阅读说明：本技术 Mcl-1抑制剂的合成 (The synthesis of MCL-1 inhibitor ) 是由 C.斯图尔特 S.哈迪 A.斯塔克 A.赫德 Q.叶 郑晓兰 C.费拉 J.克克 D 于 2018-03-29 设计创作，主要内容包括：披露了合成化合物1的中间体和方法。<Image he="754" wi="572" file="DDA0002214599730000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(Disclose the intermediate and method of synthesis compound 1.)

1. a kind of compound selected from following item:

And

Or its salt.

Background technique

Myelocytic leukemia 1 (Mcl-1) be BCL-2 protein family important anti-apoptotic member and cell survival it is main Regulatory factor.The amplification of MCL1 gene and/or the overexpression of Mcl-1 albumen have been observed in kinds cancer type, and It is usually directed to tumor development.In fact, MCL1 is one of the gene being most often amplified in human cancer.In many malignant tumours In, Mcl-1 is crucial survival factors, and has shown that it mediates the drug resistance to a variety of anticancer agents.

Mcl-1 by with pro apoptotic protein (such as Bim, Noxa, Bak and Bax) in conjunction with and neutralize its death inducing activity come Promote cell survival.Therefore, Mcl-1 inhibition can discharge these pro apoptotic proteins, typically result in dependent on Mcl-1 and survive swollen The induction of Apoptosis in oncocyte.Therefore, individually or with other therapies combining targeting Mcl-1 in the treatment is that treatment is many More malignant tumours and the promising strategy for overcoming the drug resistance in many human cancers.Entitled (the R of the compound chemistry_a)- The chloro- penta azepine ring in heptan of 5,13,14,22- tetramethyl -28- oxa- -2,9- dithia -5,6,12,13,22- of (+) -17- [27.7.1.1^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16,18,20,23,29, 31,33,35- tridecylene -23- formic acid (referred to as compound 1):

It is a kind of effective Mcl-1 inhibitor, as described in more detail below.Therefore, it is necessary to develop in an efficient way Synthesize the new method of compound 1.

Summary of the invention

Provided herein is the methods and intermediate that can be used for synthesizing compound 1:

In some embodiments, following intermediate and its any salt and its synthesis are disclosed:

And

Detailed description of the invention

Fig. 1 illustrates form A (R_a) chloro- dithia -5,6,12 5,13,14,22- tetramethyl -28- oxa- -2,9- -17-, Penta azepine ring in heptan [27.7.1.1 of 13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11, The x-ray diffractogram of powder of 14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid monohydrate.

Fig. 2 illustrates form A (R_a) chloro- dithia -5,6,12 5,13,14,22- tetramethyl -28- oxa- -2,9- -17-, Penta azepine ring in heptan [27.7.1.1 of 13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11, The differential scanning calorimetry (DSC) and thermogravimetric of 14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid monohydrate Amount analysis (TGA) trace.

Specific embodiment

In some embodiments, there is provided herein for synthesizing (R_a)-(+) the chloro- 5,13,14,22- tetramethyl -28- of -17- Penta azepine ring in heptan [27.7.1.1 of oxa- -2,9- dithia -5,6,12,13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 30 Eight carbon -1 (37), 4 (38), the side of 6,11,14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid (compound 1) Method and intermediate:

Shown in example 1 as follows, compound 1 is effective Mcl-1 inhibitor, and can be used for treating cancer, including is disliked Property blood disease such as acute myelogenous leukemia, Huppert's disease, lymphoma mantle cell, chronic lymphocytic leukemia, more Unrestrained property large B cell lymphoid tumor, Burkitt lymphoma, follicular lymphoma and solid tumor such as non-small cell lung cancer (NSCLC), Small Cell Lung Cancer (SCLC), breast cancer, neuroblastoma, prostate cancer, melanoma, cancer of pancreas, uterine cancer, son Endometrial carcinoma and colon cancer.Due to the structural complexity of compound 1, in terms of exploitation compound 1 is as potential treatment of cancer, Realization, which is effectively synthesized, can be an importance.

Intermediate

In some embodiments, (E/Z)-dimethyl 2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) adipate ester is disclosed (intermediate 1):

In some embodiments, the compound with formula (d) is disclosed:

Wherein R1 is blocking group or hydrogen.In some embodiments, R1 is to methoxy-benzyl.In some embodiments, Compound with formula (a) be the chloro- 7- of (±) -3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1, 5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid (intermediate 6):

In some embodiments, the compound with formula (e) is disclosed:

Wherein R1 is blocking group or hydrogen.In some embodiments, R1 is to methoxy-benzyl.In some embodiments, Compound with formula (b) is (R_a) the chloro- 7- of -3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1, 5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid-(1R) -1- (2- nitrobenzophenone) ethamine (1:1 salt) (intermediate 7):

In some embodiments, methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- pyrrole is disclosed Azoles -3- formic acid esters (intermediate 12):

In some embodiments, disclose 3- (((3- (methylol) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) Naphthalene -1- alcohol (intermediate 13):

In some embodiments, disclose 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) Naphthalene -1- alcohol (intermediate 14):

In some embodiments, disclose 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) Naphthalene -1- yl acetate (intermediate 15):

In some embodiments, 3- (((3- ((acetyl thio) methyl) -1- methyl-1 H- pyrazoles -5- base) first is disclosed Base) thio) naphthalene -1- yl acetate (intermediate 16):

Synthesis

In some embodiments, disclose synthesis (E/Z)-dimethyl 2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) oneself two The method of acid esters (intermediate 1):

The following steps are included: (i) making the bromo- 3- chloroaniline of 2- and methyl 2- oxo-cyclopentane -1- formic acid esters and diazotization agent It is contacted in acidic aqueous system；(ii) methyl 2- oxo-cyclopentane -1- formic acid esters and aqueous base are added to acidic aqueous body In system；(iii) isolated hydrazone；(iv) contact hydrazone with acid solution；And (v) separate (E/Z)-dimethyl 2- (2- (2- Bromo- 3- chlorphenyl) hydrazono-) adipate ester (intermediate 1).In some embodiments, which is NaNO₂、Ca(NO₂)₂ Or KNO₂.In some embodiments, which is NaNO₂.In some embodiments, which includes proton Acid and water.In some embodiments, which is hydrochloric acid.In some embodiments, which is potassium acetate, hydroxide Potassium, sodium hydroxide, potassium carbonate, sodium carbonate or potassium phosphate.In some embodiments, which is potassium acetate.In some implementations In example, which includes acid and alcohol.In some embodiments, the acid solution include the concentrated sulfuric acid and methanol, methanesulfonic acid and Methanol or p-methyl benzenesulfonic acid and methanol.In some embodiments, which includes the concentrated sulfuric acid and methanol.

In some embodiments, the chloro- 3- of the bromo- 6- of synthesizing methyl 7- (3- methoxyl group -3- oxopropyl) -1H- Yin is disclosed The method of diindyl -2- formic acid esters (intermediate 2):

The following steps are included: (i) make (E/Z)-dimethyl 2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) adipate ester (in Mesosome 1) it is contacted with acid solution；And (ii) separates the chloro- 3- of the bromo- 6- of methyl 7- (3- methoxyl group -3- oxopropyl) -1H- Yin Diindyl -2- formic acid esters (intermediate 2).In some embodiments, which includes acid and alcohol.In some embodiments, the acid Property solution include the concentrated sulfuric acid and methanol, methanesulfonic acid and methanol or p-methyl benzenesulfonic acid and methanol.In some embodiments, the acidity is molten Liquid includes the concentrated sulfuric acid and methanol.In some embodiments, the method for synthetic intermediate 2 further includes right before separation of intermediates 2 The step of acid solution heats.

In some embodiments, disclosing synthesis has the method for compound of formula (a):

Wherein R1 is blocking group or hydrogen, comprising the following steps: (i) makes 1,5- dimethyl-in the presence of the first solvent 1H- pyrazoles -3- Ethyl formate is contacted with reducing agent to form the first solution；(ii) (1,5- dimethyl -1H- pyrazoles -3- is separated Base) methanol；(iii) make in the presence of the second solvent (1,5- dimethyl -1H- pyrazole-3-yl) methanol and brominating agent with Form (the bromo- 1,5- dimethyl -1H- pyrazole-3-yl of 4-) methanol；(iv) (the bromo- 1,5- dimethyl -1H- pyrazole-3-yl of 4-) is separated Methanol；(v) make (the bromo- 1,5- dimethyl -1H- pyrazole-3-yl of 4-) methanol and alkali (optionally phase transfer catalysis (PTC) in third solvent Agent) and blocking group precursor thereof；And (v) separation has the compound of formula (a).In some embodiments, which selects From lithium aluminium hydride reduction (LAH), diisobutylaluminium hydride (DIBAL), lithium borohydride (LiBH₄), hydrogenate bis- (2- methoxy ethoxies) Aluminium sodiumWith sodium borohydride (NaBH₄).In some embodiments, which is lithium aluminium hydride reduction.In some implementations In example, which is selected from toluene, THF, 2- methyltetrahydrofuran, MTBE, methanol, ethyl alcohol and ether.In some embodiments In, which is tetrahydrofuran.In some embodiments, which is tetrahydrofuran.In some embodiments, should Bromating agent is the bromo- 5,5- dimethyl hydantoin (DBDMH) of 1,3- bis-.In some embodiments, which is N- bromo amber Amber acid imide.In some embodiments, which is lithium hydroxide, sodium hydroxide, potassium hydroxide, sodium hydride, lithium hydride or hydrogenation Potassium.In some embodiments, using phase transfer catalyst, such as Bu₄N·HSO₄Or benzyltrimethylammonium chloride.In some implementations In example, which is potassium hydroxide, and the phase transfer catalyst is 4-butyl ammonium hydrogen sulfate.In some embodiments, the protection Group precursor is 1- (chloromethyl) -4- methoxybenzene.In some embodiments, R1 is to methoxy-benzyl.In some embodiments In, with formula (a) compound be the bromo- 3- of 4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles (in Mesosome 3):

In some embodiments, disclosing synthesis has the method for compound of formula (b):

Wherein R1 is blocking group or hydrogen, comprising the following steps: (i) makes the chemical combination with formula (a) in the presence of the solvent Object is contacted with metalating reagent；(ii) 2- isopropoxy -4,4,5,5- tetramethyl -1,3,2- dioxaborolanes are added； (iii) acid flux material solution is added；And (iv) separation has the compound of formula (b).In some embodiments, the metalating agent It is n-BuLi.In some embodiments, which is tetrahydrofuran.In some embodiments, which includes Acetic acid and toluene.In some embodiments, R1 is to methoxy-benzyl.In some embodiments, the compound with formula (a) is The bromo- 3- of 4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles (intermediate 3).In some embodiments, Compound with formula (b) is 3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -4- (4,4,5,5- tetramethyl - 1,3,2- dioxaborolanes -2- base) -1H- pyrazoles (intermediate 4):

In some embodiments, disclosing synthesis has the method for compound of formula (c):

Wherein R1 is blocking group or hydrogen, comprising the following steps: (i) makes the compound with formula (b) and the bromo- 6- of methyl 7- Chloro- 3- (3- methoxyl group -3- oxopropyl) -1H- indole-2-carboxylic acid ester (intermediate 2) and palladium catalyst are deposited alkali and solvent In lower contact；And (ii) separation has the compound of formula (c).In some embodiments, palladium catalyst is selected from four (triphenyls Phosphine) palladium (0), bis- (di-t-butyl (4- dimethylaminophenyl) phosphine) palladium chlorides (II) and 1,1 '-bis- (di-t-butyl phosphines) two Luxuriant iron palladium chloride.In some embodiments, which is potassium carbonate or potassium phosphate.In some embodiments, which is 1,1 '-bis- (di-t-butyl phosphine) ferrocene palladium chlorides.In some embodiments, which is dioxanes, water or combinations thereof.In In some embodiments, R1 is to methoxy-benzyl.In some embodiments, the compound with formula (b) is 3- (((4- methoxyl group Benzyl) oxygroup) methyl) -1,5- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolanes -2- base) -1H- pyrrole Azoles (intermediate 4).In some embodiments, the compound with formula (c) is the chloro- 3- of (±)-methyl 6- (3- methoxyl group -3- oxygen For propyl) -7- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1H- indoles -2- first Acid esters (intermediate 5):

In some embodiments, disclosing synthesis has the method for compound of formula (d):

Wherein R1 is blocking group or hydrogen, comprising the following steps: (i) makes the compound with formula (c) and methylating agent exist It is contacted in solvent；(ii) methylated derivative handled with ester hydrolysis reagent, and (iii) separation has the change of formula (d) Close object.In some embodiments, the compound with formula (c) is intermediate 5.In some embodiments, which is Hydroxide bases selected from lithium hydroxide, potassium hydroxide and sodium hydroxide.In some embodiments, which is selected from first Base iodine, dimethyl disulfide acid esters and dimethylformamide-dimethylacetal (DMF-DMA).In some embodiments, which tries Agent is dimethylformamide-dimethylacetal.In some embodiments, which is toluene.

In some embodiments, R1 is to methoxy-benzyl.In some embodiments, the compound with formula (c) is The chloro- 3- of (±)-methyl 6- (3- methoxyl group -3- oxopropyl) -7- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- two Methyl-1 H- pyrazoles -4- base) -1H- indole-2-carboxylic acid ester (intermediate 5).In some embodiments, with the compound of formula (d) It is the chloro- 7- of (±) -3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles - 4- yl) -1- Methyl-1H-indole -2- formic acid (intermediate 6):

In some embodiments, disclosing synthesis has the method for compound of formula (e):

Wherein R1 is blocking group or hydrogen, comprising the following steps: (i) makes the compound with formula (d) and (1R) -1- (2- Nitrobenzophenone) ethylamine hydrochloride contacts in the presence of alkali and solvent；And (ii) separation has the compound of formula (e).One In a little embodiments, which is sodium hydroxide, potassium hydroxide, diisopropylethylamine or triethylamine.In some embodiments, which is Sodium hydroxide.In some embodiments, the solvent be water, THF, methanol, ethyl alcohol, isopropanol, n-butanol, methyl ethyl ketone or its Combination.In some embodiments, which is water, ethyl alcohol or combinations thereof.In some embodiments, R1 is to methoxy-benzyl. In some embodiments, the compound with formula (d) is (±) -3- (2- carboxyethyl) -6- chloro- 7- (3- (((4- methoxybenzyl Base) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid (intermediate 6).Some In embodiment, the compound with formula (e) is (R_a) the chloro- 7- of 3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) Methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid-(1R) -1- (2- nitrobenzophenone) ethamine (1:1 salt) (intermediate 7):

In some embodiments, synthesis (R is disclosed_aThe chloro- 7- of)-methyl -6- (3- (methylol) -1,5- dimethyl -1H- Pyrazoles -4- base) -3- (3- methoxyl group -3- oxopropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 8) method:

The following steps are included: (i) contacting the compound with formula (e) with acid in the presence of the first solvent；(ii) divide Free acid component from the compound with formula (e)；(iii) methylating agent or methyl alcohol process free acid are used in the second solvent； (iv) blocking group optionally is removed in third solvent；And (v) separate (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1, 5- dimethyl -1H- pyrazoles -4- base) -3- (3- methoxyl group -3- oxopropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 8).In some embodiments, the compound with formula (e) is R_a) 3- (2- carboxyethyl) -6- chloro- 7- (3- (((4- methoxybenzyl Base) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid-(1R) -1- (2- nitrobenzene Base) ethamine (1:1 salt) (intermediate 7).In some embodiments, which is tetrahydrofuran, toluene or combinations thereof.One In a little embodiments, which is DMF-DMA.In some embodiments, which is toluene.In some embodiments In, which is methanol.In some embodiments, which removed by chloroacetic chloride.In some embodiments, The blocking group is removed by the chloroacetic chloride in methanol.In some embodiments, which such as passed through by acid The hydrochloric acid that chloroacetic chloride is reacted with methanol and is formed in situ removes.

In some embodiments, synthesis (R is disclosed_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrrole Azoles -4- base) -3- (3- hydroxypropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 9) method:

The following steps are included: (i) making (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- Base) -3- (3- methoxyl group -3- oxopropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 8) and reducing agent be in solvent In the presence of contact；And (ii) separates (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) - 3- (3- hydroxypropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 9).In some embodiments, which is two different Butyl aluminum hydride.In some embodiments, which is THF, hexane or combinations thereof.

In some embodiments, it is (intermediate to disclose synthesizing methyl 5- (chloromethyl) -1- methyl-1 H- pyrazoles -3- formic acid esters Body 10) method:

The following steps are included: (i) making dimethyl 1- methyl-1 H- pyrazoles -3,5- dicarboxylic acid esters in the presence of the solvent and going back Former agent contact；(ii) methyl 3- (methylol) -1- methyl-1 H- pyrazoles -5- formic acid esters is separated；(iii) make methyl 3- (hydroxyl first Base) -1- methyl-1 H- pyrazoles -5- formic acid esters and chlorinating agent；And (iv) separates methyl 5- (chloromethyl) -1- methyl-1 H- Pyrazoles -3- formic acid esters (intermediate 10).In some embodiments, which is the mixture of methanol and 2- methyltetrahydrofuran. In some embodiments, which is sodium borohydride or lithium borohydride.In some embodiments, which is hydroboration Sodium.In some embodiments, which is methanol, ethyl alcohol, water, 2- methyltetrahydrofuran, dimethyl acetamide, DCM, THF, ring Or mixtures thereof amyl methyl ether, acetonitrile.In some embodiments, which is the mixing of methanol and 2- methyltetrahydrofuran Object.

In some embodiments, the method for synthesis 3- (acetyl thio) naphthalene -1- yl acetate (intermediate 11) is disclosed:

The following steps are included: contacting 4- hydroxyl naphthalene -2- sodium sulfonate in a solvent with triphenylphosphine and iodine；(ii) divide From 3- mercaptonaphthalene -1- alcohol；(iii) make 3- mercaptonaphthalene -1- alcohol and acylating agent in the presence of amine base and nucleophilic catalyst；And And (iv) separates 3- (acetyl thio) naphthalene -1- yl acetate (intermediate 11).In some embodiments, which is acetonitrile.In In some embodiments, which is acetic anhydride or chloroacetic chloride.In some embodiments, which is acetic anhydride.Some In embodiment, which is selected from triethylamine, pyridine or diisopropylethylamine.In some embodiments, which is triethylamine.In In some embodiments, which is selected from 4-dimethylaminopyridine, pyridine and N- methylimidazole.In some embodiments In, which is 4-dimethylaminopyridine.

In some embodiments, synthesizing methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- is disclosed The method of pyrazoles -3- formic acid esters (intermediate 12):

The following steps are included: (i) making 3- (acetyl thio) naphthalene -1- yl acetate (intermediate in the presence of alkali and solvent 11) it is contacted with methyl 5- (chloromethyl) -1- methyl-1 H- pyrazoles -3- formic acid esters (intermediate 10)；And (ii) separates 5- (((4- Hydroxyl naphthalene -2- base) thio) methyl) -1- methyl-1 H- pyrazoles -3- formic acid esters (intermediate 12).In some embodiments, the alkali Selected from potassium carbonate, lithium hydroxide, 11 carbon -7- alkene of 1,8- diazabicyclo [5.4.0], sodium hydroxide, potassium hydroxide and ethyl alcohol Sodium.In some embodiments, which is potassium carbonate.In some embodiments, which is selected from methanol, ethyl alcohol, water and combinations thereof. In some embodiments, which is methanol.

In some embodiments, synthesis 3- (((3- (methylol) -1- methyl-1 H- pyrazoles -5- base) methyl) sulphur is disclosed Generation) naphthalene -1- alcohol (intermediate 13) method:

The following steps are included: (i) making methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- pyrazoles -3- Formic acid esters (intermediate 12) contacts in a solvent with reducing agent；And (ii) separates 3- (((3- (methylol) -1- methyl-1 H- pyrrole Azoles -5- base) methyl) thio) naphthalene -1- alcohol (intermediate 13).In some embodiments, which is diisobutyl aluminium hydride. In some embodiments, which is tetrahydrofuran, hexane or combinations thereof.

In some embodiments, synthesis 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) sulphur is disclosed Generation) naphthalene -1- alcohol (intermediate 14) method:

The following steps are included: (i) making 3- (((3- (methylol) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- Alcohol (intermediate 13) and chlorinating agent；And (ii) separates 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) first Base) thio) naphthalene -1- alcohol (intermediate 14).In some embodiments, which is selected from THF, methylene chloride, dimethylformamide And combinations thereof.In some embodiments, which is dimethylformamide.In some embodiments, which is methylsulfonyl Chlorine or thionyl chloride.In some embodiments, which is mesyl chloride.In some embodiments, step (i) is further wrapped Include lithium chloride.

In some embodiments, synthesis 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) sulphur is disclosed Generation) naphthalene -1- yl acetate (intermediate 15) method:

The following steps are included: (i) making 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- Alcohol (intermediate 14) contacts in a solvent with acetic anhydride, optional amine base and nucleophilic catalyst；And (ii) separates 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) thio) naphthalene -1- yl acetate (intermediate 15).In some embodiments In, which is triethylamine.In some embodiments, which is selected from 4-dimethylaminopyridine, N- methylimidazole Or pyridine.In some embodiments, which is 4-dimethylaminopyridine.In some embodiments, which is Acetonitrile.

In some embodiments, synthesis 3- (((3- ((acetyl thio) methyl) -1- methyl-1 H- pyrazoles -5- base) is disclosed Methyl) thio) method of naphthalene -1- yl acetate (intermediate 16):

The following steps are included: (i) making 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- Yl acetate (intermediate 15) contacts in a solvent with thioacetic acid potassium；And (ii) separates 3- (((3- ((acetyl thio) first Base) -1- methyl-1 H- pyrazoles -5- base) methyl) thio) naphthalene -1- yl acetate (intermediate 16).In some embodiments, this is molten Agent is acetonitrile.

In some embodiments, synthesis (R is disclosed_a)-(+) the chloro- 5,13,14,22- tetramethyl -28- oxygen of-methyl 17- Miscellaneous penta azepine ring in heptan [27.7.1.1 of -2,9- dithia -5,6,12,13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 Carbon -1 (37), 4 (38), the method for 6,11,14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid esters

The following steps are included: (i) making (R_a) the chloro- 7- of methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- Base) -3- (3- hydroxypropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 9) and sulfonylation agent be in non-proton alkali and molten Contact is in the presence of agent to form the first solution；(ii) iodide salt is added into the first solution；(iii) separation methyl 6- is chloro- 7- [3- (iodomethyl) -1,5- Dimethyl-pyrazol -4- base] -1- methyl -3- (3- sulfonyloxy methyl oxygroup propyl) indole-2-carboxylic acid Ester；(iv) make 3- (((3- ((acetyl thio) methyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- yl acetate (intermediate 16) is contacted with the methanol solution of sodium methoxide to form the second solution；(v) by the chloro- 7- of methyl 6- [3- (iodomethyl) -1, 5- Dimethyl-pyrazol -4- base] -1- methyl -3- (3- sulfonyloxy methyl oxygroup propyl) indole-2-carboxylic acid ester is added to the second solution； And (vi) separates chloro- dithia -5,6,12,13 5,13,14,22- tetramethyl -28- oxa- -2,9- (Ra)-(+) methyl 17-, Penta azepine ring in heptan [27.7.1.1 of 22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14, 16,18,20,23,29,31,33,35- tridecylene -23- formic acid esters.In some embodiments, which is diisopropyl Ethamine or N-methylmorpholine.In some embodiments, which is diisopropylethylamine.In some embodiments, the sulphur Acylating agent is selected from mesyl acid anhydrides, mesyl chloride and p-toluenesulfonic anhydride.In some embodiments, which is methylsulphur Acetyl anhydride.In some embodiments, which is aprotic solvent.In some embodiments, which is tetrahydro Furans.In some embodiments, which is lithium iodide.

In some embodiments, the method for synthesis compound 1 is disclosed:

The following steps are included: (i) making (R_a)-(+) the chloro- 5,13,14,22- tetramethyl -28- oxa- -2,9- two of-methyl 17- Penta azepine ring in heptan [27.7.1.1 of thia -5,6,12,13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid esters connects in a solvent with ester hydrolysis reagent Touching, and (ii) separates (R_a)-(+) chloro- dithia -5,6,12,13 5,13,14,22- tetramethyl -28- oxa- -2,9- -17-, Penta azepine ring in heptan [27.7.1.1 of 22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14, 16,18,20,23,29,31,33,35- tridecylene -23- formic acid.In some embodiments, which is selected from hydrogen-oxygen Change the hydroxide bases of lithium, sodium hydroxide and potassium hydroxide.In some embodiments, which is sodium hydroxide.In some implementations In example, which is selected from methanol, ethyl alcohol, isopropanol, DMSO, water and combinations thereof.

Term " separation " refers to any suitable method that required compound is obtained from reaction mixture.Term " point From " it include extraction, filtering, drying, crystallization, evaporation, the removing of chromatography (such as HPLC, column chromatography), metal etc..Term " separation " include the compound for not purified and being purified from reaction mixture method.In some embodiments, extraction can be passed through Take and separate compound from reaction mixture, and even if compound is not yet purified or removes from solvent, in reaction under One step can also carry out.Those skilled in the art can determine isolates the suitable of required compound from reaction mixture Method.

Term " blocking group " includes hydroxy-protective group, for example, benzyl, to methoxy-benzyl (PMB), THP trtrahydropyranyl (THP), p-methoxyphenyl (PMP) and t-butyldimethylsilyl (TBDMS), Etc..Those skilled in the art being capable of nothing Need the excessive suitable hydroxy-protective group of experimental identification.

Term " blocking group precursor " includes the reagent that blocking group is added to required part.In some embodiments, Blocking group precursor is hydroxy-protective group precursor, such as benzyl chloride, 4- metoxyphenol, bromomethyl pyrazoles, dihydropyran, right Metoxyphenol, chloromethyl pyrazoles, tert-butyl chloro-silicane, tert-butyl chloro-silicane and 1- (chloromethyl) -4- first Oxygroup benzene.Those skilled in the art will easily identify suitable blocking group precursor without excessive experiment.

Term " reducing agent " includes lithium aluminium hydride reduction, sodium borohydride, diisobutyl aluminium hydride etc..

Term " solvent " includes nonpolar solvent, aprotic solvent and proton solvent.Term " nonpolar solvent " includes penta Alkane, pentamethylene, hexane, heptane, hexamethylene, benzene, toluene, chloroform, ether, cyclopentyl-methyl ether, t-butyl methyl ether, tertiary pentyl Methyl ether, methylene chloride and methyl ethyl ketone.Term " aprotic solvent " includes tetrahydrofuran, ethyl acetate, acetone, dimethyl Formamide, acetonitrile, 2- methyltetrahydrofuran, 1,4- dioxanes, dimethyl acetamide and dimethyl sulfoxide.Term " proton solvent " packet Include n-butanol, isopropanol, normal propyl alcohol, ethyl alcohol, methanol, acetic acid and water.In some embodiments, solvent may include any aforementioned The combination of solvent.Those skilled in the art usually can determine the suitable solvent or solvent combination for specific reaction.

Term " bromating agent " includes the bromo- 5,5- dimethyl hydantoin (DBDMH) of 1,3- bis- and N-bromosuccinimide (NBS)。

Term " diazotization agent " includes sodium nitrite (NaNO₂), calcium nitrite (Ca (NO₂)₂) and potassium nitrite (KNO₂)。

Term " metalating agent " includes n-BuLi (n-BuLi).

Term " ester hydrolysis agent " includes sodium hydroxide (NaOH), potassium hydroxide (KOH), lithium hydroxide (LiOH), sodium chloride (NaCl) and lithium iodide (LiI).

Term " alkali " includes potassium acetate (KOAc), potassium carbonate (K₂CO₃), sodium carbonate (Na₂CO₃), potassium phosphate (K₃PO₄), hydrogen Potassium oxide, sodium hydroxide, bis- (trimethyl silyl) amide sodium (NaHMDS), bis- (trimethyl silyl) amides (LiHMDS), sodium hydride (NaH), tert-butyl ammonium hydrogen sulfate, n-BuLi, tert-butyl lithium (t-BuLi), magnesium, zinc, lithium hydroxide, Lithium diisopropylamine (LDA), Sodamide (NaNH₂), potassium tert-butoxide, pyridine, triethylamine (TEA), diisopropylethylamine (DIEA), 11 carbon -7- alkene (DBU) of 1,8- diazabicyclo [5.4.0], sodium methoxide (NaOMe) and sodium ethoxide (NaOEt).Art Language " alkali " further includes alkaline phase transfer catalyst, such as 4-butyl ammonium hydrogen sulfate (Bu₄·HSO₄), it is benzyltrimethylammonium chloride, poly- Ethylene glycol and its derivative, 18- crown ether -6 and other crown ethers.

Term " acid " includes hydrochloric acid (HCl), sulfuric acid (H₂SO₄), p-methyl benzenesulfonic acid (p-TsOH), methanesulfonic acid and acetic acid.One In a little embodiments, acid is concentration.

Term " alcohol " includes methanol, ethyl alcohol, normal propyl alcohol, isopropanol, n-butanol, isobutanol and the tert-butyl alcohol.

Term " reducing agent " includes lithium aluminium hydride reduction (LAH), borane-dimethyl sulfide complex compound, borine-tetrahydrofuran complexing Object, diisobutyl aluminium hydride (DIBAL), lithium borohydride (LiBH₄), bis- (2- methoxy ethoxy) sodium aluminum hydridesWith sodium borohydride (NaBH₄)。

Term " methylating agent " includes methyl iodide, dimethyl disulfide acid esters and dimethylformamide-dimethylacetal (DMF- DMA)。

Term " acylating agent " includes acetic anhydride and chloroacetic chloride.

Term " nucleophilic catalyst " includes dimethyl aminopyridine (DMAP), pyridine and N- methylimidazole.

Term " amine base " includes triethylamine, pyridine and diisopropylethylamine.

Term " sulfonyl agent " includes toluenesulfonic acid acid anhydride, mesyl chloride, paratoluensulfonyl chloride and mesyl anhydride.

Term " iodide salt " includes lithium iodide, sodium iodide and potassium iodide.

In some embodiments, term " palladium catalyst " include 1,1 '-bis- (di-t-butyl phosphine) ferrocene palladium chlorides, [1,1 '-bis- (di-t-butyl phosphine) ferrocene] palladium chloride (II) (Pd 188, PdCl₂(dtbpf)), (R)-(-) -4,12- is double (diphenylphosphine)-[2.2]-(R-Phanephos) luxuriant to ring, (S)-(-) -4,12- bis- (diphenylphosphines)-[2.2]-luxuriant to ring (S-Phanephos), (2- dicyclohexylphosphontetrafluoroborate -2 ', 4 ', 6 '-triisopropyls -1,1 '-biphenyl) [2- (2 '-amino -1,1 '-connection Benzene)] palladium (II) mesylate (XPhos-G3- ring palladium, XPhos-Pd-G3), (2- dicyclohexylphosphontetrafluoroborate -2 ', 6 '-diisopropoxies - 1,1 '-biphenyl) [2- (2 '-amino -1,1 '-biphenyl)] palladium (II) mesylate (RuPhos-G3- ring palladium, RuPhos-Pd-G3), (2- dicyclohexylphosphontetrafluoroborate -2 ', 6 '-diisopropoxies -1,1 '-biphenyl) [2- (2 '-amino -1,1 '-biphenyl)] palladium (II) methanesulfonic acid Salt (RuPhos-G3- ring palladium, RuPhos-Pd-G3), [(2- dicyclohexylphosphontetrafluoroborate -3,6- dimethoxy -2 ', 4 ', 6 '-three isopropyls Base -1,1 '-biphenyl) -2- (2 '-amino -1,1 '-biphenyl)] palladium (II) mesylate (BrettPhos G3), tBuXPhos-Pd- G3, [(2- di-t-butyl phosphine -2 ', 4 ', 6 '-triisopropyls -1,1 '-biphenyl) -2- (2 '-amino -1,1 '-biphenyl)] palladium (II) Mesylate (tBu XPhos G3), tetrakis triphenylphosphine palladium (0) and bis- (di-t-butyl (4- dimethylamino phenyl) phosphine) dichloros Change palladium (II).

In some embodiments, compound 1 can be synthesized as shown in scheme I-VII:

Scheme I

A=i) NaNO₂, HCL aqueous solution；Ii) methyl 2- pentamethylene -1- formic acid esters, KOAc aqueous solution；iii)H₂SO₄、 MeOH

B=H₂SO₄、MeOH

Scheme II

C=i) LiAlH₄,THF；ii)NBS iii)PMBCl,KOH,Bu₄NHSO₄、THF

D=i) n-BuLi, THF；ii)i-PrOBPin

Scheme III

E=Pd (di-t-BPF) Cl₂、K₂CO₃, 1,4- dioxanes, H₂O

F=i) DMF-DMA, PhMe；Ii) NaOH aqueous solution, MeOH

G=(1R) -1- (2- nitrobenzophenone) ethylamine hydrochloride, NaOH, EtOH, H₂O

H=i) HCL aqueous solution, PhMe, THF；ii)DMF-DMA,PhMe；iii)AcCl,MeOH

I=iBu₂AlH、THF

Scheme IV

J=i) NaBH₄、MeOH、MeTHF

Plan V

K=i) PPh₃、I₂,MeCN；ii)Ac2O,Et₃N、DMAP

Plan V I

L=K₂CO₃、MeOH

M=i-Bu₂AlH、THF

N=MeSO₂Cl、LiCl、DMF

O=Ac₂O、DMAP、MeCN

P=KSAc, MeCN

Plan V II

Q=i) intermediate 9, (MeSO₂)₂O、i-Pr₂NEt；ii)LiI,MeCN；Iii) in a separate container, intermediate 16,,NaOMe,MeOH；Iv) will come from iii) solution be added to from ii) compound in the solution of MeCN；V) it adds To in hot DMSO

R=NaOH, DMSO, EtOH, H₂O

Example

The many aspects of present disclosure can further be defined by reference to following non-limiting example, these examples are retouched in detail The method for having stated the preparation and the compound using present disclosure of the certain compounds and intermediate of present disclosure.The common skill in this field Art personnel, which should be understood that, can carry out many modifications to material and method without departing from the range of present disclosure.

Unless otherwise stated:

(i) that, unless otherwise stated, under environment temperature (i.e. within the scope of 17 DEG C to 25 DEG C) and in the lazy of such as nitrogen It is synthesized under the atmosphere of property gas；

(ii) pass through rotary evaporation or be evaporated under reduced pressure using Genevac equipment or Biotage v10 evaporator；

(iii) in automation NovasepOr Teledyne IscoRf or Teledyne IscoIt is used in system60-10-SIL titanium dioxide Silicon (10 μm of particles,Aperture) or using prepackage RediSep Rf Gold^TM(20-40 μm, spherical of silica column Grain), GraceResolv^TMFilter core (Dioxide/silica gel) or (40-63 μm) progress silica gel chromatograph of Silicycle filter core Purifying.

(iv) in Waters (Waters) X5SFC-MS detected with UV or the Waters detected with UV and ELSD (Waters) chiral analysis color is carried out in UPC2 SFC-MS or Agilent (Agilent) the 1100HPLC system detected with UV Spectrum.

(v) yield (when it is present) needs not be accessible maximum value；

(vi) in general, the structure of isolated compound is confirmed by NMR spectra；Dissolvent residual peak is used to mark as inside Standard [uses the Bruker Ultrashield Avance that QCI cryoprobe is housed with δ tape measure nmr chemical shift value III 500MHz spectrometer, the Bruker Ultrashield Avance III 400MHz spectrometer equipped with BBFO probe, Bruker Avance 500(500MHz)、Bruker Avance 400(400MHz)、Bruker Avance 300(300MHz) Or Bruker DRX (300MHz) Instrument measuring proton magnetic resonance spectra]；Unless otherwise specified, it is measured at 27 DEG C；It uses It abridges below: s, it is unimodal；D, doublet；T, triplet；Q, quartet；M, multiplet；Dd, double doublet；Ddd, double doublet Doublet；Bs, bandwidth signals.

(vii) in general, also using equipped with the mass spectrometric Waters of Waters (Waters) SQ or QDa (Waters) UPLC, characterized by the mass spectrography after liquid chromatography separation compound (30 DEG C or 40 DEG C of column temperature, UV=220- 300nm or 210-400nm or 190-400nm, mass spectrum=ESI have the ESI of positive/negative switching), use 97%A+3%B to 3%A The dicyandiamide solution of+97%B, through 1.50min (total run time for returning to initial conditions etc. plus balance is 1.70min) with 1mL/ The flow velocity of min carries out, wherein A=0.1% formic acid or 0.05% trifluoroacetic acid aqueous solution (for acid operation) or 0.1% hydrogen-oxygen Change aqueous ammonium (being used for alkaline operation) and B=acetonitrile.Acidity is analyzed, the column used is Waters (Waters) Acquity HSS T3 (1.8 μm, 2.1 × 50mm or 2.1 × 30mm) or Waters (Waters) Acquity BEH C18 (1.7 μm, 2.1 × 50mm or 2.1 × 30mm), neutral-pH is analyzed, Waters (Waters) Acquity BEH C18 is used (1.7 μm, 2.1 × 50mm) column, and alkalinity is analyzed, used column is Waters (Waters) Acquity BEH C18 (1.7 μm, 2.1 × 50mm or 2.1 × 30mm).Alternatively, (returning to total fortune of initial conditions plus balance using through 1.5min The row time is the Solvent Gradient of 2min) 2% to 98%B, wherein A=0.1% aqueous formic acid, and B=0.1% formic acid is in acetonitrile Solution (for acid operation) or A=0.1% ammonium hydroxide aqueous solution and B=acetonitrile (being used for alkaline operation).Alternatively, Through 3.6min (total cycle time for returning to initial conditions etc. plus balance is 5.1min), 92%A+5%B+3%C to 7% is used The Solvent Gradient of A+90%B+3%C or 90%A+5%B+5%D to 5%A+90%B+5%D, wherein A=water, B=acetonitrile, C The aqueous solution of=1%TFA and the aqueous solution of D=250mM ammonium acetate；Unless otherwise specified, the molecular ion of report is corresponding In [M+H]+；For the molecule (Br, Cl etc.) with multiple isotopic patterns, unless otherwise specified, the value of report is institute The value with maximum intensity obtained.

(viii) in general, measuring the wt% purity of compound (relative to suitable by proton NMR under quantitative conditions Inside reference standard (such as the chloro- 3- nitrobenzene of 1,2,4,5- tetra-, maleic acid or Ergol)).

(ix) it is reacted on a large scale equipped with heat transfer jacket and in the reactor of ancillary equipment appropriate maintenance；And

(x) following abbreviation has been used:

MeCN acetonitrile

Aq. aqueous

Conc. it is concentrated

DCM methylene chloride

Bis- (the di-tert-butyl phosphino-) ferrocene of di-t-BPF 1,1 '-

DIBAH diisobutylaluminium hydride

DMAP 4-dimethylaminopyridine

DMF N,N-dimethylformamide

DMF-DMA N,N-dimethylformamide dimethylacetal

DMSO dimethyl sulfoxide

E.e. enantiomter is excessive

ES electrospray mode

HPLC high performance liquid chromatography

IPA isopropanol

LAH lithium aluminium hydride reduction

MS mass spectrography

MTBE methyl tertiary butyl ether(MTBE)

NBS N-bromosuccinimide

NMR nuclear magnetic resonance

PMB 4- methoxy-benzyl

TFA trifluoroacetic acid

THF tetrahydrofuran

UPLC ultra-performance liquid chromatography

Wt% weight percent

Intermediate 1:(E/Z)-dimethyl 2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) adipate ester

By the bromo- 3- chloroaniline (2.00kg, 9.69mol) of 2-, hydrochloric acid (36wt%, 4.85L, 58.1mol) and water (5L) Mixture stirs 1h.Acquired solution is cooled to 0 DEG C, then gradually adds NaNO in 1h at 0 DEG C -5 DEG C₂(702g, 10.2mol) the solution (2.4L) of Yu Shuizhong.After stirring 1h, at 0 DEG C -5 DEG C, 2- oxo-cyclopentane -1- carboxylic acid first is gradually added Ester (1.38kg, 9.69mol).Then the solution (20L) of KOAc (13.3kg, 136mol) Yu Shuizhong is gradually added.Allow gained Solution reacts other 45min at 0 DEG C -5 DEG C.Then the solution is extracted with DCM and (extracts 12L every time) three times.It will merge Organic extract washed with salt water (10L), be then loaded into another and contain the concentrated sulfuric acid (4.75kg, 48.5mol) in MeOH In the reactor of solution in (3.1kg).The solution allowed reacts 3h at 10 DEG C -20 DEG C.Solution is concentrated into about 8L, Then it completes addition MeOH (every circulation 18L) and two circulations of solvent (every circulation 18L) is distilled off under depressurizing.Gained is starched Liquid is cooled to 0 DEG C -10 DEG C, stirs 1h, then filters.Solid is washed with MeOH (2 × 2L), is then dried under reduced pressure in an oven, To provide (E/Z) -2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) dimethyl adipate, (intermediate 1,3.3kg, 94wt%, 82%)；M/z (ES+), [M+H] +=391.¹H NMR (500MHz, chloroform-d, 27 DEG C) δ 1.98 (m, 2H), 2.41 (t, 2H), 2.59(t,2H),3.66(s,3H),3.87(s,3H),7.05(dd,1H),7.17-7.23(m,1H),7.49(dd,1H), 12.48(bs,1H)。

Intermediate 2: the chloro- 3- of the bromo- 6- of methyl 7- (3- methoxyl group -3- oxopropyl) -1H- indole-2-carboxylic acid ester

By (E/Z) -2- (2- (the bromo- 3- chlorphenyl of 2-) hydrazono-) dimethyl adipate (intermediate 1,3.3kg, 93.7wt%, 7.9mol) solution in the concentrated sulfuric acid (8.4kg, 84mol) and MeOH (26L) stirs 72 hours at 80 DEG C.It will Reaction mixture is cooled to 0 DEG C.Obtained solid is collected by filtration, is washed with MeOH (2L), is then dried at 40 DEG C in vacuum It is dry in case, to provide the solid of 2.5kg.By the solid with identical scale prepare in the same manner it is other 3 batches merging, with Provide the non-purified product of the total 11.9kg obtained by 13.6kg (93.7wt%, 32.5mol) starting material.

The unpurified product of half is added in the MeOH (36L) of stirring.Heat the mixture to 65 DEG C, and by gained Solution keeps 1h at 65 DEG C, is cooled to 0 DEG C later.Gained slurries are stirred into 1h at 0 DEG C, are then filtered.With MeOH (3L) Wash filter cake and the drying in vacuum drying oven at 40 DEG C.The process is repeated with remaining material, merges these materials, to give The solid of 9.7kg out.3.7kg therein is mixed with active carbon (0.74kg), DCM (3.4L) and MeOH (34L), and by slurries It is heated to 65 DEG C of -70 DEG C of holding 1h.Slurries are cooled to 55 DEG C and are filtered.Filter cake is washed with DCM (10L), then by Solvent is distilled off under vacuum, combined filtrate is concentrated into about 8L.It completes to be distilled off under addition MeOH (10L) and vacuum molten Two circulations of agent (10L), then merge obtained slurries with the slurries of the other similar preparation from remaining solid.It will close And slurries be cooled to 0 DEG C and stir 1h, filter later.Filter cake is washed with MeOH (3L) and is done in 40 DEG C of vacuum drying ovens It is dry, with generate the chloro- 3- of the bromo- 6- of methyl 7- (3- methoxyl group -3- oxopropyl) -1H- indole-2-carboxylic acid ester (intermediate 2, 9.4kg, 97.7wt%, 76%)；M/z (ES+), [M+H] +=374.¹H NMR(500MHz,DMSO-d₆,27℃),δ2.60(t, 2H),3.26(t,2H),3.53(s,3H),3.89(s,3H),7.28(d,1H),7.73(d,1H),11.62(bs,1H)。

The bromo- 3- of intermediate 3:4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles

At 4 DEG C -23 DEG C, through 1.5h, LAH (1.05M in THF, 15.0kg, 17.4mol) is gradually added to 1,5- Dimethyl -1H- pyrazoles -3- carboxylic acid, ethyl ester (5.33kg, 31.7mol) then adds in the agitating solution in THF (10.7L) THF(1.0L).After 30min, solution is cooled to 15 DEG C, then while continuing cooling, gradually adds water through 20min The solution of (0.66L, 37mol) in THF (1.9L).Then a few minutes added aqueous NaOH (15wt%, 0.66L, 2.8mol), then water (2.0L) is added.Gained slurries are stirred into 20min at 4 DEG C -11 DEG C, are then filtered under suction.It will The solid of collection with THF (washing is washed four times with 10.7L) every time, to provide (1,5- dimethyl -1H- pyrazole-3-yl) methanol, As the solution in the filtrate of collection.

Reactor 1M HCl, water and THF are rinsed, are again loaded into filtrate later.Three parts of NBS are loaded into the solution (every part of 1.82kg, 99.4wt%, in total 30.6mol) is stirring 7-8min between each part loading at 19 DEG C -27 DEG C, then 45min is stirred at 21 DEG C -28 DEG C later.Then it adds by Na₂SO₃(0.81kg, 99wt%, 6.4mol), NaOH (50wt% Yu Shuizhong, 4.6kg, 57mol) and water (16L) preparation solution, and by gained mixture in 25 DEG C of -26 DEG C of stirring 10min.It will Layer separates and washs lower layer with THF (16L).Upper layer is merged to and is evaporated to drying, with provide be in solid-like (4- bromo- 1, 5- dimethyl -1H- pyrazole-3-yl) methanol.

Unpurified (the bromo- 1,5- dimethyl -1H- pyrazole-3-yl of the 4-) methanol is redissolved in THF (18.8L) simultaneously It is heated to 50 DEG C under stiring, then adds 4-butyl ammonium hydrogen sulfate (0.32kg, 0.95mol), 1- (chloromethyl) -4- methoxyl group Benzene (5.4L, 97.6wt%, 40mol) and THF (2.8L) scrubbing of pipe lines liquid.At 47 DEG C -57 DEG C, KOH is gradually added through 45min (45wt% Yu Shuizhong, 13.7L, 159mol) continues to stir 4h later at 55 DEG C -50 DEG C into the mixture being vigorously stirred.So Mixture is cooled to 20 DEG C afterwards and keeps 63h under stiring.The mixture is reheated to 50 DEG C, is added water (18.7L) And the mixture is stirred into 10min.Except sub-cloud, the solution that will be left in reactor is cooled to 20 DEG C.Addition is brilliant as crystallization The bromo- 3- of 4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles (0.01kg, 0.03mol) of kind is subsequent Heptane (32L) is gradually added through 45min, starts to crystallize during this period.Slurries are stirred into 30min at 20 DEG C, after pass through 45min gradually loads more heptane (22L) and is cooled to 0 DEG C.After 17h, more heptane (11L) is added.After 1h at 0 DEG C Afterwards, slurries are filtered by suction.Filter cake is washed with cooling (0 DEG C) mixture of heptane (17L) and THF (4.3L), then at 40 DEG C It is dry in vacuum drying oven, with provide the bromo- 3- of 4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles (in Mesosome 3,8.78kg, 96wt%, 82%)；M/z (ES+), [M+H] +=325.

¹H NMR(500MHz,DMSO-d₆,27℃)δ2.22(s,3H),3.74(s,3H),3.75(s,3H),4.32(s, 2H),4.39(s,2H),6.87-6.93(m,2H),7.22-7.27(m,2H)。

Intermediate 4:3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -4- (tetramethyl -1 4,4,5,5-, 3,2- dioxaborolanes -2- base) -1H- pyrazoles

At -73 DEG C to -66 DEG C, through 1.5h, butyl lithium (15wt% in hexane, 7.27kg, 17.3mol) is gradually added Be added to the bromo- 3- of 4- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles (intermediate 35.27kg, 96wt%, 15.6mol) in the stirring slurry of THF (43L).Acquired solution is stirred into 1.7h at -77 DEG C to -66 DEG C, then Addition 2- isopropoxy -4,4 in 15min, 5,5- tetramethyls -1,3,2- dioxaborolanes (3.4L, 16mol), then Pass through THF (3.0L) scrubbing of pipe lines.By solution in -77 DEG C to -63 DEG C stirring 1.5h, then at -77 DEG C to -58 DEG C, In Toluene (25.3L) solution of acetic acid (0.89L, 16mol) is gradually added into 15min.Mixture is heated to 20 DEG C, then 67 It is heated under DEG C -82 DEG C (atmospheric pressure) and distilling off solvent (48L).Mixture is cooled to 65 DEG C, add water (25.5L) and Stir the mixture for 10min.Except sub-cloud, then in 72 DEG C of -119 DEG C of (atmospheric pressure；Final 108 DEG C of steam temperature) under distill out More solvents (25.4L).Acquired solution is cooled to 40 DEG C, and is diluted through 10min with heptane (50.6L), is mixed during this period Object is closed to be cooled to 21 DEG C and start spontaneous crystallization.And by 3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolanes -2- base) -1H- pyrazoles (29g) is added as crystal seed.Slurries are existed 0.6h is stirred at about 21 DEG C, is cooled to about -5 DEG C through 1.5h, is then kept at such a temperature overnight (18h).It is filtered by suction slurries, Then dry in 40 DEG C of vacuum drying ovens later with cold (about 0 DEG C) heptane wash filter cake, to provide 3- (((4- methoxy-benzyl) Oxygroup) methyl) -1,5- dimethyl -4- (4,4,5,5- tetramethyl -1,3,2- dioxaborolanes -2- base) -1H- pyrazoles (in Mesosome 4,4.50kg, 99wt%, 77%)；M/z (ES+), [M+H] +=373.

¹H NMR(500MHz,DMSO-d₆,27℃)δ1.23(s,12H),2.34(s,3H),3.68(s,3H),3.73(s, 3H),4.40(s,2H),4.42(s,2H),6.85-6.91(m,2H),7.20-7.25(m,2H)。

Intermediate 5:(±)-chloro- 3- of methyl 6- (3- methoxyl group -3- oxopropyl) -7- (3- (((4- methoxy-benzyl) oxygen Base) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1H- indole-2-carboxylic acid ester

3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -4- is loaded into 100L reactor under a nitrogen (4,4,5,5- tetramethyls -1,3,2- dioxaborolanes -2- base) -1H- pyrazoles (intermediate 4,3.30kg, 99wt%, 8.78mol), the chloro- 3- of the bromo- 6- of methyl 7- (3- methoxyl group -3- oxopropyl) -1H- indole-2-carboxylic acid ester (intermediate 2, 3.20kg, 97.7wt%, 8.35mol), K₂CO₃(1.60kg, 11.6mol) and 1,1 '-bis- (di-t-butyl phosphino-) ferrocene two Palladium chloride (0.132kg, 0.203mol).Then reactor head space is vacuumized and is filled with nitrogen again three times.By 1,4- Dioxanes (26.3L) and water (3.3L) are bubbled 5-10min with nitrogen under reduced pressure, later add their sequences, and by gained Slurries are with stirring in 80 DEG C of heating 5h.Reaction mixture is cooled to 20 DEG C and keeps overnight (16h), using MTBE later The dilution of (33L) and water (33L).Addition N-acetylcystein (0.165kg, 1.01mol) simultaneously stirs the mixture for 15min.Under Layer is removed once settling.Then the hydrochloric acid (37wt%, 2.7L, 33mol) being successively used in water (30L) then uses water (32L) washs upper layer, to provide the chloro- 3- of (±)-methyl 6- (3- methoxyl group -3- oxopropyl) -7- (3- (((4- methoxybenzyl Base) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1H- indole-2-carboxylic acid ester (intermediate 5,22.9kg, 19.1wt%, 97%) MTBE solution.

Intermediate 6:(±) the chloro- 7- of -3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- two Methyl-1 H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid

By the chloro- 3- of (±)-methyl 6- (3- methoxyl group -3- oxopropyl) -7- (3- (((4- methoxy-benzyl) oxygroup) first Base) -1,5- dimethyl -1H- pyrazoles -4- base) -1H- indole-2-carboxylic acid ester (intermediate 5,22.94kg, 19.1wt%, MTBE solution 8.12mol) uses the functionalized titanium dioxide silicon substrate palladium scavenger (Quadrasil of mercapto propyl at 20 DEG C^TMMP, 1.1kg) and toluene (44L) stirs 1h.Slurries are filtered, waste solids are washed with toluene (13L).By at 75 DEG C -107 DEG C Solvent (44L) is distilled off, combined filtrate is concentrated.Then it at 91 DEG C, adds DMF-DMA (5.5L, 41mol).By solution plus Heat, in about 108 DEG C of holding 22h, distills out tetra- parts of solvents of 2L, 4.5L, 4.5L and 4.4L during this period and (is reaching reflux to flowing back Distill, then distilled after 2.0,3.4 and 4.8h respectively immediately afterwards), it is subsequently cooled to 55 DEG C.NaCl (2.2kg) is added Then water (20L) solution removes the two-phase mixture in container, and is returned by 5 μm of pot strainer filterings.It will be screened Mixture, then once settling, lower layer is removed in 50 DEG C of -53 DEG C of stirring 10min.Then MeOH (22L) He Shui is added NaOH (3.3kg, 50wt%, 41mol) aqueous solution in (18L), and by resulting two-phase mixture in 55 DEG C of stirring 3h.Sedimentation Afterwards, layer is gone divided by separation vessel.Then it sends lower layer's (containing product) back to reactor and is stirred at 55 DEG C.Then through 25min Solution of the acetic acid (4.6kg, 77mol) in water (4.4L) is gradually added, the chloro- 7- (3- of 3- (2- carboxyethyl) -6- is then added (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid (0.02kg, 0.04mol) crystalline seed.By mixture in 49 DEG C of -56 DEG C of holding 2h, 20 DEG C are cooled to through 2h, then at 20 DEG C Keep 13h.It is filtered by suction obtained slurries.Rinse reactor and filter cake using water (11L), then by ambient temperature after Continuous suction 6h and it is partially dried on filter, and be completely dried in 40 DEG C of vacuum drying ovens, to provide (±) -3- (2- carboxylic second Base) the chloro- 7- of -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- methyl-1 H- Indole-2-carboxylic acid (intermediate 6,3.78kg, 99.0wt%, 86%)；M/z (ES+), [M+H] +=526.¹H NMR(500MHz, DMSO-d₆,27℃)δ2.01(s,3H),2.50-2.57(m,2H),3.18-3.29(m,2H),3.40(s,3H),3.68(s, 3H),3.82(s,3H),4.08(d,1H),4.14(d,1H),4.19(d,1H),4.21(d,1H),6.64-6.69(m,2H), 6.69-6.74(m,2H),7.25(d,1H),7.75(d,1H),12.71(bs,1H)。

Intermediate 7:(R_a) the chloro- 7- of 3- (2- carboxyethyl) -6- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- two Methyl-1 H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid-(1R) -1- (2- nitrobenzophenone) ethamine (1:1 salt)

It is chloro- that NaOH (50wt% in water, 0.95L, 18mol) is added to (±) -3- (2- carboxyethyl) -6- at 20 DEG C 7- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- first Acid (intermediate 6,9.38kg, 97.5wt%, 17.4mol) and (1R) -1- (2- nitrobenzophenone) ethylamine hydrochloride (2.48kg, 91wt%, 11.1mol) in the stirring slurry in water (7.4kg) and ethyl alcohol (64L), then rinsed with ethyl alcohol (1.8L).Again will When acquired solution is heated to 78 DEG C, start the spontaneous crystallization of product (or lower than 41 DEG C).After being heated to 78 DEG C, complete through 1.3h Slurries are cooled to 63 DEG C, 78 DEG C are heated back to through 0.6h and are kept for three of 10min recycle.Then slurries are cooling through 1.2h To 63 DEG C, 20 DEG C then are cooled to through 1.6h, 20 DEG C is then maintained at overnight, is then filtered by suction.It is rinsed with ethyl alcohol (18L) anti- Answer device and filter cake.Once sufficiently de- liquid, filter cake return in reactor.(solid usually has needed for about 92%e.e. Diacid).Water (7.3L) and ethyl alcohol (66L) is added and reactor content is heated to 78 DEG C under stiring.Then gained is starched Liquid is cooled to 63 DEG C through 1.3h, is then cooled to 20 DEG C through 1.6h, is then maintained at 20 DEG C overnight, is then filtered by suction.Use second Alcohol (18L) rinses reactor and filter cake.Filter cake is dry in 40 DEG C of vacuum drying ovens, to provide (R_a) 3- (2- carboxyethyl) -6- Chloro- 7- (3- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- Formic acid-(1R) -1- (2- nitrobenzophenone) ethamine (1:1 salt) (intermediate 7,5.23kg, 97.5wt%, 99.0%e.e., 7.36mol) yield is 42.3%；M/z (ES+), [M+H]₊=526 (acid), 167 (amine).¹H NMR(500MHz,DMSO-d₆,27 ℃)δ1.49(d,3H),2.00(s,3H),2.61(t,2H),3.10-3.22(m,2H),3.38(s,3H),3.68(s,3H), 4.11(d,1H),4.14(d,1H),4.17(d,1H),4.22(d,1H),4.66(q,1H),6.67-6.72(m,2H),6.75- 6.80(m,2H),7.18(d,1H),7.54-7.60(m,1H),7.63(d,1H),7.76-7.82(m,1H),7.90-7.94(m, 1H),7.93-7.97(m,1H),9.16(bs,3H)。

Chiral purity analyzes HPLC method details: column=Chiralpak AD-H (4.6 × 250mm, 5 μm)；Temperature=25 ℃；Mobile phase=70:30 hexane: the ethyl alcohol (by volume) containing 0.2%TFA, flow velocity 1.0mL/min；Pass through at 254nm UV detection；Volume injected=10 μ L (it is to reach suitable detection limit that this is adjustable)；R_aAnd S_aThe retention time of enantiomter Respectively 4.8 and 13.7min.

Intermediate 8:(R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- first Oxygroup -3- oxopropyl) -1- Methyl-1H-indole -2- formic acid esters

At 45 DEG C, hydrochloric acid (37wt%, 0.77L, 9.2mol) is added to (R_a) the chloro- 7- (3- of 3- (2- carboxyethyl) -6- (((4- methoxy-benzyl) oxygroup) methyl) -1,5- dimethyl -1H- pyrazoles -4- base) -1- Methyl-1H-indole -2- formic acid - (1R) -1- (2- nitrobenzophenone) ethamine (1:1 salt) (intermediate 7,5.23kg, 97.5wt%, 7.36mol) in THF (20.5L) and In the stirring slurry of water (20.5L).After 5min, adds toluene (41L) and stir the mixture for 10min.Except sub-cloud, first is used Benzene (20.5L) dilution upper layer is distilled off solvent (60L) by decompression (590mbar) and is concentrated at 48 DEG C -92 DEG C.In DMF-DMA (3.45L, 25.8mol) is added into gained mixture at 90 DEG C, solution is obtained, solution is heated to flowing back, is kept Flow back (98 DEG C) 8h, is subsequently cooled to 49 DEG C and keeps 16h.Add DMF-DMA (1.0L, 7.5mol) and by mixture 100 Flow back 3.6h again at DEG C, is cooled to 50 DEG C later.It adds water (12.7L) and stirs the mixture for 15min.Except sub-cloud.To MeOH (20L) is added on upper layer, and chloroacetic chloride (2.15L, 29.9mol) 10min is then gradually added into the solution of stirring.Solution exists 60 DEG C of heating 21h, are then transferred into container.By in toluene (10L) He Shui (20.5L) K2CO3 (2.56kg, It 18.4mol) is packed into empty reactor, and heats the mixture to 55 DEG C.Then through 20min by the reaction solution of completion by It is gradually added back in reactor, is then rinsed with toluene (10L).Stop stirring after being stirred for 10min at 55 DEG C, once sedimentation, then Except sub-cloud.Upper layer is concentrated by depressurizing (540mbar) distillation removal solvent (21L) at 55 DEG C -93 DEG C.The solution is existed It is diluted at 50 DEG C with heptane (10L), is then inoculated with (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles - 4- yl) -3- (3- methoxyl group -3- oxopropyl) -1- Methyl-1H-indole -2- formic acid esters (16g, 0.037mol).At 50 DEG C After establishing crystallization 1h, more heptane (20L) is gradually added through 1h.Then slurries are cooled to 20 DEG C through 2h and stir 65h, it After filter.Filter cake is washed with heptane (10L), it is then dry in 40 DEG C of vacuum drying ovens, to provide (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- methoxyl group -3- oxopropyl) -1- Methyl-1H-indole - 2- formic acid esters (intermediate 8,2.85kg, 98wt%, 87%)；M/z (ES+), [M+H] +=434.¹H NMR(500MHz,DMSO- d₆,27℃)δ1.95(s,3H),2.57-2.64(m,2H),3.17-3.33(m,2H),3.44(s,3H),3.57(s,3H), 3.79(s,3H),3.84(s,3H),4.13(dd,1H),4.23(dd,1H),4.72(dd,1H),7.26(d,1H),7.72(d, 1H)。

Intermediate 9:(R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- hydroxyl Base propyl) -1- Methyl-1H-indole -2- formic acid esters

By (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- methoxyl group -3- Oxopropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 8,2.25kg, 98wt%, 5.08mol) is dissolved in THF In (13.3L), and acquired solution is cooled to -45 DEG C.Then at -41 DEG C or lower than -41 DEG C at through 1.1h add DIBAH (20.3wt% is in hexane；11.0kg, 15.7mol).Reaction mixture is kept into other 4.4h at about -45 DEG C, herein Period adds other three parts of DIBAH after 1.4h, 2.8h and 3.7h, and (20.3wt% is in hexane；1.32,0.25 and 0.07kg；1.88,0.36 and 0.10mol), IPA (2.2L, 29mol) then is added into reaction mixture, is then heated through 2h To 20 DEG C, and 2.5h is kept at such a temperature.

Meanwhile in another reactor, four hydration potassium sodium tartrate (6.47kg, 22.9mol) of filling and water (22L).In After 20 DEG C of stirring a few minutes, solution is formed, then adds isopropyl acetate (22L).Obtained two-phase mixture is heated to 50.

Ester reduction reaction mixture is transferred at 50 DEG C through 20min to the aqueous tartrate being vigorously stirred and acetic acid is different In propyl ester mixture, then rinsed with THF (1.5L).Continue to be vigorously stirred 1.9h at 50 DEG C.Except sub-cloud.By upper layer water Then (4.45L) washing removes and is returned in reactor by 5 μm of pipe line filter filterings, then uses isopropyl acetate (1.1L) carries out scrubbing of pipe lines.By distilling removal solvent (32L) under 58 DEG C -74 DEG C (atmospheric pressure) come concentrate solution, then It is cooled to 20 DEG C.It transfers the solution into lesser container, then carries out scrubbing of pipe lines with isopropyl acetate (1.1L), then lead to The distillation under 73 DEG C -85 DEG C (atmospheric pressure) is crossed to remove more solvents (15L) and be further concentrated (to about 9L).It then will stirring Solution be cooled to 70 DEG C, be inoculated with (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- - 2 formic acid esters (2g, 97.9wt%, 5mmol) of (3- hydroxypropyl) -1- Methyl-1H-indole, be cooled within 1 hour 20 DEG C and It is kept for 24 hours at 20 DEG C.It is filtered by suction obtained slurries.Filter cake is washed with isopropyl acetate (2.2L), then in 40 DEG C of vacuum It is dry in baking oven, to provide (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- hydroxyl Base propyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 9,1.76kg, 98.0wt%, 84%) m/z (ES+), [M+H]+ =406.

¹H NMR(500MHz,DMSO-d₆,27℃)δ1.68-1.77(m,2H),1.96(s,3H),2.93-3.05(m, 2H),3.43(s,3H),3.42-3.48(m,2H),3.80(s,3H),3.84(s,3H),4.14(dd,1H),4.23(dd,1H), 4.48(t,1H),4.71(dd,1H),7.25(d,1H),7.71(d,1H)。

Intermediate 10: methyl 5- (chloromethyl) -1- methyl-1 H- pyrazoles -3- formic acid esters

Sodium carbonate (2.14kg, 20.2mol) is added to 1- methyl-1 H- pyrazoles -3,5- of stirring at 20 DEG C ± 5 DEG C Dicarboxylic acid dimethyl ester (7.95kg, 40.1mol) is in the solution in MeOH (80L).It slurry is stirred for 0.5h, is then filtered.It will Filter cake is washed with 2- methyl THF (16L), then returns to filtrate in reactor, then adds 2- methyl THF (24L).It should Solution is cooled to 15 DEG C ± 3 DEG C, and point ten parts of addition sodium borohydrides (3.05kg, 80.7mol) at 15 DEG C ± 3 DEG C.Addition NaBH₄Afterwards, mixture is stirred into 2h at 18 DEG C ± 3 DEG C.Then by gradually loaded at 18 DEG C ± 3 DEG C acetone (16.4kg, It 283mol) is quenched, then stirs 1h at 20 ± 5 DEG C.Then be slowly added aqueous HCl (37wt%, about 8.6kg, 87mol), it keeps temperature to be lower than 30 DEG C, pH is adjusted to 2-3, then stirs the mixture for 1h.Then it is slowly added saturated water Property Na₂CO₃Then (about 4L) stirs the mixture for 3h so that pH is adjusted to 5-6.Mixture is filtered, and by filter cake with DCM (16L) Washing.Concentrate the filtrate to about 20L by vacuum distillation, heating is no more than 40 DEG C, then diluted with DCM (40L) and by The lower distillation of decompression removes solvent (about 40L) to be concentrated again.Filling DCM (80L) and pure water (32L) simultaneously stir gained mixture At least 10min collects lower layer (organic) phase, and is mutually extracted upper layer (aqueous) four times with more DCM (40L/ parts).By low Combined organic phase is concentrated into about 20L in 40 DEG C of vacuum distillations, is added DCM (80L), and (about by distillation removal solvent 40L) concentrate solution again.Then resulting methyl 5- (methylol) -1- methyl-1 H- pyrazoles -3- formic acid is diluted with DCM (80L) Ester solution is simultaneously cooled to 10 DEG C ± 5 DEG C, then keep temperature be lower than 15 DEG C while gradually add thionyl chloride (4.80kg, 40.4mol).Then mixture is stirred into 1h at 20 DEG C ± 5 DEG C.Mixture is concentrated by the vacuum distillation lower than 40 DEG C No more than 20L, DCM (64L) and pure water (80L) are then added.After mutually separating, by lower layer (organic) mutually with aqueous Na₂CO₃ (9wt%, every part of 80L) is washed twice, and is then washed with pure water (80L).It will be washed by the vacuum distillation lower than 40 DEG C Organic phase is concentrated into about 14L, then completes to be slowly added heptane (40L), is then being atmospherically distilled to the two of about 24L lower than 45 DEG C A circulation.It is slowly added more heptane (40L), and gained slurries is stirred at least 0.5h at 20 DEG C ± 5 DEG C, later mistake Filter.Filter cake is washed with heptane (8.0L), it is then dry in 45 DEG C of vacuum drying ovens, to provide methyl 5- (chloromethyl) -1- first Base -1H- pyrazoles -3- formic acid esters (intermediate 10,4.87kg, 64%)；M/z (ES+), [M+H] +=189.¹H NMR(500MHz, CDCl₃,27℃)δ3.91(s,3H),3.99(s,3H),4.59(s,2H),6.82(s,1H)。

Intermediate 11:3- (acetyl thio) naphthalene -1- yl acetate

By MeCN (102L), 4- hydroxyl naphthalene -2- sodium sulfonate (17.00kg, 69.05mol), triphenylphosphine (65.1kg, 248mol) and the stirring mixture of iodine (14.0kg, 55.2mol) heats 6h at 80 DEG C ± 5 DEG C, by mixture cooling and 0 At least 2h is stirred at DEG C ± 5 DEG C, filters out useless solid, is dissolved in filtrate (also containing a large amount of triphenylphosphine oxides) with providing Thick 3- mercaptonaphthalene -1- alcohol, filtrate is recharged in reactor, then loads DMAP (0.84kg, 6.9mol).Keep temperature Lower than 25 DEG C, triethylamine (21.0kg, 207mol) is gradually added, then adds acetic anhydride (17.6kg, 173mol).By mixture 2h is stirred at 15 DEG C -20 DEG C, then under reduced pressure distillation to less than 85L, not be heated beyond 45 DEG C.It adds DCM (85L), Then mixture distilled to less than 85L under reduced pressure, is not heated beyond 40 DEG C.More DCM (170L) is added, then will Mixture is washed with water (170L), is then washed with aqueous NaCl (17wt%, 170L).Lower layer's organic phase is distilled under reduced pressure To less than 51L, it not be heated beyond 40 DEG C.Then it completes to dilute the solution with MeOH (85L) and under reduced pressure lower than 40 DEG C Distillation to less than three of 51L circulations, with yield 71% provide thick 3- (acetyl thio) naphthalene -1- yl acetate (intermediate 11, 40.9kg, 31.3wt%, 49.2mol) methanol solution.The solution for also containing a large amount of triphenylphosphine oxides is directly used in system Standby intermediate 12.

Intermediate 12: methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- pyrazoles -3- formic acid esters

3- (acetyl thio) naphthalene -1- yl acetate (intermediate 11,40.9kg, 31.3wt% In are stirred at 15 DEG C -20 DEG C In methanol, 49.2mol), methanol (64L) and K₂CO₃13.6kg,98.4mol).Branch point addition methyl 5- (chloromethyl) -1- first Base -1H- pyrazoles -3- formic acid esters (intermediate 10,7.40kg, 39.2mol), while keeping temperature lower than 25 DEG C.Then by slurries 2h is stirred at 20 DEG C -25 DEG C.It is analyzed in response to HPLC, loads other four parts of methyl 5- (chloromethyl) -1- methyl-1 H- pyrazoles - 3- formic acid esters (intermediate 10,0.46kg, 2.4mol/ parts) stirs 1h at 20 DEG C -25 DEG C after every part of filling.15 DEG C- It is gradually added at 20 DEG C pure water (109L), stirring gained mixture continues at least 2h, is then allowed to stand at least 3h, is removed by pipeline Liquid stays the viscous solid of sedimentation in the reactor, then adds ethyl alcohol (25.5L) thereto.It will be mixed at 15 DEG C ± 5 DEG C It closes object and stirs at least 1h, the slurries being obtained by filtration later.Filter cake is washed with ethyl alcohol (6.4L), then in 45 DEG C of ± 5 DEG C of vacuum It is dry in baking oven, with provide methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- pyrazoles -3- formic acid esters (in Mesosome 12,13.05kg, 99.3wt%, 81%)；M/z (ES+), [M+H] +=329.¹H NMR(500MHz,DMSO,27℃)δ 3.72(s,3H),3.90(s,3H),4.39(s,2H),6.63(s,1H),6.79(d,1H),7.28-7.32(m,1H),7.38 (ddd,1H),7.45(ddd,1H),7.72(d,1H),8.07(d,1H),10.55(bs,1H)。

Intermediate 13:3- (((3- (hydroxymethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- alcohol

At 30 DEG C ± 5 DEG C by methyl 5- (((4- hydroxyl naphthalene -2- base) is thio) methyl) -1- methyl-1 H- pyrazoles -3- formic acid Ester (intermediate 12,12.7kg, 99.3wt%, 38.4mol) is dissolved in THF (254L), and solution is then cooled to 15 DEG C ± 5 DEG C. It is slowly added DIBAH (1M in hexane, 155L, 155mol), while keeping the temperature at 15 DEG C ± 5 DEG C.Then by mixture Stir 0.5h at 20 DEG C ± 5 DEG C, analyzed by HPLC, be then gradually transferred at 5 DEG C -20 DEG C aqueous hydrochloric acid (4M, 114L, In 456mol).Two-phase mixture is being atmospherically distilled to no more than about 120L lower than 40 DEG C.By gained slurries be cooled to 15 DEG C ± It 5 DEG C, then filters.Filter cake is washed with pure water (25L), is then packed into again together instead with DCM (57L) and THF (6.4L) It answers in device.After stirring mixture at least 10h at 20 DEG C ± 5 DEG C, filtered.Filter cake is washed with DCM (25L), is then existed It is dry in 45 DEG C ± 5 DEG C of vacuum drying oven, to provide 3- (((3- (methylol) -1- methyl-1 H- pyrazoles -5- base) methyl) sulphur Generation) naphthalene -1- alcohol (intermediate 13,10.45kg, 95.5wt%, 87%)；M/z (ES+), [M+H] +=301.¹H NMR (500MHz,DMSO-d₆,27℃)δ3.77(s,3H),4.28(s,2H),4.37(s,2H),4.87(bs,1H),6.14(s, 1H),6.81(d,1H),7.35(d,1H),7.39(ddd,1H),7.47(ddd,1H),7.72-7.75(m,1H),8.03-8.06 (m,1H),10.34(s,1H)。

Intermediate 14:3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- alcohol

Mesyl chloride (6.28kg, 54.8mol) is added gradually to 3- (((3- (methylol) -1- methyl-1 H- pyrazoles -5- Base) methyl) it is thio) naphthalene -1- alcohol (intermediate 13,10.30kg, 95.5wt%, 32.7mol), anhydrous lithium chloride (2.91kg, 68.6mol) and in the stirring mixture of DMF (51.5L), while keeping temperature lower than 10 DEG C.Mixture is stirred at 15 DEG C -20 DEG C Mix 2h.Then EtOAc (155L) is added, then add pure water (155L) and is sufficiently mixed mixture.Lower layer is removed and incited somebody to action Aqueous NaCl (the 17wt% in upper layer；Every part of 155L) it washes twice.Then by upper layer lower than being evaporated in vacuo at 35 DEG C to less than 50L.Then it is slowly added heptane (155L) at 30 DEG C ± 5 DEG C, mixture is cooled to 0 DEG C -5 DEG C later.Stirring slurry is at least After 1h, filtering.Filter cake is washed with heptane (10.3L), it is then dry in 30 DEG C -35 DEG C of vacuum drying oven, to provide 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- alcohol (intermediate 14,9.72kg, 95.3wt%, 89%)；M/z (ES+), [M+H] +=319.¹H NMR(500MHz,CDCl₃,27℃)δ3.80(s,3H),4.07(s,2H), 4.50(s,2H),6.11(s,1H),6.70(d,1H),7.38-7.42(m,1H),7.45-7.52(m,2H),7.52(bs,1H), 7.68-7.74(m,1H),8.16-8.20(m,1H)。

Intermediate 15:3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- yl acetate

Acetic anhydride (3.65kg, 35.8mol) is gradually packed into 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- Base) methyl) it is thio) naphthalene -1- alcohol (intermediate 14,9.51kg, 95.3wt%, 28.4mol), DMAP (360g, 2.95mol) and In the stirring mixture of MeCN (95L), while keeping temperature lower than 25 DEG C.By mixture in 15 DEG C of -20 DEG C of stirring 2h.Then add Add EtOAc (95L), then adds aqueous NaCl (10wt%, 95L.After being thoroughly mixed, lower layer is removed.By upper layer with other two Then aqueous NaCl (10wt%, the every part of 95L) washing of part is carried out lower than 40 DEG C of vacuum distillations to less than 29L, then addition Three circulations of MTBE (95L).By mixture lower than 40 DEG C of vacuum distillations to less than 48L, at least 1h is stirred at about 20 DEG C, It is subsequently cooled to 0 DEG C -5 DEG C.It is slowly added heptane (95L), and slurries is stirred at least 1h at 0 DEG C -5 DEG C, filter later.It will Filter cake is washed with heptane (17L), then dry in 35 DEG C -40 DEG C of vacuum drying oven, to provide 3- (((3- (chloromethyl) -1- Methyl-1 H- pyrazoles -5- base) methyl) thio) naphthalene -1- yl acetate (intermediate 15,8.67kg, 96.1wt%, 81%)；m/z (ES+), [M+H] +=361.¹H NMR(500MHz,CDCl₃,27℃)δ2.46(s,3H),3.82(s,3H),4.11(s,2H), 4.49(s,2H),6.10(s,1H),7.19(d,1H),7.48-7.55(m,2H),7.65-7.67(m,1H),7.73-7.79(m, 1H),7.79-7.85(m,1H)。

Intermediate 16:3- (((3- ((acetylate) methyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene - 1- yl acetate

Thioacetic acid potassium (4.15kg, 36.3mol) is added to 3- (((3- (chloromethyl) -1- methyl-1 H- pyrazoles -5- Base) methyl) thio) mixing of naphthalene -1- yl acetate (intermediate 15,8.62kg, 96.1wt%, 23.0mol) and MeCN (86L) In object, while keeping temperature lower than 25 DEG C.By mixture in 15 DEG C of -20 DEG C of stirring 3h.Then EtOAc (86L) is added, then added Add water (86L).After being thoroughly mixed, lower layer is removed.Upper layer is washed with two parts of aqueous NaCl (15wt%, every part of 86L), then Be lower than 40 DEG C of vacuum distillations to less than 29L and then adding MTBE's (86L × 3, and be 60L in last circulation) Four circulations.It by mixture in 30 DEG C of -35 DEG C of stirring at least 1h, is cooled to lower than 10 DEG C, is then being stirred at least lower than 10 DEG C Then 1h is slowly added heptane (86L).Before filtering, slurries are cooled to 0 DEG C -5 DEG C and keep at least 1h.Filter cake is used into heptan Alkane (17L) washing, it is then dry in 35 DEG C -40 DEG C of vacuum drying oven, to provide 3- (((3- ((acetyl thio) methyl) -1- Methyl-1 H- pyrazoles -5- base) methyl) thio) naphthalene -1- yl acetate (intermediate 16,8.44kg, 97wt%, 89%)；m/z(ES +), [M+H] +=401.¹H NMR(500MHz,DMSO-d₆,27℃)δ2.28(s,3H),2.45(s,3H),3.75(s,3H), 3.91(s,2H),4.42(s,2H),6.04(s,1H),7.36(d,1H),7.53(ddd,1H),7.58(ddd,1H),7.82(d, 1H),7.85(d,1H),7.90(d,1H)。

Intermediate 17:(R_a)-(+) chloro- dithia -5,6 5,13,14,22- tetramethyl -28- oxa- -2,9--methyl 17-, Penta azepine ring in heptan [27.7.1.1 of 12,13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6, 11,14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid esters

At 0 DEG C -30 DEG C, through 15min, solution of the methanesulfonic acid acid anhydride (1.20kg, 6.89mol) in MeCN (2.5L) is added It is added to (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) -3- (3- hydroxypropyl) -1- methyl - 1H- indole-2-carboxylic acid ester (intermediate 9,1.19kg, 97.6wt%, 2.86mol) and n,N-diisopropylethylamine (1.5L, 8.6mol) in the solution in THF (8.7L), scrubbing of pipe lines then is carried out with MeCN (0.3L).By acquired solution at 20 DEG C 5h is stirred, is subsequently cooled to about -5 DEG C.Meanwhile by lower than 30 DEG CXia branches point addition (every part 0.25kg) to MeCN (13L) prepares LiI (1.53kg, 11.4mol) solution.At about -5 DEG C, LiI solution is added gradually to first through 20min In sulfonylation mixture, scrubbing of pipe lines then is carried out with MeCN (1.2L).Gained slurries are warming up to 5 DEG C and in the temperature Lower stirring 5h is cooled to about -15 DEG C later and keeps 16h.Toluene (5.8L), water (11.6L) are sequentially added, salt is then added The solution of sour (37wt%, 0.23L, 2.8mol) in water (1.1L), then carries out scrubbing of pipe lines with water (0.12L).Entire In adding procedure, the temperature of reaction mixture is kept at or below -9 DEG C.Except sub-cloud, water (11.6L) is added to upper layer, stirring Obtained mixture is simultaneously heated to 0 DEG C, and lower layer is removed and (washing washs upper layer with 11.6L) every time with water at about 0 DEG C Twice, it is sufficiently mixed before sedimentation and removes sub-cloud every time.Then by the way that in decompression, (210mbar-250mbar, 30 DEG C extremely 52 DEG C) under distillation removal solvent (8L) be concentrated remaining washed solution in reactor, diluted later with MeCN (2.9L) And the 30min that flows back at 500mbar, thoroughly by solutions.Then it is cooled to and is maintained at 0 DEG C of lasting 69h (through week End), reduced-pressure backflow is repeated in the case that any air enters during keeping for a long time later under a nitrogen.Then by methyl 6- Chloro- 7- [3- (iodomethyl) -1,5- Dimethyl-pyrazol -4- base] -1- methyl -3- (3- sulfonyloxy methyl oxygroup propyl) indoles -2- first The solution of acid esters is cooled to -14 DEG C.

Meanwhile by 3- (((3- ((acetyl thio) methyl) -1- methyl-1 H- pyrazoles -5- base) methyl) is thio) naphthalene -1- base The mixture of acetic acid esters (intermediate 16,1.40kg, 97.8wt%, 3.42mol) and MeOH (7.25L) are heated to reflux and in nitrogen It maintains the reflux for continuing 30min under gas, is cooled to 0 DEG C later.Then through 10min gradually add methanolic sodium methoxide (25wt%, 1.6L, 7.0mol), acquired solution is heated up to 20 DEG C later and is kept in this constant temperature 1.7h.It is passed through at about -15 DEG C 20min is by a part of solution (about 0.35M；6.6L, about 2.3mol) it is added gradually to the above-mentioned chloro- 7- of methyl 6- [3- (iodine first Base) -1,5- Dimethyl-pyrazol -4- base] in -1- methyl -3- (3- sulfonyloxy methyl oxygroup propyl) indole-2-carboxylic acid ester solution.It will Gained mixture, which is maintained at -10 DEG C, continues 1.4h.Add 3- (((1- methyl -3- (sulphomethyl) -1H- pyrrole of another part Azoles -5- base) methyl) thio) two sodium solution (about 0.35M of naphthalene -1- alcohol；1.5L, about 0.53mol), reaction mixture is protected later It holds in -10 DEG C of lasting 18h.Mixture is heated up to 20 DEG C and is taken out from reaction vessel, is then rinsed with MeOH (0.6L).Dress It fills out DMSO and it is heated to 100 DEG C under stiring.Then midbody solution (about 15L) is pumped back at 100 DEG C through 2.8h In reactor, then rinsed with DMSO (0.6L).Reaction mixture is maintained at 100 DEG C of lasting 0.8h, is then cooled to After slightly below 60 DEG C, toluene (29L) and water (5.8L) are added.The temperature of mixture is adjusted to 50 DEG C, is then added aqueous The mixture of NaOH (50wt%, 160g, 2.02mol) and water (0.9L) are then rinsed with water (0.12L).After 10min, remove Lower layer.Upper layer is washed at 53 DEG C with solution of the NaCl (1.74kg) in water (18.6L), then by 160mbar and Distillation removal solvent (6L) is concentrated at 85 DEG C of jacket temperature, to provide crude product (19.2kg, 6.1wt%, 1.7mol) Toluene solution.By it with another kind from (R_aThe chloro- 7- of)-methyl 6- (3- (methylol) -1,5- dimethyl -1H- pyrazoles -4- base) - 3- (3- hydroxypropyl) -1- Methyl-1H-indole -2- formic acid esters (intermediate 9,1.19kg, 98.0wt%, 2.87mol) is according to phase Such solution (18.6kg, 5.7wt%, 1.5mol) of same program preparation merges.InSilica (3.0kg, 10 μm of partial sizes,Aperture) compression column (20cm diameter × 22cm long) on washed with the mixture of toluene and ethyl alcohol De- (approximate volume is than 93% toluene: 7% ethyl alcohol) come divide partial purification to merge by chromatography solution (every part 0.84L, 51 parts).Product fraction is divided into two parts and is evaporated under reduced pressure at 50 DEG C, until distillation stops, to provide foam-like product (91g And 2.40kg).Batch of material is dissolved in DMSO (148g and 3.22kg), to provide (R_a)-(+)-chloro- 5,13,14,22- of methyl 17- Penta azepine ring in heptan [27.7.1.1 of tetramethyl -28- oxa- -2,9- dithia -5,6,12,13,22-^4,7.0^11,15.0^16,21.0^{20, 24}.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid esters The DMSO solution of (intermediate 17,239g, 32.2wt%, 0.112mol and 5.63kg, 37.9wt%, 3.11mol) merges yield It is 56%；M/z (ES+), [M+H] +=686.¹H NMR(500MHz,DMSO-d₆,27℃)δ1.97(s,3H),2.16-2.27 (m,1H),2.32-2.42(m,1H),2.89(d,1H),3.08(d,1H),3.07-3.14(m,1H),3.16(d,1H),3.36- 3.42(m,1H),3.43(d,1H),3.48(s,3H),3.69(s,3H),3.76(s,3H),3.77-3.83(m,1H),3.84 (s,3H),4.13(td,1H),4.22(d,1H),4.29(d,1H),4.77(s,1H),6.65(d,1H),7.18(d,1H), 7.39-7.40(m,1H),7.44-7.48(m,1H),7.47-7.51(m,1H),7.71-7.74(m,1H),7.90(d,1H), 8.08-8.12(m,1H)。

Compound 1:(R_a)-(+) chloro- dithia -5,6,12 5,13,14,22- tetramethyl -28- oxa- -2,9- -17-, Penta azepine ring in heptan [27.7.1.1 of 13,22-^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11, 14,16,18,20,23,29,31,33,35- tridecylene -23- formic acid

(R_a)-(+) the chloro- 5,13,14,22- tetramethyl -28- oxa- -2,9- dithia -5,6,12,13,22- of-methyl 17- Penta azepine ring in heptan [27.7.1.1^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16, (intermediate 17,37.9wt% contain 57.3wt% in DMSO to 18,20,23,29,31,33,35- tridecylene -23- formic acid esters DMSO；2.81kg, 1.55mol), DMSO (2.81kg) and ethyl alcohol (1.68kg) are packed into 20L reactor, and solution is existed 50 DEG C are heated under stirring.Then NaOH (50wt% Yu Shuizhong is added；186g, 2.33mol), then add water (267mL) pipe Line cleaning solution.After 1.5h, add acetic acid (267mL, 4.66mol).Then aqueous ethanol (34.5wt% is added；3.0L), then Addition form A (R_a)-(+) the chloro- 5,13,14,22- tetramethyl -28- oxa- -2,9- dithia -5,6,12,13,22- penta of -17- Azepine ring in heptan [27.7.1.1^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16,18, 20,23,29,31,33,35- tridecylene -23- formic acid monohydrate (0.8g, 0.001mol) crystalline seeds.By mixture 50 DEG C stirring 4.5h, then gradually add other two parts (4.2 and 1.3L) aqueous ethanols (34.5wt%) (respectively through 4.1h and 0.7h).Slurries are cooled to 20 DEG C through 2h and keep 17h at 20 DEG C.Slurries are filtered by suction, by filter cake aqueous ethanol (34.5wt%；Washing is washed twice with 2.7L) every time, dry in 40 DEG C of vacuum drying ovens later, the A (R in the form of providing_a)- The chloro- penta azepine ring in heptan of 5,13,14,22- tetramethyl -28- oxa- -2,9- dithia -5,6,12,13,22- of (+) -17- [27.7.1.1^4,7.0^11,15.0^16,21.0^20,24.0^30,35] 38 carbon -1 (37), 4 (38), 6,11,14,16,18,20,23,29, 31,33,35- tridecylene -23- formic acid monohydrates (compound 1,1.01kg, 99.4wt%, >=99.9%e.e., 94%)；m/ Z (ES+), [M+H] +=672.¹H NMR(500MHz,DMSO-d₆,27℃)δ1.96(s,3H),2.16-2.28(m,1H), 2.30-2.42(m,1H),2.88(d,1H),3.06(ddd,1H),3.12(d,1H),3.18(d,1H),3.42(d,1H), 3.41-3.48(m,1H),3.51(s,3H),3.72(s,3H),3.75(s,3H),3.84(td,1H),4.09(td,1H),4.25 (d,1H),4.28(d,1H),4.75(s,1H),6.67(d,1H),7.13(d,1H),7.37-7.40(m,1H),7.43-7.47 (m,1H),7.45-7.50(m,1H),7.68-7.74(m,1H),7.84(d,1H),8.07-8.13(m,1H),13.36(bs, 1H)。

Chiral purity analyzes HPLC method details: column=ChiralPakIE-3,3 μm of 4.6x 250mm；Temperature=40 DEG C Mobile phase=50:50 ethyl alcohol: the hexane (by volume) containing 0.1%TFA, flow velocity 0.8mL/min；Pass through UV at 305nm Detection；Volume injected=10 μ L (it is to reach suitable detection limit that this is adjustable)；R_aAnd S_aThe retention time of enantiomter point It Wei not 8.5 and 11.5min.

The XRPD of form A as shown in Figure 1, and the results list it is following (table 1).

The peak XRPD of 2. form A of table

Dsc analysis shows that form A has the heat absorption thing for the desolventizing for starting and reaching at about 158 DEG C peak at about 121 DEG C Part is followed by the melting/decomposition endothermic event for starting at about 181 DEG C and reaching peak at about 194 DEG C.TGA shows form A exhibition Reveal be heated to about from about 25 DEG C after 160 DEG C about 4.0% mass loss.The representative DSC/TGA thermography of form A is illustrated in figure In 2.

Monoclinic crystal structure analytical proof form A is monohydrate form.Crystallographic data: Space group Monoclinic system P2 (1), Unit cell dimension:β=90.23 (2) °,

X-ray powder diffraction (XRPD) analysis

XRPD analysis is carried out using BrukerD4 diffractometer, and the diffractometer is available commercially from Brooker AXS company (Bruker AXS Inc^TM) (Madison, the state of Wisconsin).The XRPD spectrum, which passes through, is mounted on the material sample (about 20mg) for being used to analyze On single silicon crystal chip support (for example, zero Background X-ray diffraction sample rack of Brooker silicon) and by means of slide will The sample sprawls straticulation acquisition.By the sample with 30 revs/min of rotations (improve counting statistics) and with by 40kV and The X-ray of the wavelength with 1.5406 angstroms (that is, about 1.54 angstroms) that the long fine focusing pipe of the copper operated under 40mA generates is irradiated. In the range of the 2- θ from 2 ° to 40 °, to make every 0.02 ° of sample of 2- θ increment (continuous scanning mode) exposure 1 in θ-θ mode Second.Runing time is 31min41s.

2 θ value of XRPD can change in reasonable range, such as change in the range of ± 0.2 °, and when due to various XRPD intensity may become when the reason of various kinds (including for example preferred direction) measures substantially the same crystal form Change.The principle of XRPD is described in publication, such as Ji Kawazuo (Giacovazzo, C.) et al. (1995) crystal is basic Principle (Fundamentals of Crystallography), Oxford University Press；Charles Jenkins (Jenkins), R. and Si Nai Moral, R.L. (1996), to introduction (the Introduction to X-Ray of X-ray powder diffraction ), PowderDiffractometry John Wiley father and son publishing company (John Wiley&Sons), New York；And Ke Luge (Klug), the Alexandria H.P.& (Alexander), L.E. (1974), X-ray diffraction program (X-ray Diffraction ), Procedures John Wiley father and son publishing company (JohnWiley and Sons), New York (New York).

Dsc analysis

For the sample prepared according to standard method, using being available from TA(Newcastle, The Delaware State) Q SERIES^TMQ1000DSC calorimeter carries out dsc analysis.Sample (about 2mg) is weighed to aluminium sample disc In and be transferred to the DSC.The instrument nitrogen is purged with 50mL/min and using about 10 DEG C/min of dynamic heat speed Rate collects the data between about 22 DEG C and 300 DEG C.Dsc data is used into standard software, for example, coming from TAGeneral v.4.5A analyzed.

Thermogravimetry (TGA)

For the sample prepared according to standard method, using can be from TA instrument(Newcastle, The Delaware State) obtain Q SERIES^TMQ5000 thermogravimeter carries out TGA.Sample (about 5mg) is put into aluminium sample disc In and be transferred in TGA furnace.The instrument is purged with nitrogen with 50mL/min, and uses 10 DEG C/min of dynamic heat Data of the rate collection between 25 DEG C and 300 DEG C.Dsc data is used into standard software, for example, coming from TAGeneral v.4.5A analyzed.

Example 1: the external activity of compound 1

Caspase Activity measurement:This is (multiple in MOLP-8 for measuring after handling 6h with Mcl-1 inhibitor Property myeloma), KMS-12-BM (Huppert's disease), MV4；11 (acute myelogenous leukemias) and NCI-H23 (non-small cell Lung cancer) raji cell assay Raji that induces cell apoptosis in cell.At first day, by 3000 (MOLP-8, KMS-12-BM, MV4；11) Or 1250 (NCI-H23) a cells/well be seeded in the growth medium (IMDM+10%FBS of 50 μ L in the white microplate of the hole 384- + 2mM L-Glu is used for MV4；11, and RPMI-1640+10%FBS+2mM L-Glu is used for every other class) in, and incubate Educate overnight (37 DEG C, 5%C0₂, 80%RH).At second day, these cells are used into ECHO acoustics liquid processor (10 thirty- Logarithm serial dilution, 31.5 μm of maximum concentrations, 0.3% final DMSO concentration) it is handled with compound I.Be incubated for (37 DEG C, 5% C0₂, 80%RH) after 6h, 3/7 reagent of caspase-Glo (Pu Luomaige company (Promega)) of 25 μ L is added to In each hole, and these plates are protected from light at room temperature and are incubated for 30min.Use Infinite M200 microplate reader (Di Ken company (Tecan)) it is recorded and is shone with the 100ms time of integration.EC is calculated using GeneData analysis software₅₀It is worth and is shown in the following table 2.

The result that table 2. is measured from external Caspase Activity