil containing compound 19O. This material was subjected to enzymatic reduction, which was initiated by dissolving the colorless oil containing compound 19O in isopropyl alcohol (200 mL). This was added to a buffer solution (3 L) containing Triton® X100 (60 mL). The enzymes KRED-A6-P2D5 (0.2 g) and NADP (0.05 g) were added. The reaction was stirred at 42 °C for 28 h. Three extractions with EtOAc (500 mL) and washing with brine (200 mL) gave compound 19. The organic phases were combined, dried over NaSO, and concentrated on a rotary evaporator. Flash chromatography eluting with a gradient of hexane to 1:3 EtO / hexane gave pure compound 19 (3.8 g, 75%) as a colorless oil.

[0104] Step 16: Conversion of compound 19 to vinylstannane 20.

[0105] Exemplary procedure: PdCl(PPh) (1.55 g, 2.21 mmol) was added to a solution of compound 19 (5.01 g, 22.1 mmol) in anhydrous THF (200 mL) in a 500 mL flask. The mixture was cooled to 0° C., and n-BuSnH (17.9 mL, 66.3 mmol) was added dropwise. The mixture was stirred at 0° C. for 45 minutes, at which point the resulting mixture was concentrated to give a crude black oil. The material was extracted into hexane, filtered through a pad of Celite, and eluted with hexane. The eluate was concentrated on a rotary evaporator, and this process was repeated twice to give a clear, black solution.

[0106] Pure compound 20 (5.72 g, 50%) was obtained as a 1:5 mixture of α:β regioisomers by flash chromatography eluting with a gradient of hexane to CHCl, then CHCl to 1:20 EtO / CHCl. ​​The desired regioisomer could be obtained in 95+% purity by further flash chromatography eluting with a gradient of hexane to CHCl, then CHCl to 1:20 EtO / CHCl.

[0107] A similar procedure is carried out using Me3SnH instead of n-Bu3SnH to prepare the corresponding trimethylvinylstannane compound.

[0108] Step 17: Compound 20 and compound 14 are combined to form compound A by Stille coupling.

[0109] Exemplary procedure: Compound 20 (1.33 g, 2.57 mmol) and compound 14 (1.00 g, 2.14 mmol) were mixed in a 100 mL flask and dried by rotary evaporation of benzene. To the mixture were then added CuCl (0.425 g, 4.29 mmol), KF (0.249 g, 4.29 mmol), XPhos Pd G2 (0.169 g, 0.214 mmol), and anhydrous t-BuOH (25 mL), sequentially. The reaction vessel was purged under Ar, heated to 50 °C, and stirred overnight. Upon cessation of stirring, the solution turned into a gray, cloudy mixture. The mixture was then filtered through a plug of Celite and eluted with acetone (200 mL). The eluate was concentrated on a rotary evaporator to give a brown crude wax. Flash chromatography on neutral silica gel eluting with a gradient of hexane to 1:3 acetone / hexane gave pure compound A (1.21 g, 80%) as a pale yellow wax.

[0110] Compound A is prepared by carrying out a similar procedure using the corresponding trimethylvinylstannane compound in place of compound 20.

[0111] Example 3: Synthesis of intermediates and compound B, similar to the synthesis shown in Figure 2 for compound A Compound 19R and compound 20R are prepared as described in Example 2.

[0112] Step 18: Compound 20R is combined with compound 14 to form compound B by Stille coupling.

[0113] Exemplary procedure: Compound 20R (1.33 g, 2.57 mmol) and compound 14 (1.00 g, 2.14 mmol) are mixed in a 100 mL flask and dried by rotary evaporation of benzene. CuCl (0.425 g, 4.29 mmol), KF (0.249 g, 4.29 mmol), XPhos Pd G2 (0.169 g, 0.214 mmol), and anhydrous t-BuOH (25 mL) are then added sequentially to the mixture. The reaction vessel is purged under Ar, heated to 50 °C, and stirred overnight, at which point the solution turns into a gray, cloudy mixture. The mixture is then filtered through a plug of Celite and eluted with acetone (200 mL). The eluate is concentrated on a rotary evaporator to give a crude concentrate. Flash chromatography on neutral silica gel eluting with a gradient of hexane to 1:3 acetone / hexane gives pure compound B (1.21 g, 80%).

[0114] Compound B is prepared by carrying out a similar procedure using the corresponding trimethylvinylstannane compound in place of compound 20R.

Claims

【Request Item 1】 【Chemistry 72】 or a salt thereof, R １ is C １～６ is alkyl, R ２ is R ２ or a salt thereof, wherein the compound is an organotin moiety comprising a tin (Sn) atom covalently bonded to a carbon to which is attached 【Request Item 2】 【Chemistry 73】 2. The compound of claim 1, or a salt thereof, selected from: 【Request Item 3】 【Chemistry 74】 2. The compound of claim 1 selected from:

4. A composition comprising a compound according to one of claims 1 to 3.

5. A compound or composition according to one of claims 1 to 4 for use in the preparation of a compound to be synthesised.

6. 1. A method for preparing a compound to be synthesized, comprising: contacting the compound of claim 1 with one or more reagents to form the synthesized compound. A method comprising:

7. 10. A method for preparing a compound of claim 1, comprising: contacting a precursor compound with one or more reagents to form the compound of claim 1; A method comprising:

8. 1. A method for preparing a compound to be synthesized, comprising at least one of the following steps: Step 1) A method for preparing a compound 1 containing at least one solvent, 10-camphorsulfonic acid, and 【Chemistry 75】 and preparing a mixture containing Compound 2. 【Transformation 76】 forming a Step 2) Reacting at least one solvent, an acid, 1-(dimethoxymethyl)-4-methoxy-benzene, and Compound 2 【Chemical Formula 77】 and preparing a mixture containing Compound 3. 【Transformation 78】 forming a Step 4) Reacting at least one solvent, lithium diisopropylamide, and Compound 4 【Transformation 79】 and preparing a mixture containing Compound 5. 【Chemistry 80】 forming a Step 5) Reacting at least one solvent, 2-iodoxybenzoic acid, and Compound 5 【Chemistry 81】 and preparing a mixture containing Compound 6. 【Chemistry 82】 or At least one solvent, N,N'-dicyclohexylcarbodiimide, pyridine trifluoroacetic acid, and Compound 5 【Chemistry 83】 and preparing a mixture containing Compound 6. 【Chemical 84】 forming a Step 6) A method for producing a hydroxybenzoate comprising the steps of: at least one solvent; an enzyme having aldo-keto reductase activity; a reducing cofactor selected from NADPH or NADH; an auxiliary enzyme; and a substrate for regenerating the cofactor; and Compound 6. 【Chemical 85】 and preparing a mixture containing Compound 7. 【Chemical 86】 forming a Step 7) Reacting at least one solvent, tert-butyldimethylsilyl chloride, imidazole, tert-butyldimethylsilyl trifluoromethanesulfonate, a base, and Compound 7 【Chemistry 87】 and preparing a mixture containing Compound 8. 【Chemical 88】 forming a Step 8) Reacting at least one solvent, a base, and Compound 8 【Chemical 89】 and preparing a mixture containing Compound 9. [Chemical 90] forming a Step 15b) Reacting at least one solvent, a silanyl chloride, and compound 19 【Chemistry 91】 and compound 19R 【Chemistry 92】 to form a reaction product, and chromatographically purifying the reaction product on silica gel to form Compound 19 or Compound 19R having an EE of at least 99%; Step 15c) reacting at least one solvent with compound 19 【Chemistry 93】 and compound 19R 【Chemical 94】 and an oxidizing agent to form the compound 19O. 【Chemical 95】 forming a Step 15d) A method for the preparation of a compound 19O comprising the steps of: at least one solvent; an enzyme having aldo-keto reductase activity; a reducing cofactor selected from NADPH or NADH; a substrate for cofactor regeneration; and 【Chemistry 96】 and preparing a mixture containing Compound 19. 【Chemistry 97】 forming a Step 16) Reacting at least one solvent, tributyltin hydride, and compound 19 【Chem.98】 Or compound 19R 【Chem.99】 and preparing a mixture containing Compound 20. 【Chemistry 100】 Or compound 20R 【Chemistry 101】 (or alternatively using trimethyltin hydride to form the corresponding vinyltrimethyltin compound); Step 17) Reacting Compound 20 or Compound 20R with Compound 14 【Chemical Engineering 102】 and preparing a mixture containing Compound A. 【Chemistry 103】 Or Compound B 【Chemical 104】 forming a Step 4a) At least one solvent and compound 4 【Chemistry 105】 and Compound 21 【Chemistry 106】 and preparing a mixture containing Compound 22. 【Chemistry 107】 forming a Step 5a) Reacting Compound 22 with at least one solvent 【Chemistry 108】 and oxidizing compound 22 to form compound 23. 【Chemistry 109】 forming a Step 6a) Reacting Compound 23 with at least one solvent 【Chemical 110】 and reducing compound 23 to form compound 24. 【Chemistry 111】 forming a Step 7a) Reacting at least one solvent, tert-butyldimethylsilyl chloride, a base, and Compound 24 【Chemistry 112】 and preparing a mixture containing Compound 11. 【Chemistry 113】 forming a Step 3b) reacting at least one solvent with Compound 3 【Chemical 114】 and oxidizing compound 3 to form compound 25. 【Chemical 115】 forming a Step 4b) Reacting at least one solvent, a peptide coupling reagent, and Compound 25 【Chemistry 116】 and preparing a mixture containing Compound 26. 【Chemistry 117】 or forming Step 5b) Reacting at least one solvent, dimethylhydroxylamine, a base, and Compound 26 【Chemistry 118】 and preparing a mixture containing Compound 6. 【Chemical 119】 A process of forming

9. The method of claim 8, wherein the compound synthesized is a polyketide.

10. The polyketide 【Chemical 120】 10. The compound of claim 9, wherein:

11. The compound to be synthesized is 【Chemistry 121-1】 【Chemistry 121-2】 The method of claim 8, wherein

12. 9. A compound prepared by the method of claim 8, wherein the compound is 【Chemistry 122】 A compound.

13. A composition comprising: 1) A compound of formula selected from the following: 【Chemical 123】 wherein the composition has an enantiomeric purity at carbon 21 of the formula of greater than 99%. 【Chemistry 124】 wherein the composition has an enantiomeric purity at carbon 21 of the formula of greater than 99%. 【Chemistry 125】 wherein the composition has an enantiomeric purity at carbon 8 of the formula of greater than 99%. 【Chemistry 126】 wherein the composition has an enantiomeric purity at carbon 8 of the formula of greater than 99%. 【Chemistry 127】 wherein the composition has an enantiomeric purity at carbon 8 of the formula of greater than 99%. 【Chemistry 128】 wherein the composition has an enantiomeric purity at carbon 8 of the formula of greater than 99%. 【Chemistry 129】 wherein the composition has an enantiomeric purity at carbon 7 of the formula of greater than 99%; or 【Chemistry 130】 wherein the composition has an enantiomeric purity at carbon 7 of the formula of greater than 99%. 2) Optionally, the composition contains less than 0.5 ppm of Sn other than the Sn in the formula.

14. A composition comprising: 1) A compound of formula selected from the following: 【Chemistry 131】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 132】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 133】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 134】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 135】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Transformation 136】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 137】 wherein the composition has the formula in greater than 99% enantiomeric purity; 【Chemistry 138】 wherein the composition has the formula in greater than 99% enantiomeric purity; or 【Chemistry 139】 wherein the composition has the formula in greater than 99% enantiomeric purity; 2) Optionally, the composition contains less than 0.5 ppm of Sn other than the Sn in the formula. composition.

15. 1. A method of treating a neoplasm in a subject in need thereof, comprising: administering to said subject a therapeutically effective amount of a compound of claim 12 or a composition of claim 13 or claim 14. A method comprising:

16. 1. A method for modulating spliceosome activity in a cell, comprising: contacting said cells with an effective amount of a compound of claim 12 or a composition of claim 13 or claim 14. A method comprising:

17. 1. A method for modulating double-stranded RNA-specific adenosine deaminase (ADAR) activity in a cell, comprising: contacting said cells with an effective amount of a compound of claim 12 or a composition of claim 13 or claim 14. A method comprising:

18. 1. A method comprising the synthesis of compound 19, said synthesis comprising the synthesis of compound 19O. [Chemical 140] The compound of claim 2, which is 【Chemistry 141】 The method includes forming a

19. A method comprising the synthesis of compound 7, said synthesis comprising the synthesis of compound 6 【Chemistry 142】 The compound of claim 2, wherein the compound is 【Chemistry 143】 The method includes forming a

20. Compound A 【Chemistry 144】 20. The method according to claim 18 or claim 19, which is a method for preparing

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