Process for the synthesis of farnesyl bisdibenzoazepinone

Abstract

The present invention relates to a synthetic process for the production of farnesyl bisdibenzoazepinones, including AMO-01.

Description

BACKGROUND

[0001] Actinobacteria are a subset of a large and complex group of gram-positive bacteria known as actinomycetes. In the past few decades, these soil-rich organisms have attracted great commercial and scientific interest due to their ability to produce a large number of therapeutically useful compounds, particularly antibiotics, as secondary metabolites. Extensive research into strains capable of producing new antibiotics has led to the identification of hundreds of new species.

[0002] Many actinobacteria, particularly the genera Streptomyces and the closely related Saccharopolyspora, have been extensively studied. Both of these genera produce a remarkable diversity of biologically active metabolites. Because of the commercial significance of these compounds, much is known about the genetics and physiology of these organisms. Another representative genus of actinobacteria, Micromonospora, has also attracted commercial interest. For example, U.S. Patent No. 5,541,181 (Ohkuma et al.) discloses a dibenzodiazepinone compound, specifically 5-farnesyl-4,7,9-trihydroxy-dibenzodiazepin-11-one (named "BU-4664L"), produced by a known actinobacterial strain, Micromonospora sp. M990-6 (ATCC 55378). The Ohkuma et al. patent reports that BU-4664L and its chemically synthesized dialkoxy, trialkoxy, and acyloxy derivatives have anti-inflammatory and antitumor cell activity. In another example, U.S. Patent No. 7,101,872 (Bachmann et al.) discloses a farnesyl dibenzodiazepinone compound, specifically 10-farnesyl-4,6,8-trihydroxy-dibenzazepin-11-one (named "ECO-04601", "AMO-01" in the patent).

[0003] Pharmaceutical applications research on these compounds would be aided by a reproducible method for producing sufficient quantities of the compounds at acceptable levels of purity for in vitro and animal testing. Available methods for preparing dibenzodiazepinone compounds are primarily based on culturing the microorganism under conditions that induce production of the compound, followed by multiple rounds of extraction, concentration, and purification of the culture medium and fermentation broth. These methods are both expensive and time-consuming.

[0004] Accordingly, there is a great need for the development of synthetic methods for producing dibenzodiazepinone compounds. The present invention is directed to this end, as well as other important ends. SUMMARY

[0005] The present invention relates to a novel method for synthesizing a farnesyl dibenzodiazepinone compound, such as AMO-01 as defined herein

[0006]

[0007] As discussed in detail below, the process for synthesizing some of the farnesyl dibenzodiazepinone compounds of the invention is based on the inventors' unexpected discovery that the Ullmann coupling reaction, when carefully controlled for the amount of copper in the reaction, achieves surprising regioselectivity in the resulting compounds. In contrast, the palladium catalyzed Buchwald coupling produces the opposite regiochemical structure from the same starting materials. This difference is exploited in the process disclosed herein, allowing the production of farnesyl dibenzodiazepinone compounds with selected stereochemical structures.

[0008] In a first embodiment, the invention relates to a process for synthesizing a farnesyl dibenzodiazepinone of Formula I:

[0009]

[0010] wherein,

[0011] A is -NH-;

[0012] R 7 is -CH3, -(CH2)xCH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2 CH2CH2W 3 CH3, wherein x is an integer from 1 to 11, and wherein each of W1, W2, and W3is independently

[0013]

[0014] R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3; and

[0015] R 3 and R 4 are the same and are selected from -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3. In certain aspects, the process is carried out by a Ullmann reaction.

[0016] The method of synthesizing a farnesyl dibenzodiazepinone of Formula I comprises the following steps, wherein A, R 2 3 4 7 8 1 2 3 and x are defined as above for Formula I:

[0017] (a) preparing AP2312-A;

[0018]

[0019] (b) preparing AP2312-B;

[0020]

[0021] (c) performing a Ullmann coupling;

[0022]

[0023] (d) performing a debenzylation;

[0024]

[0025] (e) performing silylation;

[0026]

[0027] (f) preparing R 7 ;

[0028]

[0029] wherein X is Br, I or Cl,

[0030] (g) performing a farnesylation; and

[0031]

[0032] (h) desilylating

[0033]

[0034] In a second embodiment, the present application relates to a method of synthesizing a farnesyl dibenzodiazepinone of Formula II and salts thereof:

[0035]

[0036] wherein,

[0037] A is -NH-; ​​​​​​​

[0038] R 7 is -CH3, -(CH2)xCH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3or -CH2W 1 CH2CH2W 2 CH2CH2W 3 CH3, wherein x is an integer from 1 to 11, and wherein W 1 , W 2 , and W 3 each independently is

[0039]

[0040] R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3; and

[0041] R 5 and R 6 are the same and selected from -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3. In certain aspects, the method is by Buchwald coupling.

[0042] A method of synthesizing a farnesyl dibenzodiazepinone of Formula II comprises the following steps, wherein A, R 2 , R 5 , R 6 , R 7 , R 8 , W 1 , W 2 , W 3 , and x are as defined above for Formula II:

[0043] (a) preparing AP2312-A;

[0044]

[0045] (b) preparing AP2312-B;

[0046]

[0047] (c) performing Buchwald coupling;

[0048]

[0049] (d) performing de-benzylation;

[0050]

[0051] (e) performing methyl silylation;

[0052]

[0053] (f) preparing R 7 ;

[0054]

[0055] wherein X is Br, I or CI,

[0056] (g) performing farnesylation; and

[0057]

[0058] (h) performing desilylation

[0059]

[0060] In a third embodiment, the present application relates to a method of synthesizing farnesyl dibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxy-dibenzodiazepin-11-one; also known as "AP2312")

[0061]

[0062] In one aspect, the present application comprises the following steps;

[0063] (a) preparing AP2312-A;

[0064]

[0065] (b) preparing AP2312-B;

[0066]

[0067] (c) performing Ullmann coupling;

[0068]

[0069] (d) performing de-benzylation;

[0070]

[0071] (e) performing methyl silylation;

[0072]

[0073] (f) preparing farnesyl bromide;

[0074]

[0075] (g) performing farnesylation; and

[0076]

[0077] (h) performing desilylation

[0078]

[0079] In one specific aspect, synthesizing AMO-01 includes the following steps:

[0080] (a) preparing AP2312-A;

[0081]

[0082] (b) preparing AP2312-B;

[0083]

[0084] (c) performing Ullmann coupling by reacting molecular equivalents of AP2312-A and AP2312-B in the presence of CuI (0.0525 equivalents), K2CO3 (2.0 equivalents), L-proline (0.1 equivalents), and DMF to produce AP2312-3;

[0085]

[0086] (d) performing debenzyl reaction of AP2312-3 in the presence of THF, MeOH, and Pb / C under H2 to produce AP2312-4;

[0087]

[0088] (e) performing methylsilanation reaction of AP2312-4 in the presence of TIPSCl (4.0 equivalents), Et3N (5.0 equivalents), and DMF to produce AP2312-5;

[0089]

[0090] (f) reacting AP2312-C in the presence of Ms2O, LiBr (1.6 equivalents), 2,6-lutidine (1.6 equivalents), and DMF to produce AP2312-6;

[0091]

[0092] (g) farnesylation of AP2312-5 using AP2312-6 in the presence of dioxane, t BuOH and t BuOK (1.15 equivalents) to produce AP2312-8; and

[0093]

[0094] (h) desilylation of AP2312-8 in the presence of THF (1.0 equivalents), AcOH (8.0 equivalents) and TBAF (4.0 equivalents) to produce AMO-01

[0095]

[0096] The foregoing has outlined rather broadly the features and technical advantages of the present application so as to provide an overall understanding of the application's detailed description that follows. Additional features and advantages of the application will be described hereinafter that form the subject of the claims of the application. Those skilled in the art will appreciate that any of the conception and specific embodiments disclosed herein can be readily utilized as a basis for modifying or designing other structures for carrying out the same purposes of the present application. Those skilled in the art will also realize that such equivalent constructions do not depart from the spirit and scope of the application as set forth in the appended claims. The novel features which are believed to be characteristic of the application, both as to its organization and method of operation, together with further objects and advantages will be better understood from the following description when considered in connection with the accompanying figures. It is to be expressly understood, however, that any description, figure, example, etc. provided herein is merely intended to illustrate the present application and should not be construed to limit the scope of the present application. BRIEF DESCRIPTION OF DRAWINGS

[0097] The above and other objects, features and advantages of the present application will become more apparent from the following description when taken in conjunction with the accompanying drawings, in which:

[0098] Figure 1 General scheme of the method for the synthesis of farnesylidiphenyldiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxy-dibenzoazepin-11-one).

[0099] Figure 2 HPLC results show a purity of 98.3% for AMO-01. DETAILED DESCRIPTION

[0100] I. DEFINITIONS

[0101] As used herein, "a" or "an" can mean one or more. As used herein, when combined with the word "comprising," the word "a" or "an" can mean one or more than one. As used herein "another" can mean at least a second or more. Further, unless otherwise required by context, singular terms include pluralities and plural terms include the singular.

[0102] As used herein, "about" refers to a value including, for example, integers, fractions, and percentages, whether expressly stated or not. The term "about" generally refers to a range of values that a person of ordinary skill in the art would consider equivalent to the stated value (e.g., + / - 5-10% of the stated value). In certain instances, the term "about" can include values rounded to the nearest significant figure.

[0103] II. Farnesyl Dibenzazepinone Compounds

[0104] AMO-01 (10-farnesyl-4,6,8-trihydroxy-dibenzazepin-11-one; also referred to herein as "AP2312") is a member of the class of farnesyl dibenzazepinone and dibenzazepinone compounds containing a farnesyl moiety. The structure of AMO-01 is as follows:

[0105]

[0106] Farnesyl dibenzazepinone compounds can be produced by a biological process by culturing certain strains of Micromonospora, a genus of bacteria in the family of Micromonosporaceae, which are gram-positive, spore-forming, usually aerobic and branching mycelial, from which the compound is then isolated from the culture medium. Members of this genus also typically produce aminoglycoside antibiotics.

[0107] AMO-01 is produced by strain 046-ECO11 of the genus Micromonospora. Strain 046-ECO11 was deposited with the International Depository Authority of Canada (IDAC) (Health Canada Microbiology Bureau, 1015 Arlington Street, Winnipeg, Manitoba, Canada R3E 3R2) on March 7, 2003 under Accession No. 070303-01. Further details regarding strain 046-ECO11 and the biological process for producing AMO-01 can be found in International Patent Publication WO 2004 / 065591 published August 5, 2004, the contents of which are incorporated herein by reference in their entirety.

[0108] Through the persistent efforts of the inventor, a method for the total synthesis of farnesyl dibenzodiazepinone compounds, including AMO-01, has been achieved. The present invention relates to such devices, and related aspects of the present invention disclosed herein.

[0109] Accordingly, in one embodiment, the present invention relates to a method of synthesizing farnesyl dibenzodiazepinone compounds of Formula I and salts thereof:

[0110]

[0111] wherein,

[0112] A is -NH-;

[0113] R 7 is -CH3, -(CH2)xCH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3or -CH2W 1 CH2CH2W 2 CH2CH2W 3 CH3, wherein x is an integer from 1 to 11, and wherein W 1 , W 2 , and W 3 each independently is

[0114]

[0115] R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3; and

[0116] R 3 and R 4 are the same and selected from -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3. In certain aspects, the method is carried out by a Ullmann reaction.

[0117] The method of synthesizing farnesyl dibenzodiazepinones of Formula I includes the following steps, wherein A, R 2 , R 3 , R 4 , R 7 , R 8 , W 1 , W 2 , W 3and x is as defined above for Formula I:

[0118] (a) preparing AP2312-A;

[0119]

[0120] (b) preparing AP2312-B;

[0121]

[0122] (c) performing Ullmann coupling;

[0123]

[0124] (d) performing debenzyl;

[0125]

[0126] (e) performing silylation;

[0127]

[0128] (f) preparing R 7 ;

[0129]

[0130] wherein X is Br, I or Cl,

[0131] (g) performing farnesylation; and

[0132]

[0133] (h) performing desilylation

[0134]

[0135] The present application also relates to a method of synthesizing farnesyl dibenzodiazepinones of Formula II and salts thereof:

[0136]

[0137] wherein,

[0138] A is -NH-;

[0139] R 7 is -CH3, -(CH2)xCH3, -CH2CH2W 1 CH3, -CH2CH2W 1 CH2CH2W 2 CH3, or -CH2W 1 CH2CH2W 2CH2CH2W 3 CH3, wherein x is an integer from 1 to 11, and wherein W 1 , W 2 , and W 3 each independently is

[0140]

[0141] R 2 is -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3; and

[0142] R 5 and R 6 are the same and selected from -H, -OH, -OCH3, or -OP=O(OR 8 ), wherein R 8 is -Na, -CH3, or -CH2CH3. In certain aspects, the method is by Buchwald coupling.

[0143] A method of synthesizing a farnesyl dibenzodiazepinone of Formula II comprises the following steps, wherein A, R 2 , R 5 , R 6 , R 7 , R 8 , W 1 , W 2 , W 3 , and x are as defined above for Formula II:

[0144] (a) preparing AP2312-A;

[0145]

[0146] (b) preparing AP2312-B;

[0147]

[0148] (c) performing Buchwald coupling;

[0149]

[0150] (d) performing debenzylization;

[0151]

[0152] (e) performing methyl silylation;

[0153]

[0154] (f) preparing R 7 ;

[0155]

[0156] wherein X is Br, I or CI,

[0157] (g) performing farnesylation; and

[0158]

[0159] (h) performing desilylation

[0160]

[0161] The present application also relates to a method of synthesizing farnesyl bis-dibenzodiazepinone AMO-01

[0162]

[0163] In certain aspects, the method comprises the following steps;

[0164] (a) preparing AP2312-A;

[0165]

[0166] (b) preparing AP2312-B;

[0167]

[0168] (c) performing Ullmann coupling;

[0169]

[0170] (d) performing debenzyl;

[0171]

[0172] (e) performing methylsilanylation;

[0173]

[0174] (f) preparing farnesyl bromide;

[0175]

[0176] (g) performing farnesylation; and

[0177]

[0178] (h) performing desilylation

[0179]

[0180] In one particular aspect, the method of synthesizing AMO-01 comprises the following steps:

[0181] (a) preparing AP2312-A;

[0182]

[0183] (b) preparing AP2312-B;

[0184]

[0185] (c) performing a Ullmann coupling of a molecular equivalent of AP2312-A and AP2312-B in the presence of CuI (0.0525 equivalents), K2CO3 (2.0 equivalents), L-proline (0.1 equivalents), and DMF to produce AP2312-3;

[0186]

[0187] (d) performing a debenzylation of AP2312-3 in the presence of THF, MeOH, and Pb / C under H2 to produce AP2312-4;

[0188]

[0189] (e) performing a methylsilanization of AP2312-4 in the presence of TIPSCl (4.0 equivalents), Et3N (5.0 equivalents), and DMF to produce AP2312-5;

[0190]

[0191] (f) reacting AP2312-C in the presence of Ms2O, LiBr (1.6 equivalents), 2,6-lutidine (1.6 equivalents), and DMF to produce AP2312-6;

[0192]

[0193] (g) performing a farnesylation of AP2312-5 using AP2312-6 in the presence of dioxane, t BuOH, and t BuOK (1.15 equivalents) to produce AP2312-8; and

[0194]

[0195] (h) desilylation of AP2312-8 in the presence of THF (1.0 equivalent), AcOH (8.0 equivalents), and TBAF (4.0 equivalents) to produce AMO-01

[0196]

[0197] The following are exemplary compounds and specific embodiments of farnesyl dibenzodiazepinone compounds, which can be produced by the methods of the present application as defined herein:

[0198]

[0199]

[0200]

[0201]

[0202]

[0203]

[0204] With reference to the variable "x" as an integer in the formula of the present application, it is understood that x is an integer of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11. The integer x can range from 1 to 11, from 1 to 10, from 1 to 9, from 1 to 8, from 1 to 7, from 1 to 6, from 1 to 5, from 1 to 4, from 1 to 3, from 1 to 2. For the avoidance of any doubt, the above ranges include both endpoints of the range as integers.

[0205] As used herein, the term "alkyl" refers to straight chain or branched chain hydrocarbon groups. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, pentyl, hexyl, heptyl, cyclopentyl, cyclohexyl, cyclohexymethyl, and the like. The alkyl group can be optionally substituted with a substituent selected from the group consisting of acyl, amino, amido, acyloxy, carboalkoxy, carboxy, carboxyamido, cyano, halogen, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, oxo, guanidinyl, and formyl. The number of carbons in the hydrocarbon group can range from 1 to 6 carbon atoms, and includes 1 to 2 carbon atoms, 1 to 3 carbon atoms, 1 to 4 carbon atoms, and 1 to 5 carbon atoms.

[0206] As used herein, the term "alkene" refers to an unsaturated hydrocarbon group containing a carbon-carbon double bond. The number of carbons in the hydrocarbon group can range from 2 to 6 carbon atoms, and includes 2 carbon atoms, 2-3 carbon atoms, 2-4 carbon atoms, and 2-5 carbon atoms.

[0207] As used herein, the terms "aryl" and "aryl ring" refer to an aromatic group in a monocyclic or fused ring system having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring members. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, biphenyl, terphenyl. The aryl group can be optionally substituted with one or more substituents selected from acyl, amino, amido, acyloxy, azido, alkylthio, carboxyalkoxy, carboxy, carboxamido, cyano, halogen, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0208] As used herein, the terms "heteroaryl" and "heteroaryl ring" refer to an aromatic group in a monocyclic or fused ring system having 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring members and containing at least one heteroatom selected from O, N, S, SO, and SO2. Examples of heteroaryl groups include, but are not limited to, pyridyl, thiazolyl, thiadiazolyl, isoquinolyl, pyrazolyl, oxazolyl, oxadiazolyl, triazolyl, and pyrrolyl. The heteroaryl group can be optionally substituted with one or more substituents selected from acyl, amino, amido, acyloxy, carboxyalkoxy, carboxy, carboxamido, cyano, halogen, hydroxyl, nitro, thio, thioxocarbonyl, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0209] The term "alkenyl" refers to a straight chain, branched, or cyclic hydrocarbon group containing at least one carbon-carbon double bond. Examples of alkenyl groups include, but are not limited to, ethenyl, 1-propen-2-yl, 1-buten-4-yl, 2-buten-4-yl, 1-penten-5-yl, and the like. The alkenyl group can be optionally substituted with a substituent selected from acyl, amino, amido, acyloxy, carboxyalkoxy, carboxy, carboxamido, cyano, halogen, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, formyl, oxyl, and guanidinyl. The double bond portion of the unsaturated hydrocarbon chain can be in the cis or trans configuration.

[0210] The terms "cycloalkyl" and "cycloalkyl ring" refer to a ring having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 10, 11, 12, 13, 14, or 15 ring members. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclohexyl, and cycloheptyl. Cycloalkyl groups can be optionally substituted with a substituent selected from the group consisting of acyl, amino, amido, acyloxy, carboxyalkoxy, carboxy, carboxamido, cyano, halogen, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0211] The terms "heterocyclyl" and "heterocycle" refer to a saturated or partially unsaturated ring containing 1, 2, 3, or 4 heteroatoms or heterogroups selected from O, N, NH, NRx, PO2, S, SO, or SO2 in a single or fused heterocyclic ring system having 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ring members. Examples of heterocyclyl or heterocycle include, but are not limited to, morpholinyl, piperidinyl, and pyrrolidinyl. Heterocyclyl, heterocycle, or heterocyclyl ring can be optionally substituted with a substituent selected from the group consisting of acyl, amino, amido, acyloxy, oxyl, thiocarbonyl, imino, carboxyalkoxy, carboxy, carboxamido, cyano, halogen, hydroxyl, nitro, thio, alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl, alkoxy, aryloxy, sulfinyl, sulfonyl, and formyl.

[0212] 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.

[0213] The term "halogen" refers to a halogen atom, such as bromine, chlorine, fluorine, and iodine.

[0214] The terms "aralkyl" and "heteroaralkyl" refer to aryl or heteroaryl groups, respectively, directly bonded through an alkyl group, such as benzyl. Aralkyl and heteroaralkyl can be optionally substituted as aryl and heteroaryl.

[0215] Similarly, the terms "aralkenyl" and "heteroaralkenyl" refer to aryl or heteroaryl groups, respectively, directly bonded through an alkenyl group, such as benzyl. Aralkenyl and heteroaralkenyl can be optionally substituted as aryl and heteroaryl.

[0216] The compounds of the present application can have one or more asymmetric carbon atoms and can exist as optical isomers, forming mixtures of racemic or non-racemic compounds. The compounds of the present application can be used as single isomers or as mixtures of stereochemically isomeric forms. Diastereomers, i.e., non- overlapping stereochemical isomers, can be separated by conventional methods, such as chromatography, distillation, crystallization or sublimation. Optical isomers can be obtained by resolution of a racemic mixture according to conventional processes.

[0217] III. Synthetic Methods

[0218] As outlined above, the present application relates to methods of synthesizing farnesyl dibenzodiazepinones of Formula I and Formula II as defined herein. Reaction schemes for the compounds encompassed by these formulas are provided herein. Initial experiments for producing the farnesyl dibenzodiazepinones of the present application resulted in the surprising discovery that by using Buchwald coupling, compounds of Formula II were achieved, while using Ullmann coupling resulted in compounds of Formula I. Thus, while the initial steps in the synthesis of compounds of Formula I and Formula II are identical, the choice of Ullmann coupling versus Buchwald coupling drives the rejection of the formation of compounds of Formula I and Formula II, respectively.

[0219] AMO-01

[0220] In one particular embodiment, the present application relates to a method of synthesizing farnesyl dibenzodiazepinone AMO-01 (10-farnesyl-4,6,8-trihydroxy- dibenzodiazepin-11-one). Details regarding this method are provided in the following paragraphs, and a general scheme can be seen in Figure 1 AMO-01 is referred to as AP2312 in the examples.

[0221] In Step 1 of the method for synthesizing AMO-01, AP2312-A is prepared as follows:

[0222]

[0223] The preparation of AP2312-A is achieved by two alternative, highly related schemes. The first scheme includes Step 1.A, Step 1.B, and Step 1.C.

[0224] Step 1. A Ammonia gas (~240 g, 14.1 mol) was bubbled into a solution of 1,3,5-trifluoro-2-nitrobenzene (490.0 g, 2.77 mol) in THF (2.45 L) at -60 °C to -40 °C over 2 h. After stirring at 0 °C for 4 h, the reaction mixture was filtered and the filter cake was washed with EtOAc (490 mL x 4). The filtrate was concentrated to ~500 mL and petroleum ether (980 mL) was added. The mixture was reslurried at room temperature (RT) overnight, filtered and the filter cake was washed with petroleum ether (490 mL). The filter cake was dried under vacuum at 40 °C for 5 h to give 366 g of AP2312-1 as an orange solid with 76% yield and 98.0% HPLC purity.

[0225] Step 1. B A mixture of KOH (151 g, 2.7 mol) and BnEt3N + Cl - (98 g, 0.43 mol) in BnOH (1045 g, 9.7 mol) was stirred at RT for 0.5 h. AP2312-1 (188 g) was added portionwise to the reaction mixture over 0.5 h and stirred at 80 °C for 3 h. After cooling to RT, the reaction mixture was poured into water (1.5 L) and extracted with DCM (2.8 L). The organic layer was washed with water (1.5 L x 2), dried over Na2S04(94 g), filtered and concentrated. The residue was reslurried in petroleum ether (3.8 L) at RT for 1 h, filtered and the filter cake was washed with petroleum ether (0.94 L x 2) and MeOH (0.94 L x 3) sequentially. The filter cake was dried under vacuum at 50 °C for 6 h to give 348 g of AP2312-2 as an orange solid with 94% yield and 99.7% HPLC purity.

[0226] 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 dust (110.5 g, 1.7 mol) was added portionwise to the reaction mixture at room temperature. The reaction was highly exothermic, and the temperature rose to 80 °C within 1 h. The reaction mixture was stirred at 80 °C for 2 h. After cooling, the reaction mixture was brought to room temperature, the inorganic salts were filtered off and the filter cake was washed with DCM (700 mL). The filtrate was concentrated to remove the organic solvents and extracted with DCM (1.4 L). The organic layer was washed successively with water (700 mL), 3 M NaOH (350 mL x 2) and then water (700 mL). The organic layer was concentrated and purified by re-slurry in EtOH (350 mL) at 0-15 °C for 1 h. The mixture was filtered and the filter cake was washed with cold ethanol (175 mL). The filter cake was dried under vacuum at 45 °C for 7 h to give 92.5 g of AP2312-A as a yellow solid in 58% yield with 99.5% HPLC purity.

[0227] A second protocol for preparing AP2312-A includes Step 1.1, Step 1.2 and Step 1.3:

[0228] Step 1.1

[0229] (1) THF (5 L) was added to a 10-L four-necked flask equipped with a mechanical stirrer

[0230] (2) 1,3,5-trifluoro-2-nitrobenzene (1.0 kg) was added at room temperature

[0231] (3) The mixture was cooled to -60 to -40 °C in a dry ice / EtOH bath under N2protection

[0232] (4) Ammonia gas was bubbled for 1.5 h at -60 to -40 °C

[0233] Note: The volume of the reaction mixture increased, which indicated that NH3was absorbed. The escaped NH3gas was absorbed by 20% aqueous H2SO4

[0234] (5) After 2 h at -60 to -40 °C, LCMS showed 22.2% of the starting material remained

[0235] (6) The temperature was warmed to -15 to -10 °C. The mixture was masked for 2 h and stirred at -15 to -10 °C overnight (16 h); LCMS showed 0.5% of the starting material remained

[0236] (7) The mixture was warmed to 10 °C

[0237] Note: The escaped NH3gas was absorbed by 20% aqueous H2SO4

[0238] (8) Filter the salt (NH4F) under vacuum

[0239] (9) Wash the filter cake with EtOAc (500 mL x 4)

[0240] Note: Wet cake: 560 g; TLC showed no product remaining

[0241] (10) Prepare and combine another batch of product prepared using the same starting material

[0242] (11) Concentrate the combined filtrate under vacuum at room temperature for 0.5 h to remove NH3 gas

[0243] (12) Concentrate the filtrate under vacuum at 40-45 °C to a volume of 2 L

[0244] Note: A large amount of yellow to red solid precipitated out

[0245] (13) Add n-heptane (1.6 L)

[0246] (14) Concentrate the mixture under vacuum at 40-45 °C to a volume of 2 L

[0247] (15) Add n-heptane (1.0 L)

[0248] (16) Stir the mixture vigorously at room temperature for 1 h

[0249] (17) Collect the solid by filtration

[0250] (18) Wash the filter cake with n-heptane (500 mL)

[0251] (19) Dry the filter cake under vacuum at 35-40 °C to give 1350 g of red solid with 96.7% HPLC purity

[0252] (20) Concentrate the filtrate to a volume of 3 L

[0253] (21) Stir at room temperature for 0.5 h

[0254] (22) Collect the solid by filtration

[0255] (23) Wash the filter cake with n-heptane (100 mL)

[0256] (24) Dry the filter cake under vacuum at 35-40 °C to give another 390 g of red solid with 93.1% HPLC purity

[0257] (25) Total yield: 1700 g, 82%

[0258] Step 1.2

[0259] (26) Add BnOH (9732.6 g) to a 50-L reactor

[0260] (27) Add KOH (1402.7 g) and stir (150 RPM)

[0261] (28) Add BnEt3NCl (956.63 g)

[0262] (29) Cool the mixture to 15 °C with N2protection

[0263] (30) Add AP2312-1 (1740.0 g) in portions

[0264] (31) Heat the mixture to 75-80 °C for 4 h

[0265] HPLC indicated <1.0% (0.11%) of starting material remaining

[0266] (32) Cool the mixture to room temperature

[0267] (33) Add DCM (one portion 17 L)

[0268] (34) Add water (one portion 14 L)

[0269] (35) Stir the mixture for 30 min

[0270] (36) Separate the organic layer

[0271] (37) Wash the organic layer with water (9 L)

[0272] (38) Dry the solution over Na2SO4(2 kg)

[0273] (39) Filter off the salt

[0274] (40) Concentrate the filtrate to ~13 L at 35-40 °C

[0275] Note: Orange solid precipitated out, almost no distillate was observed

[0276] (41) Add PE (37 L)

[0277] (42) Stir the slurry at room temperature for 1.5 h

[0278] (43) Collect the solid by filtration

[0279] (44) Wash the filter cake with MeOH (4 L x 2)

[0280] (45) Rinse the filter cake with PE (4 L x 2)

[0281] (46) The filter cake was vacuum dried at 45 °C to give a yellow solid, 3090.1 g, 99.8% HPLC purity, 88% yield

[0282] Step 1.3

[0283] (47) EtOH (11.7 L) was added to the 50-L reactor with N2flow protection

[0284] (48) AP2312-2 (3.0 kg) was added

[0285] (49) HOAc (5.4 L) was added

[0286] (50) H2O (6.3 L) was added

[0287] (51) The solution mixture was heated to 40 °C and then the heating was stopped

[0288] (52) Zinc dust (1903.3 g) was added in portions over 2 h

[0289] Note: After 20 min, the internal temperature rose to 80 °C (exothermic, no cooling) and the mixture turned to a brown solution after the above addition was complete

[0290] (53) The mixture was stirred at ambient temperature for 2 h

[0291] Note: After 2 h, the temperature dropped to 50 °C and HPLC analysis indicated complete consumption of AP2312-2

[0292] (54) EtOH (9 L) was added to the mixture

[0293] (55) The mixture was stirred at room temperature for 1 h

[0294] (56) The solid was filtered off

[0295] (57) The filter cake was washed with DCM (15 L)

[0296] (58) The filtrate was transferred to a 100-L reactor

[0297] (59) DCM (21 L) was added

[0298] (60) Water (15 L) was added

[0299] (61) The mixture was stirred for 15 min

[0300] (62) The aqueous layer was separated (lower layer, TLC showed no residual product)

[0301] (63) The organic layer was washed with water (15 L x 2)

[0302] Note: Remove residual HOAc and Zn salts

[0303] (64) Add water (15 L) to the organic layer

[0304] (65) Add 3 M aqueous NaOH solution

[0305] Note: Adjust the pH in the organic layer to 9-10

[0306] (66) Separate the organic layer

[0307] (67) Wash with brine (15 L)

[0308] (68) Dry the organic layer using Na2SO4(1 kg)

[0309] (69) Filter the salts

[0310] (70) Concentrate the filtrate under vacuum at 45-50 °C to a volume of ~ 15 L

[0311] Note: Total volume is ~ 0.4 vol; solid product precipitates out of solution.

[0312] (71) Add EtOH (15 L)

[0313] (72) Concentrate the mixture at 40 °C to ~ 15 L

[0314] (73) Add another portion of EtOH (5 L)

[0315] (74) Stir the slurry at room temperature for 2 h

[0316] (75) Cool the mixture to 5-10 °C and stir at 5-10 °C for 1 h

[0317] (76) Collect the solid by filtration

[0318] (77) Wash the filter cake with cold EtOH (2 L x 2, 10 °C)

[0319] HPLC of wet filter cake: 99.2%

[0320] (78) Dry the filter cake under vacuum at 35 °C for 48 h to constant weight to give off-white solid 2070.2 g with HPLC purity of 99.7%, yield 75%.

[0321] In step 2 of the process of synthesizing AMO-01, AP2312-B was prepared as follows:

[0322]

[0323] The preparation of AP2312-B is achieved by two alternative, highly related protocols. The first protocol comprises Step 2.A, Step 2.B, Step 2.C and Step 2.D.

[0324] Step 2.A Boc20 (763 g, 3.5 moL) was added dropwise to a solution of methyl 3- hydroxybenzoate (486 g, 3.2 moL) and DMAP (35.4 g, 0.29 moL) in DCM (2.4 L) over 2 h. The reaction mixture was stirred at room temperature overnight, washed with 8% w / w aqueous citric acid (486 mL x 3) and water (486 mL), dried over Na2S04 (97 g), filtered and concentrated to give 727 g of AP2312-B1 as a yellow oil in 90% yield with 100% HPLC purity.

[0325] Step 2.B A mixture of TMP (367 g, 2.6 moL) and i-PrMgCl-LiCl (2.0 L, 1.3 M in THF) was stirred at room temperature for 15 h. To a solution of AP2312-B1 (327 g, 1.3 moL) in THF (2.3 L) was added the pre-synthesized TMPMgCl-LiCl dropwise at 0-10 °C over 1 h. After stirring at 0-10 °C for 3 h, the reaction mixture was quenched with 20% w / w NH4CI (1 L) at 0-10 °C over 1 h. The mixture was extracted with EtOAc (2.3 L), washed with 10% w / w aqueous Na2S203 (1.5 L x 3) and water (1.5 L), concentrated to dryness to give crude AP2312-B2 which was used directly in the next step.

[0326] Step 2.C A mixture of crude AP2312-B2 and concentrated aqueous HC1 (3.2 L, 38.4 moL) in MeOH (3.3 L) was stirred at room temperature for 48 h. The reaction mixture was poured into water (3.3) and the pH of the mixture was adjusted to 7-8 with solid NaHC03. The mixture was concentrated to remove MeOH and extracted with EA (1.5 L x 2). The combined organic layers were concentrated to dryness to give crude AP2312-B3 which was used directly in the next step.

[0327] 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 h. After cooling to room temperature, the inorganic salts were filtered off and the filter cake was washed with EA (660 mL). The filtrate was concentrated and purified by flash chromatography (PE:EtOAc = 10:1) to give 242.3 g of AP2312-B with a yield of 50% for the last three steps and a LCMS purity of 100%.

[0328] A second protocol for the preparation of AP2312-B comprises steps 2.1, 2.2, 2.3 and 2.4:

[0329] Step 2.1

[0330] (1) DCM (14.1 L) was added to a 50-L reactor

[0331] (2) Methyl 3-hydroxybenzoate (2350.0 g) was added

[0332] (3) DMAP (169.8 g) was added

[0333] (4) (Boc)20 was added dropwise at room temperature (20-25 °C)

[0334] Note: CO2 evolved

[0335] (5) The mixture was stirred at room temperature for 4 h

[0336] HPLC indicated no SM remaining (reaction end: SM / product: < 1.0%, a / a)

[0337] (6) The organic solution was washed twice with 8% aqueous citric acid solution (12 L, 4 L)

[0338] (7) The organic solution was washed with saturated NaCl solution (5 L)

[0339] (8) The organic layer was dried over anhydrous Na2SO4 (1 kg)

[0340] (9) The salts were filtered off

[0341] (10) The filtrate was concentrated in vacuo at 40 °C to a remaining volume of 5 L

[0342] (11) Anhydrous THF (10 L) was added to the residue

[0343] (12) The solution was concentrated in vacuo at 40 °C to a volume of ~ 10 L (10.10 kg)

[0344] Analysis 10.10 kg: Assay 35.07% (contains 3542.1 g); HPLC 99.5%; Moisture (KF) 0.1%; Yield: 91%

[0345] Step 2.2

[0346] (13) Set up dry, clean reactor

[0347] (14) Flush system 3 times with nitrogen

[0348] (15) Transfer iPrMgCl.LiCl (12 L, 1.3 M in THF) to reactor at room temperature (10-15 °C) under nitrogen

[0349] (16) Add TMP (2.204 kg, just distilled from CaH2) dropwise over 2 h at room temperature under N2protection

[0350] Note: Gas (propane) slowly evolved during the above dropwise addition

[0351] (17) Heat the grey solution to 30-35 °C with stirring over 1 h

[0352] Note: The amount of gas increased when the temperature reached 30 °C, but under control

[0353] (18) Stir the mixture at 30-35 °C for 22 h

[0354] Note: Gas evolution stopped. IPC by GC showed the reaction was complete.

[0355] IPC method: Quench a sample (0.1 mL of base) with 0.02 mL of PhCHO at 10 °C; add 0.5 mL of MTBE and 0.5 mL of saturated aqueous NH4C1 solution; separate the organic layer for GC analysis; the absence of 2-methyl-l-phenylpropan-l-ol indicates the Grignard reagent is fully consumed

[0356] (19) Charge AP2312-B1 (7.24 kg of THF solution, Assay 27.1%, 1.967 kg, KF: 0.11%) to a 50 L reactor under N2

[0357] (20) Cool AP2312-B1 to -5-5 °C under N2

[0358] (21) Carefully add the TMPMgCl.LiCl solution from Step 6 to the cooled AP2312-B1 solution at 0-5 °C over 1.5 h

[0359] (22) Stir the mixture at 0-5 °C for 3 h

[0360] IPC: The sample was quenched with I2 / THF and HPLC showed the exchange was complete (SM / product: 5.0 / 84.1 = 6% < 10%)

[0361] (23) A solution of I2 / THF (3.96 kg in 8 L THF) was added dropwise to the above cooled solution at 0-10 °C over 90 min

[0362] (24) The solution was stirred for another 40 min at 0-10 °C

[0363] (25) The solution was heated to 20-25 °C

[0364] (26) The mixture was stirred at 20-25 °C for 2 h

[0365] HPLC analysis showed SM was 2.6%

[0366] (27) The mixture was cooled to -10 °C

[0367] (28) A 20% NH4CI solution (5 L) was added dropwise to the reaction mixture while keeping the temperature at 0-15 °C

[0368] (29) Water (18 L) was added at room temperature

[0369] (30) EtOAc (8 L) was added

[0370] (31) The mixture was stirred at room temperature for 10 min

[0371] (32) The organic layer was separated (top)

[0372] (33) The aqueous layer was extracted with EtOAc (5 L)

[0373] (34) The combined organic layers were washed with 10% Na2S2O3 aqueous solution (10 L x 2) and the organic layer was separated to give 24.5 kg of solution, HPLC: 83.0%

[0374] Step 2.3

[0375] (35) AP2312-B2 (48.3 kg, combined solution of batch AP2312-B2-1 and AP2312-B2-2, treated solution) was added to a 100-L reactor

[0376] (36) A solution of HC1 (16 L of concentrated HC1 mixed with 24 L tap water) was added at room temperature

[0377] Note: No significant temperature increase was observed

[0378] (37) The mixture was stirred at room temperature (25-30 °C) overnight (16 h)

[0379] Note: HPLC analysis indicated no SM remaining

[0380] (38) The solution was transferred to a 200-L reactor

[0381] (39) EtOAc (50 L) was added

[0382] (40) 10% NaCl aqueous solution (50 L) was added

[0383] (41) The layers were separated

[0384] (42) The aqueous layer was extracted with EtOAc (30 L)

[0385] (43) The combined organic layers were washed with 10% NaCl aqueous solution (10 L x 2)

[0386] Note: pH after washing was 5-6

[0387] (44) It was washed with saturated NaHC03solution (10 L x 2)

[0388] Note: pH after washing was 7

[0389] (45) It was washed with saturated NaCl (10 L)

[0390] The solution was concentrated to dryness to give a thick brown oil, 3.80 kg, HPLC purity 82.1%, overall yield of AP2312-B1 was 84%

[0391] Step 2.4

[0392] (46) The crude AP2312-B3 (3700 g) was added to a 50-L reactor

[0393] (47) Acetone (37 L) was added

[0394] (48) K2C03(2758.5 g) was added

[0395] (49) BnBr (2504.2 g) was added

[0396] (50) The mixture was heated to 55 °C

[0397] (51) The mixture was stirred at 55 °C for 3 h

[0398] HPLC (210 nm) analysis indicated <0.5% starting material (0.2% remaining)

[0399] (52) The mixture was cooled to room temperature

[0400] (53) Filtered salt

[0401] (54) Wash the filter cake with acetone (3.7 L x 2)

[0402] (55) Concentrate the filtrate at 35-40 °C to get brown oil 4.5 kg

[0403] HPLC of crude residue: 82.2%

[0404] (56) Purify the crude product by silica gel chromatography (dilute 4.5 kg with DCM 1 L)

[0405] Note: Silica gel: 22.5 kg (5.0 eq, w / w), 300-400 mesh; Elution - EtAOc / PE from 50:1 to 20:1

[0406] (57) Combine product fractions (monitored by TLC)

[0407] (58) Concentrate the product fractions under vacuum at 35-40 °C to 2 L volume

[0408] Note: A lot of solid was separated

[0409] (59) Add PE (5 L)

[0410] (60) Concentrate the slurry under vacuum at 35-40 °C to 3 L volume

[0411] (61) Collect the solid by filtration

[0412] Dry the filter cake under vacuum at 30 °C to get light yellow solid, 2.4 kg, HPLC purity 99.7%, yield 49%

[0413] Note: Concentration of filtrate gave another 30 g of yellow solid, HPLC purity 84%

[0414] In step 3 of the process of synthesis of AMO-01, AP2312-3 was prepared by the following Ullmann coupling:

[0415]

[0416] Preparation of AP2312-3 was achieved by alternative, highly related protocols that used different amounts of reagents (see Table 1) to produce different amounts of the desired AP2312 product and different amounts of impurities. The general 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 by vacuum / nitrogen purging 3 times. The reaction mixture was stirred at 70 °C for 6 h and a sample was taken for IPC. After cooling to room temperature, a portion of CuI was added to the reaction mixture and degassed by vacuum / nitrogen purging 3 times. The reaction mixture was stirred at 90 °C for 15 h and a sample was taken for IPC.

[0417] Table 1

[0418]

[0419]

[0420] It was found that at high CuI loading (6thand 7th), the reaction was fast and the deiodination side product of AP2312-B (AP2312-3-IM01) was high. The 1st-6thwere run with partial CuI at 70 °C for 6 h to complete the Ullmann coupling reaction and reduce the deiodination side product, then another portion of CuI was added to speed up the cyclization at 90 °C for 15 h. The 7thwas run with 15% CuI loading at 70 °C for 6 h, then at 90 °C for 15 h.

[0421] It was found that water (compare 3rdand 4th) accelerated the reaction. Without water (4th), the Ullmann coupling reaction and cyclization were slow. After 6 h at 70 °C, 27.7% of AP2312-B remained, and after 15 h at 90 °C, 8.1% of AP2312-3J remained. AP2312-B and AP2312-3J were completely converted to AP2312-3 after 34 h at 90 °C. With 1 v of water (3rd) in the system, the Ullmann coupling reaction was complete in 6 h and the cyclization was complete in 15 h.

[0422] It was found that at low CuI loading (1% + 0.5% equivalents, 1st), the Ullmann coupling reaction and cyclization reaction were slow. After 6 h at 70 °C, 8.1% of AP2312-B remained, and after 15 h at 90 °C, 4.6% of AP2312-3J remained.

[0423] The 5th (7% + 3.5% equiv CuI) contained 79.0% AP2312-3 in the system, slightly higher than the 2nd (3% + 1.5% equiv CuI, 78.2% AP2312-3) and the 3rd (5% + 2.5% equiv CuI, 78.1% AP2312-3). However, the proportion of AP2312-3I improved with increasing CuI loading.

[0424] In one particular embodiment, AP2312-3 is prepared by the following steps.

[0425] (1) Add DMF (14.0 L)

[0426] (2) Add water (1.4 L)

[0427] (3) Add AP2312-A (1,230 g, 3.84 mol) to a 20 L 4-necked flask

[0428] (4) Add AP2312-B (1,413 g, 3.84 mol, 1.0 equiv)

[0429] (5) Add CuI (36.60 g, 0.192 mol, 0.05 equiv)

[0430] (6) Add K2CO3 (1,060 g, 7.68 mol, 2.0 equiv)

[0431] (7) Add L-proline (44.2 g, 0.384 mol, 0.1 equiv)

[0432] (8) Heat the reaction mixture to 70 °C under N2for 6 h; HPLC showed 3.0% AP2312-B in 11.7 min

[0433] (9) Add CuI (18.30 g, 0.096 mol, 0.025 equiv)

[0434] (10) Heat the reaction mixture to 90 °C overnight; HPLC showed no intermediate (Ullmann coupling product at 11.1 min) was detected. 11.4 min: deiodination side product: methyl 3- (benzyloxy)benzoate, 16.0%; 9.99 min: hydrolysis side product 3- (benzyloxy)benzoic acid: 2.4%; 13.4 min: product, 81.6%

[0435] (11) Cool the mixture to room temperature

[0436] (12) Combine batches AP2312-3-30, AP2312-3-31, and AP2312-3-33 for workup

[0437] (13) Add activated carbon (847 g)

[0438] (14) Stir slurry for 1 h

[0439] (15) Filter off solids

[0440] (16) Wash filter cake with DMF (1.4 L x 2)

[0441] (17) Add solution to a 50-L reactor

[0442] (18) Add aqueous NaOH (345.6 g in 2.1 L)

[0443] (19) Heat mixture to 70 °C for 1 h; HPLC shows complete hydrolysis of de-iodine ester to acid (within 10.0 min) within 11.4 min; 10.0 min: 3- (benzyloxy)benzoic acid, 12.7%; 13.8 min, product, 87.3%

[0444] (20) Add NH4Cl (1,540 g, 28.8 mol, 7.5 equiv)

[0445] (21) Add ethylenediamine (877 g, 14.59 mol, 3.8 equiv)

[0446] (22) Add deep purple solution dropwise to a 100-L reactor containing 75.6 L H2O

[0447] (23) Stir slurry for 2 h

[0448] (24) Filter to collect solids; HPLC shows almost no AP2312-3 in filtrate and hydrolyzed side product (3-(benzyloxy)benzoic acid) is removed into filtrate within 10.0 min; HPLC (aqueous) of filtrate:

[0449] (25) Dissolve solids in DCM (21 L)

[0450] (26) Dry solution with anhydrous Na2SO4(8 kg)

[0451] (27) Filter off salts

[0452] (28) Concentrate filtrate to ~7.5 L

[0453] (29) Add hexanes (30.0 L)

[0454] (30) Stir slurry for 1 h at room temperature

[0455] (31) Filter to collect the solids to get the crude product (2.8 kg); HPLC: 94.6%, 13.7 min; most of the impurities were removed to the filtrate

[0456] (32) Dissolve the crude solids in toluene (20 L)

[0457] (33) Add active C (420 g)

[0458] (34) Heat the mixture to 110 °C

[0459] (35) Stir the mixture at 110 °C for 2 h

[0460] (36) Cool the mixture to 70-80 °C

[0461] (37) Filter off the active C

[0462] (38) Wash the filter cake with DCM (2.1 L x 3)

[0463] (39) Concentrate the combined filtrates to ~ 20 L

[0464] (40) Cool the mixture to room temperature for 2 h

[0465] (41) Cool the mixture to 5-10 °C for 1 h

[0466] (42) Filter to collect the solids

[0467] (43) Wash the filter cake with EtOH (2.1 L)

[0468] (44) Wash the filter cake with hexane (2.1 L x 2) to get the crude product (1.7 kg); solids: 96.7%, 13.7 min

[0469] (45) Dissolve the crude product in DCM (4.28 L)

[0470] (46) Add hexane (17.0 L)

[0471] (47) Stir the slurry at room temperature for 1 h

[0472] (48) Filter to collect the solids to get the crude product

[0473] (49) Wash the filter cake with hexane (2.1 L)

[0474] (50) Dry the filter cake under vacuum at 40 °C to get a red solid, 1.58 kg, 50% yield, HPLC purity 99.8%

[0475] Table 2 Batch Record

[0476]

[0477]

[0478] In step 4 of the process for synthesizing AMO-01, AP2312-4 was prepared by debenzylating AP2312-3 as follows:

[0479]

[0480] Preparation of AP2312-4 was achieved by the following steps.

[0481] (1) AP2312-3 (870 g, 1,645.9 mmol) was charged into a 10 L four-necked flask

[0482] (2) 10% Pd-C (130.5 g, 0.15 w / w) was added

[0483] (3) THF (2.6 L) was added

[0484] (4) MeOH (2.6 L) was added

[0485] (5) The slurry was stirred under H2at room temperature for 36 h

[0486] (6) The Pd-C was filtered off

[0487] (7) The filter cake was washed with MeOH (172 mL x 3)

[0488] (8) The combined filtrate was concentrated to a black oil (660 g)

[0489] (9) Acetone (2.0 L) was added

[0490] (10) Hexanes (2.0 L) was added

[0491] (11) The slurry was stirred at room temperature for 1 h

[0492] (12) The solid was collected by filtration

[0493] (13) The filter cake was washed with hexanes (660 mL x 2)

[0494] (14) The product was dried under vacuum at 35 °C to give a light green solid 469 g, 100% HPLC purity, over-weight (110%), used directly for the next step

[0495] Table 3 Batch Record

[0496]

[0497] In step 5 of the process for synthesizing AMO-01, AP2312-5 was prepared by silylation as follows:

[0498]

[0499] Preparation of AP2312-5 was achieved by the following steps.

[0500] (1) DMF (4.0 L) was added into a 10 L four-necked flask

[0501] (2) AP2312-4 (450 g, 1,742.8 mmol) was added

[0502] (3) TEA (881.9 g, 8,717 mmol, 5.0 eq) was added

[0503] (4) The solvent was cooled to 0-5 °C

[0504] (5) TIPSCl (1,344 g, 6,971.2 mmol, 4.0 eq) was added dropwise at 0-5 °C for 1 h

[0505] (6) The mixture was stirred at room temperature for 0.5 h

[0506] (7) The mixture was poured into H2O (12.15 L)

[0507] (8) The mixture was stirred for 1 h

[0508] (9) The solid was collected by filtration

[0509] (10) The filter cake was washed with EtOH (2.9 L x 2)

[0510] (11) The product was dried under vacuum at 37 °C for 6 h to give a yellow solid 927 g, HPLC purity 100%, overall yield for last 2 steps 80.5%

[0511] Table 4 - Batch Record

[0512]

[0513] In step 6 of the process for synthesizing AMO-01, AP2312-6 was as follows:

[0514]

[0515] Preparation of AP2312-6 was achieved by the following steps.

[0516] (1) DMF (3.7 L) was added into a 10 L four-necked flask

[0517] (2) AP2312-C (409 g, 1,839 mmol) was added

[0518] (3) Add 2,6-lutidine (315.4 g, 2,942.4 mmol, 1.6 equiv)

[0519] (4) Add LiBr (255.7 g, 2,942.4 mmol, 1.6 equiv)

[0520] (5) Cool the solvent to 0-5 °C

[0521] (6) Add Ms20 in portions, keeping the internal temperature at 0-5 °C

[0522] (7) Stir the mixture at 0-5 °C for 3 h

[0523] (8) Pour the mixture into ice water (7.4 L)

[0524] (9) Add n-heptane (4.9 L)

[0525] (10) Stir the mixture for 0.5 h

[0526] (11) Separate the organic layer

[0527] (12) Pass the organic solution through a pad of silica gel (81.8 g)

[0528] (13) Wash the silica gel filter cake with n-heptane (818 ml)

[0529] (14) Concentrate the filtrate to give a yellow oil, 544 g, 90% GC purity, over weight (103%), use directly in the next step

[0530] Table 5 Batch Record

[0531]

[0532]

[0533] In step 7 of the method of synthesizing AMO-01, AP2312-8 was prepared by farnesylation as follows:

[0534]

[0535] The preparation of AP2312-8 was achieved by the following steps.

[0536] (1) Charge dioxane (8.8 L) into a 20 L four-necked flask

[0537] (2) Add tBuOH (3.8 L)

[0538] (3) Add AP2312-5 (730 g, 1,004 mmol)

[0539] (4) Cool the solvent to 10-15 °C

[0540] (5) Add 1 M t BuOK in t BuOH (1.15 L, 1,150 mmol, 1.15 equiv)

[0541] (6) Stir the mixture at 10-15 °C for 2 h

[0542] (7) Add a solution of AP2312-6 in dioxane (386.7 g, 730 mL, 1,355 mmol, 1.35 equiv)

[0543] (8) Stir the mixture at room temperature overnight

[0544] (9) Combine batches AP2312-8-8 and AP2312-8-9 for workup

[0545] (10) Add MTBE (12.96 L)

[0546] (11) Cool the solvent to 0-5 °C

[0547] (12) Slowly add H2O (19.4 L, containing 16.1 g NH4CI) to the solution

[0548] (13) Stir the mixture for 15 min

[0549] (14) Separate the organic layer

[0550] (15) Wash with H2O (16.2 L x 3, containing 1,620 g NaCI)

[0551] (16) Concentrate the organic layer to a black oil

[0552] (17) Dissolve the residue in THF (20 L)

[0553] (18) Concentrate the solution to a black oil

[0554] (19) Dissolve the residue in THF (14.6 L)

[0555] (20) Use the solution directly for the next step

[0556] Table 6 Batch Record

[0557]

[0558] In step 8 of the method of synthesizing AMO-01, AMO-01 (AP2312) was prepared by demethylsilation as follows:

[0559]

[0560] Preparation of AP2312 was achieved by the following steps.

[0561] (1) A solution of AP2312-8 in THF (1,037 g, 1,113 mmol, 1.0 equiv, 14.6 L) was added to a 20 L four-necked flask

[0562] (2) The solvent was cooled to -5-5 °C

[0563] (3) AcOH (535.1 g, 8,904 mmol, 8.0 equiv) was added

[0564] (4) TBAF.3H2O (1,404.6 g, 4,452 mmol, 4.0 equiv) was added

[0565] (5) The mixture was stirred at room temperature overnight

[0566] (6) The solution was concentrated to a black oil

[0567] (7) The black oil was dissolved in EA (10.37 L)

[0568] (8) The solution was washed with H2O (10.37 L x 3)

[0569] (9) The organic layer was separated

[0570] (10) The organic layer was concentrated to ~2 L

[0571] (11) n-Heptane (20,740 ml) was added to the solution over 1 h

[0572] (12) The mixture was stirred overnight

[0573] (13) The solid was collected by filtration

[0574] (14) The solid was dissolved in MeOH (4,148 ml) and H2O (519 ml)

[0575] (15) The solution was washed with n-Heptane (4,148 ml x 2)

[0576] (16) The MeOH-H2O layer was separated

[0577] (17) Activated carbon (104 g) was added

[0578] (18) The mixture was stirred for 1 h

[0579] (19) The activated carbon was filtered off

[0580] (20) The activated carbon filter cake was washed with MeOH (1,037 ml)

[0581] (21) H20 (6,222 ml) was added to the combined filtrates over 1 h

[0582] (22) The mixture was stirred for 2 h

[0583] (23) The solid was collected by filtration

[0584] (24) The solid was dissolved in AcOH (2,074 ml)

[0585] (25) H20 (2,593 ml) was added to the solution over 1 h

[0586] (26) The mixture was stirred overnight

[0587] (27) The solid was collected by filtration

[0588] (28) The filter cake was washed with AcOH / H20 (519 ml / 519 ml)

[0589] (29) The filter cake was washed with water (1,037 ml x 2)

[0590] (30) The filter cake was dried under vacuum at 37 °C to give a light grey solid 330 g, 64% yield over the last two steps, 98.3% HPLC purity

[0591] Table 7 Batch Record

[0592]

[0593] In total, 330 g of AP2312 was isolated, 98.3% HPLC purity Figure 2 LCMS [M+H] 463; 1 HNMR (400 MHz; d6 DMSO) d 9.99 (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). Several impurities were present in the isolated product (RRT 0.93 = 0.23%, RRT 0.98 = 0.19%, RRT 1.09 = 0.40%, RRT 1.11 = 0.36%, RRT 1.12 = 0.17%, RRT 1.14 = 0.12%). This reaction scheme, based on Ullmann coupling, was optimized and confirmed on > 1 kg scale.

[0594] Compounds of the formula I

[0595] It is apparent that the specific steps provided above for the production of AMO-01 can also be used to produce the compounds of Formula I and Formula II with only slight modifications.

[0596] For the compounds of Formula I, when one or more of the variables A, R 2 , R 3 , R 4 , R 7 , R 8 , W 1 , W 2 , W 3 and x of the compound (see Formula I) are different from the corresponding variables in AMO-01, then the specific steps provided above for the production of AMO-01 need to be changed.

[0597] AP2312M-1

[0598] The following examples are illustrative. In the first example, Step 4 is changed to produce AP2312M-1, where R 7 is -CH 3 .

[0599]

[0600] To a solution of AP2312-3 (15.9, 30 mmol) in 1,4-dioxane (192 mL) and t BuOH (90 mL) was added t BuOK (5.0 g, 45 mmol). The reaction mixture was stirred at 30 °C for 2 h. Then Mel (10.7 g, 75 mmol) was added and the flask was sealed. The reaction mixture was stirred at 30 °C for 24 h. The solvent was removed by concentration under vacuum, the residue was dissolved in water (160 mL) and then extracted with DCM (160 mL x 2). The combined organic layers were washed with water (160 mL), concentrated and purified by re-slurry in petroleum ether (160 mL) and EtOAc (16 mL) to give AP2312-11A as a yellow solid, 15 g, 99.4% HPLC purity, 92% yield.

[0601] A mixture of AP2312M-11A (15.0 g, 27.6 mmol) and 10% Pd / C (50% wet, 2.4 g) in THF (45 mL) and MeOH (45 mL) was stirred at 40 °C and 0.1 MPa hydrogen pressure for 24 h. The reaction mixture was cooled to room temperature and the catalyst was filtered off. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 20:1) to give AP2312M-1 as a yellow solid, 6.5 g, 99.1% HPLC purity, 87% yield. LCMS [M+H] 273; 1H NMR (d6-DMSO, 500 MHz) δ 10.08 (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).

[0602] AP2312M-2

[0603] In the second example, step 6 is changed to produce AP2312M-2, where R 7 is 1-bromo-3-methyl-2-butene.

[0604]

[0605] To a solution of AP2312M-21 (86.0 g, 1.0 mol) in DCM (430 mL) was added PBr3 (108.4 g, 0.4 mol) dropwise at 0-10 °C over 1 h. The reaction mixture was stirred at room temperature overnight and purified by distillation (-50 °C / -0.1 MPa) to give 35.6 g of AP2312M-22, 24% yield, which was used directly in the next step.

[0606] To a solution of AP2312-5 (21.8 g, 30 mmol) in 1,4-dioxane (262 mL) and t BuOH (110 mL) was added t BuOK (5.0 g, 45 mmol). The reaction mixture was stirred at 30 °C for 2 h. Then AP2312M-22 (11.2 g, 75 mmol) was added and the reaction mixture was stirred at 30 °C for 2 h. After the solvent was evaporated, water (220 mL) was added to the residue and extracted with EtOAc (110 mL x 2). The combined organic layers were washed with water (220 mL) and concentrated to give crude AP2312M-23, which was used directly in the next step.

[0607] The crude AP2312M-23 was dissolved in THF (220 mL), then TBAF (120 mL, 1 M in THF), AcOH (14.4 g, 240 mmoL) was added. The reaction mixture was stirred at 30 °C for 6 h. The reaction mixture was poured into water (440 mL) and extracted with EtOAc (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) to give 4.2 g AP2312M-2 as a gray solid, the yield of the last two steps was 43%, the purity of HPLC was 99.4%, confirmed by 1 H NMR and LCMS. LCMS [M+H] 327; 1 H NMR (d6-DMSO, 500 MHz) δ 10.03 (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).

[0608] AP2312M-3

[0609] In the third example, step 6 was changed to produce AP2312M-3, where R was changed again 7 .

[0610]

[0611] To a solution of AP2312M-31 (9.2 g, 60 mmoL), 2,6-lutidine (10.3 g, 96 mmoL) and LiBr (8.4 g, 96 mmoL) in DMF (92 mL) was added (Ms)20 (15.7 g) in portions at 0-10 °C. The reaction mixture was stirred at 0-10 °C for 2 h, poured into water (276 mL) and extracted with petroleum ether (92 mL x 2). The combined organic layers were washed with water (92 mL) and concentrated to give 11.0 g AP2312M-32 in 85% yield, which was used directly in the next step.

[0612] To a solution of AP2312-5 (14.5 g, 20 mmoL) in 1,4-dioxane (174 mL) and t BuOH (73 mL) was added tBuOK (3.4 g, 30 mmoL). The reaction mixture was stirred at 30 °C for 2 h. Then AP2312M-32 (6.5 g, 30 mmoL) was added and the reaction mixture was stirred at 30 °C for 2 h. After evaporation of the solvent, water (145 mL) was added to the residue and extracted with EtOAc (145 mL x 2). The combined organic layers were washed with water (145 mL) and concentrated to give crude AP2312M-33, which was used directly for the next step.

[0613] The crude AP2312M-33 was dissolved in THF (145 mL), then TBAF (80 mL, 1 M in THF), AcOH (9.6 g, 160 mmoL) was added. The reaction mixture was stirred at 30 °C for 6 h, poured into water (440 mL) and extracted with EtOAc (290 mL). The organic layer was washed with water (145 mL x 6), concentrated and purified by flash chromatography (DCM:MeOH = 40: 1) to give 5.5 g of AP2312M-3 as a gray solid, with a yield of 70% for the last two steps, HPLC purity of 98.2%, confirmed by H NMR and LCMS. LCMS [M+H] 395, 1 H NMR and LCMS. LCMS [M+H] 395, 1 H NMR (d6-DMSO, 500 MHz) δ 10.04 (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).

[0614] AP2312M-4

[0615] In the fourth example, step 6 was changed to produce AP2312M-4, where R was changed again 7 .

[0616]

[0617] At 0–10 °C, HCOOH (41.4 g, 0.9 mol), TEA (39.5 g, 0.39 mol), and Meldrum acid (43.2 g, 0.3 mol) were added sequentially to DMF (100 mL). The reaction mixture was stirred at 0–10 °C for 0.5 h, and then AP2312M-41 (44.5 g, 0.3 mol) was added. The reaction mixture was stirred overnight at 80 °C. After cooling to room temperature, the reaction mixture was poured into ice water (1.2 L), and the pH of the mixture was adjusted to 1–2 with concentrated HCl aqueous solution at 0–10 °C. The mixture was filtered, and the filter cake was washed with water (100 mL). The filter cake was dissolved in DCM (300 mL) and dried with Na2SO4 (90 g). After removing inorganic salts, the filtrate was concentrated to dryness to obtain crude AP2312M-42, which was used directly in the next step.

[0618] At 0–10 °C, NaBH4 (22.8 g, 0.6 mol) was added in portions to a solution of crude AP2312M-42 in THF (845 mL) over 0.5 h. BF3·Et2O (110.7 g, 0.78 mol) was added dropwise to the reaction mixture at 0–10 °C over 1.5 h. After stirring at room temperature for 3 h, 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 with DCM (600 mL × 2). The combined organic layers were washed with saturated NaHCO3 (500 mL) and brine (500 mL), concentrated, and purified by rapid chromatography (PE:EA = 5:1) to obtain 46.0 g of AP2312M-43 as a colorless oil, with a two-step yield of 86%.

[0619] At 0–10 °C, PBr3 (17.9 g, 66 mmol) was added dropwise to a solution of AP2312M-43 (29.4 g, 165 mmol) in DCM (294 mL). The reaction mixture was stirred at room temperature for 15 h, washed with water (210 mL), concentrated, and purified by rapid chromatography (PE:EA = 20:1) to give 19.5 g of AP2312M-53 as a grayish-white solid, with a yield of 49%.

[0620] AP2312-3 (21.2 g, 40 mmol / L) was added to 1,4-dioxane (254 mL) and t Add to the solution of BuOH (106 mL) tBuOK (5.8 g, 52 mmol). The reaction mixture was stirred at 30 °C for 2 h. Then AP2312M-44 (19.3 g, 80 mmol) was added and the reaction mixture was stirred at 30 °C for 24 h. After evaporation of the solvent, water (212 mL) was added to the residue and extracted with EtOAc (106 mL x 2). The combined organic layers were washed with water (106 mL) and concentrated to give crude AP2312M-45 as a yellow solid, which was used directly for the next step.

[0621] The crude AP2312M-45 was dissolved in THF (212 mL) and MeOH (106 mL), then 10% Pd / C, 50% water wet (3.2 g) was added. The reaction mixture was degassed by vacuum / hydrogen purging three times and stirred at 40 °C under hydrogen pressure of 0.1 MPa for 24 h. After the reaction mixture was cooled to room temperature, the catalyst was filtered off. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 40:1) to give 6.7 g of AP2312M-4 as a gray solid, with a yield of 40% for the last two steps, HPLC purity of 98.0%, confirmed by 1 HNMR and LCMS. LCMS [M+H] 419; 1 HNMR (d6-DMSO, 500 MHz) δ 10.06 (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).

[0622] AP2312M-5

[0623] In the fifth example, step 6 was changed to produce AP2312M-5, where R was changed again 7 .

[0624]

[0625] 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 by vacuum / nitrogen purging for three times. After stirring at 70 °C for 15 h, the reaction mixture was concentrated to remove MeOH and extracted with EtOAc (100 mL x 2). The combined organic layers were concentrated and purified by flash chromatography (PE:EA = 10:1) to give 22.0 g of AP2312M-52 as off-white solid in 74% yield.

[0626] To a solution of AP2312M-52 (22.0 g, 86.6 mmol) in DCM (220 mL) was added PBr3 (11.7 g, 43.3 mmol) dropwise at 0-10 °C. The reaction mixture was stirred at room temperature for 15 h, washed with water (220 mL), concentrated and purified by flash chromatography (PE:EA = 30:1) to give 11.3 g of AP2312M-53 as off-white solid in 41% yield.

[0627] To a solution of AP2312-3 (12.2 g, 23 mmol) in 1,4-dioxane (146 mL) and t BuOH (61 mL) was added t BuOK (3.4 g, 30 mmol). The reaction mixture was stirred at 30 °C for 2 h. Then AP2312M-53 (11.1 g, 35 mmol) was added and the reaction mixture was stirred at 30 °C for 24 h. After evaporation of the solvent, water (122 mL) was added to the residue and extracted with EtOAc (61 mL x 2). The combined organic layers were washed with water (61 mL) and concentrated to give a residue of AP2312M-54A as yellow solid, which was used directly for the next step.

[0628] The crude AP2312M-54A was dissolved in THF (122 mL) and MeOH (61 mL), then 10% Pd / C, 50% water wet (1.8 g) was added. The reaction mixture was degassed by vacuum / hydrogen purging for three times and stirred at 40 °C under hydrogen pressure of 0.1 MPa for 24 h. After the reaction mixture was cooled to room temperature, the catalyst was filtered off. The filtrate was concentrated and purified by flash chromatography (DCM:MeOH = 40:1) to give 8.0 g of AP2312M-5 as off-white solid in 70% yield for the last two steps with 99.0% HPLC purity, confirmed by 1 HNMR and LCMS. LCMS [M+H] 495;1 HNMR (d6-DMSO, 500 MHz) δ 10.07 (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).

[0629] Compounds of the formula II

[0630] As suggested above, the specific steps provided for the production of AMO-01 can be used with only slight modification to produce the compound of Formula II. The compound of Formula II is produced in Step 3 using a Buchwald coupling instead of a Ullmann coupling.

[0631] Initial experiments for the production of AMO-01 provided the surprising finding that by using a Buchwald coupling, the compound of Formula II was achieved, while using a Ullmann coupling produced the compound of Formula I.

[0632] Experiments were conducted to confirm that the Buchwald chemistry reaction gave isomer AP2312-3I instead of AP2312-3.

[0633]

[0634] 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 by vacuum / nitrogen purging three times. The reaction mixture was stirred at 110 °C for 48 h. The reaction mixture was poured into water (210 mL) and extracted with EA (210 mL x 2). The combined organic layers were washed with water (210 mL), concentrated and purified by flash chromatography (PE:EA = 10:1) to give 26.0 g of AP2312-14 with a yield of 73% and HPLC purity of 98.0%.

[0635] 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 portions at room temperature. The reaction was highly exothermic and the temperature rose to 80 °C within 1 h. The reaction mixture was stirred at 80 °C for 2 h. After the reaction mixture was cooled to room temperature, the inorganic salts were filtered off and the filter cake was washed with DCM (200 mL). The filtrate was concentrated to remove the organic solvents and extracted with DCM (250 mL x 1). The organic layer was washed with water (100 mL x 3), concentrated and purified by re-slurry in EtOH (100 mL) to give 21.0 g of AP2312-3I as a yellow solid with 100% purity by LCMS, 94% yield.

[0636] AP2312-3I was then produced using Buchwald coupling as follows.

[0637]

[0638] A mixture of AP2312-A (1.3 g, 4 mmol), AP2312-B (1.5 g, 4 mmol), PdCl2(dppf) (146 mg, 0.2 mmol) and Cs2CO3(1.8 g, 5.6 mmol) in DMF (26 mL) was degassed by vacuum / nitrogen purging three times. The reaction mixture was stirred at 100 °C for 15 h. A sample was taken for IPC and HPLC indicated 47.0% of AP2312-3I (20.7 min) in the system and no AP2312-3 (19.0 min). LCMS [M+H] 529; 1 HNMR (d6-DMSO, 500 MHz) d 8.76 (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).

[0639] While the application has been described with reference to certain specific embodiments thereof, a person of ordinary skill in the art will understand that various modifications can be made therein without departing from the spirit and scope of the application. The scope of the following claims is not limited to the specific embodiments described.

Claims

1. A method for synthesizing farnesyldibenzodiazepine one AMO-01 (10-farnesyl-4,6,8-trihydroxy-dibenzodiazepine-11-one), The method includes: (a) Preparation of AP2312-A; (b) Preparation of AP2312-B; (c) Perform Ullmann coupling; (d) Perform debenzylation; (e) Perform methylsilanization; (f) Preparation of farnesyl bromide; (g) Perform farinilation; and (h) Perform demethylation of silicon-based 2. The method for synthesizing farnesyldibenzodiazepine ketone AMO-01 according to claim 1, wherein the method comprises: (a) Preparation of AP2312-A; (b) Preparation of AP2312-B; (c) In the presence of 0.0525 equivalents of CuI, 2.0 equivalents of K2CO3, 0.1 equivalents of L-proline and DMF, molecular equivalents of AP2312-A and AP2312-B were reacted to carry out Ullmann coupling to produce AP2312-3. (d) AP2312-3 was debenzylated in H2 in the presence of THF, MeOH and Pb / C to produce AP2312-4; (e) AP2312-4 was methylsilanized in the presence of 4.0 equivalents of TIPSCl, 5.0 equivalents of Et3N and DMF to produce AP2312-5; (f) In the presence of Ms2O, 1.6 equivalents of LiBr, 1.6 equivalents of 2,6-rutidine and DMF, AP2312-C was reacted to produce AP2312-6; (g) In the presence of dioxane, t BuOH and 1.15 equivalent t In the case of BuOK, AP2312-5 is farnesylated using AP2312-6 to produce AP2312-8; and (h) The desilylation reaction of AP2312-8 was carried out in the presence of 1.0 equivalent of THF, 8.0 equivalent of AcOH and 4.0 equivalent of TBAF to produce AMO-01.

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