CN1993342A

CN1993342A - Intermediates for the preparation of halichondrin b

Info

Publication number: CN1993342A
Application number: CN 200580026228
Authority: CN
Inventors: B·奥斯塔; C·E·蔡斯; F·G·房
Original assignee: Eisai Co Ltd
Current assignee: Eisai R&D Management Co Ltd
Priority date: 2004-06-03
Filing date: 2005-06-03
Publication date: 2007-07-04
Also published as: CA2822994A1; CN109180615A; CA2822994C; CN108997267A

Abstract

The present invention provides macrocyclic compounds of formula (F-4), synthesis of the same and intermediates thereto. Such compounds, and compositions thereof, are useful for treating or preventing proliferative disorders.

Description

It is used to prepare the intermediate of halichondrin B

The cross reference of related application

The U.S. Provisional Patent Application 60/576 submitted this application claims on June 3rd, 2004,642,60/626 submitted on November 10th, 2004, above-mentioned each application full content is incorporated herein by reference by 60/663,300 priority that on March 18th, 769 and 2005 submits.

Technical field

The present invention relates to synthesis medical active Macrocyclic lactone compounds in, the compound as intermediate.

Background of invention

The present invention relates to medical active macrolide, intermediate used in synthesis and its synthesis.Halichondrin B (Halichondrin B) is a kind of effective anticancer agent, it is initially separated from the soft sponge of marine sponge black (Halichondria okadai)), the then discovery in small axis Sponge (Axinella sp.), Phakellia carteri and flat lance Sponge (Lissonderndryx sp.).Disclose within 1992 fully synthetic (Aicher, T.D. etc., the J.Am.Chem.Soc.114:3162-3164) of halichondrin B.It is proved halichondrin B and inhibits tubulin polymerization, micro-pipe assembly, β in vitro^sThe GTP of tubulin crosslinking, GTP and vinblastine with tubulin binding and dependent on tubulin is hydrolyzed, and has shown that anticancer property in vitro and in vivo.Therefore, it is necessary to be used as the synthetic method of the halichondrin b analogs of anticancer agent to preparation to develop.

Summary of the invention

As described herein, the present invention provides the method that preparation has the halichondrin b analogs of such as anticancer activity or antimitotic (blocking mitosis) activity medical active.These compounds include the compound of formula B-1939:

These compounds are used for treating cancer and other proliferative diseases, the including but not limited to fibroblast of melanoma, fibrosarcoma, leukaemia, colon cancer, oophoroma, breast cancer, osteosarcoma, prostate cancer, lung cancer and ras conversion.The present invention also provides the synthetic intermediates for being used to prepare the halichondrin b analogs.

The detailed description of certain embodiments of the present invention

Method and intermediate of the invention is used to prepare the various halichondrin b analogs described in such as United States Patent (USP) 6,365,759 and United States Patent (USP) 6,469,182 (being incorporated herein by reference).As shown in following scheme I, halichondrin b analogs are generally prepared by the assembling to three segments F-1, F-2 and F-3.

Scheme I

1, segment F-1

According to a kind of embodiment, the present invention provides compound F-1:

Wherein: PG¹And PG²Respectively stand alone as hydrogen or suitable hydroxyl protection base；

R¹For R or OR；

R²For CHO or-CH=CH₂；And

Each R stands alone as hydrogen, C_1-4Halogenated aliphatic base, benzyl or C_1-4Aliphatic group, condition are to work as R¹When for OMe, PG¹And PG²Do not form acetonide group.

In certain embodiments, R¹For OR.In other embodiment, R¹For OR, wherein R is hydrogen, methyl or benzyl.

In certain embodiments, PG¹And PG²For hydrogen.In other embodiment, PG¹And PG²In one be hydrogen.

Suitable hydroxyl protection base is known in the art; it include " blocking group (protecting Groups in organic synthesis) in organic synthesis " (T.W.Greene and P.G.M.Wuts; 3rd edition; John Wiley & Sons; 1999, be incorporated herein by reference) in group those of is described in detail.In certain embodiments, PG¹And PG²Respectively ester, ether, silyl ether, alkyl ether, aryl alkyl ethers and alkoxyalkyl ether are independently selected from together with oxygen atom in combination.The example of this ester includes formic acid rouge, acetic acid esters, carbonic ester and sulphonic acid ester.Specific example includes formic acid esters, benzoyl formiate, chloracetate, trifluoro-acetate, methoxyacetic acid ester, triphenylmethoxy acetic acid esters, parachlorophen-oxyacetic acid ester, 3- phenylpropionic acid ester, 4-oxopentanoic acid ester, 4; 4- (ethylene is thio) valerate, pivalate (pivaloyl group), crotonates, 4- methoxyl group crotonates, benzoic ether, to benzyl benzoate, 2; 4,6- trimethylbenzoic acid esters；Or carbonic ester, such as methyl, 9- fluorenyl methyl, ethyl, 2,2,2- trichloroethyls, 2- (trimethyl silyl) ethyl, 2- (benzene sulfonyl) ethyl, vinyl, allyl and to nitrobenzyl carbonic ester.The example of this silyl ether includes trimethyl silyl ether, triethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, triisopropylsilyl ether and other trialkylsilyl ethers.Alkyl ether includes methyl ether, benzylic ether, to methoxy-benzyl ether, 3,4- dimethoxy-benzyl ether, trityl ether, tertbutyl ether, allyl ether and allyloxycarbonyl ether or derivative.Alkoxyalkyl ether includes acetal, such as methoxy ether, methylthiomethyl ether, (2- methoxy ethoxy) methyl ether, benzyloxymethyl ether, β-(trimethyl silyl) ethoxyl methyl ether and THP trtrahydropyranyl ether.The example of aryl alkyl ethers includes benzylic ether, to methoxy-benzyl ether (MPM), 3,4- dimethoxy-benzyl ether, O- nitrobenzyl ether, to nitrobenzyl ether, to halogeno-benzyl ether, 2,6- dichloro benzyl ether, to cyanobenzyls ether, 2- and 4- picolyl ether.

In certain embodiments, the PG of F-1¹And PG²One or two of part is silyl ether or aryl alkyl ethers.In other embodiments, the PG of F-1¹And PG²One or two of part is t-butyldimethylsilyl (t-butyldimethylsilyl) or benzoyl.In other embodiment, the PG of F-1¹And PG²Part is t-butyldimethylsilyl.

According to another embodiment, PG¹And PG²It is formed together glycerol protection base, such as cyclic ketal or ketal together with the oxygen atom in conjunction with them, such group includes methylene, ethylidene, benzal, isopropylidene, cyclohexylidene and cyclopentylene；Silylene derivative, such as di-t-butyl silylene and 1, the sub- siloxy group derivative of 1,3,3- tetra isopropyl two；Cyclic carbonate ester and ring borate.The method for being added and removing such hydroxyl protection base and other protecting groups is known in the art; and it is available from such as " protecting group (protecting Groups) " (P.J.Kocienski; Thieme; and " blocking group (protective Groups in organicsynthesis) in organic synthesis " (T.W.Greene and P.G.M.Wuts 1994); 3rd edition; John Wiley & Sons, 1999).According to another embodiment, PG¹And PG²Acetonide base is collectively formed.

According to a kind of embodiment, R²For CHO.

According to another embodiment, R²For-CH=CH₂。

In certain embodiments, the present invention provides the compound of formula F-1, has spatial chemistry shown in compound F-1 ':

The wherein classification and subclass description of each variable as hereinbefore defined and above and herein.

In certain embodiments, following compound F-1a and F-1b is provided:

Wherein, " TBS " refers to t-butyldimethylsilyl.

The detailed synthesis of F-1a and F-1b is illustrated in the following embodiments.

2, segment F-2

According to another embodiment, the present invention provides compound F-2:

Wherein: each

Singly-bound or double bond are stood alone as, condition is two Base is not double bond simultaneously；

LG¹For suitable leaving group；

X is halogen or-OSO₂(R^y)；

R^yFor C_1-6Aliphatic group or saturation, part is unsaturated or complete unsaturated 5-7 member ring, wherein R^yOptionally halogen, R, NO are selected from by most 3₂, CN, OR, SR or N (R)₂Group replace；

Each R stands alone as hydrogen, C_1-6Halogenated aliphatic base or C_1-4Aliphatic group；And

PG³For suitable hydroxyl protection base.

Suitable leaving group used herein is to be easy to be required the chemical group that the chemical group of introducing replaces.Suitable leaving group is known in the art, for example, see " Advanced Organic Chemistry (Advanced Organic Chemistry) (Jerry March, the 4th edition, pp.351-357; John Wiley and Sons, N.Y. (1992)).Such leaving group includes but is not limited to halogen, alkoxy, sulfonyloxy, the alkylsulfonyloxy optionally replaced, the alkenyl sulfonyloxy optionally replaced, the aryl-sulfonyl oxygen and diazo optionally replaced.The example of suitable leaving group includes chlorine, iodine, bromine, fluorine, mesyl, tosyl (mesyl), trifluoromethanesulfonic acid root (triflate), nitrobenzenesulfonyl (nosyl) and bromobenzenesulfonyl (brosyl).In certain embodiments, the LG of F-2¹Partially (moiety) is sulfonyloxy, the alkylsulfonyloxy optionally replaced, the alkenyl sulfonyloxy optionally replaced or the aryl-sulfonyl oxygen optionally replaced.In other embodiment, the LG of F-2¹Part is the alkylsulfonyloxy optionally replaced.In other embodiments, the LG of F-2¹Part is mesyl or tosyl.

In certain embodiments, the X section of F-2 is halogen.In other embodiment, the X section of F-2 is sulfonyloxy, the alkyl sulphonyl optionally replaced, the alkenylsufonyl optionally replaced or the aryl sulfonyl optionally replaced.In other embodiment, the X section of F-2 is trifluoromethanesulfonic acid root.

In certain embodiments, the PG of F-2³Part is silyl ether together with oxygen atom in combination.In other embodiment, the PG of F-2³Part is ester group together with oxygen atom in combination.According to an aspect of the present invention, the PG of F-2³Part is t-butyldimethylsilyl together with oxygen atom in combination.According to another aspect of the present invention, the PG of F-2³Part is valeryl or benzoyl together with oxygen atom in combination.

In certain embodiments, the present invention provides the compound of formula F-2, has spatial chemistry shown in compound F-2 ':

In certain embodiments, compound F-2a or F-2b are provided:

Wherein " MsO " nail sulfonate radical, " TfO " refer to trifluoro sulfonate radical, and " Opv " refers to neopentanoic acid root (pivaloate), and " OBz " refers to that benzoate anion and " TsO " refer to tosylate.

In other embodiment, the present invention provides the compound of formula F-2b, wherein the compound is crystallization.According to another embodiment, the compound of formula F-2b is provided, wherein the compound is crystallized from alkanes solvent.In certain embodiments, F-2b crystallization is provided, wherein the compound is crystallized from pentane or heptane.In other embodiment, F-2b crystallization is provided, wherein the compound is crystallized at about 0 DEG C.

As shown in following scheme A, the compound of formula F-2 is generally prepared by intermediate F-2d and F-2e.

Option A

Therefore, another aspect of the present invention is to provide the compound of formula F-2d:

Wherein: R ' is-CH=CH₂Or-C (O) H；

AlK is the C of linear chain or branched chain_1-4Aliphatic group；And

PG⁵For suitable hydroxyl protection base.

Suitable hydroxyl protection base PG⁵With above to the PG of compound F-2³Partial description and definition.In certain embodiments, PG⁵It is silyl ether together with oxygen atom in combination.In other embodiment, PG⁵For t-butyldimethylsilyl.

According to a kind of embodiment, the part AlK of compound F-2d is methyl.

In certain embodiments, the compound of formula F-2d ' is provided:

The compound of another aspect of the present invention offer formula F-2e:

Wherein: R " is OH, OPG³Or LG⁴；

LG⁴For suitable leaving group；And

Each PG³Suitable hydroxyl protection base is stood alone as, condition is to work as PG³When for t-butyldiphenylsilyl, R " is not OMs.

Those skilled in the art will appreciate that the R " of compound F-2e can be partially transformed into hydroxyl, the OPG of protection by OH³Or it is directly converted to LG⁴.It is this to be transformed to known to one skilled in the art, it especially include those of described herein.R " is OH or LG in certain embodiments⁴.The leaving group LG of formula F-2e⁴With above to the LG of compound F-2¹Partial description and definition.In certain embodiments, LG⁴For tosyl or mesyl.

The PG of compound F-2e³Part is the same as above to the PG of compound F-2³Partial definition and description.In certain embodiments, PG³Silyl ether is constituted together with oxygen atom in combination.In other embodiment, PG³For t-butyldiphenylsilyl.

The compound of an additional aspect of the present invention offer formula F-2f:

Wherein AlK, PG³And PG⁵Classification and subclass description with general definition and above and herein.By method described herein and method those of known in the art, with the compound of the preparation of compounds of formula F-2 of formula F-2f.

The detailed synthesis of F-2a is illustrated in the following embodiments.

Alternatively, as shown in following scheme II, by the compound of D- chinic acid preparation formula F-2.The detailed preparation of the compound of formula F-2 is illustrated in the following embodiments.

Scheme II

Another route that intermediate 17 is converted into as intermediate 12 is provided from another method of the compound of D- chinic acid preparation formula F-2 (as shown in following scheme III).

Scheme III

Above scheme III indicates another method for preparing intermediate 17 by intermediate 12 by Eschenmoser-Tanabe fragmentation reaction, wherein each Rx stands alone as OPG^xOr CN, wherein PG^xFor suitable hydroxyl protection base as described herein.Then according to above scheme II, with the compound of 17 preparation formula F-2 of intermediate.

The another route that intermediate 17 is converted into as intermediate 9 is provided from another method of the compound of D- chinic acid preparation formula F-2 (as shown in following scheme IV).

Scheme IV

Wherein PG^yFor suitable carboxyl-protecting group as described herein, each PG^xStand alone as suitable hydroxyl protection base as described herein.

Plan V indicates another preparation method of the intermediate for the compound by D- chinic acid preparation formula F-2.

Plan V

In above scheme V, intermediate 7 (derives from scheme II), and wherein R is that methyl esters is used to prepare crystallization of intermediate ER-817664.As described in above scheme V, PG⁵And PG⁶Respectively stand alone as suitable hydroxyl protection base.In certain embodiments, PG⁵And PG⁶Ring glycerol protection base is collectively formed.In other embodiment, PG⁵And PG⁶Cyclohexylidene protecting group is collectively formed.As described in above scheme V, LG⁵For suitable leaving group.This suitable leaving group is known in the art, including those described herein group.In certain embodiments, LG⁵For mesyl or tosyl.

According to another embodiment, the present invention provides the compound of formula A:

Wherein: Indicate singly-bound or double bond；

N is 1,2 or 3；

PG⁵And PG⁶Respectively stand alone as suitable hydroxyl protection base；

W is CH-A or C (O)；

A is oxo or C_1-6Aliphatic group, wherein A is optionally by one or more Q¹Group replaces；

Each Q¹It is independently selected from cyano, halogen, azido, oxo, OR, SR, SO₂R、OSO₂R、N(R)₂、NR(CO)R、NR(CO)(CO)R、NR(CO)N(R)₂、NR(CO)OR、(CO)OR、O(CO)R、(CO)N(R)₂、O(CO)N(R)₂Or OPG¹, wherein PG¹For suitable hydroxyl protection base, in which:

Two Q on A¹Optionally in combination with together to form saturated rings, part unsaturated ring or the aromatic rings with the 0-4 heteroatomic 3-8 members for being independently selected from nitrogen, oxygen or sulphur；And

Each R is independently selected from hydrogen or what is optionally replaced is selected from C_1-6Aliphatic group；Saturated carbon ring, part unsaturated carbocyclic or the aromatic carbocyclic of 5-10 member；Or the group of the saturated rings of the heteroatomic 4-10 member of nitrogen, oxygen or sulphur, part insatiable hunger or ring or aromatic rings is independently selected from 0-4, in which:

Two R bases on same nitrogen-atoms are optionally combined together with the nitrogen-atoms, to form saturated rings, part unsaturated ring or the aromatic rings with the 1-4 heteroatomic 3-8 members for being independently selected from nitrogen, oxygen or sulphur.

In certain embodiments, the present invention provides the compound of formula A, has spatial chemistry shown in formula A ':

In certain embodiments, the present invention provides the compound of formula A ', and wherein A is C (O) and the compound has -1 structural formula of A ':

Such as institute's general definition above, the A base of formula A and A ' are C_1-6Aliphatic group, wherein A is optionally by Q¹Replace.In certain embodiments, the A base of formula A and A ' are C_2-5Aliphatic group, wherein A is by one or more Q¹Group replaces.

Such as institute's general definition above, each Q of formula A and A '¹Base is independently selected from cyano, halogen, azido, oxo, OR, SR, SO₂R、OSO₂R、N(R)₂、NR(CO)R、NR(CO)(CO)R、NR(CO)N(R)₂、NR(CO)OR、(CO)OR、O(CO)R、(CO)N(R)₂、O(CO)N(R)₂Or OPG¹, wherein PG¹For suitable hydroxyl protection base.In certain embodiments, each Q of formula A and A '¹Base is independently selected from cyano, halogen, azido, oxo, N (R)₂、OR、SR、SO₂R or OSO₂R.In other embodiment, each Q of formula A and A '¹Base is independently selected from cyano, halogen, azido, oxo, OR, SR, SO₂R、OSO₂R、N(R)₂, NR (CO) OR, NR (CO) R and O (CO) N (R)₂.In other embodiments, exemplary Q¹Base includes NH (CO) (CO)-(heterocycle or heteroaryl), OSO₂(aryl or substituted aryl), O (CO) NH- (aryl or substituted aryl), aminoalkyl, hydroxy alkyl, NH (CO) (CO)-(aryl or substituted aryl), NH (CO)-(alkyl) (heteroaryl or heterocycle), O (substituted or unsubstituted alkyl) (substituted or unsubstituted aryl) and NH (CO) (alkyl) (aryl or substituted aryl).

In certain embodiments, the A base of formula A and A ' have one of following characteristics:

(1) there is A at least one to be selected from the substituent group of hydroxyl, amino, azido, halogen and oxo；

(2) A is C_1-6Alkyl is selected from the substituent group of hydroxyl, amino and azido at least one；

(3) A has at least two substituent groups for being independently selected from hydroxyl, amino and azido；

(4) A has at least two substituent groups for being independently selected from hydroxyl and amino；

(5) A has at least one hydroxyl substituent and at least one amino-substituent；

(6) A has at least two hydroxyl substituents；

(7) A is the C replaced_2-4Aliphatic group；

(8) A is the C replaced₃Aliphatic group；

(9) A has (S)-hydroxyl or (R)-hydroxyl on the position α relative to the carbon atom that A is connected to the ring containing G；And

(10) A is C_1-6Radical of saturated aliphatic base is selected from the substituent group of hydroxyl and cyano at least one.

What term " (S)-hydroxyl " referred to the carbon atom with hydroxyl is configured as (S).Embodiments of the present invention further include compound, and wherein A is at least substituted primary on each carbon atom relative to (1) α and γ of the carbon atom being connected with A, (2) β and γ or (3) α and β.Each α, β and gamma carbon independently have (R) or (S) configuration.In certain embodiments, the present invention provides the compound, and wherein A is at least substituted primary on each carbon atom relative to α and β of the carbon atom being connected with A.

The exemplary A base of formula A and A ' include 2,3- dihydroxypropyl, 2- ethoxy, 3- hydroxyl -4- perfluoro butyl, 2,4, tri- hydroxyl amyl of 5-, 3- amino -2- hydroxypropyl, 1,2- dihydroxy ethyl, 2,3- dihydroxy -4- perfluoro butyl, 3- cyano -2- hydroxypropyl, 2- amino-1-hydroxyethyl, 3- azido -2- hydroxypropyl, 3,3- bis- fluoro- 2,4- dihydroxy butyl, 2,4- dihydroxy butyl, 2- hydroxyl -2- p-fluorophenyl-ethyl,-CH₂(CO) (substituted or unsubstituted aryl) ,-CH₂(CO) (alkyl replaces alkyl, such as halogenated alkyl or hydroxyalkyl) and 3, the bis- amyl- 4- alkenyl of fluoro- 2- hydroxyl of 3-.

In certain embodiments, the A base of one of A and A ' are 3- amino -2- hydroxypropyl.

According on one side, the present invention provides the compound of one of formula A and A ' a kind of, wherein Q¹For OPG¹, wherein PG¹For suitable hydroxyl protection base.Suitable hydroxyl protection base is known in the art; include " blocking group in organic synthesis " (T.W.Greene and P.G.M.Wuts, the 3rd edition, John Wiley & Sons; 1999, be incorporated herein by reference) in group those of is described in detail.In certain embodiments, the PG of one of formula A and A '¹Part is selected from ester, ether, silyl ether, alkyl ether, aryl alkyl ethers and alkoxyalkyl ether together with oxygen atom in combination.The example of this ester includes formic acid esters, acetic acid esters, carbonic ester and sulphonic acid ester.Specific example includes formic acid esters, benzoyl formiate, chloracetate, trifluoro-acetate, methoxyacetic acid ester, triphenylmethoxy acetic acid esters, parachlorophen-oxyacetic acid ester, 3- phenylpropionic acid ester, 4-oxopentanoic acid ester, 4, 4- (ethylene is thio) valerate, pivalate (pivaloyl group), crotonates, 4- methoxyl group crotonates, benzoic ether, to benzyl benzoate, 2, 4, 6- trimethylbenzoic acid ester, or carbonic ester, such as methyl, 9- fluorenyl methyl, ethyl, 2, 2, 2- trichloroethyl, 2- (trimethyl silyl) ethyl, 2- (benzene sulfonyl) ethyl, vinyl, allyl and to nitrobenzyl carbonic ester.The example of this silyl ether includes trimethyl silyl ether, triethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, triisopropylsilyl ether and other trialkylsilyl ethers.Alkyl ether includes methyl ether, benzylic ether, to methoxy-benzyl ether, 3,4- dimethoxy-benzyl ether, trityl ether, tertbutyl ether, allyl ether and allyloxycarbonyl ether or derivative.Alkoxyalkyl ether includes acetal, such as methoxy ether, methylthiomethyl ether, (2- methoxy ethoxy) methyl ether, benzyloxymethyl ether, β-(trimethyl silyl) ethoxyl methyl ether and THP trtrahydropyranyl ether.The example of aryl alkyl ethers includes benzylic ether, to methoxy-benzyl ether (MPM), 3,4- dimethoxy-benzyl ether, O- nitrobenzyl ether, to nitrobenzyl ether, to halogeno-benzyl ether, 2,6- dichloro benzyl ether, to cyanobenzyls ether, 2- and 4- picolyl ether.

In certain embodiments, the PG of one of formula A and A '¹Part is silyl ether or aryl alkyl ethers together with oxygen atom in combination.In other embodiments, the PG of one of formula A and A '¹Part is t-butyldimethylsilyi or benzoyl.In other embodiment, the PG of one of formula A and A '¹Part is t-butyldimethylsilyl (" TBS ").

Two Q such as institute's general definition above, on A¹Optionally in combination with together to form saturated rings, part unsaturated ring or the aromatic rings with the 0-4 heteroatomic 3-8 members for being independently selected from nitrogen, oxygen or sulphur.Two Q in certain embodiments, on A¹Epoxidation ring is collectively formed.

In certain embodiments, the PG of formula A and A '⁵And PG⁶Group is independently selected from above to the PG of formula A and A '¹Those suitable protecting groups described in group.In other embodiment, the PG of formula A and A '⁵And PG⁶Group is combined together to form ring glycerol protection base.This kind of glycerol protection base is known in the art, including GreeneThose described groups, and including cyclohexylidene and benzal glycerol protection base.

In certain embodiments, the present invention provides a kind of method of the compound of preparation formula F-2 according to following scheme V-a, V-b and V-c.

Plan V-a

In other embodiment, the present invention provides ER-817664 crystallization.

Plan V-b

Plan V-c

Using ER-817664 as crystallization of intermediate, plan V I-a indicates a kind of universal method that the intermediate of the compound of formula F-2 is used to prepare with compound preparation.

Plan V I-a

As shown in following scheme VI-b, in the method that another preparation is used to prepare the intermediate of the compound of formula F-2, three alcohol intermediates described in above scheme VI-a are used.

Plan V I-b

In plan V I-b as shown above, three alcohol intermediates are handled to generate aldehyde with periodate.Make the compound and methyl Wittig reagent homologization (homologated), by resulting olefin reduction at ester type compound.Remaining free hydroxyl group is handled to generate iodo intermediate with N- N-iodosuccinimide, and above-mentioned hydroxy compounds is generated with Ester Reduction with Sodium Borohydride.It will be appreciated by those of ordinary skill in the art that resulting iodo compound is consistent with compound 21 described in above scheme II, wherein compound 21 has protecting group in hydroxy position.Finally handle to obtain above-mentioned lactone by zinc.It will be appreciated by those of ordinary skill in the art that resulting lactone compound is consistent with compound 22 described in above scheme II, wherein compound 22 has protecting group in hydroxy position.

As shown in following scheme VII, in another preparation method of the intermediate for the compound from D- chinic acid preparation formula F-2, another route for originating in intermediate 2 described in scheme II is provided.

Plan V II

In above scheme VII, the ER-812829 of three-dimensional selection form is prepared with intermediate 2 (deriving from scheme II).Preferably, other blocking groups can be used for protecting glycol ER-812829.This kind of group is including cyclohexylidene and benzal glycerol protection base known to those of ordinary skill in the art.Firstly, in step (a), with the hydroxyl of 2- bromochloro-acetic acid processing ER-811510 to generate ER-812771.With the substantially similar method of method (Tettrahedron Letters, 40, (1991) 3455-3456) with the descriptions such as Murphy, which is handled with triphenyl phasphine, Wittig reagent is generated with (in situ) on the spot.Then the Wittig reagent forms lactone ER-812772.In step (b), three-dimensional selection is carried out to double bond plus hydrogen obtains ER-812829.

As shown in following scheme VII-a, the present invention also provides a kind of preparation method of intermediate for the compound from D- chinic acid preparation formula F-2, this method originates in intermediate E R-812829 shown in above scheme VII.

Plan V II-a

3, segment F-3

According to another embodiment, the present invention provides compound F-3:

Wherein, each PG⁴It is independently selected from suitable hydroxyl protection base；

R³For CHO or C (O) OR⁴；

R⁴For suitable carboxyl-protecting group；And

LG²For suitable leaving group.

Suitable carboxylate protecting group be it is known in the art, be described in detail in " blocking group in organic synthesis " (T.W.Greene and P.G.M.Wuts, the 3rd edition, John Wiley & Sons, 1999).In certain embodiments, the R of F-3⁴Base is the C optionally replaced_1-6Aliphatic group or the aryl optionally replaced.Suitable R⁴The example of base includes methyl, ethyl, propyl, isopropyl, butyl, isobutyl group, benzyl and phenyl, wherein each group is optionally substituted.

As described above, suitable leaving group is known in the art, for example, see " Advanced Organic Chemistry (Advanced organic Chemistry) (Jerry March; the 4th edition; pp.351-357, John Wiley and Sons, N.Y. (1992)).Such leaving group includes but is not limited to halogen, alkoxy, sulfonyloxy, the alkylsulfonyloxy optionally replaced, the alkenyl sulfonyloxy optionally replaced, the aryl-sulfonyl oxygen optionally replaced, silicyl and diazo (diazonium moieties).The example of suitable leaving group includes chlorine, iodine, bromine, fluorine, mesyl (mesyl), tosyl, trifluoromethanesulfonic acid root (triflate), nitrobenzenesulfonyl (nosyl) and bromobenzenesulfonyl (brosyl).In certain embodiments, the LG of F-3²Part is iodine.

According to another embodiment, suitable leaving group can in reaction medium in-situ generation.For example, the LG in the compound of formula F-3²It can be easy wherein the precursor contains by LG by the precursor in-situ generation of the compound of formula F-3²The group replaced on the spot.In a this substituted specific example, the precursor of the compound of the formula F-3 contains by LG such as such as iodine²The group (for example, trimethyl silyl) replaced on the spot.The source of iodine can be N- N-iodosuccinimide etc..This kind of in-situ generation of suitable leaving groups is well-known in the art, for example, see Id.

As described above, suitable hydroxyl protection base is known in the art, includes " blocking group in organic synthesis " (T.W.Greene and P.G.M.Wuts; 3rd edition; John Wiley& Sons, 1999, is incorporated herein by reference) in group those of is described in detail.In certain embodiments, each PG⁴Ester, ether, silyl ether, alkyl ether, aryl alkyl ethers and alkoxyalkyl ether are independently selected from together with oxygen atom in combination.The example of this kind of ester includes formic acid esters, acetic acid esters, carbonic ester and sulphonic acid ester.Specific example includes formic acid esters, benzoyl formiate, chloracetate, trifluoro-acetate, methoxyacetic acid ester, triphenylmethoxy acetic acid esters, p-chlorophenoxyacetic acid ester, 3- phenylpropionic acid ester, 4-oxopentanoic acid ester, 4, 4- (ethylene is thio) valerate, pivalate (pivaloyl group), crotonates, 4- methoxyl group crotonates, benzoic ether, to benzyl benzoate, 2, 4, 6- trimethylbenzoic acid ester, or carbonic ester, such as methyl, 9- fluorenyl methyl, ethyl, 2, 2, 2- trichloroethyl, 2- (trimethyl silyl) ethyl, 2- (benzene sulfonyl) ethyl, vinyl, allyl and to nitrobenzyl carbonic ester.Such silyl ether includes trimethyl silyl ether, triethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, triisopropylsilyl ether and other trialkylsilyl ethers.Alkyl ether includes methyl ether, benzylic ether, to methoxy-benzyl ether, 3,4- dimethoxy-benzyl ether, trityl ether, tertbutyl ether, allyl ether and allyloxycarbonyl ether or derivative.Alkoxyalkyl ether includes acetal, such as methoxy ether, methylthiomethyl ether, (2- methoxy ethoxy) methyl ether, benzyloxymethyl ether, β-(trimethyl silyl) ethoxyl methyl ether and THP trtrahydropyranyl ether.The example of aryl alkyl ethers includes benzylic ether, to methoxy-benzyl ether (MPM), 3,4- dimethoxy-benzyl ether, O- nitrobenzyl ether, to nitrobenzyl ether, to halogeno-benzyl ether, 2,6- dichloro benzyl ether, to cyanobenzyls ether, 2- and 4- picolyl ether.

In certain embodiments, the PG of F-3⁴One, two or three in part is silyl ether or aryl alkyl ethers together with the oxygen atom in conjunction with them.In other embodiment, the PG of F-3⁴One, two or three in part is t-butyldimethylsilyl or benzyl.All three PG in other embodiment, in F-3⁴Part is t-butyldimethylsilyl.

According to another embodiment, the compound of formula F-3 is provided, wherein the compound has spatial chemistry shown in formula F-3 ':

In certain embodiments, compound F-3a is provided:

Wherein " TBS " refers to t-butyldimethylsilyl.

The detailed synthesis of F-3a is illustrated in the following embodiments.

4, group is carried out to F-1, F-2 and F-3 and is filled with prepare compound I

Generally, as shown in following scheme VIII, segment F-1 and F-2 are coupled.Plan V III

Above scheme VIII indicates a kind of universal method that intermediate F-5a is prepared by segment F-1 and F-2.Firstly, being coupled segment F-1 and F-2 to obtain intermediate F-4 with the substantially similar method of method (Org Lett, 4:25, p 4431 (2002)) with the descriptions such as Kishi.The coupling carries out in the presence of chiral azoles (ER-807363), or carries out under the conditions of ER-807363 is not present.But the coupling reaction of the F-1 and F-2 carried out in the presence of ER-807363 has higher selectivity.F-4 is handled with potassium hexamethyldisilazide (potassiumhexamethyldisilazide), forms the intramolecular Williamson ether (williamsonether) of F-4, then the oxinane F-5 of stereoisomer mixture form is provided.Then the stereoisomer is separated, obtains F-5a.The detailed content of these three steps is described in the following embodiments.

According to another embodiment, the present invention provides compound F-4:

Wherein PG¹、PG²、PG³、LG¹And R¹Subclass definition with general definition and above and herein.

In certain embodiments, the present invention provides the compound of formula F-4, wherein the compound has spatial chemistry shown in formula F-4 ':

The present invention also provides compound F-4a:

Wherein " MsO " nail sulfonate radical, " TBS " refer to t-butyldimethylsilyl, and " OPv " refers to neopentanoic acid root.

The detailed synthesis of F-4a is described in the following embodiments.

According to another embodiment, the present invention provides compound F-5.

Wherein each PG¹、PG²、PG³、LG¹And R¹Subclass definition with general definition and above and herein.

In certain embodiments, the present invention provides the compound of formula F-5, has spatial chemistry shown in formula F-5 ' or F-5a.

As shown in following scheme IX, the PG of intermediate F-5a is removed³Group, then by gained hydroxy compounds F-6 and compound F-3a (wherein R³It is coupled for CHO), to generate F-7.

Scheme IX

Above scheme IX indicates a kind of universal method that intermediate F-9 is prepared by F-3a and F-6.First sulfone intermediate F-6 is handled with aldehyde F-3a with n-BuLi and then.Then SmI is then used to generate ketone-aldehyde intermediate F-8 with the resulting diol intermediates F-7 of Dess-Martin reagent oxidation₂F-8 is handled, intermediate F-9 is obtained.The detailed content of these steps is described in the following embodiments.

Scheme X

Above scheme X describes one kind by F-9a (LG²For iodine) universal methods of preparation halichondrin b analogs of the present invention.Firstly, carrying out intramolecular coupling under the conditions of with condition is substantially similar those of described in above scheme V to generate hydroxy compounds F-10.In another method, intramolecular coupling is carried out in the presence of chirality azoles ligand as described herein.Chirality azoles ligand is added and improves reaction yield, increases reaction efficiency.The detailed content of the reaction is described in the following embodiments.Then compound F-10 is oxidized to F-11.F-12 is obtained with the hydroxyl protection base of method appropriate removing F-11.It will be appreciated by those of ordinary skill in the art that the proper method of the protecting group of removing compound F-11 depends on the protecting group of actual use, including GreeneThose of described group.For example, can realize this removing by the tetrabutyl ammonium fluoride processing optionally buffered when each hydroxyl protection base of F-11 is TBS base.The detailed content of these steps is described in the following embodiments.

Intermediate F-12 is used to prepare various halichondrin b analogs described in such as United States Patent (USP) 6,365,759 and United States Patent (USP) 6,469,182 (being incorporated herein in its full text by quoting).

Embodiment

By taking the preparation of halichondrin b analogs B-1939 as an example, following examples set forth with the method for the present invention and compound synthesis halichondrin b analogs.

Those skilled in the art will be recognized, by the method for the invention, a variety of halichondrin b analogs as prepared by the compounds of this invention include but is not limited to United States Patent (USP) 6,214,865 and 6, halichondrin b analogs those of described in 365,759 (being incorporated by reference in its entirety).It is to be understood, therefore, that the synthetic method described in the form of embodiment below is not intended to limit the scope of the invention that appended claims define.

Embodiment 1

The preparation of F-1a:

In appropriately sized container, mix D- glucuronic acid -6,3- lactone (1 part of weight, 1 equivalent) with acetonitrile (3 parts of volumes) and acetone (9 parts of volumes).Concentrated sulfuric acid catalyst is added, system is kept to flow back 3 hours.The dissolution situation of D- glucuronic acid -6,3- lactone in inspection system.Cooling reaction solution stirs 15 hours to 25 DEG C.It is added solid sodium bicarbonate (0.5 part of weight), is further stirred for reaction solution 3 hours.Filtering removal solid, partial concentration organic matter, and with other acetonitrile (2 parts of weight) azeotropic.ER-806045 is used to react in next step without separation.

At -20 DEG C, crude product ER-806045 (1 part of weight, 1 equivalent) is dissolved in acetonitrile (6.5 parts of volumes).Under the conditions of temperature is lower than 5 DEG C in keeping, pyridine (1.5 parts of volumes, 4.0 equivalents) are added and are slowly added to SO₂Cl₂(0.38 part of volume, 1.02 equivalents).Temperature is lower than under the conditions of 10 DEG C in keeping, and acetonitrile (0.5 part of volume) is added in cold water (28 parts of volumes) under rinsing by reaction solution is counter, is quenched.Pass through heptane (2 parts of volumes) washing and filtering and dry isolated white solid ER-806410 (0.87 part of weight, 79% theoretical value).

ER-806410 (1 part of weight, 1 equivalent) and THF (10 parts of volumes) are added in appropriately sized container, is subsequently cooled to 10 DEG C.It is added wet palladium carbon (5%, 0.5 part of weight), stirs multi-phase solution 10 minutes.Reaction solution is buffered through pyridine (0.44 part of weight, 1.3 equivalents), is placed 3 hours under a hydrogen atmosphere.Filtering reacting liquid rinses solid with water (2 parts of volumes) and ethyl acetate (10 parts of volumes).Acquired solution is acidified through 1N hydrochloric acid (2.1 parts of volumes), is sufficiently mixed, separating obtained each layer.Successively organic layer is washed with sodium bicarbonate aqueous solution (5 parts of volumes) and water (5 parts of volumes).Organic matter is concentrated under reduced pressure, products therefrom is recrystallized with IPA (3.4 parts of volumes), and heptane (3.4 parts of volumes) is added at 15 DEG C and further harvests product, isolated white solid ER-806047 (yield 67%).

ER-806047 (1 part of weight, 1 equivalent) and toluene (8 parts of volumes) are added in appropriately sized container, is subsequently cooled to -40 DEG C.Under the conditions of temperature is lower than -35 DEG C in keeping, the 17 weight % toluene solutions (4.6 parts of weight, 1.1 equivalents) of DIBAL are added.After being measured to reaction solution, temperature is kept to be lower than under the conditions of 10 DEG C, be added acetone (0.15 part of weight, 0.5 equivalent), quenches excessive reagent.Lower than at 10 DEG C, reaction solution ethyl acetate (7 parts of volumes) and 15% aqueous citric acid solution (8 parts of weight) dilution, stirring is until obtain clear solution at 20 DEG C.Each layer is separated, aqueous layer with ethyl acetate (2 × 10 parts of volumes) is stripped twice.Combined organic matter is dry through magnesium sulfate (0.2 part of weight) after successively being washed with sodium bicarbonate aqueous solution (5 parts of volumes) and salt water (5 parts of volumes).After filtering, subtract pressure part concentration of organic layers, and form azeotropic mixture with toluene (4 parts of volumes).Product is saved in the form of THF solution, for reacting in next step.

TMSCH is added in appropriately sized container₂The ethereal solution (20 weight %, 2.04 parts of weight, 3.0 equivalents) of MgCl, it is cooling at 5 DEG C or less.Under the conditions of temperature is lower than 15 DEG C in keeping, THF (the 7 parts of volumes) solution of ER-806048 (1 part of weight, 1 equivalent) is added into reaction vessel.So that reaction solution is warming up to 35 DEG C, is kept for 1.5 hours.Cooling reaction solution is lower than at 20 DEG C and is diluted with toluene (7 parts of volumes), quenched with acetic acid (3 parts of volumes).Reaction solution further uses 10% aqueous ammonium chloride solution (6 parts of weight) to dilute, and is sufficiently mixed, separates each layer.Organic layer is successively washed with sodium bicarbonate aqueous solution (5 parts of volumes) and salt water (5 parts of volumes).It is dried, filtered through magnesium sulfate (0.2 part of weight), then decompression concentrated solution, the toluene concentrated solution (yield 90%) of isolated ER-807114.

ER-807114 (1 part of weight, 1 equivalent) and THF (20 parts of volumes) are sequentially added in appropriately sized container, and solution is cooled to 5 DEG C or less.The 15 weight % toluene solutions (9.16 parts of weight, 2.0 equivalents) of KHMDS are added.It is quenched and is reacted with 10% aqueous ammonium chloride solution (5 parts of volumes).Each layer is separated, organic layer is successively washed with ammonium chloride (5 parts of volumes), 2N hydrochloric acid (8.5 parts of volumes), sodium bicarbonate aqueous solution (5 parts of volumes) and water (5 parts of volumes).Organic matter is transferred in inspissator with ethyl acetate, is concentrated into viscous oil (yield 90%).At 35 DEG C, the substance is recrystallized with toluene (4 parts of volumes) and heptane (4 parts of volumes), (15 DEG C and 10 DEG C) at lower temperatures further harvest product (yield 94%) with heptane (2 × 4 parts of volumes).

KOtBu (0.67 part of weight, 1.2 equivalents) and THF (7.7 parts of volumes) are added in appropriately sized container, being cooled to interior temperature is -20 DEG C.Under the conditions of temperature is lower than -7 DEG C in keeping, THF (the 2.3 parts of volumes) solution of ER-806049 (1 part of weight, 1 equivalent) is added.Maximum temperature is maintained under conditions of -7 DEG C, and pure BnBr is added.It is stirred to react liquid at -20 DEG C 2 hours, is stirred 10 hours at 10 DEG C.Reaction solution is quenched with 10% aqueous ammonium chloride solution (4 parts of volumes), is diluted, is sufficiently mixed with toluene (4 parts of volumes).Each layer is separated, organic layer is washed with 10% salt water (4 parts of volumes), dry through magnesium sulfate (0.15 part of weight).The tBuOH solution (2.5 parts of volumes) (yield 95%) of isolated ER-806050 after reduced pressure.

K is sequentially added in appropriately sized container₃Fe (CN) 6 (3.5 parts of weight, 3.4 equivalents), K₂CO₃(1.5 parts of weight, 3.4 equivalents), (DHQ)₂AQN (0.0134 part of weight, 0.005 equivalent), water (18 parts of volumes), tBuOH (13 parts of volumes) and ER-806050 tBuOH solution (1 part of weight, 1 equivalent are dissolved in 5 parts of volumes).It is 0 DEG C that multiphase mixture, which is cooled to interior temperature, and K is added₂OsO₄·2H₂O (0.0029 part of weight, 0.22 mole of %).After 36 hours, at 0 DEG C, Na is used₂S₂O₃(3.5 equivalents, 1.7 parts of weight) quenching reaction, makes flask heating overnight to room temperature.After 15 hours, mixture is transferred in post-processing container (workupvessel), is diluted with toluene (15 parts of volumes) and water (4 parts of volumes).It is vigorously stirred and separates two-phase mixture.Organic layer is washed with salt water (10 parts of volumes), and concentrated and exchange of solvent obtains 10% toluene solution (yield 92%) of glycol ER-806051 and ER-806052 crude mixture.

It is further diluted with toluene solution (10.1 weight %s, 9.9 part weight) of the other toluene (3 parts of weight) to ER-806051/52 (1 part of weight, 1 equivalent).N-methylmorpholine (0.94 part of weight, 3.0 equivalents) and DMAP (0.075 part of weight, 0.2 equivalent) are added in toluene solution, gained mixture is cooled to 15 DEG C or less.Under the conditions of temperature is lower than 25 DEG C in keeping, chlorobenzoyl chloride is added.Then it is stirred to react at 75 DEG C liquid 12 hours.Cooling reaction solution keeps temperature to be lower than under the conditions of 25 DEG C, is quenched with 1N hydrochloric acid (5 parts of volumes) to 15 DEG C.It is sufficiently mixed and separates each layer.Organic layer is successively washed with salt water (3 parts of weight), sodium bicarbonate aqueous solution (3 parts of weight) and salt water (3 parts of weight).Dry organic layer (magnesium sulfate, 0.25 part of weight) is handled with activated carbon (0.1 part of weight), filters (Celite , 0.3 part of weight) through toluene (1 part of weight).Subtract pressure part enriched product, forms azeotropic mixture with toluene (3 parts of weight).The toluene solution (5 parts of volumes) (yield 95%) of isolated double benzoic ether ER-806053/54.

Under an inert atmosphere, by 20 weight %TiCl₄The toluene solution of (6.42 parts of weight, 3.6 equivalents) is cooled to 15 DEG C.Under the conditions of temperature is lower than 30 DEG C in keeping, it is added Ti (OiPr)₄(0.64 part of weight, 1.2 equivalents) stir acquired solution 15 minutes.By allyl TMS (1.03 parts of weight, 4.8 equivalents) ER-806053/54 (1 part of weight for obtaining with previous step, 1 equivalent) 22 weight % toluene solutions (4.55 parts of weight, 1 equivalent) premixing, be added to Ti (OiPr) Cl of fresh generation₃In.Temperature is lower than 30 DEG C in keeping during sample-adding.At 20-30 DEG C, it is stirred to react liquid 2 hours.Cooling reaction solution is quenched to -5 DEG C with 1N hydrochloric acid (6 parts of volumes), and interior temperature is during which kept to be lower than 30 DEG C.After being sufficiently stirred, each layer is separated, organic layer is successively washed with 1N hydrochloric acid (3 parts of volumes) and salt water (2 × 3 parts of volumes).Organic layer stirs together with magnesium sulfate (0.3 part of weight) and activated carbon (0.15 part of weight), under toluene (1 volume) rinses, is filtered through CeliteIt fills in (0.2 part of weight).The product of 3: 1 form of mixtures of isolated C-34, yield 83% after concentration.It is recrystallized through IPA/ normal heptane, obtains ER-806055 (C-34 > 99.5%d.e., yield 71%)

At room temperature, alcohol ER-806055 (1 part of weight, 1 equivalent), toluene (7 parts of volumes), DMSO (0.31 part of weight, 2.0 equivalents) and triethylamine (0.78 part of weight, 4.0 equivalents) are added in appropriately sized container.Acquired solution is cooled to -19 DEG C.Under the conditions of temperature is lower than -10 DEG C in keeping, it is added dropwise TCAA (0.84 part of weight, 1.4 equivalents).It is further stirred for reaction solution 10 minutes.Under the conditions of temperature is lower than 10 DEG C in keeping, reaction solution is diluted with IPA (0.5 part of volume), is quenched with 1N hydrochloric acid (5 parts of volumes).Each layer is separated, successively washs organic layer with sodium bicarbonate aqueous solution (5 parts of weight) and water (3 parts of volumes).Subtract pressure part concentration of organic layers (crude product yield 100%), further forms azeotropic mixture with other toluene (4 parts of volumes).Resulting ketone (ER-806058) is dissolved in the toluene of last 4 parts of volumes, water content is detected, for reacting in next step.

THF (2.7 parts of volumes) solution of ER-107446 (1 part of weight, 1.5 equivalents) is cooled to 10 DEG C, interior temperature is kept to be lower than under the conditions of 15 DEG C, is handled with the THF solution (5.2 parts of weight, 1.4 equivalents) of 25.2 weight %LHMDS.In second container, the toluene solution (21.9 weight %, 5.4 parts of volumes) of crude product ER-806058 is cooled to 10 DEG C.Under the conditions of temperature is lower than 20 DEG C in keeping, the content in first container is transferred in the solution containing matrix (substrate).It is stirred to react liquid 30 minutes, 1M hydrochloric acid (6.5 parts of volumes) is then added and is quenched, interior temperature is during which kept to be lower than 20 DEG C.Each layer is separated, organic layer is washed after washing four times with 1: 1 methanol/water (4 × 5 parts of volumes) with sodium bicarbonate aqueous solution (5 parts of volumes) and saline solution (2 × 5 parts of volumes).Product is dry through magnesium sulfate (0.52 part of weight), filters and (is rinsed with 0.7 part of volumes toluene), is concentrated under reduced pressure into heavy oil.

At room temperature, ER-806059 is dissolved in 1: 1 toluene/acetonitrile (5 parts of volumes).It keeps initial temperature to be lower than under the conditions of 40 DEG C, filtering TMSI (1.23 parts of weight, 4 equivalents) is added.Reaction solution is heated to 60 DEG C, is kept for 2 hours.Cooling reaction solution is lower than at 30 DEG C, is quenched with 25% ammonium hydroxide aqueous solution to -15 DEG C.It is stirred to react content to stay overnight, separates each layer.Toluene (5 parts of volumes) and water (2 parts of volumes) are separately added in organic layer.It is sufficiently mixed and separates each layer.Then organic layer successively is washed with 10% sodium sulfite aqueous solution (5 parts of volumes), 1N hydrochloric acid (5 parts of volumes), 5% sodium bicarbonate aqueous solution (5 parts of volumes) and salt water (5 parts of volumes).Organic layer is dry through magnesium sulfate (0.2 part of weight), filters, partial concentration, 50% toluene solution is obtained, for reacting in next step.

In the container for being suitble to size, NaBH (OAc) is mixed at 65 DEG C₃(1.19 parts of weight, 3.15 equivalents), Bu₄NCl (1.04 parts of weight, 2.1 equivalents), DME (8.2 parts of volumes) and toluene (4 parts of volumes), and stir at room temperature.75 DEG C are heated the mixture to, is kept for 1 hour.The toluene solution of 50 weight %ER-806060 (1 part of weight, 1 equivalent) is added at 75 DEG C, is separately rinsed with toluene (0.3 part of volume).Reaction temperature is increased to 85 DEG C, is stirred to react liquid 2-4 hours.Cooling reaction solution keeps interior temperature to be lower than under the conditions of 20 DEG C, is quenched with water (3.2 parts of volumes) to 10 DEG C of <.It is sufficiently mixed and separates each layer.Organic layer is successively washed with sodium bicarbonate aqueous solution (2 × 5 parts of volumes) and water (2 × 5 parts of volumes).Organic layer obtains the methanol solution of 40 weight %ER-806061 by concentration and exchange of solvent.

The methanol solution (40 weight %) of ER-806061 (1 part of weight, 1.0 equivalents) is dissolved in other methanol (1.6 parts of volumes).It is added potassium carbonate (0.24 part of weight, 1.0 equivalents), reaction temperature is increased to 50 DEG C, is kept for 1 hour.To 15 DEG C, interior temperature is lower than under the conditions of 30 DEG C cooling reaction solution, is quenched with 1N hydrochloric acid (3.5 parts of volumes, 2 equivalents).With water (3.9 parts of volumes) and toluene (3 parts of volumes) dilute reaction solution.Each layer is separated, water layer is stripped with toluene (1.5 parts of volumes).Sodium bicarbonate (0.3 part of weight) and sodium chloride (0.6 part of weight) are added in water phase, is stripped with nBuOH (3 parts of volumes).Merge three parts of organic phases, and be concentrated to dryness, obtains crude product triol ER-806064 and inorganic salts.At 80 DEG C, product is dissolved in 7: 1 toluene/nBuOH, through heat filtering, cooling recrystallization and is stirred overnight.The isolated ER-806064 (F-1b) (five step gross production rates 57%) after filtering and being rinsed with methanol.FAB(+)-MS M/z357(M+H).96.2 DEG C of fusing point.

At 25 DEG C, it is scattered in the triol ER-806064 (1 part of weight, 1 equivalent) of purifying in acetone (2 parts of volumes), with 2,2- dimethoxy propane (1 part of volume) dilution, is handled with the concentrated sulfuric acid (0.0086 part of weight, 0.03 equivalent).Liquid is stirred to react until uniformly.Reaction solution is diluted with toluene (5 parts of volumes), is added in 5% potassium carbonate (2 parts of volumes) and is quenched.It is sufficiently mixed and separates each layer.10% salt water washing of organic layer, it is dry through sodium sulphate (0.5 part of weight).Solution is filtered (toluene rinse) and the ER-806126 for being concentrated under reduced pressure to give yellow oily.The substance is used for next step.

10 DEG C or less are cooled to after solid NaOtBu (0.34 part of weight, 1.4 equivalents) is dissolved in THF (2.7 parts of volumes) and DMF (0.3 part of volume).Under the conditions of temperature is lower than 15 DEG C in keeping, under THF (0.5 part of volume) is rinsed, THF (2.5 parts of volumes) solution of ER-806126 (1 part of weight, 1 equivalent) is added in NaOtBu solution.After stirring 30 minutes, temperature is kept to be lower than under the conditions of 15 DEG C, is added methyl iodide (0.204 part of volume, 1.3 equivalents) (heat release).Reaction solution is set to be warming up to 25 DEG C, reaction solution is quenched with water (5 parts of volumes), is diluted with toluene (7 parts of volumes).It is sufficiently mixed and separates each layer.Organic layer is washed twice with salt water (2 × 5 parts of volumes), filtering dry through sodium sulphate (0.5 part of weight) and reduced pressure.

ER-806068 (1 part of weight, 1 equivalent) is dissolved in the methanol of 1 part of volume.Water (1.5 parts of volumes) and 2N hydrochloric acid (1.25 parts of volumes, 1 equivalent) is added, and is stirred to react liquid at 25 DEG C.At 10 DEG C, counter be added in 2M NaOH (1.34 parts of volumes) of reaction solution is quenched.Reaction solution is diluted with isopropyl acetate (5 parts of volumes).It is sufficiently mixed and separates each layer.The isopropyl acetate of 5 parts of volumes of water layer is stripped, and combined organic layer is dry through magnesium sulfate (0.5 part of weight), filters and be concentrated under reduced pressure to give crude product glycol ER-806063.

To 25 DEG C of crude product ER-806063 (1 part of weight, 1.0 equivalents) DMF (4 parts of volumes) solution in imidazoles (0.62 part of weight, 3.4 equivalents) are added, then lower than under 30 DEG C of interior temperature, it is added TBSCl (1.02 parts of weight, 2.53 equivalents).Liquid is stirred to react at 25 DEG C.Reaction solution is diluted with MTBE (10 parts of volumes), is washed with water (4 parts of volumes).Organic layer is successively washed with 1M hydrochloric acid (3 parts of volumes), water (3 parts of volumes), sodium bicarbonate aqueous solution (3 parts of volumes) and salt water (3 parts of volumes).Organic layer is dry through magnesium sulfate (0.5 part of weight), with the MTBE washing and filtering of 1 part of volume, is concentrated under reduced pressure, carries out exchange of solvent with heptane (4 parts of volumes).

ER-806065 (1 part of weight, 1 equivalent) is dissolved in heptane, isooctane or IPA (10 parts of volumes).Extremely -60 DEG C (± 10 DEG C) of cooling solution is below.It under low temperature, is bubbled in the solution through ozone, until the solution keeps blue.With nitrogen purged solution 15-30 minutes, while continuing to be purged with nitrogen, while reaction solution is made to be warming up to 5 DEG C.It is added 7-15 weight %Lindlar catalyst (5% palladium of lead poisoning/calcium carbonate, 0.1 part of weight).Reactor head (reactor head) is purged for several times with nitrogen, is evacuated, is placed under a nitrogen atmosphere (g).Then reaction solution is made to be warming up to room temperature (20-25 DEG C).It is stirred to react liquid 2.5 hours.Resulting multi-phase solution is filtered through Celite(1.0 parts of weight, using MTBE (2 parts of volumes) as flushing liquor).Concentrate solution is to dry, the crude product ER-806067 of isolated 1.0 parts of weight.Crude separation object is recrystallized through heptane or isooctane, obtains ER-806067 (F-1a), is white crystalline solid (five step yields 68%).FAB(+)-MS M/z601(M+H).64.5 DEG C of fusing point.

Embodiment 2

The preparation of F-2a

The Amberlyst15 (0.05 part of weight) and water (4.63 parts of volumes) of pre-flush are added in the reactor, being cooled to interior temperature is 0-5 DEG C.In keeping under the conditions of 5 DEG C of Wen Weiyue, 2,3-dihydrofuran (1 part of weight, 1 equivalent) is added in the reactor, stirs 1.5 hours.Water (4.63 parts of volumes) is added in a second reactor, being heated to interior temperature is 35 DEG C.Glass putty (2.2 parts of weight, 1.3 equivalents) are added in same reactor, are diluted respectively with 2,3- propylene bromide (3.71 parts of weight, 1.3 equivalents) and 48% hydrobromic acid (0.002 part of volume).After observing that reaction starts and (reaches 36-38 DEG C of instruction with temperature tracer value), keeps interior temperature to be lower than under the conditions of 45 DEG C, 2,3- propylene bromide (9 × 0.37 parts of weight) is added portionwise in a second reactor.After sample-adding, in 35 DEG C under temperature, it is further stirred for the content of second reactor 60 minutes.So that interior temperature is no more than 45 DEG C of speed, the filtering content of first reactor is added in second reactor.After sample-adding, heat source is removed, Celite is added in a second reactor545 (2.0 parts of weight), stirring gained mixture 30 minutes.Multiphase mixture is filtered through Celite545 pads (2.0 parts of weight), another exterior-applied liquid medicine (5 parts of volumes) wash filter cake.All filtrate is merged into a reactor, concentrated hydrochloric acid (1.5 parts of volumes) is added until cloudy solution becomes clarification.It is vigorously stirred down, sodium chloride (3.6 parts of weight) is added in the reactor, layering.It separates and shelves organic layer.With n-butanol (20 parts of volumes) aqueous layer extracted.Water layer is discharged, organic matter isolated for the first time is added in the reactor.Organic matter is washed with dense sodium bicarbonate (24 parts of volumes), is then stripped water layer with n-butanol (20 parts of volumes).Merge all organic matters, and is concentrated in vacuo.Concentrate is dissolved in MTBE (10 parts of volumes), is filtered, filtrate is concentrated into 2 parts of volumes.Under stirring, so that concentrate is cooled to interior temperature is 0 DEG C, and normal heptane (4 parts of volumes) then is added.It in 0 DEG C under temperature, stirs multiphase mixture 2 hours, is isolated by filtration to obtain purpose product, vacuum drying obtains the ER-806909 (1.34 parts of weight, 0.45 equivalent) of white powder.

Imidazoles (0.65 part of weight, 2 equivalents), ER-806909 (1 part of weight, 1 equivalent) and anhydrous DMF (4.04 parts of volumes) are added in the reactor.Under stirring, it is 0 DEG C that reactor, which is cooled to interior temperature, then so that tert-butyl diphenyl chlorosilane (1.25 parts of weight, 0.95 equivalent) is added in speed of the interior temperature no more than 15 DEG C.Under the conditions of temperature is 15 DEG C of < in keeping, it is further stirred for reaction solution 1 hour.Water (3.2 parts of volumes) and normal heptane (6.4 parts of volumes) are added in the reactor.Stirring mixture 5-15 minutes, layering.It separates and shelves organic layer.With normal heptane (3.2 parts of volumes) aqueous layer extracted.Merge all organic matters, washed with salt water (3.2 parts of volumes), be concentrated in vacuo to constant weight, obtains the ER-806545 (2.13 parts of weight, 0.95 equivalent) of yellow oily.The product is used for next step without being further purified.

With Simulation moving bed (SMB) chromatographic isolation enantiomter ER-806909, the ER-808373 (0.55 part of weight, 0.55 equivalent) and ER-806721 (0.45 part of weight, 0.45 equivalent) of yellow oily are obtained.SMB chromatography regulation for separating enantiomter ER-806909 is as follows:

Column and medium: 20 μm of 30mm of Chiracel OD column ×

150mm (12 columns)

Dicyandiamide solution: 96: 4 (volume/volume) heptane: the tert-butyl alcohol (mobile phase)

Simulated Moving Bed Chromatography instrument: Knauer SMB system CSEP C912

Thermoisopleth (Lang Gemiaoer is determined by frontal analysis)

Non-targeted isomers: Qi^*=2.8768 × Ci/ (1+0.02327*Ci)

Target isomer: Qi=4.5320 × Ci/ (1+0.0034595*Ci)

^*Wherein Qi=solid concentration (g/L), Ci=liquid concentration (g/L)

Column porosity: 0.658

Temperature: 27 DEG C

With EuroChrom 2000 for windows, SMB guide 1.2 editions(Berlin Wissenschaftliche Geratebau Dr.-Ing.Herbert Knauer GmbH, D-14163；Author H.Kniep and A.Seidel-Morgenstern) simulation calculating is carried out to flow velocity etc.:

Sample introduction (feed) concentration: 36g/L (ER-806909)

Sample introduction flow velocity (No. 1 pump) 15mL/min

Eluent flow rate (No. 2 pumps) 76.4mL/min

The area IV (No. 3 pumps) flow velocity 107mL/min

The area II (No. 4 pumps) flow velocity 134.3mL/min (actual flow velocity=

143.5mL/min)

The area I flow velocity 183.4mL/min

The area III flow velocity 149.3mL/min

Raffinate^*Flow velocity 42.3mL/min^*Raffinate=weak binding isomery

Body

Extract liquor flow velocity 49.1mL/min^*Extract liquor=combine isomery by force

Body

Intermittent time (tact time)

(port switching time) 0.8864 minute (53.18 seconds, the practical intermittent time

=54 seconds)

With the above regulation, enantiomter ER-806909 is separated in the following manner.11 hours are operated, it is 142mm that 10L mobile phase (ER-806909 containing 36g/L) pump (SilogmodelChemtech), which is sent through diameter, the nylon filter (Cole-Parmer # 2916-48) that aperture is 0.45 μm, into sample introduction tank (feed tank).36L mobile phase is filled in eluant groove, which has passed through 1 μm of glass fibre filter (Whatman GFC) that the diameter in pipeline is 45mm and filtered, 1 part of volume being separately added during operation.The interior temperature of SMB instrument is adjusted to 27 DEG C.

When initial starting up, injection port and eluent import are connected with eluant groove.Fluid injection and cleaning are carried out to sampling pump and elution liquid pump with mobile phase solvent.The column switching for starting SMB instrument, opens pump, and flow velocity is gradually increased at full speed, while keeping the absolute flow velocity between each pump poor.Once reaching at full speed, raffinate flow velocity and extract liquor flow velocity are measured, adjusts flow rate pump to be corrected to the deviation in pump technical specification.Sampling pump (No. 1 pump) is decelerated to 0mL/min, and import is connected with sample introduction tank again, injects sample introduction solution to the pump, then flow velocity is gradually adjusted back to the running speed to full speed.Raffinate and extract liquor outlet are connected into separating tank, obtain each sample within every 2 hours.With following HPLC methods, the chiral purity of sample is monitored by analysis HPLC.The flow velocity and the intermittent time of No. 2, No. 3 and No. 4 pumps are adjusted, to obtain required outlet purity.

At the end of operating, the flow velocity of sampling pump is reduced to 0 again, and is allowed to be connected with eluant groove.Make sampling pump restore at full speed, washing system 20 minutes.During washing, raffinate and extract liquor outlet are kept it turning on 10 minutes (10 intervals), after washing, and outlet is connected into separating tank.Gradually column liquid is washed in concentration, and is added in sample in subsequent operating.

Collected extract liquor (ER-806721) merges with the substance collected from same a collection of starting material at the end of operating every time, and the analysis HPLC method described in the following table 1 analyzes the chiral purity of last batch consolidated material again.Collected raffinate (ER-808373) is similarly operated.

The HPLC of 1 ER-806721 chiral purity of table is analyzed

Column: flow velocity: temperature (DEG C): sample volume: instrument: flowing phase composition ABCD	10 μm of 250 × 4.6mm of Chiracel OD column, DAICEL chemical industry Co., Ltd (DAICEL Chemical Industries, Ltd.) the usual 10 μ L of catalog number 140250.8mL/min27 (preferably) -35, sample is dissolved in solvent A, 5mg/mLWaters Alliance W2690, band UV W2487 (also having advanced laser polarimeter) (PDR-Chiral, Inc.) 99: 1 heptane: 2- propyl alcohol
						Gradient table:	(%)				Gradient
Time (min)	A	B	C	D		Gradient table:	(%)
Time (min)	A	B	C	D	0	100	0	0	0	It is isocratic
The duration of runs	30min					100	0	0	0	It is isocratic

Detection: the ultraviolet absorption value at 254nm

Triphenyl phasphine (0.7 part of weight, 1.2 equivalents), paranitrobenzoic acid (0.45 part of weight, 1.2 equivalents), ER-808373 (1 part of weight, 1 equivalent) and dry toluene (8 parts of volumes) are added in the reactor.Cooling reaction solution to interior temperature is 0 DEG C, so that interior temperature is no more than 7 DEG C of speed, is slowly added to DEAD (1.17 parts of weight, 1.2 equivalents).It is added normal heptane (3.3 parts of volumes), it is 10 DEG C that mixture, which is cooled to interior temperature, is then stirred 30-40 minutes.Filtering removes resulting sediment.Filter cake is washed with normal heptane (3.3 parts of volumes), TBME (0.55 part of volume), normal heptane (1.1 parts of volumes) and MTBE (0.55 part of volume) respectively.Merge all filtrate and is concentrated in vacuo.Thick concentrate is dissolved in THF (8 parts of volumes), water (0.8 part of volume) and two water lithium hydroxides (0.18 part of weight, 2 equivalents) are then added.Normal heptane (3.3 parts of volumes) is added after stirring mixture at room temperature, stirs 5 minutes.Water (2.2 parts of volumes) and normal heptane (3.3 parts of volumes) is added, stirs two-phase mixture 5 minutes, makes to be layered.Water layer is separated, is stripped when necessary with normal heptane.Merge organic layer and is concentrated in vacuo.Crude product is through SiO₂Column chromatography purifies to obtain the ER-806721 (0.74-0.85 parts of weight, 0.74-0.85 equivalent) of pale yellowish oil.

ER-806721 (1 part of weight, 1 equivalent) and anhydrous methylene chloride (4.2 parts of volumes) are added in the reactor.It is 0-5 DEG C that reaction solution, which is cooled to interior temperature, then triethylamine (0.34 part of weight is added, 1.5 equivalents), paratoluensulfonyl chloride (0.51 part of weight, 1.2 equivalents) and 4- (dimethylamino) pyridine (0.001 part of weight, 0.25 equivalent).Resulting mixture is stirred at room temperature 48 hours, water (1.8 parts of volumes) and methylene chloride (1.8 parts of volumes) is then added.After being sufficiently stirred, organic matter, and condensed organic are separated.Concentrate is dissolved in MTBE (1.8 parts of volumes), is washed with salt water (1.8 parts of volumes).It separates and shelves organic layer.Water layer is stripped with MTBE (1.8 parts of volumes), is then combined with all organic matters and is concentrated in vacuo.Under MTBE (7 parts of volumes) elution, crude product oil is filtered through SiO₂It fills in (70-230 mesh, 1 part of weight), is concentrated in vacuo filtrate.Concentrate is dissolved in IPA (5 parts of volumes), is added water (0.25 part of volume).It is 15 DEG C that gained mixture, which is cooled to interior temperature, and ER-807204 crystal seed is then added.After being put into crystal seed, it is 0 DEG C that mixture, which is cooled to interior temperature, is stirred 4-5 hours.Suspension is filtered, washs filter cake with cold IPA (1 part of volume), Vacuum dry filter cake to constant weight obtains the ER-807204 (1.05 parts of weight, 0.78 equivalent) of white powder.2597,1633,1363,1177,907,729.LRMS m/z 602 (M+H) of IR (film, cm-1) λ

The ethanol solution (2.97 parts of weight, 0.9 equivalent) of 21% sodium ethoxide is added in the reactor.Heating the solution to interior temperature is 65 DEG C, then so that diethyl malonate (3.24 parts of weight, 2 equivalents) are added in speed of the interior temperature no more than 70 DEG C.Stirring mixture 30 minutes, then through 3-5 hours addition ER-806906 (1 part of weight, 1 equivalent).After sample-adding, it is stirred to react liquid 60 minutes, being subsequently cooled to interior temperature is 50 DEG C.So that concentrated hydrochloric acid (0.84 weight, 1.05 equivalents) are added in speed of the interior temperature no more than 65 DEG C.Distilled water (0.25 part of volume, the 1.4 equivalents) solution of magnesium chloride hexahydrate (0.21 part of weight, 0.1 equivalent) is added after distillation removing DMF (3 parts of volumes) and ethyl alcohol.It is 135 DEG C that gained mixture, which is heated to interior temperature, while removing distillate.It is heated at reflux mixture, is then cooled to room temperature, salt water (12 parts of volumes) and TBME (16 parts of volumes) is added.Organic layer is separated, is concentrated in vacuo after being washed with water (1.3 parts of volumes) and salt water (1.2 parts of volumes).Product obtains ER-805552 (0.95-1.09 parts of weight, 0.71 equivalent) through distillation purifying.

The toluene solution (6.61 parts of weight, 1.04 equivalents) of 1.0M LHMDS is added in the reactor, being cooled to interior temperature is -75 DEG C.ER-805552 (1 part of weight, 1 equivalent) is dissolved in anhydrous THF, so that speed of the interior temperature no more than -70 DEG C is added in reactor.After sample-adding, anhydrous THF (2.5 parts of volumes) and methyl iodide (1.27 parts of weight, 1.25 equivalents) is added in stirring gained mixture 30 minutes in a second reactor, and being cooled to interior temperature is -75 DEG C.So that interior temperature is no more than -65 DEG C of speed, the THF solution of ER-805552 is added into methyl iodide solution.After sample-adding, it is stirred to react under temperature liquid 30 minutes in -78 DEG C.It is vigorously stirred down, by the inverse quenching (inverse quenched) of reaction solution into 1N hydrochloric acid solution (1 0 parts of volumes) and MTBE (8 parts of volumes).After sample-adding, separates and discard water layer.Organic layer is washed with saline solution (3 parts of volumes), is concentrated in vacuo.Product obtains about 6/1 non-enantiomer mixture of ER-806724 (0.75 part of weight) through distillation purifying.

Hydrochloric acid N, O- dimethyl hydroxylamine (1.05 parts of weight, 1.5 equivalents) and anhydrous CH are added in the reactor₂Cl₂(8.1 parts of volumes), being cooled to interior temperature is 0 DEG C.So that the toluene solution (3.93 parts of weight, 1.5 equivalents) of 2M trimethyl aluminium is added in speed of the interior temperature no more than 5 DEG C.Reaction solution is further stirred for 10 minutes, so that ER-806724 is added in speed of the interior temperature no more than 5 DEG C.Reaction solution CH₂Cl₂(15 parts of volumes) dilution, then so that speed of the interior temperature no more than 10 DEG C, which is added in the 1.3M sodium tartrate (20 parts of volumes) that interior temperature is 0 DEG C, carries out inverse quenching.After sample-adding, make to be layered, separates and shelve water layer.Organic matter is washed with water (1 part of volume), and, filtering dry through sodium sulphate (1 part of weight) is concentrated in vacuo to the methylene chloride for removing denier.Anhydrous DMF (6.3 parts of volumes), imidazoles (0.64 part of weight, 1.5 equivalents) and tert-butyl chloro-silicane (0.94 part of weight, 0.97 equivalent) are separately added into concentrate.Water (5 parts of volumes) and MTBE (10 parts of volumes) is added, stirring gained mixture makes to be layered.It separates and discards water layer.Organic layer is washed with water (5 parts of volumes), separates each layer.1N sodium hydroxide (2.5 parts of volumes) and methanol (2.5 parts of volumes), stirring gained mixture is added.Water layer is separated, vacuum concentration obtains the ER-806753 (1.94 parts of weight, 0.91 equivalent) of brown oil after organic layer is washed with salt water (2.5 parts of volumes).

ER-806753 (1 part of weight, 1 equivalent), CH are added in the reactor₂Cl₂50% aqueous solution (0.8 part of weight, 1.1 equivalents) of (5 parts of volumes) and NMO.Mixture is cooled to after interior temperature is 10 DEG C, 0.197M OsO is added₄Toluene (0.06 part of volume, 0.004 equivalent) solution.Sodium sulfite (0.1 part of weight, 0.25 equivalent) and water (0.85 part of volume) is added, is stirred to react liquid 1 hour.Mixture is diluted with salt water (0.85 part of volume), vacuum concentration organic matter to about 1/3 volume.Periodate sodium (1.3 parts of weight, 2 equivalents) and THF (2.5 parts of volumes) are successively added in a second reactor.Mixture is diluted with the phosphate buffer (3.0 parts of volumes) of pH=7, and being cooled to interior temperature is 20 DEG C.So that dense glycol is added no more than 30 DEG C of speed in interior temperature.After sample-adding, gained mixture is stirred at room temperature.Water (1.25 parts of volumes), MTBE (7 parts of volumes) and saline solution (1.25 parts of volumes) is added, separates each layer.The mixture secondary washing of organic matter saline solution (1 part of volume) and saturated sodium bicarbonate (1 part of volume).Finally, the mixture of organic matter and salt water (1 part of volume) and 10% (weight/volume) hypo solution (1 part of volume) stirs 1 hour, then it is concentrated in vacuo.Thick material is through SiO₂Column chromatography purifies to obtain 2953,2856,1725,1664,1463,1254,1092, the 833.LRMS m/z 332 (M+H) of ER-806754 (0.93 part of weight, 0.93 equivalent) IR (film, cm-1) λ of yellow oily.

ER-806629 (1.53 parts of weight, 3.1 equivalents) and THF (10.5 parts of volumes) are added in the reactor, sprays nitrogen 60 minutes, solution is made to deaerate.ER-807204 (1 part of weight, 1.0 equivalents), ER-806754 (0.66 part of weight, 1.2 equivalents) and THF (2.7 parts of volumes) are added in second inerted reactor, sprays argon gas 45 minutes, the solution is made to deaerate.CrCl is successively added in the reactor containing ER-806906₂(0.63 part of weight, 3.1 equivalents) and triethylamine (0.52 part of weight, 3.1 equivalents).In 30-35 DEG C under temperature, the bottle green suspension is stirred 1 hour, be cooled to 0-5 DEG C, NiCl is then added₂(0.1 equivalent).The content of second reactor was slowly added into first reactor through 0.5 hour, reaction solution is made to be warming up to room temperature.Cooling reaction solution to interior temperature is 0 DEG C, then through 30 minutes addition ethylenediamines (1.0 parts of weight, 10 equivalents), in 25 DEG C under temperature, is stirred to react liquid at least 30 minutes.Water (4 parts of volumes), TBME (10 parts of volumes) and normal heptane (1 part of volume) are added in reaction solution, stirring gained mixture 15 minutes makes each phase separate (about 30min).Water layer is separated, is stripped with TBME (about 7.5 parts of volumes).Merge organic layer, is washed with water (5 parts of volumes), salt water (3 parts of volumes), be concentrated in vacuo to minimum volume.IPA (10 parts of volumes) and SiO are added in crude mixture₂(1 part of weight), in 25 DEG C under temperature, stirring gained mixture at most 4 days.Suspension is filtered, filter cake is washed with IPA (2 × 1 parts of volumes).Normal heptane (6.6 parts of volumes) is added in filtrate, vacuum concentrated mixture is until form suspension.Mixture is filtered, filter cake is washed with normal heptane, then vacuum concentrated mixture.Crude product is through SiO₂Column chromatography purifies to obtain ER-807524 (0.54 part of weight, 0.48 equivalent) IR (film, cm-1) λ 2934 of glassy yellow (clear yellow) oily, 1668,1471,1108,833.LRMS m/z 704 (M+Na).

ER-807524 (1 part of weight, 1 equivalent) and anhydrous THF (1.25 parts of volumes) are added in the reactor.It is -20 DEG C that mixture, which is cooled to interior temperature, so that 3M methyl-magnesium-chloride (0.59 part of volume, 1.2 equivalents) are added in speed of the interior temperature no more than 0 DEG C.After sample-adding, so that mixture is warming up to interior temperature is 0 DEG C.Reaction mixture is added in half saturated ammonium chloride (2.62 parts of volumes) and carries out inverse quenching, is vigorously mixed down, dilutes gained mixture with TBME (2 parts of volumes).Water layer is discarded, organic matter is concentrated in vacuo after being washed with salt water (2 parts of volumes).Crude product is through SiO₂Column chromatography purifies to obtain the ER-807525 (0.79-0.82 parts of weight, 0.85-0.88 equivalent) of yellow oily.

ER-807525 (1 part of weight, 1 equivalent), the double trifluoro Methanesulfomides (0.59 part of weight, 1.1 equivalents) of N- phenyl and anhydrous THF (4.1 parts of volumes) are added in the reactor, it is -75 DEG C that mixture, which is cooled to interior temperature,.So that the toluene solution (2.75 parts of weight, 1 equivalent) of 0.5MKHMDS is added in speed of the interior temperature no more than -60 DEG C, reaction solution is then made to be warming up to -20 DEG C through 2 hours.So that speed of the interior temperature no more than 0 DEG C is quenched with half saturated ammonium chloride (2.4 parts of volumes) and is reacted.So that mixture is warming up to interior temperature is 20 DEG C, is added normal heptane (2.4 parts of volumes).Mixture is stirred, separates and discards water layer.Organic layer is washed three times with saturated sodium bicarbonate (2.3 parts of volumes every time), then obtains ER-807526 (1.2-1.4 parts of weight, 1.0-1.2 equivalent) through vacuum concentration.The substance without further purification, is used for next step.

At 20 DEG C, ER-807526 (1 part of weight, 1 equivalent) and anhydrous methanol (3 parts of volumes) are added in the reactor.The IPA solution (4 parts of volumes, 5 equivalents) of 1.25M hydrochloric acid is added, stirs mixture 3 hours.Solid sodium bicarbonate (0.42 part of weight, 5 equivalents) are added portionwise under stirring, until the pH of reaction mixture reaches 6-7.Under methanol (3 × 2 parts of volumes) washing, reaction mixture is filtered.It is concentrated in vacuo all filtrate, then through SiO₂Column chromatography purifies to obtain ER-807527 (0.43 part of weight, 0.79-0.85 equivalent).

Non-enantiomer mixture ER-807527 is separated with HPLC chromatogram is prepared, is concentrated target fraction (desired fraction), obtains the ER-806730 (0.56 part of weight, 0.56 equivalent) of glassy yellow oily.For separate ER-806730 to prepare HPLC chromatogram regulation as follows:

Column and medium: Kromasil spherical silica gel column, 60A, partial size 10

μm l, in the Varian of 7.7cm × 60cm

It fills in Dynamax Rampak column to 7.7cm

(diameter) × 30cm (length)

HPLC packing station: Varian (Rainin) Dynamax Rampak

41/77mm column packing station

HPLC pump: Varian (Rainin) SD-1 titanium pump head

Main (primary) HPLC detector: Waters R403 refractive index detector

Secondary (secondary) HPLC detector: Varian (Rainin) UV-1 detector (has

Prepare flow cell)

Chromatography control and acquisition software: Varian (Rainin) Dynamax DA 1.4.6 editions

Chromatographic data processing software: Varian (Rainin) Dynamax R4.3 editions

Mobile phase: 28.5: 63.7: 7.85 (volume/volume) normal heptanes: first

The tertiary butyl ether of base: 2- propyl alcohol

Flow velocity: 140mL/min

Column temperature: room temperature (25 DEG C)

Detection: refractive index, cathode (are decayed) at 16X, 215nm

Locate ultraviolet absorption value

Eluent gradient: isocratic

The duration of runs: 40 minutes

Sample volume: 10ml, the ER-807527 containing 0.8g/mL

With the above regulation, diastereoisomer ER-807527 is separated in the following manner.Each crowd of ER-807527 is diluted to 0.1g/ml with mobile phase first, and is filtered through the 47mm glass fibre filter (Whatman GFC) that aperture is 1 μm under vacuum.Then filtrate is concentrated with rotavapor under vacuum.Start to flow on SD-1 HPLC pump A (with mobile phase fluid injection and cleaning), flow velocity is gradually increased to 140mL/min.Washing is carried out to system until UV and RI detector reaches baseline stability.It is rinsed with reference flow cell of the fresh mobile phase to RI detector.

ER-807527 ought be criticized with mobile phase and is diluted to 0.8g/mL concentration, to obtain the chromatography of 8gER-807527 injection sample.The aliquot of 10mL solute is injected, corresponding ER-807527 peak value (about starting in 24 minutes summits, continue to 35 minutes) collects eluent.Continue subsequent note sample and fraction collector, until exhausting starting material.

The fraction of corresponding ER-806730 is merged, is concentrated in vacuo with Rotary Evaporators.The HPLC analysis method described in table 2 evaluates diastereomeric purity and area % purity area.

The HPLC of 2 ER-806730 diastereomeric purity of table is analyzed

Column: flow velocity: temperature (DEG C): sample volume: instrument: flowing phase composition A B C D	250 × 4.6mm of Kromasil silicagel column, 5 μm, MetaChem catalog number 0475-250 × 0461mL/min2710 μ L, the sample in solvent A, 2.5mg/mLWaters Alliance W2690, band W248730: 67: 3 normal heptane of UV: methyl tertiary butyl ether(MTBE): 2- propyl alcohol 2- propyl alcohol
						Gradient table: time (min)	(%)				Gradient
A	B	C	D				(%)
A	B	C	D	0 22 26 32	1001009090		0 0 10 10	0 0 0 0	0 0 0 0	It is isocratic linear isocratic
The duration of runs	32min, wherein under primary condition, rebalancing 18min						0 0 10 10	0 0 0 0	0 0 0 0	It is isocratic linear isocratic

Detection: ultraviolet absorption value at 205nm

ER-806730 (1 part of weight, 1 equivalent) and anhydrous methylene chloride (4.8 parts of volumes) are added in the reactor, being cooled to interior temperature is 0 DEG C.2,4,6- trimethylpyridine (1.16 parts of weight are added, 4 equivalents) and DMAP (0.03 part of weight, 0.1 equivalent), stirring gained mixture 15 minutes, then so that trimethyl-aceyl chloride (0.3 part of weight, 1.05 equivalents) are added in speed of the interior temperature no more than 10 DEG C.It is added water (3 parts of volumes), stirs mixture 15 minutes.It is added TBME (10 parts of volumes), is further stirred for mixture 10 minutes.Organic layer is washed with 1N hydrochloric acid (10 parts of volumes) until 2,4,6- trimethylpyridine is in elecrtonegativity, is then washed respectively with water (5 parts of volumes), saturated sodium bicarbonate (5 parts of volumes) and saturated brine (5 parts of volumes).It is concentrated in vacuo organic layer, concentrate is through SiO₂Column chromatography purifies to obtain the ER-806732 (1.02 parts of weight, 0.85 equivalent) of yellow oily.

ER-806732 (1 part of weight, 1 equivalent) and anhydrous THF (2.35 parts of volumes) are added in the reactor, being cooled to interior temperature is 0 DEG C.So that triethylamine (0.22 part of weight, 1.1 equivalents) and mesyl chloride (0.24 part of weight, 1.05 equivalents) is successively added in speed of the interior temperature no more than 10 DEG C.Liquid is stirred to react under temperature in 0 DEG C, lower addition normal heptane (3.4 parts of volumes) is then vigorously stirred, makes to be layered.Organic matter is washed with saturated brine (3.4 parts of volumes), filtering dry through saturation sodium sulphate (2 parts of weight), and filter cake is washed with normal heptane until ER-805973 (F-2a) is in elecrtonegativity.It is concentrated in vacuo filtrate, obtains ER-805973 (1.12 parts of weight, 0.97 equivalent).Crude product ER-805973 (F-2a) is used for next step without being further purified.2961,1725,1413,1208,926.LRMS m/z 579 (M+H) of IR (film, cm-1) λ.

Embodiment 3

Another preparation method of ER-806730

Chinic acid (1 part of weight), cyclohexanone (2.11 equivalents, 1.08 weight) and the concentrated sulfuric acid (0.011 equivalent, 0.0056 part of weight) are added in the reactor.Reaction mixture is heated to 160 DEG C, moisture (in 100 DEG C of beginning azeotropic) are removed by azeotropic distillation.Reaction solution is cooled to 90 DEG C -100 DEG C, sodium bicarbonate (0.0096 part of weight) and toluene (3.6 parts of weight) is added.It is cooled to room temperature reaction solution through 4-6 hours, gained sediment washs and be dried to obtain 1 (0.97 part of weight) of white powder through filtering, toluene (2 × 0.9 parts of weight).

Mixed compound 1 (1 equivalent, 1 part of weight) and imidazoles (2.5 equivalents, 0.80 part of weight), are purged with nitrogen, are allowed to be suspended in anhydrous THF (10 parts of volumes).TMSCl (1.2 equivalents, 0.61 part of weight) is added with the speed for keeping the temperature below 30 DEG C.Reaction solution is cooled to room temperature, is added heptane (10 parts of volumes), resulting suspension is filtered.Filter cake is washed with 1: 1 heptane/THF (10 parts of volumes), by air-distillation, is exchanged filtrate solvent with toluene, is obtained toluene (about 5 parts of weight) solution of 2a (1.34 parts of weight, calculated value).

DIBAL-H (1.5M toluene solution, 1.2 equivalents, 2.1 parts of weight) are added to -78 DEG C, with the speed for keeping the temperature below -65 DEG C in cooling 2a solution.Excessive DIBAL-H is quenched with methanol (0.3 equivalent, 0.034 part of weight), solution is made to be warming up to 0 DEG C.To keep the temperature below 25 DEG C of speed, solution is transferred in the solution of 30 weight % Rochelle salt (RochelleSalt) aqueous solutions (10 parts of weight) and sodium bicarbonate (1 part of weight).It is vigorously stirred mixture, obtains two phase liquid.Each layer is separated, with MTBE (5 parts of volumes) aqueous layer extracted.Combined organic layer is successively washed with water (2.5 parts of weight) and saturated brine (2.5 parts of weight).Condensed organic carries out exchange of solvent with TFH to obtain TFH (5 parts of volumes) solution of 2b (0.98 part of weight, calculated value).

Cooling 2b solution is added acetic acid (2.9 equivalents, 0.51 part of volume) to 5 DEG C.It is added water (1.0 equivalents, 0.055 part of volume), the agitating solution at 0 DEG C -5 DEG C.As needed, at most two equal part acetic acid and an equal part water are additionally incorporated to promote silicyl to be deprotected.Triethylamine (12 equivalents, 3.6 parts of weight) and DMAP (0.05 equivalent, 0.02 part of weight) is added with the speed for keeping the temperature below 20 DEG C.It is added acetic anhydride (6 equivalents, 2.0 parts of weight), is stirred to react liquid at room temperature.Reaction solution is cooled to 5 DEG C, saturated sodium bicarbonate aqueous solution (10 parts of volumes) is added with the speed for keeping the temperature below 30 DEG C.Stirring gained mixture 3-4 hours, separates each layer.With MTBE (5 parts of volumes) aqueous layer extracted.Combined organic matter is washed with water (5 parts of volumes).Exchange of solvent is carried out to extract liquor using IPA by distillation, obtains IPA (3 parts of volumes) solution of 2c.The solution is cooled to 5 DEG C, resulting crystallization is filtered.Concentrated mother liquor obtains aftercrop after recrystallization, obtains the 2c (0.87 part of weight) of white crystalline solid shape.

2c (1 part of weight) is dissolved in acetonitrile (6 parts of volumes), the amyl- obtusilic acid methyl esters of 3- trimethyl silyl (3.0 equivalents, 1.86 parts of weight) and TFAA (0.2 equivalent, 0.083 part of volume) is successively added.Then BF is added into solution with the speed for keeping the temperature below 25 DEG C₃OEt₂(1.0 equivalents, 0.42 part of volume).Saturated sodium bicarbonate aqueous solution (10 parts of volumes) is added in reaction solution, stirring gained mixture 15 minutes.Mixture successively uses heptane (10 parts of volumes) and MTBE (5 parts of volumes) to extract, and is concentrated to combined extract liquor, obtains orange oily 3 (0.72 part of weight, calculated values).

To keep the temperature below 25 DEG C of speed, THF (9 parts of volumes) solution of 3 (1 part of weight) is handled with sodium methoxide (25% w/w methanol solution (1.5 equivalents, 2.2 parts of weight)).1N hydrochloric acid (10 parts of volumes), which is added, quenches reaction.Organic layer is separated, water layer is extracted with MTBE (10 parts of volumes).Combined organic matter water (2.5 parts of volumes), saturated sodium bicarbonate (2.5 parts of volumes) and water (2.5 parts of volumes) wash.Concentrate solution obtains THF (2.5 parts of volumes) solution of 4 (0.88 part of weight, calculated values).The solution is directly used in next step.

Methanol (5 parts of volumes) is added in tetrahydrofuran (2.5 parts of volumes) solution of 4 (1 part of weight).It is added 1N hydrochloric acid (0.75 equivalent, 2 parts of volumes), reaction solution is made to be warming up to 60-80 DEG C.Saturated bicarbonate aqueous solution is added to room temperature in cooling reaction solution.Mixture is extracted with DCM (3 × 2.5 parts of volumes), and combined DCM extract liquor and ethyl acetate carry out exchange of solvent, obtains 5 ethyl acetate (3 parts of volumes) solution.Heptane (2 parts of volumes) is added to induce crystallization, gained suspension is cooled to 0 DEG C.Solid is collected by filtration, filter cake is washed with cold ethyl acetate/heptane (1: 1 volume ratio), is dried to obtain the 5 of white powder (0.55 part of weight).

Compound 5 (1 part of weight) is dissolved in acetonitrile (10 parts of volumes), then 2- acetate -2- methyl N-Propyl Bromide (2-acetoxy-2-methylpropanyl bromide) (4.0 equivalents are sequentially added, 2.2 parts of weight) and water (1 equivalent, 0.067 part of weight).Stirring gained mixture at room temperature, is subsequently cooled to 5-10 DEG C.It is added NaOMe (25% w/w methanol solution, 8 equivalents, 6.2 parts of weight), reaction solution is made to be warming up to room temperature.Saturated sodium bicarbonate (10 parts of volumes) is added and quenches reaction solution, is extracted with MTBE (2 × 10 parts of volumes).Exchange of solvent is carried out by air-distillation and methanol, obtains methanol (10 parts of volumes) solution of 6 (0.91 part of weight, calculated values).

Methanol (10 parts of volumes) solution of 6 (1 part of weight) is heated to 55 DEG C.Divide 6 addition sodium borohydrides (5 equivalents, 0.68 part of weight), cooling reaction solution is quenched to 5 DEG C with 1N hydrochloric acid (10 parts of volumes).It is added salt water (5 parts of volumes), extracts reaction solution with ethyl acetate (2 × 10 parts of volumes).Combining extraction liquid, it is concentrated to obtain 7, it is brown residue.

Compound 7 (1 part of weight) is dissolved in CH₂Cl₂In (10 parts of volumes), DMAP (0.1 equivalent, 0.054 part of weight), triethylamine (3.0 equivalents, 1.85 parts of volumes) and TBDPSCl (1.2 equivalents, 1.38 parts of volumes) are then added at room temperature.It is added sodium bicarbonate (10 parts of volumes), separates organic layer.Water layer uses CH again₂Cl₂(10 parts of volumes) extraction.Merge organic extract liquid, concentrated 8 (1.8 parts of weight, the calculated values) for obtaining colorless oil.

At room temperature, LDA (1.5M cyclohexane solution, 4 equivalents, 6 parts of volumes) are added in THF (10 volume) solution of 8 (1 part of weight).It is quenched after so that the solution is warming up to 50 DEG C with 1N hydrochloric acid (5 parts of volumes), is extracted with MTBE (10 parts of volumes).Concentrated extract obtains 9 (0.9 part of weight) of oily.

The methylene chloride (5 parts of volumes) of 9 (1 part of weight) and methanol (5 parts of volumes) solution are cooled to -60 DEG C, keeps temperature to be lower than under the conditions of -50 DEG C, uses O₃It is handled.Reaction solution is purged with nitrogen, NaBH is added₄(0.5 equivalent, 0.04 part of weight), makes mixture be warming up to 0 DEG C.In addition NaBH is added portionwise₄(1 equivalent, 0.08 part of weight), makes reaction solution be warming up to room temperature.After 3 hours, mixture is quenched with 1N hydrochloric acid (10 parts of volumes), is added methylene chloride (5 parts of volumes), makes to be layered.Water layer is extracted again with methylene chloride (10 parts of volumes), merges organic extract liquid, concentrated 10 (the 0.97 part of weight) for obtaining colorless oil.

Compound 10 is dissolved in THF (10 parts of volumes), is added phosphate buffer (pH=7,5 parts of volumes).NaIO is added₄(2 equivalents, 0.854 part of weight), makes reaction solution be warming up to room temperature.Water (5 parts of volumes) and MTBE (10 parts of volumes) is added, is vigorously stirred gained mixture 10 minutes.Separate organic layer, it is washed with 10% weight/volume sodium thiosulfate solution (5 parts of volumes), water (5 parts of volumes) and salt water (5 parts of volumes), then dry (about aqueous 200ppm) with THF azeotropic distillation, obtain THF (10 parts of volumes) solution of 11 (0.93 part of weight, calculated values).The solution is directly used in next step.

(methyl esters methylene) triphenyl phasphine (1 part of weight) is added in THF (10 parts of volumes) solution of 11 (1 part of weight), is heated to 65 DEG C.It is added heptane (40 parts of volumes), stirring gained mixture 30 minutes.Gained sediment is filtered, concentration filtrate to total volume is 10 parts of volumes.SiO is added₂(5 parts of weight), suspension is filtered through SiO₂Pad (is eluted) with MTBE (20-40 parts of volumes).Solvent is exchanged with methanol, methanol (10 parts of volumes) solution of 12 (0.95 part of weight, calculated values) is obtained, is directly used in next step.

10% w/w palladium carbon (0.23 equivalent, 0.37 part of weight) is added in methanol (10 parts of volumes) solution of 12 (1 part of weight), and uses hydrogen treat.Suspension is filtered, while rinsing filter cake with THF (10 parts of volumes).Solvent is exchanged with THF, THF (10 parts of volumes) solution of 13 (0.95 part of weight, calculated values) is obtained, is directly used in next step.

THF (10 parts of volumes) solution of 13 (1 part of weight) is cooled to 0-5 DEG C, to keep speed of the temperature lower than 10 DEG C that LAH (1M THF solution, 0.78 equivalent, 1.5 parts of volumes) are added.Then to keep speed of the temperature lower than 10 DEG C that water (1.7 equivalents, 0.06 part of volume) is added.To keep speed of the temperature lower than 10 DEG C that 10% weight of sodium hydroxide/aqueous solution (0.16 equivalent, 0.06 part of volume) and water (4.98 equivalents, 0.17 part of volume) is successively added, gained mixture is warming up to room temperature with vigorous stirring.Suspension is filtered, filter cake is rinsed with THF (5 parts of volumes).Partial concentration is carried out to filtrate, THF (10 parts of volumes) solution of 14 (0.9 part of weight, calculated values) is obtained, is directly used in next step.

THF (10 parts of volumes) solution of 14 (1 part of weight) is cooled to 0 DEG C, imidazoles and TrCl (1.5 equivalents, 0.59 part of weight) is then added.It is added saturated sodium bicarbonate aqueous solution (5 parts of volumes), extracts mixture with heptane (10 parts of volumes).Extract liquor is washed with salt water (10 parts of volumes), concentrated to obtain 15 (1.35 parts of weight).

Compound 15 (1 part of weight) is dissolved in THF (10 parts of volumes), is handled with TBAF (1M, 1.2 equivalents, 1.6 parts of volumes).Then heptane (5 parts of volumes) and SiO is added to 2 parts of volumes in concentrated reaction mixture₂(5 parts of volumes).Filtering gained suspension, is successively eluted with heptane (5 parts of volumes) and THF (10 parts of volumes).THF eluent is collected, THF (10 parts of volumes) solution of 16 (0.61 part of weight, calculated values) is obtained, is directly used in next step.

PPh is added in THF (10 parts of volumes) solution of 16 (1 part of weight)₃(5 equivalents, 2.3 parts of weight), pyridine (10 equivalents, 1 part of volume) and NIS (3.0 equivalents, 1.1 parts of weight).Then it is added 20% w/w aqueous citric acid solution (10 equivalents, 14 parts of weight), stirring gained mixture 10 minutes.Reaction solution is diluted with heptane (10 parts of volumes), separates water layer.Organic layer water (5 parts of volumes), 10% weight/volume sodium thiosulfate solution (5 parts of volumes), water (5 parts of volumes) and salt water (5 parts of volumes) wash.Solvent is exchanged with ethyl alcohol, is concentrated into 5 parts of volumes.It is added water (10 parts of volumes), collection is filtered to gained sediment, obtains 17 (0.65 part of weight) of white solid.

Compound 17 (1 part of weight) and potassium cyanide (6 equivalents, 0.54 part of volume) are suspended in ethyl alcohol (5 parts of volumes) and water (10 parts of volumes), gained suspension is heated to 80 DEG C.Reaction solution water (5 parts of volumes) and ethyl acetate (10 parts of volumes) dilution, mix 10 minutes.Water layer is removed, organic layer is washed with water (5 parts of volumes) and salt water (5 parts of volumes).Solvent is exchanged with ethyl alcohol, ethyl alcohol (10 parts of volumes) solution of 18 (0.75 part of weight) is obtained, is directly used in next step.

Zinc (37 equivalents, 3.9 parts of weight) are added in ethyl alcohol (10 parts of volumes) solution of 18 (1 part of weight), heat mixture to 75-80 DEG C.It by reaction solution partial concentration to 2-3 parts of volumes, is cooled to room temperature, is allowed to be layered with MTBE (10 parts of volumes) and water (5 parts of volumes).Water layer is removed, organic layer saturated sodium bicarbonate (5 parts of volumes), water (5 parts of volumes) and salt water (5 parts of volumes) wash.Then dry to aqueous about 200ppm through THF azeotropic distillation, obtain THF (10 parts of volumes) solution of 19 (0.81 part of weight).Acquired solution is directly used in next step.

At -78 DEG C, LDA (1.0M THF solution, 1.2 equivalents, 2.4 parts of volumes) are added in THF (10 parts of volumes) solution of 19 (1 part of weight).Stirring gained mixture 10 minutes, then at -78 DEG C, is added the enolate solution into THF (5 parts of volumes) solution of methyl iodide (1.5 equivalents, 0.19 part of volume).Reaction solution is added in saturated sodium bicarbonate (10 parts of volumes) and carries out inverse quenching, is extracted with MTBE (15 parts of volumes).Extract liquor is washed with salt water (5 parts of volumes), obtains 20 (0.86 part of weight) through chromatogram purification after concentration.

At 0 DEG C, AlMe3 (2M toluene solution, 1.5 equivalents, 1.5 parts of volumes) are added in methylene chloride (2.5 parts of volumes) suspension of hydrochloride base azanol (1 part of weight).To keep speed of the reaction temperature lower than 5 DEG C that methylene chloride (5 parts of volumes) solution of 20 (1 part of weight) is added.Then under the conditions of keeping temperature to be lower than 10 DEG C, to addition reaction mixture in sodium tartrate aqueous solution (1.3M, 20 parts of volumes).Make to be layered and be separated, organic layer is dry through sodium sulphate (5 parts of weight).Gained suspension is filtered, filtrate is concentrated.Residue is dissolved in DMF (2 parts of volumes), imidazoles (0.19 part of weight) and TBSCl (0.29 part of weight) is then added.Reaction solution water (5 parts of volumes) and MTBE (10 parts of volumes) dilution, are stirred 10 minutes.Water layer is removed, organic layer is washed with water (5 parts of volumes).Extract liquor is added in the solution of sodium hydrate aqueous solution (1N, 0.78 part of volume) and methanol (0.7 part of volume).Water layer is removed after being stirred to react liquid, organic layer is washed with salt water (2.5 parts of volumes), concentrated to obtain 21 (1.2 parts of weight).

To keep reaction temperature to be lower than 0 DEG C of speed, methyl-magnesium-chloride (3.0M, 59 parts of weight, 1.2 equivalents) are added in anhydrous THF (1.11 parts of weight, 1.25 parts of volumes) solution of 21 (1 part of weight).After being stirred at 0 DEG C, reaction solution is added in saturated ammonium chloride (2.5 parts of volumes) and water (2.3 parts of volumes) and carries out inverse quenching.Gained mixture is diluted with MTBE (10 parts of volumes), is vigorously stirred.Water layer is separated, organic layer is washed with salt water (2.5 parts of volumes), concentrated to obtain 22 (0.84 part of weight).

Compound 22 (1 part of weight) is dissolved in THF (4 parts of volumes), is cooled to -78 DEG C.It keeps temperature to be lower than under the conditions of -60 DEG C, is added KHMDS (1.5M toluene solution, 1.01 equivalents, 2.78 parts of weight).Tf is added₂THF (1.5 parts of volumes) solution of NPh (0.62 part of weight, 1.1 equivalents), makes reaction solution be warming up to -20 DEG C.Saturated ammonium chloride (2.5 parts of volumes), water (2.5 parts of volumes) and normal heptane (2.5 parts of volumes) is added, mixture is warming up to room temperature.Make to be layered, removes water layer.Organic extract liquid is washed with saturated sodium bicarbonate aqueous solution (3 × 2.5 parts of volumes) and salt water (2.5 parts of volumes), then obtains 23 (1.1 parts of weight) through vacuum concentration.

Compound 23 is dissolved in methanol (2.5 parts of volumes), is cooled to 15 DEG C.It is added hydrochloric acid (5N IPA solution, 1.30 equivalents, 1.18 parts of weight), acquired solution is warming up to 25 DEG C.Cooling reaction solution is added sodium bicarbonate (3 equivalents, 0.33 part of weight) to 0 DEG C.It is stirred to react liquid 15 minutes, filtering removal gained sediment.Filter cake is washed with ACS grades of methanol (1 part of volume), and merging filtrate is simultaneously concentrated.Thick concentrate obtains ER-806730 (24) (0.5 part of weight) through chromatogram purification.

Embodiment 4a

Under the general scheme illustrated using above scheme V, embodiment 4a provides the method for the compound (intermediate for preparing F2) of another preparation formula A.This method (is prepared) by intermediate of ER-812935 according to embodiment 3 (compound 4).

ER-812935 (1 part of weight) is dissolved in THF (10 parts of volumes), is cooled to 0 DEG C.It keeps temperature to be lower than under the conditions of 5 DEG C, is added LAH (1.0M THF solution, 0.70 equivalent, 2.0 parts of volumes).It is vigorously stirred down, quenches excessive reagent with water (0.078 part of volume), during which temperature is kept to be lower than 5 DEG C.Side keeps being vigorously stirred, and sodium hydroxide (15% w/w aqueous solution (0.078 part of volume)) and water (0.18 part of volume) is successively added in side.Celite is added(2 parts of weight) filters suspension afterwards, rinses filter cake with THF (5 parts of volumes).ER-817633 (0.92 part of weight, the calculated value based on 100% conversion ratio) is concentrated into 5 parts of volumes, is directly used in next step.

The ER-816961 solution (1 part of weight is dissolved in 5 parts of volume THF) prepared by THF (5 parts of volumes) dilution front, is cooled to 5 DEG C, is added triethylamine (3 equivalents, 0.94 part of weight).To keep speed of the temperature lower than 10 DEG C that MsCl (1.05 equivalents, 0.25 part of volume) is added.It is added water (5 parts of weight), quenches reaction.It is added heptane (8 parts of volumes), makes demixing.It separates water phase and is extracted with MTBE (2 parts of volumes).Combined organic extract liquid is washed with saturated sodium bicarbonate (5 parts of volumes) and water (1.9 parts of volumes).Concentration of organic layers exchanges solvent with ethyl alcohol to prepare ethyl alcohol (1 part of volume) solution of ER-818937 (1.23 parts of weight, the calculated value based on 100% conversion ratio), is directly used in next step.

The ER-818937 solution prepared by ethyl alcohol (190 normal intensities, 9 parts of volumes) dilution front (1 part of weight is dissolved in 0.8 part of volume ethanol).It is added potassium cyanide (3 equivalents, 0.41 part of weight), heats suspension to 70-80 DEG C.Water (10 parts of volumes) and MTBE (10 parts of volumes) is successively added to room temperature in cooling reaction solution.Each layer is separated, water layer is extracted with MTBE (5 parts of volumes).Combined organic matter is washed with water (2 parts of volumes) and saturated brine (4 parts of weight).Concentrated extract is directly used in next step.

ER-818950 is dissolved in acetic acid (5 parts of volumes), is added hydrochloric acid (1.0M, 1 equivalent, 3 parts of volumes), is stirred to react liquid at room temperature.Cooling reaction solution is to 0 DEG C, to keep speed of the temperature lower than 10 DEG C that sodium hydroxide (50 weight %, 30 equivalents, 7 parts of weight) are added.With heptane (2 × 10 parts of volumes) extraction solution.Water phase is saturated with sodium chloride, is extracted with acetonitrile (2 × 10 parts of volumes).Combined acetonitrile extraction liquid is concentrated, by air-distillation, carries out exchange of solvent with ethyl acetate, obtains ethyl acetate (3 parts of volumes) solution of ER-817664.0 DEG C is cooled to after filtering out salt in hot solution.Filtering suspension obtains the ER-817664 of white crystalline solid shape.

Embodiment 4b

Embodiment 4b provides the method for the compound of another general scheme preparation formula F-2 described in above scheme Vb and Vc.This method is using ER-817664 (being prepared according to above embodiments 4a) as intermediate.

ER-817664 (1 part of weight) is dissolved in acetonitrile (10 parts of volumes), cooling suspension is to 0 DEG C, 2- acetate -2- methyl N-Propyl Bromide (4.0 equivalents, 2.4 parts of volumes) and water (1.0 equivalents, 0.07 part of volume) is successively added.Stirring gained mixture 2 hours at 0 DEG C.At 0 DEG C, it is slowly added to sodium bicarbonate (saturated aqueous solution, 8.0 equivalents, 40 parts of volumes).Stirring gained mixture 30 minutes at room temperature, is then extracted with MTBE (2 × 20 parts of volumes).Organic layer is washed with salt water (5 parts of volumes), concentrated to obtain colorless oil as product.

Bromine starting material (1 part of weight) described immediately above is dissolved in toluene (10 parts of volumes).It is added DBU (1.8 equivalents, 0.73 part of volume), at 80 DEG C, mixture is heated.Cooling mixture is diluted with MTBE (20 parts of volumes), is washed with water (5 parts of volumes) and salt water (5 parts of volumes) to room temperature.Then concentration of organic layers obtains gray powdery product.

Starting olefinic compound (1 part of weight) (describing just above) is dissolved in methylene chloride (5 parts of volumes) and methanol (5 parts of volumes), is cooled to -40 DEG C to -45 DEG C.Use O₃Processing solution.Excessive O is removed by nitrogen purging₃, solution is made to be warming up to -15 DEG C.NaBH is added₄(1.0 equivalents, 0.18 part of weight), makes mixture be warming up to 0 DEG C.It is added potassium carbonate (1.3 equivalent), at room temperature stirred suspension.At 0 DEG C, with 1N hydrochloric acid (about 4 equivalents, about 20 parts of volumes) neutralization reaction liquid, solution is extracted with MTBE (10 parts of volumes) to remove lipophilic substance.Water layer is concentrated to remove methylene chloride and methanol.THF (4 parts of volumes) and NaIO is successively added₄(2 equivalents, 2 parts of weight).Reaction solution is extracted with MTBE (10 parts of volumes) and n-BuOH (10 parts of volumes).Combined organic extract liquid is concentrated, gained powder is through ethyl acetate development.The inner hemiacetal of isolated light yellow powder after filtering.

ER-818638 (1 part of weight) and LiCl (2 equivalents, 0.35 part of weight) is stirred in acetonitrile (8.7 parts of volumes).At 25 DEG C, it is added Hunigs alkali (1.5 equivalent).It is added 1N hydrochloric acid (5 parts of volumes), extracts mixture with MTBE (10 parts of volumes).Condensed organic obtains ER-818640, is used for next step.

Starting α-enester compound (1 part of weight) (describing just above) is dissolved in methanol (10 parts of volumes), 10 weight % palladium carbons (0.09 equivalent, about 0.33 part of weight) is added under nitrogen.Then the suspension is stirred under hydrogen.Suspension is filtered through CeliteIt pads (20 parts of weight), rinses filter cake with methanol (20 parts of volumes).Filtrate is concentrated, obtains colorless oil as product (yield 94.3%) through purification by flash chromatography.

Pyridine (10 equivalent), Ph are separately added into THF (15 parts of volumes) solution of ester (1 part of weight)₃P (7 equivalent) and NIS (4 equivalent).It is stirred to react mixture at room temperature.It is added aqueous citric acid solution (20 weight %, 10 equivalents), with TBME (30 parts of volumes) diluted mixture.Separate water layer, organic layer water (5 parts of volumes), sodium thiosulfate solution (10% weight/volume, 5 parts of volumes), water (5 parts of volumes) and salt water (5 parts of volumes) washing.Concentration of organic layers obtains colorless oil as product through purification by flash chromatography.

Iodine starting material (1 part of weight) is dissolved in methanol (30 parts of volumes), is heated to 55 DEG C.At 55 DEG C, points 6 times through 80 minutes addition NaBH₄(47 equivalent).Cooling reaction solution is quenched to 0 DEG C with 1N hydrochloric acid (30 parts of volumes).After five minutes, mixture is diluted with salt water (30 partial volume) for stirring, is extracted with DCM (50 parts of volume × 2).Organic layer is dried over sodium sulfate, is concentrated.Crude product is directly used in next step.

Alcohol starting material (1 part of weight) (describing just above) is dissolved in ethyl alcohol (70 parts of volumes), is added Zn (165 equivalent).At 75-80 DEG C, flow back to suspension.Cooling reaction mixture is added 1N hydrochloric acid (70 parts of volumes) to room temperature.Mixture is extracted with DCM (3 × 100 parts of volumes), organic layer is washed with brine and is concentrated.

Lactone starting material (describing just above) is dissolved in DCM (50 parts of volumes), under a nitrogen, it is separately added into triethylamine (5.0 equivalent), DMAP (0.3 equivalent) and TBDPSCl (1.5 equivalent) at room temperature, stirs acquired solution 2-3 hours at room temperature.After fully reacting, mixture is diluted with TBME (100 parts of volumes), is washed with saturated sodium bicarbonate aqueous solution (10 parts of volumes), water (10 parts of volumes) and salt water (10 parts of volumes).Concentration of organic layers obtains colorless oil as product through purification by flash chromatography.

Embodiment 4c

Using general scheme described in above scheme VII, embodiment 4c provides the method for the compound of another preparation formula F-2.This method is with ER-811510 (preparing according to embodiment 3, wherein acetone is used to replace cyclohexanone) as intermediate.

ER-811510 (1 part of weight, 1 equivalent) is dissolved in methylene chloride (6.3 parts of volumes), is cooled to -5 DEG C.It keeps temperature to be lower than under the conditions of 0 DEG C, pyridine (0.41 part of volume, 1.1 equivalents) and bromoacetyl bromide (0.44 part of volume, 1.1 equivalents) is successively added.It is stirred to react liquid 1 hour, is warming up to room temperature.It is added water (8 parts of volumes), makes to be layered.Organic layer successively uses cupric sulfate pentahydrate aqueous solution (1.0M, 10 parts of volumes), water (8 parts of volumes) and salt water (10 parts of volumes) washing, it is dried and filtered with magnesium sulfate and is concentrated in vacuo, obtain the ER-812771 of brown solid.

ER-812771 (1 part of weight, 1 equivalent) is dissolved in acetonitrile (6 parts of volumes), triphenyl phasphine is added, reaction solution is heated to 50 DEG C, is kept for 45 minutes.1,8- diazabicyclo [5.4.0], 11 carbon -7- alkene (DBU, 0.35 part of volume, 0.8 equivalent) is added after extremely -10 DEG C of cooling reaction solution.It is stirred to react liquid 15 minutes, is heated to 80 DEG C, kept for 45 minutes, then cooled to room temperature.It is added ammonium chloride (saturated aqueous solution, 10 parts of volumes), aqueous layer with ethyl acetate (3 × 10 parts of volumes) extraction.Combined organic layer is dried over magnesium sulfate, vacuum concentration.Crude product obtains the ER-812772 of white solid through chromatogram purification.

ER-812772 (1 part of weight, 1 equivalent) is dissolved in ethyl acetate (8 parts of volumes), is added 10% palladium carbon (0.05 part of weight, 0.01 equivalent), is stirred under a hydrogen atmosphere 2 hours after purging reaction solution with nitrogen.Under ethyl acetate washing, pass through diatomite filtering and removing catalyst.Combined filtrate is concentrated in vacuo to obtain the ER-812829 of white solid.

Embodiment 5

The preparation of F-3a

D- gulonolactone (1 part of weight, 1 equivalent), cyclohexanone (2-3 equivalent), toluene (6 parts of volumes) and p-methyl benzenesulfonic acid (0.021 part of weight, 0.02 equivalent) are added in the reaction vessel.Under stirring, heating reaction solution to reflux.Azeotropic removes fully reacting after moisture.Reaction mixture is cooled to 85-90 DEG C, stirring aggravation.Under stirring, through 20-30 minutes addition heptane (5.2 parts of volumes).It is cooled to 65-70 DEG C, is stirred 30 minutes at 65-70 DEG C.Solid product is filtered at 65-70 DEG C, while keeping 35 DEG C of mother liquor temperature >.It is refiltered at 35-40 DEG C, mother liquor is kept for room temperature 30 minutes.Refilter mother liquor.Filter cake is dried to obtain ER-805715 (yield 84%, 1.6 parts of weight) after being washed twice with heptane (2 × 1.7 parts of volumes).

It prepares in another method of ER-805715, by D- gulonolactone (1 part of weight), cyclohexanone (1.32 parts of weight, 2.4 equivalents), p-methyl benzenesulfonic acid monohydrate (0.02 part of weight, 0.02 equivalent) and toluene (12 parts of volumes) flow back 19 hours together, while azeotropic remove moisture.Mixture successively uses 5% sodium bicarbonate aqueous solution (4 parts of volumes) and saturated sodium-chloride water solution (2 parts of volumes × 2, pH=7) to wash.Distillation and concentration organic phase (remaining 4.5 parts of calculating volumes toluenes), after being cooled to 100 DEG C, is added heptane (10 parts of volumes) under the conditions of 80 DEG C of > of temperature in keeping.Mixture is heated to flowing back, is kept at least 1 hour, is subsequently cooled to 85 DEG C of agings 3 hours, until 80 DEG C are cooled to 40 DEG C of agings 12 hours after aging 3 hours.Product is collected by filtration, filter cake is washed with heptane (2 parts of volumes).Pneumatic conveying drying filter cake obtains ER-805715 (yield 78%, 1.48 parts of weight).

ER-805715 (1 part of weight, 1 equivalent) is added in the reaction vessel, is allowed to be dissolved in anhydrous THF (3.34 parts of volumes) and dry toluene (2.5 parts of volumes).Cooling mixture is to -15 DEG C -- and 10 DEG C.- 15 DEG C -- at 10 DEG C, through 1 hour addition DIBALH (1.5M toluene solution, 2.4 parts of volumes, 1.2 equivalents) and stir mixture 15-30 minutes.At 10 DEG C, reaction solution is added in potassium sodium tartrate solution (1 part of weight sodium potassium tartrate tetrahydrate is dissolved in 2.9 parts of weight waters) and carries out inverse quenching, so that gained mixture is warming up to room temperature, stir 4 hours.It is layered after filtering mixture, is extracted with MTBE (2 parts of volumes).Merge organic layer, solvent removed by vacuum obtains ER-805814 (1.02 parts of weight, yield 100%).

ER-805814 (1 part of weight) is dissolved in anhydrous THF (3.3 parts of volumes), is handled with (methoxy) triphenyl phosphine dichloride (2.11 parts of weight, 2.1 equivalents).Heat reaction mixture to 28-32 DEG C, then keep reaction temperature be 30-35 DEG C under the conditions of, through 100-140 minutes addition KOtBu (0.66 part of weight, 2 equivalents) anhydrous THF (2.64 parts of volumes) solution.Cooling reaction solution is added MTBE (5.11 parts of volumes) to 20-25 DEG C after 5 hours, stirs mixture.Salt water (3 parts of weight) and water (3 parts of weight) is added (charging starts to occur as soon as heat release, is controlled with water-bath at 20-25 DEG C).Organic layer is separated, is handled with MTBE/THF (1/1 volume ratio, 1.78 parts of volumes) solution of maleic anhydride (0.27 part of weight).Sodium hydroxide solution is slowly added into reaction mixture (0.88 part of weight is dissolved in 2.5 parts of volume of water).Concentration of organic layers obtains crude product ER-805815 (0.985 part of weight).Residue is developed three times with MTBE/ heptane (1/4 volume ratio, 6.6 parts of volumes).Under MTBE/ heptane (1/2 volume ratio, 45 parts of volumes) elution, extract liquor is filtered through SiO₂(3 parts of weight).Concentration filtrate obtains ER-805815 (0.88 part of weight, yield 81%).

In another method for preparing ER-805815, under the conditions of holding reaction temperature is 0-10 DEG C, in (methoxy) triphenyl phosphine dichloride (3.12 parts of weight, 3.1 equivalents) THF (1.78 parts of weight) suspension in be added t-BuOK (0.989 part of weight, 3 equivalents) THF (4 parts of weight) solution.The feed container (additionvessel) is rinsed with THF (2 × 0.7 parts of weight).It keeps at 0-10 DEG C, THF (the 1.42 parts of weight) solution of ER-805814 (1 part of weight, 1 equivalent) is added into reaction solution.The feed container is rinsed with THF (2 × 0.7 parts of weight).Mixture is stirred at 20-30 DEG C to stay overnight, and is stirred 3 hours at 30-35 DEG C.Cooling reaction solution is to 30 DEG C hereinafter, successively being diluted with MTBE (3.7 parts of weight) and 10 weight % sodium-chloride water solutions (4 parts of weight).Stirring mixture 30 minutes, separates each layer.It is added maleic anhydride (0.63 part of weight, 2.2 equivalents), stirs mixture 30 minutes at room temperature.It keeps reaction solution to be lower than under the conditions of 15 DEG C, water (6 parts of weight) and sodium hydroxide solution (48 weight %, 0.64 part of weight, 2.6 equivalents) is added dropwise.After stirring at lower than 15 DEG C, lower layer is separated.Water (6 parts of weight) and sodium hydroxide solution (48 weight %, 0.64 part of weight, 2.6 equivalents) is successively added, maintains the mixture below 15 DEG C during sample-adding.Lower layer is separated after stirring at lower than 15 DEG C.Organic layer is washed three times with 15 weight % sodium-chloride water solutions (3 × 4 parts of weight).It is concentrated in vacuo organic layer.Residue is diluted and is concentrated in vacuo with MTBE (1 part of weight).At 40-50 DEG C, through 30 minutes dropwise addition IPE (3 parts of weight) residue is diluted.At 40-50 DEG C, stirred suspension 1 hour, 0-10 DEG C is slowly cooled to, is stirred 1 hour.Filter solid is crossed, filter cake is washed with IPE (2 parts of weight).It is concentrated in vacuo filtrate and cleaning solution.Residue methanol (2.37 parts of weight) and water (0.4 part of weight) processing, are extracted with heptane (2.74 parts of weight).Lower layer is extracted 9 times with (2.05 parts) of heptane.Combining extraction liquid obtains ER-805815 (1.07 parts of weight, 98.6%) through vacuum concentration.

In another post-processing approach of ER-805815, obtained thick organic layer is handled with MTBE (2.86 parts of weight) and diatomite (0.5 part of weight) after salt water washing and concentration.After stirring 2.5 hours, through 2 hours addition heptane (1.46 parts of weight), stirs mixture and stay overnight.Filtering precipitate.Filter cake is washed with MTBE/ heptane (1: 1,5 parts of weight).Vacuum concentration filtrate is until volume is decreased to about 3 parts of volumes.Residue methanol (2 parts of weight) and water (6 parts of weight) dissolution, extract mixture with heptane/MTBE (5: 1,3 × 6 parts of weight).Organic layer is separated, it is concentrated to obtain ER-805815, it is used for next step.

ER-805815 (1 part of weight) is dissolved in acetone (2.4 parts of volumes) and water (0.4 part of volume).It is added N-methylmorpholine N- oxide (0.62 part of weight, 2 equivalents), cooling mixture is to 0-5 DEG C.OsO is added₄(0.15M aqueous solution, 0.065 part of volume), reaction solution is maintained at 0-5 DEG C.Mixture is stirred to react at 0-5 DEG C 12 hours.At 0-2 DEG C, through 1 hour addition water (0.2 part of volume).It is stirred mixture 1 hour at 0-5 DEG C.Filtration product washs solid twice with the acetone/water (1/1, volume ratio, 2 × 0.7 parts of volumes) of pre-cooling (0-5 DEG C).Desciccate obtains ER-805816 (0.526 part of weight, 52% yield remain Os < 17ppm).

It prepares in another method of ER-805816, acetone (the 4 parts of volumes) solution of ER-805815 (1 part of weight, 1 equivalent) is added in four-neck flask, water (0.5 part of weight) then is added at room temperature.Anhydrous N-methyl methylmorpholine-N-oxide (0.38 part of weight, 1.2 equivalents) are added in the mixture.At 25-35 DEG C, while being water-cooled, while two water potassium osmates (0.003 part of weight, 0.003 equivalent) is added portionwise.Mixture is kept at such a temperature 4 hours.Water (the 0.5 part of weight) solution of sodium thiosulfate (0.075 part of weight, 0.49 equivalent) is added at room temperature, then stirs mixture 0.5 hour.Mixture is cooled to 0-5 DEG C, is stirred 2 hours.Resulting sediment is collected, wet filter cake methanol (0.6 part of weight) and water (1.5 parts of weight) wash to obtain crude product (1.25 parts of weight).(0.611 part of weight) is dried to crude product sample.Crude product ER-805816 (1.25 parts of weight) is added in water (3.05 parts of weight), is stirred 2 hours at about 25 DEG C.Filtering precipitate is washed with water (1.53 parts of weight), obtains crude product wet cake (1.05 parts of weight).Crude product sample is dried and is sampled, ICP Os=37ppm.Crude product ER-805816 (1.05 parts of weight) is added in water (2.81 parts of weight), is stirred 2 hours at about 25 DEG C.Filtering precipitate is washed with water (1.4 parts of weight) and methanol (0.45 part of weight), obtains crude product ER-805816 (0.736 part of weight).Wet filter cake is dried to obtain crude product (0.56 part of weight, ICP (Os)=28ppm).At 45-55 DEG C, ER-80581 6 (0.56 part of weight) is dissolved in acetone (1.76 parts of weight).Activated carbon (0.027 part of weight) is added in the solution, is stirred 0.5 hour at identical temperature.Mixture is filtered, filter cake is washed with hot acetone (0.214 part of weight).Filtrate is maintained at 45-50 DEG C, through 10 minutes addition water (0.83 part of weight), temperature during water is added to be maintained at 40-50 DEG C.Cooling mixture stirs 1.5 hours to 0-5 DEG C.White depositions are filtered, it is dry after being washed with the solution of acetone (0.17 part of weight) and water (0.22 part of weight), obtain ER-805816 (0.508 part of weight, 0.49 equivalent, KF5.0%, ICP (Os) 9.6ppm).

ER-805816 (1 part of weight) is suspended in acetic acid (0.89 part of volume, 5.8 equivalents) and acetic anhydride (3.57 parts of weight, 13 equivalents).It is added anhydrous zinc chloride (0.2 part of weight, 0.54 equivalent).Mixture is stirred to react at 18-22 DEG C 24 hours.Reaction solution is quenched in ice (5 parts of weight) and water (5 parts of volumes).It is added with stirring ethyl acetate (10 parts of volumes), separates water layer.Aqueous layer with ethyl acetate (10 parts of volumes) back extraction.Combined organic layer is successively washed with salt water (10 parts of volumes), 5% ethyl alcohol sodium water solution (6 parts of volumes) and salt water (6 parts of volumes).Concentration of organic layers.Thick concentrate is dissolved in 25%EtOAc/hex (4 parts of volumes), through SiO₂Filtering.Filter bed is washed with 25%EtOAc/hex (2 × 12 parts of volumes), is further washed with 25%EtOAc/hex (48 parts of volumes).Concentration of organic layers obtains ER-805819 (1 part of weight, 81%).

It prepares in another method of ER-805819, zinc chloride (0.2 part of weight, 0.54 equivalent), acetic anhydride (2.75 parts of weight, 10 equivalents) and acetic acid (1 part of weight, 6 equivalents) is mixed.Cooling mixture is to 15-20 DEG C.It keeps being added ER-805816 (1 part of weight, 1 equivalent) in 15-30 DEG C under the conditions of temperature.Then it is stirred mixture 6 hours at 35-40 DEG C.Cooling reaction mixture is to 25 DEG C or less.It keeps reaction temperature to be lower than under the conditions of 25 DEG C, is added dropwise methanol (3.2 parts of weight, 4 parts of volumes).It is added heptane (2.7 parts of weight, 4 parts of volumes).It keeps reaction temperature to be lower than under the conditions of 25 DEG C, is added water (4 parts of weight, 4 parts of volumes).Stirring mixture 15 minutes, then separates each phase.Lower layer is washed twice with heptane (2.7 parts of weight, 4 parts of volumes), discards heptane layer.Lower layer is extracted twice with toluene (6.1 parts of weight, 8 parts of volumes).Combined toluene layer is washed twice with 17 weight % potassium bicarbonate aqueous solutions (0.82 part of weight saleratus is dissolved in 3.98 parts by weight water, 4.36 parts of volumes), is washed twice and is concentrated with water (4 parts of weight).Methanol (3.95 parts of weight, 5 parts of volumes) are added at 25-30 DEG C, stir mixture 10 minutes.Water (0.3 part of weight) is added at 25-30 DEG C.Crystal seed is added to 0 DEG C in cooling mixture.It is stirred mixture 1 hour at 0 DEG C, through 1 hour dropwise addition water (0.7 part of weight).Through 1 hour dropwise addition water (4 parts of weight).Resulting sediment is filtered, filter cake is washed twice with 0 DEG C of methanol (1.03 parts of weight) and the solution of water (0.7 part of weight).Dry cake obtains ER-805819 (0.99 part of weight, 0.84 equivalent).

ER-805819 (1 part of weight) is dissolved in anhydrous acetonitrile (15 parts of volumes), is handled with ER-803055 (0.93 part of volume, 2 equivalents).Cooling reaction mixture was to 0-5 DEG C, through 5 minutes addition BF3-OEt₂(0.54 part of volume, 1.95 equivalents), during which keeping reaction temperature is 0-5 DEG C.Mixture is stirred to react at 0-5 DEG C 12 hours.It is vigorously stirred down, reaction solution is added in saturated sodium bicarbonate (20 parts of volumes), is quenched.It is extracted twice with ethyl acetate (2 × 8 parts of volumes).Combined organic matter is washed with salt water (12 parts of volumes), concentrated to obtain ER-805821 (1 part of weight, 88% yield directly use).

It prepares in another method of ER-805821, ER-805819 (1 part of weight, 1 equivalent) and ER-803055 (0.93 part of volume, 2 equivalents) is dissolved in anhydrous acetonitrile (5.46 parts of weight, 7 parts of volumes).Cooling reaction mixture was to 0-5 DEG C, through 5 minutes addition BF₃-OEt₂(0.54 part of volume, 1.95 equivalents), during which keeping reaction temperature is 0-5 DEG C.It is stirred to react mixture at 0-5 DEG C 20 hours, heptane (5.47 parts of weight, 8 parts of volumes) then are added at 0-5 DEG C.Each phase is separated, uses heptane (5.47 parts of weight, 8 parts of volumes) to handle lower layer at 0-5 DEG C.It under the conditions of holding reaction temperature is 0-15 DEG C, is added dropwise 7.4% potassium bicarbonate aqueous solution (0.64 part of weight saleratus is dissolved in 8 parts of weight waters), quenches reaction.It is added toluene (8.65 parts of weight, 10 parts of volumes), stirs mixture 30 minutes.Lower layer is separated, upper organic layer is washed twice with water (10 parts of volumes), concentrated to obtain ER-805821 crude product oil (1.05 parts of weight, 0.935 equivalent).

ER-805821 (1 part of weight) is dissolved in anhydrous THF (8.4 parts of volumes) and anhydrous acetic acid methyl esters (2 parts of volumes).It maintains under 17-23 DEG C of reaction, through 2 minutes addition TritonB (OH) (3.6 parts of volumes).It is stirred to react liquid 1.5 hours.Filter reaction mixture.Filtrate is concentrated, passes through SiO₂Pad (5 parts of weight, 20 parts of volume of ethylacetate elutions).Filtrate is washed with salt water (2.2 parts of volumes), obtains ER-805822 (0.54 part of weight, 72% yield) through evaporation.

It prepares in another method of ER-805822, ER-805821 (1 part of weight, 1 equivalent, 11.18g, 21.81mmol) is dissolved in anhydrous MTBE (4.4 parts of weight, 6 parts of volumes), is cooled to 0-5 DEG C.At 0-5 DEG C, into mixture through 1 hour addition sodium methoxide (28 weight % methanol solutions, 0.564 part of weight, 1.5 equivalents), stirred 3 hours under same temperature ranges stated.Acetic acid (0.188 part of weight, 1.6 equivalents) quenching reaction is added, is kept for 0-5 DEG C during sample-adding.It stirs mixture to stay overnight, then at 0-5 DEG C, is handled, be subsequently agitated for 15 minutes with 5 weight % potassium bicarbonate aqueous solutions (3 parts of weight) and ethyl acetate (3.6 parts of weight, 4 parts of volumes).After mutually separating, lower layer is extracted twice with ethyl acetate (3.6 parts of weight, 4 parts of volumes).Combined organic layer is concentrated.Acetone (2 parts of weight, 2.5 parts of volumes) and IPE (2 parts of weight, 2.7 parts of volumes) are added in residue, is stirred overnight at 0-5 DEG C.Mixture is filtered through Celite(0.25 part of weight) is washed with acetone (2 parts of weight).It is oily (0.55 part of weight) that concentration filtrate obtains crude product.It is added at acetone (0.2 part of weight, 0.25 part of volume) and IPE (0.54 part of weight, 0.75 part of volume) and 40-50 DEG C and stirs 1 hour in residue.ER-805822 crystal seed is added in the solution at room temperature, is stirred overnight at room temperature.At room temperature, IPE (1.27 parts of weight, 1.75 parts of volumes) were added into suspension through 2 hours.After stirring 5 hours at room temperature, sediment is collected by filtration, filter cake is washed with acetone/IPE (1/10) (2 parts of volumes).Gained filter cake is dried overnight in disc type chamber (tray-type chamber) in 30-40 DEG C, and obtaining target product ER-805822, (38.0%) 0.286 part of weight, 0.38 equivalent, the yield based on ER-805819 are.

In another post-processing approach of crude product ER-805822, obtained residue is dissolved in IPA (2 parts of weight) after final ethyl acetate solution will be concentrated, and heated solution is to 50 DEG C.It is added heptane (5 parts of weight), crystal seed is added to 20 DEG C in cooling mixture.Mixture is stirred at 20 DEG C to stay overnight.It is added heptane (10 parts of weight), mixture is cooled to -5 DEG C in 30 minutes, is stirred 5 hours at -5 DEG C.Mixture is filtered, washs filter cake with heptane (2 parts of weight).Pneumatic conveying drying is carried out to filter cake under vacuum, obtains ER-805822 (60%).

ER-805822 (1 part of weight) is dissolved in ethyl acetate or other solvents appropriate (5 parts of volumes) and water (5 parts of volumes).Under the conditions of holding reaction temperature is 0-10 DEG C, NaIO was added portionwise through -1 hour 30 minutes₄(0.58 part of weight, 1.05 equivalents).It is stirred to react liquid at most 2 hours.With sodium chloride (1 part of weight) reaction mixture and stirring 30 minutes at 0-10 DEG C.Reaction mixture is filtered, filter cake is rinsed with ethyl acetate (2 parts of volumes).Each phase is separated, lower layer is extracted three times with ethyl acetate (5 parts of volumes).Combined organic layer is washed with 20% sodium-chloride water solution (5 parts of weight).Concentration of organic layers obtains ER804697 (1 part of weight).Residue is dissolved in toluene (2 parts of volumes), concentrate solution.Residue is dissolved in acetonitrile (7 parts of volumes), next step is used for.

Under inert atmosphere, NiCl is added in the reaction vessel₂(0.025 part of weight) and CrCl₂(2.5 parts of weight).It is added anhydrous methylene chloride (5 parts of volumes).Start to stir, cooling mixture is to 0-3 DEG C.It keeps temperature to be lower than under the conditions of 20 DEG C, anhydrous DMSO (6.7 parts of volumes) is added, while being vigorously stirred 45 minutes.ER-804697 (1 part of weight) is dissolved in anhydrous methylene chloride (1 part of volume) and is added in reaction vessel.Gained mixture is set to be warming up to 25 DEG C, through 20 minutes addition ER-806643 (2.58 parts of weight).Reaction temperature is kept to be lower than 45 DEG C.After sample-adding, liquid is stirred to react at 25-35 DEG C 30 minutes.It is added methanol (5 parts of volumes), stirs mixture 10 minutes.It is added MTBE (33 parts of volumes), suspension is transferred in 1N hydrochloric acid (25 parts of volumes) and water (10 parts of volumes).Stirring mixture 5 minutes.Water layer is stripped with MTBE (10 parts of volumes), and combined organic layer successively uses 0.2N hydrochloric acid (17 parts of volumes), 1% sodium chloride solution (2 × 17 parts of volumes, wash twice) and salt water (13 parts of volumes) washing.Concentration of organic layers, purified (SiO₂, 25 parts of weight, the ethyl acetate/hexane of 10 column volumes, 1/3.5 volume ratio) and obtain ER-804698 (0.53 part of weight, 61%).

In another method, which carries out in the presence of chiral ligand ER-807363, and method and ER-118047 preparation method described below are substantially similar.

It prepares in another method of ER-804698, DMSO (7 parts of volumes) and acetonitrile (7 parts of volumes) are de-gassed and are cooled to 0-10 DEG C.It is no more than interior temperature under the conditions of 20 DEG C, uses CrCl₂(10 equivalents, 3.47 parts of weight) and NiCl₂(0.1 equivalent, 0.037 part of weight) the batch processing solution.At 0-10 DEG C, be added dropwise ER-804697 (1 part of weight, 1 equivalent) acetonitrile (7 parts of volumes) solution and ER-806643 (5 equivalents, 2.5 parts of weight), during temperature must not exceed 15 DEG C.Mixture is stirred to react at 5-15 DEG C to stay overnight.Methanol (5.5 parts of weight), water (7 parts of weight) and MTBE (5.2 parts of weight) are added in the mixture.It is stirred to react liquid 1 hour, separates lower layer (layer 1).The premixed solution of sodium chloride (1.5 parts of weight) and water (13.5 parts of weight) is added in the upper layer.Stirring mixture 1 hour separates lower layer (layer 2).The premixed solution of heptane (4.8 parts of weight), methanol (2.8 parts of weight) and sodium chloride (1.5 parts of weight) and water (13.5 parts of weight) is added in the upper layer.Stirring mixture 1 hour separates lower layer (layer 3).Upper layer saves (organic matter 1) by dehydration.Layer 1, methanol (2.8 parts of weight) and MTBE (2.8 parts of weight) are added in the reactor.Mixture is stirred to stay overnight.It separates and discards lower layer.Upper layer is handled with layer 2.Stirring mixture 1 hour, separates and discards lower layer.Upper layer is handled with layer 3 and heptane (4.8 parts of weight).Stirring mixture 1 hour, separates and discards lower layer.Upper layer saves (organic matter 2) by dehydration.Layer 3, MTBE (0.8 part of weight) and heptane (2.7 parts of weight) are added in the reactor.Stirring mixture 1 hour, separates and discards lower layer.Upper layer is merged with organic matter 1 and organic matter 2.Combined organic matter is filtered and is concentrated under reduced pressure, crude product ER-804698 is obtained, through chromatogram purification (SiO₂, 25 parts of weight, the ethyl acetate/hexane of 10 column volumes, volume ratio 1/3.5) and obtain ER-804698 (0.67 part of weight, 57% yield).

It prepares in another method of ER-804698, thick material is directly used in next step without purifying.

With acetic acid (4.2 parts of weight) and water (4.2 parts of weight) processing ER804698 (1 part of weight, 1 equivalent).Mixture is heated to 90-97 DEG C, is kept for 100 minutes.Cooling mixture is to 15 DEG C hereinafter, then being washed twice at 15 DEG C or less with heptane (2 × 2.7 parts of weight).After mutually separating, under the conditions of making temperature be no more than 15 DEG C, the mixture of 20 weight % potassium bicarbonate aqueous solutions (7.7 parts of weight, 35 equivalents) and MTBE (5.95 parts of weight) is added dropwise to lower layer.After mutually separating, upper layer is successively washed with 5 weight % potassium bicarbonate aqueous solutions (0.2 part of weight), is washed twice with 5 weight % sodium-chloride water solutions (2 × 0.2 parts of weight).Organic layer is concentrated under reduced pressure, is added MTBE (1.49 parts of weight).55 DEG C are heated the mixture to, stirring to dissolution.Heptane (1.00 parts of weight) is added in the solution, cooling solution is to 40-45 DEG C.It additionally incorporates in the solution heptane (4.47 parts of weight), then cooling solution is stirred overnight to 5-15 DEG C.Filtering for crystallizing body, is rinsed with heptane, obtains ER-807023 (0.58 part of weight, 71% yield).

Under nitrogen atmosphere, ER-807023 (1 part of weight, 1 equivalent) and MTBE (7.43 parts of weight) are added in the reactor.2,6- lutidines (2.15 parts of weight, 7.5 equivalents) are added in reaction solution.At 0 DEG C, TBSOTf (2.47 parts of weight, 3.5 equivalents) are added dropwise in the mixture.It is stirred to react mixture at 0-10 DEG C 30 minutes, was then warming up to 23 DEG C through 1 hour, is saved 16 hours at 23 DEG C.It keeps temperature to be lower than under the conditions of 30 DEG C, methanol (0.21 part of weight, 2.5 equivalents) and water (14.8 parts of weight) is successively added dropwise in the reactive mixture.After mutually separating, upper layer is washed with 1N aqueous hydrochloric acid solution (16.2 parts of weight), 5% sodium-chloride water solution (14.8 parts of weight), 5% sodium bicarbonate aqueous solution (14.8 parts of weight), 5% sodium-chloride water solution (14.8 parts of weight) and 5% sodium-chloride water solution (14.8 parts of weight) respectively.Upper organic layer is concentrated by vacuum distillation, obtains crude product ER-804699.It is added methanol (7.91 parts of weight), heats mixture to 50 DEG C, kept for 30 minutes.Mixture is cooled to 0 DEG C through 5 hours, is then stirred overnight at 0 DEG C.Filter solid is crossed, filter cake is washed with cold methanol (4 parts of weight), is dried to obtain ER-804699 (1.42 parts of weight, 74% yield).

Under nitrogen atmosphere, toluene (the 2.60 parts of weight) solution of ER-804699 (1 part of weight, 1 equivalent) is added in the reactor.It is added acetonitrile (4.72 parts of weight).It is added TBSCl (0.011 part of weight, 0.05 equivalent).So that reaction mixture is warming up to 30 DEG C, is added NIS (1.25 parts of weight, 4 equivalents).Mixture is stirred to react at 30 DEG C 22 hours.Cooling reaction solution keeps interior temperature to be lower than at 30 DEG C, the mixture (10.35 parts of weight) of sodium thiosulfate solution and sodium bicarbonate was added through 10 minutes to 25 DEG C.At 25 DEG C, it is stirred to react liquid 30 minutes.Separate water layer.Upper layer is washed twice with 10% sodium-chloride water solution (2 × 9.9 parts of weight).Organic layer is concentrated under reduced pressure, obtains crude product ER-803895, obtains ER-803895 (0.96 part of weight, 89.5% yield) through silica gel chromatography.

Embodiment 6

The assembling of F-1a, F-2a and F-3a and the preparation of B-1939

A. the preparation of (R) or (S) N- [2- (4- isopropyl -4,5- dihydroxy- azoles -2- base) -6- methylphenyl]-Methanesulfomide

Triphosgene (1 part of weight, 1 equivalent) and anhydrous THF (2 parts of volumes) are added in pre-dried glass-lined reactor, being cooled to interior temperature is -10 DEG C.ER-807244 (1.27 parts of weight, 2.5 equivalents) and anhydrous THF (3 parts of volumes) are added in second pre-dried glass-lined reactor, being cooled to interior temperature is -10 DEG C.So that the content of first reactor is transferred in second reactor by speed of the interior temperature no more than 15 DEG C.After sample-adding, it is stirred to react under temperature liquid 1 hour in 0 DEG C, is then gradually heated to 25 DEG C.Injection nitrogen 18 hours to remove excessive phosgene (collecting exhaust gas with 2N sodium hydroxide solution).It is added MTBE (3 parts of volumes), more MTBE are added in the lower distillation removing solvent of 40-46 DEG C of nitrogen purging if necessary.After completing removing phosgene, it is 5-10 DEG C that mixture, which is cooled to interior temperature, is filtered under MTBE (3 parts of volumes) washing to solution, obtains white crystalline solid shape ER-807245 (1.12 parts of weight, 0.97 equivalent).

In pre-dried inerted reactor 1, ER-807245 (1 part of weight, 1 equivalent) and anhydrous DMF (4 parts of volumes) is added.Under stirring, heating the mixture to interior temperature is 95 DEG C.Heating response device 2 to interior temperature is 90 DEG C, and in the reactor, D or L- valerian ammonia alcohol (1.05 equivalents, 0.61 part of weight) is dissolved in anhydrous DMF (1.3 parts of volumes).The content of reactor 2 is transferred in the reactor 1 that interior temperature is 90 DEG C.It was found that there is carbon dioxide evolution, discharge reaction solution is lost by nitrogen.After being stirred to react solution at 90 DEG C 3 hours, being cooled to interior temperature is 65 DEG C.Then, water (the 2 parts of volumes) suspension of lithium hydroxide (0.47 part of weight, 2 equivalents) is added in reactor 1, temperature is stirred suspension 1 hour lower in 65 DEG C.Water (5 parts of volumes) is added in the reactor, is cooled within 3 hours 5 DEG C of interior Wen Weiyue.In about 5 DEG C under temperature, stirs mixture 8 hours, in the case where water (2 × 4 parts of volumes) and normal heptane (2 × 3 parts of volumes) successively wash, target product is collected by filtration.Nitrogen stream desciccate is used under vacuum, at 35 DEG C 24 hours or until KF≤250ppm, obtains the ER-806628 or ER-808056 (0.80 part of weight, 0.60 equivalent) of solid crystal shape.

Under nitrogen, ER-806628 or ER-808056 (1 part of weight, 1 equivalent), pyridine (3 parts of weight, 11.4 equivalents) and DMAP (0.03 part of weight, 0.05 equivalent) are added in pre-dried inerted reactor.By reaction solution be cooled to interior temperature be -10 DEG C after so that interior temperature lower than 15 DEG C speed addition mesyl chloride (1.46 parts of weight, 3 equivalents).After sample-adding, it is stirred to react under temperature liquid 1 hour in 0-15 DEG C, was then to slowly warm up to 25 DEG C through 2 hours.So that MTBE (2.6 parts of volumes) and water (2 parts of volumes) is successively added in speed of the interior temperature no more than 35 DEG C.Two-phase mixture is titrated in batches with 6N hydrochloric acid (about 1.9 parts of volumes), until the pH of water layer is about 3-5.If pH is lower than 3,30 weight % aqueous sodium carbonates are added and carry out back titration to obtain required pH.It is each mutually to separate, separate water phase.All organic matters are mixed with water (0.7 part of volume), discard water phase.By MTBE air-distillation to about 2 parts of volumes, to obtain 55 DEG C of constant boiling points, and KF < 500ppm.MTBE is additionally incorporated if necessary.Cooling solution to interior temperature is 5-10 DEG C, and crystal seed induction crystallization is added when needing.It is added normal heptane (0.5 part of volume), is stirred mixture 18 hours at 5 DEG C, under normal heptane (2 × 3 parts of volumes) washing, ER-806629 or ER-807363 is collected by filtration.It concentrates the filtrate to 1/2 volume and is cooled to 0 DEG C, obtain second batch crystallization.Dry cake 18 hours under nitrogen.The ER-806629 slightly to weigh is added in pre-dried reactor, is added MTBE (3 parts of volumes).It is 45-50 DEG C that gained mixture, which is heated to interior temperature, is kept for 45 minutes, then slowly cooled to 5 DEG C through 3 hours, crystal seed can be added when needing.It is added normal heptane (0.5 part of volume), lower stirring mixture 18 hours of temperature in 5 DEG C.Solid product is collected by filtration under normal heptane (2 × 3 parts of volumes) washing, is then dried in vacuo 24 hours at 35 DEG C, obtains the ER-806629 or ER-807363 (1.7 parts of weight, 0.57 equivalent) of solid crystal shape.

The assembling of B.F-1a and F-2a and the formation of molecule inner ether:

ER-807363 (1.82 parts of weight, 3.55 equivalents) are added in appropriately sized reactor 1, carry out nitrogen displacement.Anhydrous THF (15 parts of volumes) is added.In reactor 2, ER-806067 (F-1a, 1.14 parts of weight, 1.1 equivalents) and ER-805973 (F-2a, 1 part of weight, 1 equivalent) are mixed, are allowed to be dissolved in anhydrous THF (6.3 parts of volumes).Under stirring, which is sprayed nitrogen 30-45 minutes.Under inert atmosphere, CrCl is added in reactor 2₂(0.75 part of weight, 3.55 equivalents), being then heated to interior temperature is 30 DEG C.So that triethylamine (0.62 part of weight, 3.55 equivalents) are added in speed of the interior temperature no more than 45 DEG C in reactor 2.After sample-adding, temperature 1 hour in 30 DEG C is kept.After 1 hour, reactor 2 is cooled to 0 DEG C, NiCl is added₂(0.02 part of weight, 0.1 equivalent) forms inert environments, so that reaction solution is warming up to room temperature after the content of reactor 1 is added.Cooling reactor 2 to interior temperature is 0 DEG C, then so that ethylenediamine (1.2 parts of volumes, 10 equivalents) are added in speed of the interior temperature no more than 10 DEG C.Note: discovery heat release.It is stirred to react liquid 1 hour, water (8 parts of volumes) and normal heptane (20 parts of volumes) is then added, stir two-phase mixture 4 minutes, layering.Organic layer is separated, water layer is stripped with MTBE (20 parts of volumes).Combined organic layer in vacuo is concentrated into crude product oil, then forms azeotropic mixture with anhydrous THF (2 × 10.5 parts of volumes).Crude product is dissolved in anhydrous THF (4.5 parts of volumes), is then saved at -20 DEG C, until being used for next step.

ER-808227/TFH solution obtained to previous step carries out KF analysis.If KF < 1000ppm, the reaction was continued.If KF > 1000ppm, azeotroped in vacuo is carried out with anhydrous THF (4.1 parts of volumes).Azeotropic operation is repeated until meeting the requirements.Contain the crude product ER-808227 for being dissolved in anhydrous THF (4.1 parts of volumes) in satisfactory end solution.Once meeting the requirements, anhydrous THF (106 parts of volumes) and the obtained ER-808227/TFH solution of previous step are added in appropriately sized inerted reactor.Cooling reactor to interior temperature is -15 DEG C to -20 DEG C, then so that toluene (9.1 parts of weight, 3.0 equivalents) solution of 0.5M KHMDS is added in speed of the interior temperature no more than -12 DEG C.Reaction completely is promoted to need the KHMDS of about 4.5 equivalents.In 0 DEG C under temperature, reaction solution is added in half saturated ammonium chloride (40 parts of volumes) and carries out inverse quenching.It is added normal heptane (80 parts of volumes), stirring is layered after 2-5 minutes.Organic layer is separated, water layer is stripped with MTBE (70 parts of volumes), is then combined with organic layer and is washed with saturated sodium chloride solution (70 parts of volumes).Separation organic layer is simultaneously concentrated in vacuo.Normal heptane (60 parts of volumes) is added in thick concentrate.Note: ER-808373 is precipitated out from solution.Resulting suspension is filtered, washs solid with normal heptane (20 parts of volumes).It is concentrated in vacuo filtrate, obtains the crude product ER-806746 (about 4 parts of weight) of brown oil.Note: when other ER-808373 is precipitated out from solution, repeating filter operation.Crude product ER-806746 is through SiO₂Column chromatography purifies to obtain the ER-804027 (1.16 parts of weight, 0.55 equivalent) of glassy yellow (clear yellowish) oily.Chromatographic run is as follows: rinsing pillar with enough MTBE first to remove moisture, then rinses pillar with heptane to remove MTBE.ER-806746 is filled into column in the form of n-heptane solution, is successively eluted out it from column with heptane/MTBE (5: 1) and heptane/MTBE (4: 1), with the fraction at UV detector monitoring 230nm.

ER-804027 (1 part of weight, 1 equivalent) and anhydrous methylene chloride (7.6 parts of volumes) are added in the reactor.Cooling reactor to interior temperature is -78 DEG C, then so that methylene chloride (3.0 parts of weight, 2.25 equivalents) solution of 1M DIBALH is added in speed of the interior temperature no more than -60 DEG C.So that methanol (0.1 part of volume) is added in speed of the interior temperature no more than -60 DEG C.Note: selecting hydrogen, and be diluted with nitrogen stream.After sample-adding, 1N hydrochloric acid (10.6 parts of volumes) and MTBE (25 parts of volumes) are added after so that mixture is warming up to room temperature.Stirring mixture 20 minutes, layering.Organic layer is separated, water layer is stripped with MTBE (15.3 parts of volumes).Merge organic layer, is concentrated in vacuo after being washed respectively with water (3 parts of volumes), saturated sodium bicarbonate (3 parts of volumes) and saturated sodium-chloride (3 parts of volumes).Thick concentrate is through SiO₂Column chromatography purifies to obtain the ER-804028 (0.84 part of weight, 0.93 equivalent) of white foam.

C.F-3a's is incorporated to and is converted into B-1939

Under argon atmospher, ER-803895 (F-3a) is dissolved in dry toluene (14 parts of weight) and is cooled to < -75 DEG C.Then so that DIBALH (1.5M toluene solution, 0.95 part of weight, 1.3 equivalents) is added in < -70 DEG C of internal reaction temperature of speed.It stirs resulting mixture to be quenched after 30 minutes with anhydrous methanol (0.13 part of weight, 3.2 equivalents), during which keeps < -65 DEG C of internal reaction temperature.So that reaction mixture is warming up to -10 DEG C, is transferred in the post-processing container containing 1N salt water (10.2 parts of weight) under MTBE (3.74 parts of weight) flushing.Stirring mixture 30 minutes excludes water layer.Organic phase is successively washed with 1N hydrochloric acid (10.2 parts of weight), water (10 parts of weight), saturated sodium bicarbonate aqueous solution (10 parts of weight) and salt water (10 parts of weight), is then concentrated under reduced pressure.Concentrate obtains ER-803896 (0.96 part of weight, 93% yield) through silica gel chromatography.The product is stored under argon gas at -20 DEG C.

At 0 DEG C, with n-BuLi (1.6M hexane solution, 1.02 part weight, 1.5 parts of volumes, 2.05 equivalents) so that speed of the interior temperature no more than 5 DEG C handles sulfone ER-804028 (1.0 parts of weight of azeotropic drying, 1 equivalent) anhydrous tetrahydro furan (5 parts of volumes, 4.45 parts of weight) solution.Lower stirring mixture 10 minutes of temperature, are subsequently cooled to < -75 DEG C in 0-5 DEG C.< -75 DEG C are cooled to after the aldehyde ER-803896 (1.07 parts of weight, 1.23 equivalents) of azeotropic drying is dissolved in anhydrous hexane (3.53 parts of weight, 5.35 parts of volumes).Inside temperature≤- 65 DEG C under the conditions of, aldehyde solution is added in ER-804028 anion liquid by casing.It in -78 DEG C under temperature, stirs mixture 45 minutes, saturated ammonium chloride (5 parts of volumes), methyl tertiary butyl ether(MTBE) (10 parts of volumes) and water (5 parts of volumes) is then added and is quenched.Water layer is discarded, organic layer is concentrated under reduced pressure.Thick material obtains ER-804029 (84%, 1.57 parts of weight) through C-18 Reverse phase chromatography.

Sulfone-glycol ER-804029 (1 part of weight, 1 equivalent) is dissolved in wet methylene chloride (7.4 parts of volumes, aqueous 0.04 weight %), is placed in 20-25 DEG C of water-bath.It is primary that Dess-Martin reagent (0.67 part of weight, 2.5 equivalents) are added.Reaction mixture is quenched with saturated sodium bicarbonate (10 parts of volumes) and 10 weight % sodium sulfite aqueous solutions (10 parts of volumes), is stirred 30 minutes.Mixture is diluted with saturated sodium-chloride (10 parts of volumes), is extracted with MTBE (25 parts of volumes).Water layer is discarded, concentration of organic layers obtains ER-804030 (0.9 part of weight, 90%) through silica gel chromatography.At -20 DEG C, which is stored under inert atmosphere.

Under inert atmosphere, samarium diodide solution (2.5 equivalent) is added in pre-dried reactor, which is cooled to < -70 DEG C of interior temperature.ER-804030 (1 part of weight) is dissolved in anhydrous methanol (4.1 parts of weight) and anhydrous THF (2.3 parts of weight), is subsequently cooled to < -70 DEG C.So that ER-804030 is added in cold samarium solution in speed of the interior temperature no more than -70 DEG C.Interior temperature is no more than under the conditions of -65 DEG C, with potassium carbonate/Rochelle salt/water (1/10/100；W/w/volume, 15 parts of volumes) and MTBE (5 parts of volumes) quenching reaction.After post-treatment solution adds, reaction solution is made to be warming up to room temperature, mixture is transferred in another container with post-treatment solution (20 parts of volumes are rinsed) and MTBE (20 parts of volumes are rinsed).Water layer is discarded, organic layer is evaporated, residue obtains ER-118049 (0.77 part of weight, 85%) through silica gel chromatography.The product is stored under inert atmosphere at -20 DEG C.

(S)-ligand ER-807363 (2.05 parts of weight) is added in pre-dried reactor, carries out nitrogen displacement.It is primary that CrCl is added₂Anhydrous acetonitrile (21.5 parts of weight) is added afterwards in (0.85 part of weight, 10 equivalents), is warming up to mixture and is maintained at 30 DEG C -35 DEG C.It is primary that triethylamine (0.7 part of weight, 0.96 part of volume, 10 equivalents) is added, it stirs mixture 1 hour.It is primary that NiCl is added₂(0.09 part of weight, 1 equivalent) anhydrous THF (2.43 parts of weight, 2.73 parts of volumes) solution of keto-aldehyde ER-118049 was added by 30 minutes.Heat source is removed, heptane (20.5 parts of weight, 30 parts of volumes) and Celite is added(1.5 parts of weight).Stirring mixture 5 minutes, is filtered through CeliteIt pads (15 parts of weight) and rinses Celite with heptane (7.3 parts of volumes) and acetonitrile (5 parts of volumes)Pad.Filtrate is transferred in another funnel, sub-cloud is removed.When needing, combined heptane layer is washed with acetonitrile (most 47.2 parts of weight, most 60 parts of volumes).Heptane layer is evaporated under reduced pressure, product obtains ER-118047/048 (0.64 part of weight, 70%) through silica gel chromatography.

Allyl alcohol ER-118047/048 is dissolved in methylene chloride (containing 0.04 weight % water, 9 volumes), by reactor as in water-bath (20 DEG C), with Dess-Martin reagent (0.48 part of weight, 1.5 equivalents) processing solution.It stirs 20 minutes after reaction mixture saturated sodium bicarbonate aqueous solution (9 parts of volumes) and 10% sodium sulfite aqueous solution (9 parts of volumes) processing, is transferred in another funnel by DCM (10 parts of volumes).Discard water layer, evaporation organic layer to residue.Thick material (is pre-processed, substance is filled column with 1: 1DCM/ heptane, is then eluted with 10/10/1 heptane/DCM/MTBE) through purification by flash chromatography with the DCM/ heptane (1: 1 volume ratio) of 3 column volumes.The fraction containing product is concentrated, is stored under inert atmosphere at -20 DEG C.

Alternatively, ER-118047/48 is oxidized to diketone ER-118046 as follows.ER-118047/48 (1 part of weight, 1.0 equivalents) are added in flask, toluene (10 parts of volumes) and DMSO (0.15 part of weight, 2.5 equivalents) are added at room temperature.It is added triethylamine (0.31 part of weight, 4.0 equivalents), solution is cooled to -15 DEG C.Pure TCAA (0.33 part of weight, 1.4 equivalents) are added, reaction solution is made to be warming up to 0 DEG C.Then it is stirred 10 minutes at 0 DEG C.Reaction solution is again stirring for be quenched with IPA (0.15 part of volume) after ten minutes.Liquid is stirred to react at 0 DEG C 10 minutes.Through 2 minutes addition 1N hydrochloric acid (5 parts of volumes), reaction solution is made to be warming up to room temperature and be diluted with MTBE (5 parts of volumes).Two hyaline layers are formed, water layer is removed.Organic layer is washed with 5% bicarbonate (aqueous solution) of 5 parts of volumes, is concentrated into deep yellow oil with Rotary Evaporators, through silica gel chromatography (91% separation yield).

At room temperature, salt imidazole acid (0.39 part of weight, 5 equivalents) and the THF solution (7.6 parts of volumes, 10 equivalents) of 1M TBAF are successively added in appropriately sized reaction vessel (container A).Resulting mixture is stirred until uniform (15-30 minutes).ER-118046 (1 part of weight, 1 equivalent) and THF (33 parts of volumes) are added in second reaction vessel (container B).The content of container B is placed under inert atmosphere and is stirred until ER-118046 is completely dissolved.The content (TBAF/ imidazoles) of flask A is once added in flask B (ER-118046/THF).After 3-4 days, reaction solution is filled into column, carries out silica gel chromatography.

Under nitrogen atmosphere, by dry ER-118064 (F-12, wherein R¹For methoxyl group) residue is dissolved in anhydrous methylene chloride (28 parts of volumes), with PPTS (1.0 parts of weight, 5.2 equivalents) single treatment.After 30-90 minutes, reaction mixture is directly entered in suitable pillar from top loading, carries out silica gel chromatography.It is concentrated in vacuo target fraction ER-076349.All pure fractions are concentrated, azeotropic is twice in toluene (20 parts of volumes) for obtained substance, obtain frangible colorless solid/foam-like ER-076349 (0.44 part of weight, 0.79 equivalent (after residual toluene correction)).

In the dry reaction vessel (flask C) of cleaning, ER-076349 (1 part of weight, 1 equivalent) is dissolved in dry toluene (20 parts of volumes), is concentrated to dryness.The substance is dissolved in again in dry toluene (20 parts of volumes), is concentrated to dryness.The substance is dissolved in DCM (5 parts of volumes), solution is placed under argon atmospher.It is primary that trimethylpyridine (0.66 part of weight, 4.0 equivalents) are added.The primary DCM solution (in flask B) (5 moles of %) that pyridine is added.It is -20 to -25 DEG C that gained mixture in flask C, which is cooled to interior temperature,.Ts is added dropwise at temperature is lower than -16 DEG C in keeping₂The DCM solution (1.02 equivalent) of O.It is stirred to react at -20 to -25 DEG C after liquid 80 minutes and was warming up to 0 DEG C through 20 minutes, futher stirred 20 minutes.It is quenched and is reacted with water (2 parts of volumes).Water-bath is removed, so that reaction solution is warming up to room temperature (15-20 DEG C) and is stirred (20 minutes).Reaction solution is rinsed into bigger container with IPA (100 parts of volumes), ammonium hydroxide aqueous solution (100 parts of volumes) is added in reaction solution.It is stirred to react liquid at room temperature 15-36 hours, monitors the tosylate (ER-082892) of in-situ generation and the Disappearance Scenarios of epoxides (ER-809681).Under decompression, concentration of reaction solution is dry to doing or approaching.Obtained material is diluted with DCM (25-40 parts of volumes), is washed with the buffer (sodium bicarbonate/sodium carbonate (aqueous solution), 10 parts of volumes) of pH10.The DCM of 25 parts of volumes of water phase is stripped, and combined organic layer is concentrated to dryness.Gained unhindered amina is purified through silica gel chromatograph (using acetonitrile/water buffer as mobile phase).Combined fraction is concentrated under reduced pressure to remove acetonitrile.Gained water layer is diluted with DCM (40 parts of volumes) and the buffer stock liquid (pH10, sodium bicarbonate/sodium carbonate) of 30 parts of volumes.It is sufficiently mixed each layer and is separated.The DCM of 25 parts of volumes of water phase is stripped, and combined organic layer is concentrated to dryness.Gained unhindered amina is filtered into 3: 1DCM/ pentane solution through exquisiteness, is concentrated to dryness (0.80 part of weight) and obtains B-1939.

Although a large amount of embodiments of the invention are described in we, it is clear that can be converted to our basic embodiment to provide the other embodiment using the compounds of this invention and method.It is to be understood, therefore, that the scope of the present invention is defined by the attached claims, rather than it is limited by the specific embodiment that way of example is stated.

Claims

1. a kind of compound of formula F-4:

Wherein: PG¹、PG²And PG³Respectively stand alone as hydrogen or suitable hydroxyl protection base；

R¹For R or-OR；

Each R stands alone as hydrogen, C_1-4Halogenated aliphatic base, benzyl or C_1-4Aliphatic group；And

LG¹For suitable leaving group.

2, compound according to claim 1, wherein the compound is the compound of formula F-4 ':

3, compound according to claim 2, wherein R¹For OR, wherein R is hydrogen, methyl or benzyl.

4, compound according to claim 2, wherein PG¹And PG²It is hydrogen.

5, compound according to claim 2, wherein PG¹And PG²Respectively stand alone as suitable hydroxyl protection base.

6, compound according to claim 5, wherein PG¹And PG²One or two of together with the oxygen atom of each self-bonding be silyl ether or aryl alkyl ethers.

7, compound according to claim 6, wherein PG¹And PG²It is t-butyldimethylsilyl.

8, compound according to claim 5, wherein PG¹And PG²Glycerol protection base is formed together together with the oxygen atom in conjunction with them.

9, compound according to claim 8, wherein the glycerol protection base is cyclic ketal or ketal, silylene derivative, cyclic carbonate ester or ring borate.

10, compound according to claim 2, wherein LG¹For sulfonyloxy, the alkylsulfonyloxy optionally replaced, the alkenyl sulfonyloxy optionally replaced or the aryl-sulfonyl oxygen optionally replaced.

11, compound according to claim 10, wherein LG¹For mesyl or tosyl.

12, compound according to claim 2, wherein the compound are as follows: