Synthesis method for farnesyl dibenzodiazepinones

Abstract

The present invention relates to a means for preparing farnesyl dibenzodiazepinone compounds, including AMO-01. [Formula 1] JPEG2023527866000143.jpg45134

Description

[Background technology]

[0001] Euactinomycetes are a subset of a large and complex group of Gram-positive bacteria known as actinomycetes. Abundant in soil, these organisms have attracted considerable commercial and scientific interest over the past few decades due to their production of numerous therapeutically useful compounds, particularly antibiotics, as secondary metabolites. Intensive research into strains capable of producing new antibiotics has led to the identification of hundreds of new species.

[0002] Many true actinomycetes, particularly the genera *Streptomyces* and its close relative *Saccharopolyspora*, are widely studied. These genera all produce remarkably diverse biologically active metabolites. Because these compounds are commercially important, much is known about the genetics and physiology of these organisms. Another representative genus of true actinomycetes, *Micromonospora*, also attracts commercial interest. For example, U.S. Patent No. 5,541,181 (Ohkuma et al.) discloses a dibenzodiazepinone compound produced by the known actinomycete strain Micromonospora M990-6 (ATCC55378), specifically 5-farnesyl-4,7,9-trihydroxy-dibenzodiazepin-11-one (named "BU-4664L"). The patent by Ohkuma et al. reports that BU-4664L and its chemically synthesized di- and tri-alkoxy and acyloxy derivatives have anti-inflammatory and antitumor cell activity. As another example, U.S. Patent No. 7,101,872 (Bachmann et al.) discloses farnesyldibenzodiazepinone compounds, specifically 10-farnesyl-4,6,8-trihydroxy-dibenzodiazepin-11-one (named "ECO-04601" and "AMO-01").

[0003] Research into the pharmaceutical applications of these compounds will be supported by reproducible means of producing acceptable levels of purity and sufficient quantities of the compounds in both in vitro and animal experiments. The methods available for preparing dibenzodiazepinone compounds primarily rely on culturing microorganisms under conditions that induce compound production, followed by multiple extractions, concentrations, and purifications of the culture medium and fermentation broth. These methods are costly and time-consuming. [Overview of the Initiative] [Problems that the invention aims to solve]

[0004] Therefore, there is a strong desire to develop synthetic methods for producing dibenzodiazepinone compounds. This invention addresses this objective and other important objectives. [Means for solving the problem]

[0005] The present invention relates to novel means for the synthesis of farnesyl dibenzodiazepinone compounds, for example, AMO-01 as defined below. [ka]

[0006] As will be described later, the synthesis methods for several farnesyldibenzodiazepinone compounds based on the present invention are based on the remarkable discovery by the inventors that the regioselectivity of the resulting compounds is dramatically increased by using the Ullmann coupling reaction while carefully controlling the amount of copper in the reaction. In contrast, Buchwald coupling with a palladium catalyst yields regiochemically opposite compounds from the same starting materials. The method disclosed herein utilizes this difference to enable the production of farnesyldibenzodiazepinone compounds with selective stereochemical properties.

[0007] In a first embodiment, the present invention relates to a method for synthesizing farnesyldibenzodiazepinone of formula (Formula) I and a salt thereof. [ka] In formula I, A is -NH-; R 7 -CH3, -(CH2) x CH3, -CH2CH2W 1CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2 CH2CH2W 3 is CH3, where x is an integer from 1 to 11, and W 1 , W 2 , and W 3 each is, independently, [Chemical formula] ; R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ), where R 8 is -Na, -CH3, or -CH2CH3; R 3 and R 4 are the same and are selected from -H, -OH, -OCH3, or -OP=O(OR 8 ), where R 8 is -Na, -CH3, or -CH2CH3. In certain aspects, the method proceeds via an Ullmann reaction.

[0008] A method for synthesizing farnesyl dibenzodiazepinone of formula I and its salts comprises the following steps, where A, R 2 , R 4 , R) 5 , R 7 , R 8 , W s 1 , W 2 , W 3 and x are as defined above for formula I. (a) Preparing AP2312 - A; [Chemical formula] (b) Preparing AP2312 - B; [Chemical formula] (c) Perform Ullmann coupling; [ka] (d) Debenzylation; [ka] (e) Silylation; [ka] (f)R 7 To prepare; [ka] Here, X is Br, I, or Cl. (g) Perform farnesylation; [ka] Furthermore, (h) desilylation is performed. [ka]

[0009] In a second embodiment, the present invention relates to a method for synthesizing farnesyldibenzodiazepinone of formula II and its salts. [ka] In formula II, A is -NH-, R 7 -CH3, -(CH2) x CH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2 CH2CH2W 3CH3, where x is an integer from 1 to 11, W 1 , W 2 , and W 3 Each of them is independent, [ka] and; R 2 -H, -OH, -OCH3, or -OP=O(OR 8 ) and here R 8 is -Na, -CH3, or -CH2CH3; R 5 and R 6 These are the same, -H, -OH, -OCH3, or -OP=O(OR 8 ) is selected, and here R 8 The group is -Na, -CH3, or -CH2CH3. In some cases, the method involves Buchwald coupling.

[0010] A method for synthesizing farnesyldibenzodiazepinone of formula II and its salts comprises the following steps: A, R 2 , R 5 , R 6 , R 7 , R 8 , W 1 , W 2 , W 3 , and x are as defined above for Equation II. (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) Perform a Buchwald coupling; [ka] (d) Debenzylate; [ka] (e) Perform silylation; [ka] (f)R 7 To prepare; [ka] Here, X is Br, I, or Cl. (g) Perform farnesylation; [ka] Furthermore, (h) desilylation is performed. [ka]

[0011] In a third embodiment, the present invention relates to a method for synthesizing farnesyldibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxy-dibenzodiazepin-11-one; also referred to as "AP2312"). [ka]

[0012] In a given situation, the method includes the following steps: (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) Perform Ullmann coupling; [ka] (d) Debenzylate; [ka] (e) Perform silylation; [ka] (f) Prepare farnesyl bromide; [ka] (g) Perform farnesylation; [ka] Furthermore, (h) desilylation is performed. [ka]

[0013] In a particular context, the synthesis method for AMO-01 includes the following steps: (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) In the presence of CuI (0.0525 equivalents), K2CO3 (2.0 equivalents), L-proline (0.1 equivalent), and DMF, molecular equivalents of AP2312-A and AP2312-B are reacted to perform Ullmann coupling to obtain AP2312-3; [ka] (d) Debenzylate AP2312-3 in the presence of THF, MeOH, and Pd / C under H2 to obtain AP2312-4; [ka] (e) Silylate AP2312-4 in the presence of TIPSCl (4.0 equivalents), Et3N (5.0 equivalents), and DMF to obtain AP2312-5; [ka] (f) React AP23132-C in the presence of Ms2O, LiBr (1.6 equivalents), 2,6-lutidine (1.6 equivalents), and DMF to obtain AP2312-6; [ka] (g) dioxane, t BuOH, and t AP2312-5 is farnesylated with AP2312-6 in the presence of BuOK (1.15 equivalents) to obtain AP2312-8; [ka] Furthermore, AP2312-8 is desilylated in the presence of (h)THF (1.0 equivalent), AcOH (8.0 equivalents), and TBAF (4.0 equivalents) to obtain AMO-01. [ka]

[0014] The above description provides a comprehensive overview of the features and technical advantages of the present invention in order to clarify the detailed description of the present invention below. Further features and advantages of the present invention, which constitute the subject matter of the claims, are described below. It will be apparent to those skilled in the art that other means for achieving the same objectives as the present invention may be easily designed or modified using all the concepts and specific embodiments disclosed herein as a basis. It will also be apparent to those skilled in the art that such equivalent configurations do not depart from the spirit and scope of the present invention as described in the claims. Further objectives and advantages of novel features, as well as their configuration and operation, will become clear through the following description with reference to the accompanying drawings. Note that the following description, drawings, and specific examples are provided for illustrative and explanatory purposes only and are not intended to limit the scope of the present invention. [Brief explanation of the drawing]

[0015] [Figure 1] This figure shows a comprehensive scheme for the synthesis of farnesyldibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxydibenzodiazepine-11-one).

[0016] [Figure 2] This figure shows that the purity of AMO-01 was 98.3% based on HPLC results. [Modes for carrying out the invention]

[0017] I. Definition The indefinite article "a" or "an" used here may mean one or more. When used in conjunction with the words "comprising," the indefinite article "a" or "an" may mean one or more. "Another" here may mean at least two or more. Furthermore, unless the context specifically requires otherwise, the singular form includes the plural, and the plural form includes the singular.

[0018] As used herein, “approximately” refers, whether express or implied, to numerical values, such as integers, fractions, and percentages. The term “approximately” broadly means a range of numerical values ​​(e.g., + / - 5 to 10% of the indicated value) that a person skilled in the art could consider equivalent to (e.g., having the same function or result as) the indicated value. In some cases, the term “approximately” may include numerical values ​​rounded to the nearest significant figure. II. Farnesyl dibenzodiazepinone compounds

[0019] AMO-01 (10-farnesyl-4,6,8-trihydroxydibenzodiazepine-11-one; also referred to here as "AP2312") is a farnesyl dibenzodiazepinone, a type of dibenzodiazepinone compound containing a farnesyl moiety. The structure of AMO-01 is as follows: [ka]

[0020] Farnesyl dibenzodiazepinone compounds may also be produced by biological means, such as culturing specific strains of the genus Micromonospora, a genus of bacteria in the order Micromonosporales that are Gram-positive, spore-forming, generally aerobic, and form branched mycelia, and then isolating the compounds from the culture medium. Members of this genus also generally produce aminoglycoside antibiotics.

[0021] AMO-01 is produced by Micromonospora sp. strain 046-ECO11. Strain 046-ECO11 was deposited on March 7, 2003, with the International Depositary Authority of Canada (IDAC) at the Bureau of Microbiology, Health Canada, 1015 Arlington Street, Winnipeg, Manitoba, Canada R3E 3R2, under registration number 070303-01. Details of strain 046-ECO11 and the biological means for producing AMO-01 are described in International Patent Publication WO2004 / 065591, issued on August 5, 2004, which is incorporated herein by reference in its entirety.

[0022] The inventors have, through diligent efforts, achieved a complete synthetic means for the production of farnesyldibenzodiazepinone compounds, including AMO-01. The present invention relates to such means, along with the relevant aspects of the present invention disclosed herein.

[0023] Therefore, in one embodiment, the present invention relates to the farnesyldibenzodiazepinone group of formula I and a method for synthesizing salts thereof. [ka] In formula I, A is -NH-, R 7 -CH3, -(CH2) x CH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2 CH2CH2W 3 CH3, where x is an integer from 1 to 11, W 1 , W2 , and W 3 Each of them is independent, [ka] And, R 2 -H, -OH, -OCH3, or -OP=O(OR 8 ) and here, R 8 is -Na,-CH3, or -CH2CH3, R 3 and R 4 These are the same, -H, -OH, -OCH3, or -OP=O(OR 8 ) is selected, and here R 8 is -Na, -CH3, or -CH2CH3. In some cases, this method proceeds via the Ullmann reaction.

[0024] The synthesis method for farnesyldibenzodiazepinone of formula I includes the following steps, where A, R 2 , R 3 , R 4 , R 7 , R 8 , W 1 , W 2 , W 3 x is defined above for equation I. (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) Perform Ullmann coupling; [ka] (d) Debenzylate; [ka] (e) Perform silylation; [Chemical formula] (f) Preparing R 7 ; [Chemical formula] where X is Br, I, or Cl. (g) Performing farnesylation; [Chemical formula] and (h) performing desilylation. [Chemical formula]

[0025] The present invention also relates to a method for synthesizing a group of farnesyl dibenzodiazepinones of formula II and their salts. [Chemical formula] <()000513> In formula II, <00005!5>A is -NH-; R 7 is -CH3, -(CH2) x CH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2 CH2CH2W 3 CH3, where x is an integer from 1 to 11, and W 1 , W 2 , and W 3 are each independently [Chemical formula] ; [[ID=?4]] R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ) where R 8 is -Na, -CH3, or -CH2CH3; R5 and R 6 These are the same, -H, -OH, -OCH3, or -OP=O(OR 8 ) is selected, and here R 8 It is -Na, -CH3, or -CH2CH3. In certain situations, this method proceeds via Buchwald coupling.

[0026] The synthesis method for farnesyldibenzodiazepinone of formula II includes the following steps, where A,R 2 ,R 5 ,R 6 ,R 7 ,R 8 ,W 1 ,W 2 ,W 3 x is defined above for equation II. (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) Perform a Buchwald coupling; [ka] (d) Debenzylate; [ka] (e) Perform silylation; [ka] (f)R 7 To prepare; [ka] Here, X is Br, I, or Cl. (g) Perform farnesylation; [ka] Furthermore, (h) desilylation is performed. [ka]

[0027] Furthermore, the present invention relates to a method for synthesizing farnesyldibenzodiazepinone AMO-01. [ka]

[0028] In certain situations, this method involves the following steps: (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) Perform Ullmann coupling; [ka] (d) Debenzylate; [ka] (e) Perform silylation; [ka] (f) Prepare farnesyl bromide; [ka] (g) Perform farnesylation; [ka] Furthermore, (h) desilylation is performed. [ka]

[0029] In a particular context, the synthesis method for AMO-01 includes the following steps: (a) Prepare AP2312-A; [ka] (b) Prepare AP2312-B; [ka] (c) React molecular equivalents of AP2312-A and AP2312-B in the presence of CuI (0.0525 equivalents), K2CO3 (2.0 equivalents), L-proline (0.1 equivalent), and DMF to perform Ullmann coupling to obtain AP2312-3; [ka] (d) Debenzylate AP2312-3 in the presence of THF, MeOH, and Pd / C under H2 to obtain AP2312-4; [ka] (e) Silylate AP2312-4 in the presence of TIPSCl (4.0 equivalents), Et3N (5.0 equivalents), and DMF to obtain AP2312-5; [ka] (f) React AP23132-C in the presence of Ms2O, LiBr (1.6 equivalents), 2,6-lutidine (1.6 equivalents), and DMF to obtain AP2312-6; [ka] (g) dioxane, t BuOH, and t AP2312-5 is farnesylated with AP2312-6 in the presence of BuOK (1.15 equivalents) to obtain AP2312-8; [ka] Furthermore, AP2312-8 is desilylated in the presence of (h)THF (1.0 equivalent), AcOH (8.0 equivalents), and TBAF (4.0 equivalents) to obtain AMO-01. [ka]

[0030] The following are exemplary compounds and specific examples of farnesyldibenzodiazepinone compounds that can be produced by the method of the present invention as defined herein. [ka] [ka] [ka] [ka] [ka] [ka]

[0031] If the variable "x" in the formula of this invention is an integer, then it is understood that x is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11. The range of integer x may be 1 to 11, 1 to 10, 1 to 9, 1 to 8, 1 to 7, 1 to 6, 1 to 5, 1 to 4, 1 to 3, and 1 to 2. To avoid misunderstanding, the ranges shown above also include the maximum and minimum values ​​as integers within the range.

[0032] The term "alkyl" as used herein refers to a linear or branched hydrocarbon group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, pentyl, hexyl, heptyl, cyclopentyl, cyclohexyl, and cyclohexymethyl. Alkyl groups may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, carboalkoxy, carboxy, carboxamide, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, oxo, guanidino, and formyl. The number of carbon atoms in the hydrocarbon group may be 1 to 6 carbon atoms, including 1 to 2 carbon atoms, 1 to 3 carbon atoms, 1 to 4 carbon atoms, and 1 to 5 carbon atoms.

[0033] The term "alkene" used here refers to an unsaturated hydrocarbon group containing a carbon-carbon double bond. The number of carbon atoms in the hydrocarbon group may be 2 to 6, including groups with 2 carbon atoms, 2 to 3 carbon atoms, 2 to 4 carbon atoms, and 2 to 5 carbon atoms.

[0034] The terms “aryl” and “aryl ring” as used herein refer to a single or fused ring aromatic group having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring-member atoms. Examples of aryls include, but are not limited to, phenyl, naphthyl, biphenyl, and terphenyl. Aryls may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, azide, alkylthio, carboalkoxy, carboxy, carboxyamide, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0035] The terms "heteroaryl" and "heteroaryl ring" as used herein refer to single or fused ring aromatic groups having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring-member atoms and containing at least one heteroatom selected from O, N, S, SO, and SO2. Examples of heteroaryl groups include, but are not limited to, pyridinyl, thiazolyl, thiadiazoyl, isoquinolinyl, pyrazolyl, oxazolyl, oxadiazoyl, triazolyl, and pyrrolyl groups. The heteroaryl group may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, carboxoxy, carboxy, carboxamide, cyano, halo, hydroxyl, nitro, thio, thiocarbonyl, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0036] The term "alkenyl" refers to a linear, branched, or cyclic hydrocarbon group containing at least one carbon-carbon double bond. Examples of alkenyl groups include, but are not limited to, vinyl, 1-propen-2-yl, 1-buten-4-yl, 2-buten-4-yl, and 1-penten-5-yl groups. Alkenyls may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, carboalkoxy, carboxy, carboxamide, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, formyl, oxo, and guanidino. The double bond portion of the unsaturated hydrocarbon chain can be in either a cis or trans configuration.

[0037] The terms "cycloalkyl" and "cycloalkyl ring" refer to saturated or partially unsaturated carbon rings in single or condensed carbocyclic systems having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring-member atoms. Examples of cycloalkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclohexyl, and cycloheptyl. Cycloalkyls may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, carboalkoxy, carboxy, carboxamide, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0038] The terms "heterocyclyl" and "heterocyclic" refer to saturated or partially unsaturated rings containing 1, 2, 3, or 4 heteroatoms or heterogroups selected from O, N, NH, NRx, PO2, S, SO, or SO, in single or fused heterocyclic systems having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring-member atoms. Examples of heterocyclyls or heterocyclics include, but are not limited to, morpholinyl, piperidinyl, and pyrrolidinyl. A heterocyclyl, heterocyclic, or heterocyclyl ring may optionally have one or more substituents, which can be selected from acyl, amino, acylamino, acyloxy, oxo, thiocarbonyl, imino, carboalkoxy, carboxy, carboxamide, cyano, halo, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0039] The term "amino acid" refers to any natural amino acid, such as alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine.

[0040] The term "halo" refers to halogen atoms, such as bromine, chlorine, fluorine, and iodine.

[0041] The terms "aralkyl" and "heteroaralkyl" refer to aryl and heteroaryl groups directly bonded via alkyl groups, respectively, such as benzyl. The aralkyl and heteroaralkyl groups may optionally have substituents, as with the aryl and heteroaryl groups described above.

[0042] Similarly, the terms "aralkenyl" and "heteroaralkenyl" refer to aryl and heteroaryl groups directly bonded via an alkene group, respectively, such as benzyl. The aralkenyl and heteroaralkenyl groups may optionally have substituents, as with the aryl and heteroaryl groups described above.

[0043] The compounds of the present invention may have one or more chiral carbon atoms and may exist as optical isomers forming racemic or non-racemic compounds. The compounds of the present invention are useful as single isomers or as mixtures of stereoisomers. Diastereoisomers, i.e., non-superimal stereoisomers, can be separated by conventional means such as chromatography, distillation, crystallization, or sublimation. Optical isomers can be obtained by decomposing racemic mixtures according to conventional processes. III.Synthesis method

[0044] As summarized above, the present invention provides a method for synthesizing the farnesyldibenzodiazepinone compounds of formulas I and II as defined herein. The reaction schemes for the compounds contained in these formulas are shown here. Initial experiments used in the production of the farnesyldibenzodiazepinone compounds of the present invention yielded the surprising result that the compound of formula II was realized by using the Buchwald coupling, and the compound of formula I was obtained by using the Ullmann coupling. Thus, although the initial steps in the synthesis of compounds of formulas I and II are similar, the choice between the Ullmann coupling and the Buchwald coupling results in rejection, producing compounds of formulas I and II, respectively. AMO-01

[0045] In a specific embodiment, the present invention provides a method for synthesizing farnesyldibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxydibenzodiazepine-11-one). The details of this method will be described in the following section, but the general scheme is shown in Figure 1. In this example, AMO-01 is referred to as AP2312.

[0046] In step 1 of the synthesis method for AMO-01, AP2312-A is prepared as follows. [ka]

[0047] The preparation of AP2312-A was achieved through two binary, highly related schemes. The first scheme included steps 1.A, 1.B, and 1.C.

[0048] Step 1.A: A solution of 1,3,5-trifluoro-2-nitrobenzene (490.0 g, 2.77 mol) in THF (2.45 L) was bubbling with ammonia gas (~240 g, 14.1 mol) at -60 to -40°C for 2 hours. After stirring at 0°C for 4 hours, the reaction mixture was filtered and the filter cake was washed with ELISA (490 mL x 4). The filtrate was concentrated to 500 mL and petroleum ether (980 mL) was added. The mixture was re-slurred overnight at RT, filtered, and the filter cake was washed with petroleum ether (490 mL). The filter cake was dried under vacuum at 40°C for 5 hours to obtain 366 g of AP2312-1 as an orange solid with HPLC purity of 98.0% in 76% yield.

[0049] Step 1.B: KOH (151g, 2.7mol) and BnEt3N in BnOH (1045g, 9.7mol) + Cl - A mixture of (98g, 0.43mol) was stirred at RT for 0.5 hours. AP2312-1 (188g) was added to the reaction mixture in small portions over 0.5 hours, and the mixture was stirred at 80°C for 3 hours. After cooling to RT, the reaction mixture was poured into water (1.5L) and extracted with DCM (2.8L). The organic layer was washed with water (1.5L x 2), dried over Na2SO4 (94g), filtered, and concentrated. The residue was re-slurred in petroleum ether (3.8L) at RT for 1 hour, filtered, and the filter cake was sequentially washed with petroleum ether (0.94L x 2) and MeOH (0.94L x 3). The filter cake was dried under vacuum at 50°C for 6 hours to obtain 348g of AP2312-2 as an orange solid with 94% yield and HPLC purity of 99.7%.

[0050] Step 1.C: AP2312-2 (175.0 g, 0.5 mol) was suspended in EtOH (700 mL), H2O (350 mL), and AcOH (315 mL). Zinc powder (110.5 g, 1.7 mol) was added to the reaction mixture in small portions by RT. A vigorous exothermic reaction occurred, and the temperature rose to 80°C in 1 hour. The reaction mixture was stirred at 80°C for 2 hours. After the reaction mixture was cooled to room temperature, the inorganic salts were removed by filtration, and the filter cake was washed with DCM (700 mL). The filtrate was concentrated to remove the organic solvent and extracted with DCM (1.4 L). The organic layer was sequentially washed with water (700 mL), 3M NaOH (350 mL x 2), and water (700 mL). The organic layer was concentrated and purified by restrush in EtOH (350 mL) at 0-15°C for 1 hour. The mixture was filtered, and the filter cake was washed with chilled ethanol (175 mL). The filter cake was dried under vacuum at 45°C for 7 hours to obtain 92.5 g of AP2312-A as a yellow solid with HPLC purity of 99.5% in 58% yield.

[0051] A second scheme for preparing AP2312-A included steps 1.1, 1.2, and 1.3. Step 1.1 (1) THF (5L) was added to a 10L four-necked flask fitted with a mechanical stirrer. (2) 1,3,5-trifluoro-2-nitrobenzene (1.0 kg) was added via RT. (3) Under N2 protection, the mixture was cooled to -60 to -40°C in a dry ice / EtOH bath. (4) Ammonia gas was introduced at -60 to -40°C for 1.5 hours. Note: The volume of the reaction mixture increased, indicating that NH3 was absorbed. The generated NH3 gas was absorbed by 20% H2SO4 water. (5) After 2 hours at -60 to -40°C, LC-MS showed that 22.2% of the starting material remained. (6) The mixture was heated to -15°C to -10°C. After covering it for 2 hours, it was stirred overnight (16 hours) at -15°C to -10°C. The LCMS showed that 0.5% of the starting material remained. (7) The mixture was heated to 10°C. Note: The generated NH3 gas was absorbed by 20% H2SO4 water. (8) The salt (NH4F) was filtered under vacuum. (9) The cake was washed with methoxy (500 mL x 4). Note: Wet cake: 560g; TLC showed no remaining product. (10) Another batch prepared using the same starting material was prepared and merged. (11) The combined filtrate was vacuum concentrated in RT for 0.5 hours to remove NH3 gas. (12) The filtrate was concentrated to a volume of 2 L under vacuum at 40-45°C. Note: A large amount of yellow to red solid precipitated. (13) n-heptane (1.6L) was added. (14) The mixture was concentrated to a volume of 2 L under vacuum at 40-45°C. (15) n-heptane (1.0 L) was added. (16) The mixture was stirred vigorously in RT for 1 hour. (17) Solid matter was collected by filtration. (18) The cake was washed with n-heptane (500 mL). (19) This cake was dried under vacuum at 35-40°C to obtain 1350 g of a red solid with an HPLC purity of 96.7%. (20) The filtrate was concentrated to a volume of 3 L. (21) Stirred in RT for 0.5 hours. (22) Solid matter was collected by filtration. (23) The cake was washed with n-heptane (100 mL). (24) The cake was dried under vacuum at 35-40°C to obtain a further 390 g of red solid with an HPLC purity of 93.1%. (25) Total yield: 1700 g, 82%. Step 1.2 (26) BnOH (9732.6g) was added to the 50L reactor. (27) KOH (1402.7g) was added while stirring (150 RPM). (28) BnEt3NCl (956.63g) was added. (29) The mixture was cooled to 15°C under N2 protection. (30) AP2312-1 (1740.0g) was added in small amounts. (31) The mixture was heated at 75-80°C for 4 hours. HPLC analysis revealed that less than 1.0% (0.11%) of the starting material remained. (32) The mixture was cooled to RT. (33) DCM (17L at a time) was added. (34) Water (14L at a time) was added. (35) The mixture was stirred for 30 minutes. (36) The soil was isolated. (37) The organic layer was washed with water (9L). (38) The solution was dried with Na2SO4 (2 kg). (39) The salt was removed by filtration. (40) The filtrate was concentrated to 13 L at 35-40°C. Note: An orange solid precipitate formed, and very little distillate was observed. (41) PE (37L) was added. (42) The slurry was stirred with RT for 1.5 hours. (43) Solid matter was collected by filtration. (44) The cake was washed with MeOH (4L x 2). (45) The cake was washed with PE (4L x 2). (46) The cake was dried under vacuum at 45°C to obtain 3090.1 g of a yellow solid with HPLC purity of 99.8% in 88% yield. Step 1.3 (47) Under N2 protection, EtOH (11.7 L) was added to the 50 L reactor. (48) AP2312-2 (3.0 kg) was added. (49) HOAc (5.4L) was added. (50) H2O (6.3 L) was added. (51) The solution mixture was heated to 40°C and then the heating was stopped. (52) Zn powder (1903.3g) was added in several portions over a period of 2 hours. Note: After 20 minutes, the internal temperature rose to 80°C (without exothermic reaction or cooling), and upon completion of the addition, the mixture turned into a brown solution. (53) The mixture was stirred at ambient temperature for 2 hours. Note: The temperature dropped to 50°C after 2 hours, and HPLC analysis showed that AP2312-2 was completely consumed. (54) EtOH (9L) was added to the mixture. (55) The mixture was stirred in RT for 1 hour. (56) The solid was removed by filtration. (57) The cake was washed with DCM (15L). (58) The filtrate was transferred to a 100L reactor. (59) DCM (21L) was added. (60) Water (15L) was added. (61) The mixture was stirred for 15 minutes. (62) The aqueous layer was separated (the lower layer, TLC, showed no residue). (63) The organic layer was washed with water (15L x 2). Note: To remove residual HOAc and Zn salts. (64) Water (15L) was added to the organic layer. (65) 3M NaOH solution was added. Note: The pH of the organic layer was adjusted to 9-10. (66) The organic layer was separated. (67) Washed in brine (15L). (68) The organic layer was dried with Na2SO4 (1 kg). (69) The salt was filtered out. (70) The filtrate was concentrated to a volume of 15 L under vacuum at 45-50°C. Up to a total volume of 0.4 vol; solid matter precipitated from a solution. (71) EtOH (15L) was added. (72) The mixture was concentrated to 15 L at 40°C. (73) A further 5L of EtOH was added. (74) The slurry was stirred with RT for 2 hours. (75) The mixture was cooled to 5-10°C and stirred at 5-10°C for 1 hour. (76) Solid matter was collected by filtration. (77) The cake was washed with cooled EtOH (2L x 2, 10°C). HPLC of wet cake: 99.2%. (78) The cake was dried at 35°C under vacuum for 48 hours to a constant weight, yielding 2070.2 g of an off-white solid with HPLC purity of 99.7% in 75% yield.

[0052] In step 2 of the AMO-01 synthesis method, AP2312-B is prepared as follows. [ka]

[0053] The preparation of AP2312-B was achieved through two binary, highly related schemes. The first scheme included steps 2.A, 2.B, 2.C, and 2.D.

[0054] Step 2.A: Boc2O (763g, 3.5mol) was added dropwise over 2 hours to a DCM (2.4L) solution of methyl 3-hydroxybenzoate (486g, 3.2mol) and DMAP (35.4g, 0.29mol). The reaction mixture was stirred overnight in RT, washed with 8% w / w citric acid solution (486mL x 3) and water (486mL), dried over Na2SO4 (97g), filtered and concentrated to obtain 727g of AP2312-B1 as a yellow oil with HPLC purity 100 in 90% yield.

[0055] Step 2.B: The mixture of TMP (367g, 2.6mol) and i-PrMgCl·LiCl (2.0L in THF, 1.3M) was stirred at RT for 15 hours. The pre-synthesized TMPMgCl·LiCl was added dropwise to a solution of AP2312-B1 (327g, 1.3mol) in THF (2.3L) over 1 hour at 0-10°C. After stirring at 0-10°C for 3 hours, the reaction mixture was added dropwise to a solution of I2 (658g, 2.6mol) in THF (1.3L) over 1 hour at 0-10°C. The reaction mixture was stirred at RT for 1 hour and then rapidly cooled at 0-10°C with 20% w / w NH4Cl (1L). The mixture was extracted with toluene (2.3 L), washed with 10% w / w Na2S2O3 water (1.5 L x 3) and water (1.5 L), and concentrated to a dry state to obtain crude AP2312-B2, which was used directly in the next step.

[0056] Step 2.C: The mixture of crude AP2312-B2 in MeOH (3.3 L) and concentrated HCl water (3.2 L, 38.4 mol) was stirred at RT for 48 hours. The reaction mixture was poured into water (3.3 L) and the pH of the mixture was adjusted to 7-8 with solid NaHCO3. The mixture was concentrated to remove the MeOH and extracted with EA (1.5 L x 2). The combined organic layer was concentrated and dried to obtain crude AP2312-B3, which was used directly in the next step.

[0057] Step 2.D: A mixture of crude AP2312-B3, BnBr (393 g, 2.3 mol), and K2CO3 (290 g, 2.1 mol) in acetone (3.3 L) was stirred at 65°C for 5 hours. After cooling to RT, the inorganic salts were removed by filtration, and the filter cake was washed with EA (660 mL). This filtrate was concentrated and purified by flash chromatography (PE:siRNA = 10:1) to obtain 242.3 g of AP2312-B with 100% LCMS purity and a yield of 50% in the last three steps.

[0058] A second scheme for preparing AP2312-B included steps 2.1, 2.2, 2.3, and 2.4. Step 2.1 (1) DCM (14.1 L) was charged into a 50 L reactor. (2) Methyl 3-hydroxybenzoate (2350.0 g) was charged. (3) DMAP (169.8 g) was charged. (4) (Boc)2O was added dropwise at RT (20 - 25°C). Note: CO2 was generated. (5) The mixture was stirred at RT for 4 h. HPLC showed that there was no residue of SM (reaction completed: SM / Product: ≤1.0%, a / a). (6) The organic solution was washed twice with 8% aqueous citric acid (12 L, 4 L). (7) The organic solution was washed with saturated NaCl solution (5 L). (8) The organic layer was dried over anhydrous Na2SO4 (1 kg). (9) The salt was removed by filtration. (10) The filtrate was concentrated under vacuum at 40°C to obtain a residue with a volume of 5 L. (11) Anhydrous THF (10 L) was charged into the residue. (12) The solution was concentrated under vacuum at 40°C to a volume of 10 L (10.10 kg). Analysis of 10.10 kg: Quantitative 35.07% (containing 3542.1 g); HPLC 99.5%; moisture (KF) 0.1%; yield: 91%. Step 2.2 (13) A dried and clean reactor was set up. (14) The system was flushed three times with N2. (15) At RT (10 - 15°C) and under N2, iPrMgCl·LiCl (12 L, 1.3 M in THF) was transferred into the reactor. (16) Under N2 protection and at RT, TMP (2.204 kg, freshly distilled from CaH2) was added dropwise over 2 h. Note: Gas (propane) was slowly generated during the addition. (17) The grey solution was warmed to 30 - 35°C and stirred for 1 h. Note: The amount of gas increased when the temperature reached 30°C, but it was controllable. (18) The mixture was stirred at 30-35°C for 22 hours. Note: Gas generation has stopped. IPC by GC indicated that the reaction was complete. IPC method: Aliquots (0.1 mL base) were quenched at 10°C with 0.02 mL of PhCHO, and 0.5 mL of saturated NH4Cl aq was added. The organic layer was separated for GC analysis, and the absence of 2-methyl-1-phenylpropan-1-ol indicated complete consumption of the Grignard reagent. (19) AP2312-B1 (7.24 kg THF solution, 27.1% concentration, 1.967 kg, KF: 0.11%) was added to a 50 L reactor under N2. (20) AP2312-B1 was cooled to -5~5°C under N2. (21) The TMPMgCl·LiCl solution from step 6 was carefully added dropwise to the AP2312-B1 solution cooled to 0-5°C over a period of 1.5 hours. (22) The mixture was stirred at 0-5°C for 3 hours. IPC: The sample was rapidly cooled with I2 / THF, and HPLC showed that the exchange was complete (SM / Product: 5.0 / 84.1 = 6% < 10%). (23) The I2 / THF solution (3.96 kg in 8 L of THF) was added dropwise to a cooling solution at 0-10°C over 90 minutes. (24) The solution was stirred for a further 40 minutes at 0-10°C. (25) The solution was heated to 20-25°C. (26) The mixture was stirred at 20-25°C for 2 hours. HPLC analysis revealed that the SM (sulfate mass) was 2.6%. (27) The mixture was cooled to -10°C. (28) While maintaining the temperature at 0-15°C, 5 L of 20% NH4Cl solution was added dropwise to the reaction mixture. (29) Water (18L) was added to RT. (30) 8L of Filtrate was added. (31) The mixture was stirred at RT for 10 minutes. (32) The organic layer (top) was separated. (33) The aqueous layer was extracted with phenyl(5L). (34) The combined organic layers were washed with 10% Na2S2O3 water (10L x 2). The organic layer was separated, and HPLC was performed to obtain a 24.5 kg solution with an 83.0% concentration. Step 2.3 (35) AP2312-B2 (48.3 kg, a solution obtained by combining batches AP2312-B2-1 and AP2312-B2-2, the work-up solution) was added to a 100 L reactor. (36) In RT, an HCl solution (a mixture of 16 L of concentrated HCl and 24 L of tap water) was added to obtain the solution. Note: No significant temperature increase was observed. (37) The mixture was stirred overnight (16 hours) at room temperature (25-30°C). Note: HPLC analysis revealed no residual SM. (38) The solution was transferred to a 200L reactor. (39) 50L of butyl was added. (40) 10% NaCl solution (50L) was added. (41) Layers were separated. (42) The aqueous layer was extracted using phenyl(30L). (43) The combined organic layers were washed with 10% NaCl water (10 L x 2). Note: pH 5-6 after washing. (44) This was washed with saturated NaHCO3 solution (10 L x 2). Note: pH 7 after washing. (45) This was washed with saturated NaCl (10 L). The solution was concentrated to a dry state, yielding 3.80 kg of dark brown oil with an overall yield of 84% from AP2312-B1 and an HPLC purity of 82.1%. Step 2.4 (46) Crude AP2312-B3 (3700g) was added to a 50L reactor. (47) Acetone (37L) was added. (48) K2CO3 (2758.5g) was added. (49) BnBr (2504.2 g) was charged. (50) The mixture was warmed to 55 °C. (51) The mixture was stirred at 55 °C for 3 hours. In the HPLC (210 nm) analysis, the starting material was less than 0.5% (0.2% remaining). (52) The mixture was cooled to RT. (53) The salt was filtered off. (54) The cake was washed with acetone (3.7 L × 2). (55) The filtrate was concentrated at 35 - 40 °C to give 4.5 kg of a brown oil. HPLC of the crude residue: 82.2%. (56) The crude product (4.5 kg diluted with 1 L of DCM) was purified by silica gel chromatography. Note: Silica gel: 22.5 kg (5.0 equivalents, w / w), 300 - 400 mesh; elution EtOAc / PE from 50:1 to 20:1. (57) The product fractions were combined (monitored by TLC). (58) The product fractions were concentrated to a volume of 2 L under vacuum at 35 - 40 °C. Note: A lot of solids were separated out. (59) PE (5 L) was charged. (60) The slurry was concentrated to a volume of 3 L under vacuum at 35 - 40 °C. (61) The solid was collected by filtration. The cake was dried under vacuum at 30 °C to give 2.4 kg of a pale yellow solid with an HPLC purity of 99.7% in a 49% yield. Note: The filtrate was concentrated to give an additional 30 g of a yellow solid with an HPLC purity of 84%.

[0059] In step 3 of the method for synthesizing AMO - 01, AP2312 - 3 is prepared via an Ullmann coupling reaction as follows. [[ID=4②]]

Chemical formula

[0060] The preparation of AP2312-3 was achieved by a binary, highly related scheme using different amounts of reagent (see Table 1), resulting in different amounts of the desired AP2312 product and impurities. The comprehensive procedure was as follows: A mixture of AP2312-B (5.0 g, 13.6 mmol), AP2312-A (4.4 g, 13.6 mmol), CuI, L-proline, and K2CO3 (3.8 g, 27.2 mmol) in DMF (50 mL) and H2O (5 mL) was degassed three times by vacuum / nitrogen purging. The reaction mixture was stirred at 70°C for 6 hours, and the sample was extracted for IPC. After cooling to room temperature, a fixed amount of CuI was added to the reaction mixture, and it was degassed three times by vacuum / nitrogen purging. The reaction mixture was stirred at 90°C for 15 hours, and the sample was extracted for IPC. [Table 1]

[0061] It was found that increasing the CuI loading (No. 6 and 7) accelerated the reaction and increased the amount of deiodination byproducts of AP2312-B (AP2312-3-IM01). In Nos. 1-6, the Ullmann coupling reaction was carried out at 70°C for 6 hours with a fixed amount of CuI to reduce the deiodination byproducts, and then a fixed amount of CuI was added again to promote cyclization at 90°C for 15 hours. In No. 7, the reaction was carried out at 70°C for 6 hours with a 15% CuI loading, followed by 15 hours at 90°C.

[0062] It was found that water accelerates the reaction (comparing No. 3 and No. 4). Without water (No. 4), the Ullmann coupling reaction and cyclization were slow. After 6 hours of reaction at 70°C, 27.7% of AP2312-B remained, and after 15 hours of reaction at 90°C, 8.1% of AP2312-3J remained. AP2312-B and AP2312-3J were completely converted to AP2312-3 after 34 hours at 90°C. When 1v of water was added to the system (No. 3), the Ullmann coupling reaction was completed in 6 hours, and the cyclization reaction was completed in 15 hours.

[0063] Under low CuI loading (1% + 0.5% equivalent, No. 1), the Ullmann coupling reaction and cyclization were found to be slow. After 6 hours of reaction at 70°C, 8.1% of AP2312-B remained, and after 15 hours of reaction at 90°C, 4.6% of AP2312-3J remained.

[0064] System No. 5 (7% + 3.5% equivalent CuI) contained 79.0% AP2312-3, which was slightly higher than systems No. 2 (3% + 1.5% equivalent CuI, 78.2% AP2312-3) and No. 3 (5% + 2.5% equivalent CuI, 78.1% AP2312-3). However, the proportion of AP2312-3 increased under high CuI loads.

[0065] In a specific example, AP2312-3 was prepared by the following steps. (1) DMF (14.0L) was added. (2) Water (1.4L) was added. (3) AP2312-A (1,230 g, 3.84 mol) was added to a 20 L four-necked flask. (4) AP2312-B (1,413 g, 3.84 mol, 1.0 equivalent) was added. (5) CuI (36.60 g, 0.192 mol, 0.05 equivalents) was added. (6) K2CO3 (1,060 g, 7.68 mol, 2.0 equivalents) was added. (7) L-proline (44.2 g, 0.384 mol, 0.1 equivalent) was added. (8) The reaction mixture was heated under N2 at 70°C for 6 hours; by HPLC, AP2312-B was 3.0% at 11.7 minutes. (9) CuI (18.30 g, 0.096 mol, 0.025 equivalents) was added. (10) Reaction mixture heated overnight at 90°C; no intermediate (Ullmann coupling product at 11.1 min) was detected by HPLC. 11.4 min: Deiodination byproduct: Methyl 3-(benzyloxy)benzoate, 16.0%; 9.99 min: Hydrolysis byproduct 3-(benzyloxy)benzoic acid: 2.4%; 13.4 min: Product 81.6% (11) The mixture was cooled to room temperature. (12) The AP2312-3-30, AP2312-3-31, and AP2312-3-33 batches were merged for workup. (13) Activated carbon (847g) was added. (14) The slurry was stirred for 1 hour. (15) The solid was removed by filtration. (16) The cake was washed with DMF (1.4L x 2). (17) The solution was poured into the 50L reactor. (18) NaOH solution (345.6g in 2.1L) was added. (19) The mixture was heated at 70°C for 1 hour; HPLC showed that all deiodized esters were hydrolyzed at 11.4 min to produce acid (in 10.0 min); 10.0 min: 3-(benzyloxy)benzoic acid, 12.7%; 13.8 min, product 87.3%. (20) NH4Cl (1,540 g, 28.8 mol, 7.5 equivalents) was added. (21) Ethylenediamine (877 g, 14.59 mol, 3.8 equivalents) was added. (22) A deep purple solution was added dropwise to a 100 L reactor containing 75.6 L of H2O. (23) The slurry was stirred for 2 hours. (24) When solid matter was collected by filtration, HPLC showed that AP2312-3 was hardly present in the filtrate, and the by-product hydrolyzed at 10.0 mins was purged into the filtrate (3-(benzyloxy)benzoic acid) (HPLC of the filtrate (aqueous solution)). (25) The solid was dissolved in DCM (21 L). (26) Dried with anhydrous Na2SO4 (8 kg). (27) The salt was removed by filtration. (28) The filtrate was concentrated to 7.5 L. (29) Hexane (30.0 L) was added. (30) The slurry was stirred for 1 hour in RT. (31) Solid matter was collected by filtration to obtain the crude product (2.8 kg); HPLC: 94.6%, 13.7 min; most impurities were purged into the filtrate. (32) The crude solid was dissolved in toluene (20 L). (33) Activated carbon (420g) was added. (34) The mixture was heated to 110°C. (35) The mixture was stirred at 110°C for 2 hours. (36) The mixture was cooled to 70-80°C. (37) The activated carbon was removed by filtration. (38) The cake was washed with DCM (2.1L x 3). (39) The combined filtrate was concentrated to 20 L. (40) The mixture was cooled to RT for 2 hours. (41) The mixture was cooled to 5-10°C for 1 hour. (42) Solid matter was collected by filtration. (43) The cake was washed with EtOH (2.1 L). (44) The cake was washed with hexane (2.1 L x 2) to obtain the crude product (1.7 kg); solid: 96.7%, 13.7 min. (45) The crude product was dissolved in DCM (4.28 L). (46) Hexane (17.0 L) was added. (47) The slurry was stirred for 1 hour in RT. (48) Solid matter was collected by filtration to obtain the crude product. (49) The cake was washed with hexane (2.1 L). (50) The cake was dried under vacuum at 40°C to obtain 1.58 kg of red solid with a yield of 50% and HPLC purity of 99.8%. [Table 2]

[0066] In step 4 of the synthesis method for AMO-01, AP2312-4 is prepared by debenzylation as follows. [ka]

[0067] The preparation of AP2312-4 was achieved by the following steps. (1) AP2312-3 (870g, 1,645.9 mmol) was added to a 10L four-necked flask. (2) 10% Pd-C (130.5g, 0.15w / w) was added. (3) THF (2.6L) was added. (4) MeOH (2.6L) was added. (5) The slurry was stirred under H2 conditions with RT for 36 hours. (6) Pd-C was removed by filtration. (7) The cake was washed with MeOH (172 mL x 3). (8) The combined filtrate was concentrated into black oil (660g). (9) Acetone (2.0L) was added. (10) Hexane (2.0 L) was added. (11) The slurry was stirred with RT for 1 hour. (12) Solid matter was collected by filtration. (13) The cake was washed with hexane (660 mL x 2). (14) The mixture was dried under vacuum at 35°C to obtain 469 g of a pale green solid with 100% HPLC purity and 110% excess weight, which was used directly in the next step. [Table 3]

[0068] In step 5 of the method for synthesizing AMO-01, AP2312-5 is prepared by silylation as follows. [ka]

[0069] The preparation of AP2312-5 was achieved by the following steps. (1) DMF (4.0 L) was added to a 10 L four-necked flask. (2) AP2312-4 (450g, 1,742.8 mmol) was added. (3) TEA (881.9g, 8,717mmol, 5.0 equivalents) was added. (4) The solvent was cooled to 0-5°C. (5) TIPSCl (1,344 g, 6,971.2 mmol, 4.0 equivalents) was added dropwise over 1 hour at 0-5°C. (6) The mixture was stirred at RT for 0.5 hours. (7) The mixture was poured into H2O (12.15 L). (8) The mixture was stirred for 1 hour. (9) Solid matter was collected by filtration. (10) The cake was washed with EtOH (2.9L x 2). (11) The product was dried under vacuum at 37°C for 6 hours to obtain 927 g of a yellow solid with 100% HPLC purity and a yield of 80.5% in the last two steps. [Table 4]

[0070] In step 6 of the AMO-01 synthesis method, AP2312-6 is as follows: [ka]

[0071] The preparation of AP2312-6 was achieved by the following steps. (1) DMF (3.7 L) was added to a 10 L four-necked flask. (2) AP2312-C (409g, 1,839 mmol) was administered. (3) 2,6-Lutidine (315.4 g, 2,942.4 mmol, 1.6 equivalents) was added. (4) LiBr (255.7g, 2,942.4 mmol, 1.6 equivalents) was added. (5) The solvent was cooled to 0-5°C. (6) Ms2O was added in several batches while maintaining the internal temperature at 0-5°C. (7) The mixture was stirred at 0-5°C for 3 hours. (8) Pour the mixture into ice water (7.4L). (9) n-heptane (4.9L) was added. (10) The mixture was stirred for 0.5 hours. (11) The organic layer was isolated. (12) The organic solution was passed through a silica gel (81.8g) pad. (13) The silica gel cake was washed with n-heptane (818 ml). (14) The filtrate was concentrated to obtain 544 g of yellow oil with a GC purity of 90% and excess weight (103%), which was used directly in the next step. [Table 5]

[0072] In step 7 of the AMO-01 synthesis method, AP2312-8 is prepared by farnesylation as follows. [ka]

[0073] The preparation of AP2312-8 was achieved by the following steps. (1) Dioxane (8.8 L) was added to a 20 L four-necked flask. (2) t BuOH (3.8L) was added. (3) AP2312-5 (730g, 1,004 mmol) was added. (4) The solvent was cooled to 10-15°C. (5) t 1M in BuOH (1.15 L, 1,150 mmol, 1.15 equivalents) t I installed BuOK. (6) The mixture was stirred at 10-15°C for 2 hours. (7) AP2312-6 was added to dioxane (386.7 g, 730 mL, 1,355 mmol, 1.35 equivalents). (8) The mixture was stirred overnight in RT. (9) Batch AP2312-8-8 and AP2312-8-9 were merged for workup. (10) MTBE (12.96L) was added. (11) The solvent was cooled to 0-5°C. (12) H2O (19.4 L, containing 16.1 g of NH4Cl) was gradually added to the solution. (13) The mixture was stirred for 15 minutes. (14) The organic layer was isolated. (15) Washed with H2O (16.2L x 3 bottles, containing 1,620g of NaCl). (16) The organic layer was concentrated into black oil. (17) The residue was dissolved in THF (20L). (18) The solution was concentrated in black oil. (19) The residue was dissolved in THF (14.6 L). (20) The solution was used directly in the next step. [Table 6]

[0074] In step 8 of the synthesis method for AMO-01, AMO-01 (AP2312) was prepared by desilylation as follows. [ka]

[0075] AP2312 was prepared by the following steps. (1) AP2312-8 in THF (1,037 g, 1,113 mmol, 1.0 equivalent, 14.6 L) was added to a 20 L four-necked flask. (2) The solvent was cooled to -5°C to 5°C. (3) AcOH (535.1 g, 8,904 mmol, 8.0 equivalents) was added. (4) TBAF.3H2O (1,404.6 g, 4,452 mmol, 4.0 equivalents) was added. (5) The mixture was stirred overnight in RT. (6) The solution was concentrated into black oil. (7) Black oil was dissolved in EA (10.37 L). (8) The solution was washed with H2O (10.37 L x 3). (9) The organic layer was isolated. (10) The organic layer was concentrated to 2L. (11) n-heptane (20,740 ml) was added to the solution over 1 hour. (12) The mixture was stirred overnight. (13) Solid matter was collected by filtration. (14) The solid was dissolved in MeOH (4,148 ml) and H2O (519 ml). (15) The solution was washed with n-heptane (4,148 ml x 2). (16) The MeOH-H2O layer was isolated. (17) Activated carbon (104g) was added. (18) The mixture was stirred for 1 hour. (19) Activated carbon was removed by filtration. (20) The activated carbon cake was washed with MeOH (1,037 ml). (21) H2O (6,222 ml) was added to the combined filtrate over a period of 1 hour. (22) The mixture was stirred for 2 hours. (23) Solid matter was collected by filtration. (24) The solid was dissolved in AcOH (2,074 ml). (25) H2O (2,593 ml) was added to the solution over a period of 1 hour. (26) The mixture was stirred overnight. (27) Solid matter was collected by filtration. (28) The cake was washed with AcOH / H2O (519 ml / 519 ml). (29) The cake was washed with H2O (1,037 mL x 2). (30) The cake was dried under vacuum at 37°C, yielding 330 g of a pale gray solid with a yield of 64% in the last two steps and an HPLC purity of 98.3%. [Table 7]

[0076] In summary, a total of 330 g of AP2312 was isolated with HPLC purity of 98.3% (Figure 2). LCMS[M+H]463; 1 HNMR(400MHz;d6DMSO)δ9.99(br.s,1H),9.10(br.s,1H),9.00(br.s,1H),7.16(m,1H),6.78(m,1H),6.68(m,2H),6. 14(m,2H),5.20(m,1H),5.01(m,2H),4.35(m,2H),1.94(m,8H),1.61(s,3H),1.57(s,3H),1.51(s,3H),1.48(s,3H). The isolated products (RRT0.93=0.23%, RRT0.98=0.19%, RRT1.09=0.40%, RRT1.11=0.36%, RRT1.12=0.17%, RRT1.14=0.12%) contained several impurities. This reaction scheme, based on Ullmann coupling, was optimized and verified on a >1 kg scale. Compound of formula I

[0077] It is clear that the specific steps presented above for the production of AMO-01 may also be used for the production of compounds of formulas I and II with only minor modifications.

[0078] For the compound of formula I, the specific steps for the production of AMO-01 described above are performed by using one or more variable parts (or variables) A, R of the compound. 2 , R 3 , R 4 , R 7 , R 8 , W 1 , W 2 , W 3And x (see Equation I) only needs to be changed if it differs from the corresponding variable (or variable) of AMO-01. AP2312M-1

[0079] The following examples are illustrative. In the first example, step 4 is modified, and R 7 Using -CH3, AP2312M-1 is generated. [ka]

[0080] AP2312-3 (15.9, 30 mmol) 1,4-dioxane (192 mL) and t In a BuOH (90 mL) solution, t BuOK (5.0 g, 45 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours. Next, MeI (10.7 g, 75 mmol) was added, and the flask was sealed. The reaction mixture was stirred at 30°C for 24 hours. The solvent was removed by concentration under vacuum, and the residue was dissolved in water (160 mL) and extracted with DCM (160 mL x 2). The combined organic layer was washed with water (160 mL), concentrated, and then purified by reslurrying in petroleum ether (160 mL) and ethyl acetate (16 mL) to obtain 15 g of yellow solid AP2312-11A with HPLC purity of 99.4% in 92% yield.

[0081] A mixture of AP2312M-11A (15.0 g, 27.6 mmol) and 10% Pd / C (wet 50%, 2.4 g) was stirred at 40°C for 24 hours under a hydrogen pressure of 0.1 MPa in THF (45 mL) and MeOH (45 mL). The reaction mixture was cooled to room temperature, and the catalyst was removed by filtration. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 20:1) to obtain 6.5 g of a yellow solid, AP2312M-1, with HPLC purity of 99.1% in 87% yield. LCMS[M+H]273; 1HNMR(d6-DMSO,500MHz)δ10.08(s,1H),9.99(s,1H),9.12(s,1H),7.10(m,1H), 6.85(m,1H),6.76(s,1H),6.71(m,1H),6.20(m,1H),6.12(m,1H),3.29(s,3H). AP2312M-2

[0082] In the second example, step 6 is modified, R 7 AP2312M-2 is produced by using 1-bromo-3-methyl-2-butene. [ka]

[0083] PBr3 (108.4g, 0.4mol) was added dropwise to a DCM solution (430mL) of AP2312M-21 (86.0g, 1.0mol) over 1 hour at 0-10°C. The reaction mixture was stirred overnight in RT and purified by distillation (~50°C / -0.1MPa) to obtain 35.6g of AP2312M-22 in 24% yield, which was used directly in the next step.

[0084] AP2312-5 (21.8g, 30 mmol) 1,4-dioxane (262 mL) and t In a BuOH (110 mL) solution, t BuOK (5.0 g, 45 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours. Next, AP2312M-22 (11.2 g, 75 mmol) was added, and the reaction mixture was stirred at 30°C for 2 hours. After evaporating the solvent, water (220 mL) was added to the residue, and it was extracted with ELISA (110 mL x 2). The combined organic layers were washed with water (220 mL), concentrated to obtain crude AP2312M-23, which was used directly in the next step.

[0085] Crude AP2312M-23 was dissolved in THF (220 mL), then TBAF (120 mL, 1 M in THF) and AcOH (14.4 g, 240 mmol) were added. The reaction mixture was stirred at 30°C for 6 hours. The reaction mixture was poured into water (440 mL) and extracted with SiO2 (440 mL x 1). The organic layer was washed with water (110 mL x 6), concentrated, and purified by flash chromatography (DCM:MeOH = 30:1), resulting in a yield of 43% in the final two steps. 1 4.2 g of AP2312M-2 was obtained as a gray solid with 99.4% HPLC purity, as confirmed by HNMR and LC-MS. LC-MS[M+H]327; 1 HNMR(d6-DMSO,500MHz)σ10.03(s,1H),9.96(s,1H),9.07(s,1H),7.07(d,1H),6.83( d,1H),6.72(m,2H),6.17(s,2H),5.26(m,1H),4.39(m,2H),1.68(s,3H),1.65(s,3H). AP2312M-3

[0086] In the third example, we modify step 6 and again R 7 Modify the code to generate AP2312M-3. [ka]

[0087] To a 92 mL DMF solution of AP2312M-31 (9.2 g, 60 mmol), 2,6-lutidine (10.3 g, 96 mmol), and LiBr (8.4 g, 96 mmol), (Ms)2O (15.7 g) was added in small quantities at 0-10°C. The reaction mixture was stirred at 0-10°C for 2 hours, poured into water (276 mL), and extracted with petroleum ether (92 mL x 2). The combined organic layer was washed with water (92 mL), concentrated, and 11.0 g of AP2312M-32 was obtained in 85% yield, which was used directly in the next step.

[0088] AP2312-5 (14.5g, 20 mmol) contains 1,4-dioxane (174 mL) andt BuOH (73 mL) t BuOK (3.4 g, 30 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours. Next, AP2312M-32 (6.5 g, 30 mmol) was added, and the reaction mixture was stirred at 30°C for 2 hours. After evaporating the solvent, water (145 mL) was added to the residue, and it was extracted with pharmaceutically acceptable phosphate (145 mL x 2). The combined organic layers were washed with water (145 mL), concentrated to obtain crude AP2312M-33, which was used directly in the next step.

[0089] Crude AP2312M-33 was dissolved in THF (145 mL), then TBAF (80 mL in THF, 1 M) and AcOH (9.6 g, 160 mmol) were added. The reaction mixture was stirred at 30°C for 6 hours, poured into water (440 mL), and extracted with toluene (290 mL). The organic layer was washed with water (145 mL x 6), concentrated, and purified by flash chromatography (DCM:MeOH = 40:1), resulting in a yield of 70% in the final two steps. 1 5.5 g of AP2312M-3 was obtained as a gray solid with HPLC purity of 98.2%, as confirmed by HNMR and LC-MS. LC-MS[M+H]395, 1 HNMR(d6-DMSO,500MHz)σ10.04(s,1H),9.95(s,1H),9.05(s,1H),7.07(m,1H),6.83(m,1H),6.72(m,2H), 6.17(m,2H),5.24(m,1H),5.03(m,1H),4.40(m,2H),2.24(m,4H),1.65(s,3H),1.61(s,3H),1.55(s,3H). AP2312M-4

[0090] In the fourth example, we modify step 6 and again R 7 Modify the code to generate AP2312M-4. [ka]

[0091] At 0-10°C, HCOOH (41.4g, 0.9mol), TEA (39.5g, 0.39mol), and meldramic acid (43.2g, 0.3mol) were sequentially added to DMF (100mL). After stirring the reaction mixture at 0-10°C for 0.5 hours, AP2312M-41 (44.5g, 0.3mol) was added. The reaction mixture was stirred overnight at 80°C. After cooling to RT, the reaction mixture was poured into ice water (1.2L) and the pH of the mixture was adjusted to 1-2 with concentrated hydrochloric acid water at 0-10°C. The mixture was filtered, and the filter cake was washed with water (100mL). The filter cake was dissolved in DCM (300mL) and dried with Na2SO4 (90g). After filtering out the inorganic salts, the filtrate was concentrated and dried to obtain crude AP2312M-42, which was used directly in the next step.

[0092] To a solution of crude AP2312M-42 in THF (845 mL), NaBH4 (22.8 g, 0.6 mol) was added in small portions over 0.5 hours at 0-10°C. BF3.Et2O (110.7 g, 0.78 mol) was added dropwise to the reaction mixture over 1.5 hours at 0-10°C. After stirring at RT for 3 hours, the reaction mixture was poured into ice water (300 mL) and the pH of the mixture was adjusted to 2-3 with 2 M HCl at 0-10°C. The mixture was extracted using DCM (600 mL x 2). The combined organic layer was washed with saturated NaHCO3 (500 mL) and brine (500 mL), concentrated, and purified by flash chromatography (PE:EA = 5:1) to obtain 46.0 g of colorless oil AP2312M-43 in a two-step yield of 86%.

[0093] PBr3 (17.9 g, 66 mmol) was added dropwise to a DCM solution (294 mL) of AP2312M-43 (29.4 g, 165 mmol) at 0-10°C. The reaction mixture was stirred in RT for 15 hours, washed with water (210 mL), concentrated, and purified by flash chromatography (PE:EA = 20:1) to obtain 19.5 g of AP2312M-53 as an off-white solid in 49% yield.

[0094] AP2312-3 (21.2g, 40 mmol) 1,4-dioxane (254 mL) andt In a BuOH (106 mL) solution, t BuOK (5.8 g, 52 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours. Next, AP2312M-44 (19.3 g, 80 mmol) was added, and the reaction mixture was stirred at 30°C for 24 hours. After evaporating the solvent, water (212 mL) was added to the residue, and it was extracted with ELISA (106 mL x 2). The combined organic layers were washed with water (106 mL), concentrated, and crude AP2312M-45 was obtained as a yellow solid, which was used directly in the next step.

[0095] Crude AP2312M-45 was dissolved in THF (212 mL) and MeOH (106 mL), and then 10% Pd / C and 50% water-wet product (3.2 g) were added. The reaction mixture was degassed three times by vacuum / hydrogen purging and stirred at 40°C for 24 hours at a hydrogen pressure of 0.1 MPa. After the reaction mixture cooled to room temperature, the catalyst was removed by filtration. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 40:1), resulting in a yield of 40% in the last two steps. 1 6.7 g of AP2312M-4 was obtained as a gray solid with 98.0% HPLC purity, as confirmed by HNMR and LC-MS. LC-MS[M+H]419; 1 HNMR(d6-DMSO,500MHz)σ10.06(s,1H),10.01(s,1H),9.09(s,1H),7.06(m,3H),6.99(m,2H),6.83(m,1H) ,6.78(s,1H),6.73(m,1H),6.19(m,2H),3.91(m,2H),2.81(m,1H),2.53(m,2H),1.76(m,2H),1.15(d,6H). AP2312M-5

[0096] In the fifth example, we modify step 6 and again R 7 Modify the code to generate AP2312M-5. [ka]

[0097] A mixture of AP2312M-51 (25.0 g, 116 mmol), 4-isopropylphenylboronic acid (22.8 g, 139 mmol), PdCl2 (dppf) (878 mg, 1.2 mmol), and K2CO3 (32.0 g, 232 mmol) in MeOH (300 mL) and H2O (100 mL) was degassed three times by vacuum / nitrogen purging. After stirring at 70°C for 15 hours, the reaction mixture was concentrated to remove the MeOH and extracted with siRNA (100 mL x 2). The combined organic layer was concentrated and purified by flash chromatography (PE:EA = 10:1) to obtain 22.0 g of AP2312M-52 as an off-white solid in 74% yield.

[0098] PBr3 (11.7 g, 43.3 mmol) was added dropwise to a solution of AP2312M-52 (22.0 g, 86.6 mmol) in DCM (220 mL) at 0-10°C. The reaction mixture was stirred at RT for 15 hours, washed with water (220 mL), concentrated, and purified by flash chromatography (PE:EA = 30:1) to obtain 11.3 g of AP2312M-53 as an off-white solid in 41% yield.

[0099] AP2312-3 (12.2g, 23 mmol) contains 1,4-dioxane (146 mL) and t BuOH (61 mL) solution t BuOK (3.4 g, 30 mmol) was added. The reaction mixture was stirred at 30°C for 2 hours. Next, AP2312M-53 (11.1 g, 35 mmol) was added, and the reaction mixture was stirred at 30°C for 24 hours. After evaporating the solvent, water (122 mL) was added to the residue, and it was extracted with ELISA (61 mL x 2). The combined organic layers were washed with water (61 mL), concentrated, and a yellow solid, the residue AP2312M-54A, was obtained and used directly in the next step.

[0100] Crude AP2312M-54A was dissolved in THF (122 mL) and MeOH (61 mL), and then 10% Pd / C and 50% water-wet product (1.8 g) were added. The reaction mixture was degassed three times by vacuum / hydrogen purging and stirred at 40°C for 24 hours at a hydrogen pressure of 0.1 MPa. After the reaction mixture cooled to room temperature, the catalyst was removed by filtration. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 40:1), resulting in a final two-step yield of 70%. 1 8.0 g of AP2312M-5 was obtained as an off-white solid with 99.0% HPLC purity, as confirmed by HNMR and LC-MS. LC-MS[M+H]495; 1 HNMR(d6-DMSO,500MHz)σ10.07(s,1H),10.02(s,1H),9.10(s,1H),7.50(m,4H),7.30(m,2H),7.16(m,2H),7.07(m,1H) ,6.83(m,1H),6.80(s,1H),6.73(m,1H),6.20(m,2H),3.94(m,2H),2.90(m,1H),2.62(m,2H),1.82(m,2H),1.22(d,6H). Compound of formula II

[0101] As suggested above, the specific steps used to produce AMO-01 can also be used to produce the compound of formula II with only minor modifications. The compound of formula II is the result of using a Buchwald coupling instead of an Ullmann coupling in step 3.

[0102] In the initial experiments used to produce AMO-01, the surprising result was obtained that the compound of formula II was realized using Buchwald coupling, and the compound of formula I was obtained using Ullmann coupling.

[0103] Experiments were conducted to confirm that the Buchwald chemical reaction yields the AP2312-3I isomer, rather than the AP2312-3 isomer. [ka]

[0104] A mixture of AP2312-2 (21.0 g, 60 mmol), AP2312-B (28.8 g, 78 mmol), Pd2(Dba)3 (1.1 g, 1.2 mmol), X-Phos (2.8 g, 3.6 mmol), and Cs2CO3 (49.2 g, 150 mmol) in Tol (210 mL) was degassed three times by vacuum / nitrogen purging. The reaction mixture was stirred at 110 °C for 48 hours. The reaction mixture was poured into water (210 mL) and extracted with EA (210 mL x 2). The combined organic layer was washed with water (210 mL), concentrated, and purified by flash chromatography (PE:EA = 10:1) to obtain 26.0 g of AP2312-14 with HPLC purity of 98.0% in 73% yield.

[0105] AP2312-14 (25.0 g, 42.4 mmol) was suspended in EtOH (100 mL), H2O (50 mL), and AcOH (45 mL). Zinc powder (9.4 g, 144.2 mmol) was added to the reaction mixture in fractions by RT. A vigorous exothermic reaction occurred, and the temperature rose to 80°C in 1 hour. The reaction mixture was stirred at 80°C for 2 hours. After the reaction mixture cooled to room temperature, the inorganic salts were removed by filtration, and the filter cake was washed with DCM (200 mL). The filtrate was concentrated to remove the organic solvent and extracted with DCM (250 mL × 1). The organic layer was washed with water (100 mL × 3), concentrated, and purified by reslurrying in EtOH (100 mL) to obtain 21.0 g of AP2312-3I as a yellow solid with 100% LCMS purity in 94% yield.

[0106] Next, using Buchwald coupling, AP2312-3I was generated as follows. [ka]

[0107] A mixture of AP2312-A (1.3g, 4 mmol), AP2312-B (1.5g, 4 mmol), PdCl2 (dppf) (146mg, 0.2 mmol), and Cs2CO3 (1.8g, 5.6 mmol) in DMF (26 mL) was degassed three times by vacuum / nitrogen purging. The reaction mixture was stirred at 100°C for 15 hours. Samples were taken for IPC, and HPLC showed that 47.0% of AP2312-3I was present in the system (20.7 min), while AP2312-3 was absent (19.0 min). LCMS[M+H]529; 1 HNMR(d6-DMSO,500MHz)σ8.76(s,1H),7.53(m,4H),7.35(m,11H),7.22(m,1H),7.13(m,1H), 7.08(s,1H),6.85(m,1H),6.49(m,1H),6.40(m,1H),5.26(s,2H),5.11(s,2H),5.03(s,2H).

[0108] Although the present invention has been described with reference to specific embodiments, it will be apparent to those skilled in the art that various modifications may be made without departing from the spirit and scope of the invention. The claims are not limited to the specific embodiments described herein.

Claims

1. A method for synthesizing the compound of formula I and its salt, 【Chemistry 1-1】 In formula I, A is -NH- and R 7 is -CH 3 , -(CH 2 ) x CH 3 , -CH 2 CH 2 W 1 CH 3 , -CH 2 CH 2 W 1 CH 2 CH 2 W 2 CH 3 , or -CH 2 W 1 CH 2 CH 2 W 2 CH 2 CH 2 W 3 CH 3 where x is an integer from 1 to 11, and each of W 1 , W 2 , and W 3 is independently [Chemistry 1-2] And; The aforementioned method, (a) Prepare AP2312-A; [Chemistry 1-3] (b) Prepare AP2312-B; [Chemistry 1-4] (c) Perform Ullmann coupling; [Chemistry 1-5] (d) Debenzylation; [Chemistry 1-6] (e) Perform silylation; [Chemistry 1-7] (f) Prepare R7-X from R7-OH (where R 7 This is the same definition as in formula I above, and also the same in formulas (g) and (h) below; [Chemistry 1-8] Here, X is Br, I, or Cl, (g) Add R7-X; [Chemistry 1-9] Furthermore, (h) desilylation; 【Chemistry 1-10】 The method, including the method described above.

2. A method for synthesizing farnesyldibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxydibenzodiazepine-11-one), 【Chemistry 2-1】 (a) Prepare AP2312-A; 【Chemistry 2-2】 (b) Prepare AP2312-B; [Chemistry 2-3] (c) Perform Ullmann coupling; 【Chemistry 2-4】 (d) Debenzylation; 【Chemistry 2-5】 (e) Perform silylation; 【Chemistry 2-6】 (f) Prepare farnesyl bromide; 【Chemistry 2-7】 (g) Perform farnesylation; 【Chemistry 2-8】 Furthermore, (h) desilylation; 【Chemistry 2-9】 The method, including the method described above.

3. A method for synthesizing farnesyl dibenzodiazepinone AMO-01, (a) Prepare AP2312-A; 【Chemistry 3-1】 (b) Prepare AP2312-B; 【Chemistry 3-2】 (c) CuI (0.0525 equivalent), K 2 CO 3 In the presence of (2.0 equivalents), L-proline (0.1 equivalent), and DMF, molecular equivalents of AP2312-A and AP2312-B are reacted to perform Ullmann coupling to obtain AP2312-3; 【Chemistry 3-3】 (d) In the presence of THF, MeOH, and Pd / C, H 2 Below, debenzylate AP2312-3 to obtain AP2312-4; [Chemistry 3-4] (e) TIPSCl (4.0 equivalents) Et 3 AP2312-4 is silylated in the presence of N (5.0 equivalents) and DMF to obtain AP2312-5; [Transformation 3-5] (f) Ms 2 AP23132-C is reacted in the presence of O, LiBr (1.6 equivalents), 2,6-lutidine (1.6 equivalents), and DMF to obtain AP2312-6; [Chemistry 3-6] (g) dioxane, t BuOH, and t AP2312-5 is farnesylated with AP2312-6 in the presence of BuOK (1.15 equivalents) to obtain AP2312-8; 【Chemistry 3-7】 Furthermore, (h) desilylation of AP2312-8 in the presence of THF (1.0 equivalent), AcOH (8.0 equivalent), and TBAF (4.0 equivalent) to obtain AMO-01; 【Transformation 3-8】 The method, including the method described above.

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