CN1869008A - Preparation method of chiral non alpha amino acid derivative of simultaneously protected by hydroxyl group and amino group - Google Patents

Abstract

A (S or R)-2(or 3)-tert-butyloxy-(3+n)-(9-fluorenyl methoxy) carbonylamino fatty-chain carboxylic acid with protected hydroxy and amino is prepared from (S or R)-2(or 3)-hydroxy-(3+n)-amino fatty-chain carboxylic acid through reaction on dichlorosulfoxide in solvent to generate the hydrochloride of relative methylester, reacting on FmocONSu in solvent to generate (S or R)-2(or 3)-hydroxy-(3+n)-(9- fluorenylmethoxy) carbonylamino fatty-chain methyl carboxylate, reacting on isobutene in solvent to generate (S or R)-2(or 3)-tert-butyloxy-(3+n)-(9-fluorenyl methoxy) carbonylamino fatty-chain methyl carboxylate, hydrolyzing, acidifying to generate (S or R)-2(or 3)-tert-butyloxy-(3+n)-amino fatty-chain carboxylic acid, and reacting of FmocONSu in solvent.

Description

Hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection

Technical field

What the present invention relates to is the synthesizing of derivative that is connected with the chiral non alpha-amino-acid compound of amino and hydroxyl at same aliphatic chain. The amino of this compound is connected with the carbon atom of non-ring carbon atom or the ring except aromatic ring, and two on hydroxyl and the aliphatic chain is connected with three carbon atoms, and carboxyl is in the first place of carbochain. Has optically pure chiral non alpha-amino acid derivativges by the protection preparation to amino and hydroxyl. Particularly this invention is the preparation method of hydroxyl and the amino non-alpha-amino acid derivatives that can protect simultaneously.

Background technology

In polypeptide is synthetic, during especially the solid phase of polypeptide is synthesized, often use the non-a-amino acid of hydroxyl, and amino and hydroxyl to be protected simultaneously by other group temporarily usually. At present, in the solid phase of polypeptide was synthetic, what blocking group amino and hydroxyl was used often was respectively: 9-fluorenylmethyloxycarbonyl (9-fluorenylmethoxycarbonyl; be called for short Fmoc; descend together) and the tert-butyl group (tert-butyl is called for short t-Bu, and is lower same). Because; the amino of Fmoc protection is unstable and the hydroxyl t-Bu protection is unstable under sour condition under the alkali condition; so; difficult the realization protected amino and the hydroxyl of non-a-amino acid simultaneously with Fmoc and t-Bu; especially to containing the non-a-amino acid that replaces with hydroxyl of chirality, more difficult realization is protected simultaneously amino and hydroxyl with Fmoc and t-Bu and is not produced racemization. The synthetic method that has been reported and technology:

1. although have an alpha-aminobetahydroxypropionic acid d amino and that hydroxyl is protected simultaneously with what following method can be synthesized^1，2, its synthetic route is:

But, in this way and experiment condition, can not or be difficult to synthesize following target compound:

Because intermediate (c) is very unstable, at excessive silylating reagent CF₃SO ₃Si(CH ₃) ₃The strong acid CF that hydrolysis produces₃SO ₃The H effect is lower, and tertbutyl ether and trimethylsilyl group almost all are hydrolyzed into corresponding hydroxyl and carboxyl and can not obtain target compound among the present invention. In addition, CF₃SO ₃Si(CH ₃) ₃Relatively more expensive, very easily the moisture absorption is decomposed, so difficult the preservation and operating difficulties, this goes on foot severe reaction conditions; In reaction, must use excessive CF₃SO ₃Si(CH ₃) ₃So,, make the complicated and difficult control of last handling process, be difficult to obtain the product of high yield.

2. use the method for isobutene/concentrated sulfuric acid/carrene, prepare the hydroxy-amino-acid of other tert-butyl group protection³Reach the amino acid that hydroxyl and carboxyl are protected by the tert-butyl group simultaneously⁴, its synthetic route is respectively:

But, in these reports, be not product and the corresponding intermediate among synthetic the present invention.

3. also useful FmocONSu reagent is protected amino report in amino acid under alkali condition³Its synthetic route is:

But obviously, their reaction raw materials is different from the present invention with product, reacts detailed process conditions and also is not quite similar. In sum, prior art is not yet found preparation method among hydroxyl and the amino target chipal compounds (comprising the intermediate that it is corresponding) that can protect simultaneously and the present invention thereof.

List of references

1.A.Trzeciak，W.Bannwarth，Synthesis，1996，1433.

2.D.R.Bolin et al，In.J.Peptide Protein Res.1989，33，353.

3.J.G.Adamson et al，J.Org.Chem.1991，56，3447.

4.H.C.Beyerman，J.S.Bontekoe，Rec.trav.Chim.1962，81，691.

5.B.S.Axelsson et al，J.Chem.Soc.Perkin Trans.I，1994，807.

Summary of the invention

For the defective that prior art exists, the invention provides one can be from 2 (or 3)-hydroxyls with optically active S or R configuration-(the 3+n)-synthetic corresponding hydroxyl with optically active S or R configuration and amino simultaneously process route and method of the amino acid derivativges of protection of amino fatty chain carboxylic acid.

The general formula of target chipal compounds of the present invention is:

Wherein: the carbon of asterisk is asymmetric carbon atom, i.e. S or R configuration; N represents natural number, n=1～10; The hydrogen atom of the carbon atom combination on the main chain can be replaced with D atom; Fmoc is that 9-fluorenylmethyloxycarbonyl (9-fluorenylmethoxycarbonyl) and t-Bu are the tert-butyl group (tert-butyl).

Its synthetic route is:

Synthesizing of synthetic route I[general formula (1) compound]:

Synthesizing of synthetic route II[general formula (2) compound]:

1. in general formula (1), the 2-tert-butoxy of S or R configuration-(3+n)-(9-fluorenes methoxyl group) carbonyl amino butyric acid and in general formula (2), the carbonyl amino butyric acid of the 3-tert-butoxy of S or R configuration-(3+n)-(9-fluorenes methoxyl group), n 〉=1.

2. in general formula (1), when n=1, (S)-2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1a) and (R)-2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1b) and their intermediate (see Table 1 and synthetic route I): (S)-synthetic (4a) of 2-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (S)-synthetic (5a) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (S)-synthetic (6a) of 2-tert-butoxy-4-Aminobutanoicacid, (R)-synthetic (4b) of 2-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (R)-synthetic (5b) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, and (R)-synthetic (6b) of 2-tert-butoxy-4-Aminobutanoicacid. In general formula (2), when n=1, (S)-3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1c) and (R)-3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1d) and their intermediate (see Table 1 and synthetic route II): (S)-synthetic (3c) of 3-hydroxyl-4-Aminobutanoicacid methyl ester hydrochloride, (S)-synthetic (4c) of 3-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (S)-synthetic (5c) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (S)-synthetic (6c) of 3-tert-butoxy-4-Aminobutanoicacid, (R)-synthetic (3d) of 3-hydroxyl-4-Aminobutanoicacid methyl ester hydrochloride, (R)-synthetic (4d) of 3-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, (R)-synthetic (5d) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate, and (R)-synthetic (6d) of 3-tert-butoxy-4-Aminobutanoicacid.

3. in general formula (1), the 2-tert-butoxy of S or R configuration-(3+n)-(9-fluorenes methoxyl group) carbonyl amino butyric acid and in general formula (2), the preparation method of the 3-tert-butoxy of S or R configuration-(3+n)-(9-fluorenes methoxyl group) carbonyl amino butyric acid and their intermediate. General step is: (i) take 2 (or 3)-hydroxyls of S or R configuration-(3+n)-amino fatty chain carboxylic acid as initiation material, reaction generates the hydrochloride of corresponding (S or R)-2 (or 3)-hydroxyls-(3+n)-amino fatty chain carboxylic acid's methyl esters in the methanol solution of thionyl chloride; (ii) above-mentioned product is 1 of carbonic acid amber uncle acid imide 9-fluorene methyl ester, in the solution of 6-dioxane, be preferably under the existence of sodium acid carbonate, at room temperature stir generate the carbonyl amino aliphatic chain carboxylate methyl ester of (S or R)-2 (or 3)-hydroxyls-(3+n)-(9-fluorenes methoxyl group); (iii) above-mentioned product is in the solution of the carrene of isobutene, be preferably under the existence of the concentrated sulfuric acid, at room temperature stir generate the carbonyl amino aliphatic chain carboxylate methyl ester of (S or R)-2 (or 3)-tert-butoxies-(3+n)-(9-fluorenes methoxyl group); (iv) after above-mentioned product is hydrolyzed, generate (S or R)-2 (or 3)-tert-butoxies-(3+n)-amino fatty chain carboxylic acid through acidifying under alkali condition; (v) last, above-mentioned product is in the solution of the glycol dimethyl ether of carbonic acid amber uncle acid imide 9-fluorene methyl ester, under the alkali condition, stir to generate the carbonyl amino aliphatic chain carboxylic acid of final products (S or R)-2 (or 3)-tert-butoxies-(3+n)-(9-fluorenes methoxyl group).

4. in general formula (1), when n=1, (S)-preparation method of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1a) and (R)-2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1b) and their intermediate (seeing synthetic route I); In general formula (2), when n=1, (S)-preparation method's (seeing synthetic route II) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1c) and (R)-3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1d) and their intermediate.

In said synthesis route: 1) a and b represent respectively the compound of S and R configuration, such as: 2a represents (S)-2-hydroxy-4-amino-butyrate; 2b represents (R)-2-hydroxy-4-amino-butyrate. 2) FmocONSu is: carbonic acid amber uncle acid imide 9-fluorene methyl ester. 3) Fmoc is: 9-fluorenylmethyloxycarbonyl. 4) t-Bu is: the tert-butyl group. 5) in the first step of present technique route, the thionyl chloride molar equivalent between 1.8～3.5, preferably 2.5～2.8; The amount of methyl alcohol is at 1.2～3.0mL/mmol raw material, preferably 1.5～1.8mL/mmol; Reaction temperature was preferably reacted 15 minutes at zero centigrade first between zero centigrade and 25 degree, then in 22 degree reactions Celsius, until till the raw material complete reaction. 6) in the second step of present technique route, by 3a, 3b, 3c and 3d be synthetic 4a respectively under alkali condition, 4b, and 4c and 4d, this alkali is NaOH, sodium carbonate, sodium acid carbonate, but sodium acid carbonate preferably; Solvent is DMF, acetonitrile, Isosorbide-5-Nitrae-dioxane, glycol dimethyl ether, but Isosorbide-5-Nitrae-dioxane preferably. The FmocONSu molar equivalent between 1.0～2.0, preferably 1.0～1.1; NaHCO₃Molar equivalent between 1.0～3.0, preferably 1.2; The amount of solvent is at 1.5～6.0mL/mmol raw material, preferably 2.5～3.0mL/mmol; Reaction temperature is between zero centigrade and 45 degree, and first reaction raw material in 30 minutes when zero centigrade preferably is then 22 degree reactions Celsius 25～30 hours. 7) in the 3rd step of present technique route, by 4a, 4b, 4c and 4d be synthetic 5a respectively under acid condition, 5b, and 5c and 5d, this acid is the concentrated sulfuric acid preferably; Solvent is carrene, Isosorbide-5-Nitrae-dioxane, glycol dimethyl ether, but carrene preferably. The amount of the concentrated sulfuric acid is at 20～300 μ L/mmol raw materials, preferably 120～125 μ L/mmol; The amount of carrene is at 4～80mL/mmol raw material, preferably 50mL/mmol; The amount of isobutene is in the amount of the volume of 0.5～10 times carrene, preferably 1.5～2.0 times; At the following reaction raw material of zero centigrade, then between zero centigrade and 25 degree, react first, preferably first at the following reaction raw material of zero centigrade, stirred 30 minutes, then continue 22 degree reactions Celsius 25-70 hour. 8) the present technique route the 4th the step in, alkalescence and acid condition under by 5a, 5b, 5c and 5d be synthetic 6a respectively, 6b, 6c and 6d, this alkali is NaOH, sodium carbonate, but NaOH preferably, and this acid is inorganic acid, such as, dilute sulfuric acid, watery hydrochloric acid, phosphoric acid,diluted, but 10% hydrochloric acid preferably; Solvent is DMF, Isosorbide-5-Nitrae-dioxane, glycol dimethyl ether, but Isosorbide-5-Nitrae-dioxane preferably. The concentration of NaOH: 1～5M, but 2M preferably; The amount of NaOH is at 0.2～20mL/mmol raw material, preferably 0.5～3.0mL/mmol; The amount of solvent is at 5～10mL/mmol; Reaction temperature is between 15 degree Celsius and 60 degree, and preferably 40 to 45 degree reactions are 1 hour. 9) in the 5th step of present technique route, by 6a, 6b, 6c and 6d be synthetic 1a respectively under alkali condition, 1b, and 1c and 1d, this alkali is inorganic base, such as, NaOH, sodium carbonate, sodium acid carbonate, but sodium acid carbonate preferably. Solvent is DMF, acetonitrile, Isosorbide-5-Nitrae-dioxane, glycol dimethyl ether, but glycol dimethyl ether preferably. The FmocONSu molar equivalent between 1.0～2.0, preferably 1.2; NaHCO₃Molar equivalent between 1.0～3.0, preferably 1.2; The amount of solvent is at 1～15 mL/mmol raw material, preferably 4～6mL/mmol; Reaction temperature is between zero centigrade and 45 degree, and first reaction raw material in a hour when zero centigrade preferably is then 22 degree reactions Celsius ten hours.

Compared with prior art the invention solves following problem: 1; can comprise 1a from 2 (or 3)-hydroxyls with optically active S or R configuration-(the 3+n)-synthetic corresponding hydroxyl with optically active S or R configuration and amino simultaneously amino acid derived compounds (seeing general formula (1) and (2)) of protection of amino fatty chain carboxylic acid; 1b; 1c and 1d, the optical purity of product is higher than 97%. 2, by the enforcement of the inventive method, can in a chiral non alpha-amino acid, protect simultaneously with the t-Bu group with Fmoc and hydroxyl amino. The particularly important is: this target compound synthetic be the hydroxylic moiety of and t-Bu radical protection unstable to alkali in the amino part of Fmoc radical protection to acid unstable and carboxyl, amino is realized under the hydroxyl coordinating protection optimal conditions. 3, synthetic method of the present invention has each step reaction and easily realizes the easy purifying of intermediate, productive rate high; And in synthetic target product, several very useful high optically pure chiral intermediate compounds have also been synthesized. 4, synthetic method of the present invention is to synthetic other hydroxyls and amino simultaneously respectively with the chiral amino acid derivative with hydroxyl modified of t-Bu group and Fmoc radical protection or to the synthetic tangible meaning of similar chiral amino acid derivative.

The specific embodiment

The compound that synthesizes in the embodiment of the invention and structure and the English name of raw material are as shown in table 1 below:

Example 1:(S)-preparation of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1a)

(1) (S)-synthetic (3a) of 2-hydroxy-4-amino-butyrate methyl ester hydrochloride⁵ With 4g (33.58mmol) (S)-2-hydroxyl-4-Aminobutanoicacid 2a, (95%, thionyl chloride 93.35mmol) and 60mL methyl alcohol were 0 ℃ of lower mix and blend 15 minutes for 7mL, then, continue stirring reaction under the room temperature, until raw material complete reaction (TLC detection, CH₂Cl ₂/ MeOH/HOAc=2/2/1). Then, the solvent of this reactant mixture is removed by Rotary Evaporators. Then, with the mixed solvent of methyl alcohol and carbon tetrachloride (1: 1, v/v) behind the resulting solid of dissolving, solvent is rotated again evaporimeter and removes, same operation is carried out several times. At last, provide the solid product 3a (productive rate: 99.2%) .TLC R of 5.66g white_f 0.50(CH ₂Cl ₂/MeOH/HOAc，2/2/1，v/v). ¹H-NMR (400MHz，D ₂O)：δ4.33(dd， ³J _H，H＝8.4Hz， ³J _H，H＝4.0Hz，1H，2-H)，3.66(s，3H， 1-OCH ₃)，3.21(d， ³J _H，H＝1.2Hz，1H，2-OH)，3.04(m，2H，4-H)，2.08 & 1.91(m，2H， 3-H)。ESI Mass：C ₅H ₁₂ClNO ₃Calculated value m/z, 169.15; Measured value, 133.10[M-HCl+H]⁺。

(2) (S)-synthetic (4a) of 2-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. 3a with 212mg (1.25 mmol), the carbonic acid amber uncle acid imide 9-fluorene methyl ester of 430mg (1.28mmol), 3.75mL 1,4-dioxane and 2.50mL sodium bicarbonate solution (5% aqueous solution) were 0 ℃ of lower mix and blend 30 minutes, then, continued stirring reaction 30 hours under the room temperature, (TLC detects, CH until raw material almost reacts completely₂Cl ₂/ MeOH=19/1). Then, add frozen water in this mixture, after 2 hours, precipitation generates, and filters, and reclaims filter cake also by silica gel column chromatography products of separated (eluent: CH₂Cl ₂/ MeOH=50/1-19/1, v/v). The final solid product 4a (productive rate: 75.2%) .TLC R that obtains 333.5mg white_f 0.30 (CH ₂Cl ₂/ MeOH, 19/1, v/v). analyze HPLC:t_R=22.70min (150 * 10mm ChemcoPak Chemcoband 5-ODS-H reversed-phase column, elution requirement: 0.1% aqueous acetic acid and acetonitrile are done eluent, and with 0～100% acetonitrile gradient, 30 minutes, wavelength 254nm detected. The testing conditions of other compound Hs PLC is identical with it, unless Special Statement. ).¹H-NMR(400MHz，DMSO-d ₆)：δ7.87(d， ³J _H，H＝7.6 Hz，2H，Fluoren.-H)，7.67(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.6Hz， 2H，Fluoren.-H)，7.31(t， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.28(t， ³J _H，H＝6.4Hz，1H， 4-NH)，4.27(d， ³J _H，H＝6.4Hz，2H，CH ₂-O)，4.19(t， ³J _H，H＝6.4Hz，1H，Fluoren.-H)， 4.05(dd， ³J _H，H＝8.4Hz， ³J _H，H＝4.0Hz，1H，2-H)，3.62(s，3H，1-OCH ₃)，3.07(quartet， ³J _H，H＝6.4Hz，2H，4-H)，1.80 & 1.63(m，2H，3-H)。ESI Mass：C ₂₀H ₂₁NO ₅Calculated value m/z, 355.14; Measured value, 356.12[M+H]⁺。

(3) (S)-synthetic (5a) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. With the 4a of 71.0mg (0.20mmol), 20mL isobutene, the concentrated sulfuric acid of 25 μ L and 10mL carrene are at the following mix and blend 30min of zero centigrade, and then, at room temperature, after reaction was carried out 20 hours, TLC detected (CH₂Cl ₂/ MeOH=100/1) reaction is until after raw material almost reacts completely, carefully remove excessive isobutene. Then, this mixed liquor is through 10% sodium bicarbonate aqueous solution, and saturated salt is washed, dried over sodium sulfate, (eluent: CH after silica gel column chromatography separates₂Cl ₂/ MeOH=150/1-100/1, v/v), finally obtain the milky product 5a (productive rate: 88.3%) .TLC R of 72.7mg oily_f 0.33(CH ₂Cl ₂/ MeOH, 100/1, v/v). analyze HPLC:t_R＝25.9min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.87(d， ³J _H，H＝7.2Hz，2H， Fluoren.-H)，7.66(d， ³J _H，H＝6.8Hz，2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.6Hz，2H， Fluoren.-H)，7.32(td， ³J _H，H＝7.6Hz， ⁴J _H，H＝1.2Hz，2H，Fluoren.-H)，7.25(t， ³J _H，H＝ 5.6Hz，1H，4-NH)，4.28(d， ³J _H，H＝6.8Hz，2H，CH ₂-O)，4.19(t， ³J _H，H＝6.8Hz，1H， Fluoren.-H)，4.07(dd， ³J _H，H＝8.8Hz， ³J _H，H＝4.0Hz，1H，2-H)，3.62(s，3H，1-OCH ₃)， 3.06(m，2H，4-H)，1.70-1.61(m，2H，3-H)，1.08(s，9H，2-OC(CH ₃) ₃)。ESI Mass： C ₂₄H ₂₉NO ₅Calculated value m/z, 411.20; Measured value, 412.16[M+H]⁺。

(4) (S)-synthetic (6a) of 2-tert-butoxy-4-Aminobutanoicacid. With the 5a of 176.6mg (0.43mmol), 1.25mL sodium hydrate aqueous solution (2M), 3mL 1, after the 4-dioxane mixed, after 1 hour, this reactant liquor was extremely neutral through 10% salt acid for adjusting pH 45 ℃ of lower stirrings, then, solvent is removed by rotary evaporation in vacuo. Resulting solid dissolves repeatedly through methyl alcohol, remove by filter insoluble matter after, separate this product (eluent: CH with silica gel column chromatography₂Cl ₂/ MeOH/HOAc=4/1/0.1-4/1/0.2, v/v), finally obtain 73.8mg white solid 6a (productive rate: 98.0%) .TLC R_f 0.52(CH ₂Cl ₂/MeOH/HOAc，2/2/0.5，v/v). ¹H-NMR(400 MHz，DMSO-d ₆)：δ4.20(dd， ³J _H，H＝8.4Hz， ³J _H，H＝4.0Hz，1H，2-H)，2.89(m，2H， 4-H)，1.78 & 1.65(m，2H，3-H)，1.09(s，9H，2-OC(CH ₃) ₃)。ESI Mass：C ₈H ₁₇NO ₃Calculated value m/z, 175.12; Measured value, 176.10[M+H]⁺；198.16[M+Na] ⁺。

(5) (S)-synthetic (1a) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid. 6a with 206.7mg (1.18 mmol), 477.6mg carbonic acid amber (1.42mmol) uncle acid imide 9-fluorene methyl ester, the glycol dimethyl ether of 6.6mL, 119.3mg sodium acid carbonate and the 2.0mL aqueous solution are at 0 ℃ of lower mix and blend after 60 minutes, at room temperature, continue reaction 10 hours. Then, this reactant liquor is through being acidified to pH=5, solvent is removed by rotary evaporation in vacuo, resulting solid dissolves repeatedly through ethyl acetate, after removing by filter insoluble matter, separate this product (eluent: ethyl acetate/cyclohexane/acetic acid=4/1/0-4/1/0.01, v/v), finally obtain 375.2mg white solid 1a (productive rate: 80.0%) .R with silica gel column chromatography_f0.28 (ethyl acetate/cyclohexane/acetic acid, 2/1/0.01, v/v). Analyze HPLC:t_R＝24.02min。 ¹H-NMR(400MHz，DMSO-d ₆)：δ12.3 (br，1H，1-COOH)，7.87(d， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.67(d， ³J _H，H＝7.2Hz， 2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.30(t， ³J _H，H＝7.6Hz，2H， Fluoren.-H)，7.25(t， ³J _H，H＝4.8Hz，1H，4-NH)，4.27(d， ³J _H，H＝6.4Hz，2H， CH ₂-O)，4.19(t， ³J _H，H＝6.8Hz，1H，Fluoren.-H)，3.94(dd， ³J _H，H＝8.4Hz， ³J _H，H＝3.6 Hz，1H，2-H)，3.04(quartet， ³J _H，H＝6.4Hz，2H，4-H)，1.71 & 1.60(m，2H，3-H)，1.13 (s，9H，2-OCMe ₃)。ESI-Mass：C ₂₃H ₂₇NO ₅Calculated value m/z, 397.19; Measured value, 398.26 [M+H]⁺，420.17[M+Na] ⁺。

Example 2:(R)-preparation of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1b)

Preparation 1b adopts the synthetic route of 1a equally, each intermediate 3b, and 4b, the data characterization of 5b and 6b and final products 1b is as follows:

(1) (R)-synthetic (3b) of 2-hydroxy-4-amino-butyrate methyl ester hydrochloride⁵ White solid. Productive rate: 97.8%. TLC R_f 0.50(CH ₂Cl ₂/MeOH/HOAc，2/2/1，v/v). ¹H-NMR(400MHz，D ₂O)：δ4.32 (dd， ³J _H，H＝8.4Hz， ³J _H，H＝4.0Hz，1H，2-H)，3.67(s，3H，1-OCH ₃)，3.22(d， ³J _H，H＝ 1.2Hz，1H，2-OH)，3.04(m，2H，4-H)，2.07 & 1.90(m，2H，3-H)。ESI Mass： C ₅H ₁₂ClNO ₃Calculated value m/z, 169.15; Measured value, 133.10[M-HCl+H]⁺。

(2) (R)-synthetic (4b) of 2-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. White solid. Productive rate: 78.1%.TLC R_f 0.30(CH ₂Cl ₂/ MeOH, 19/1, v/v). analyze HPLC:t_R＝22.70min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.87(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.67(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.30(t， ³J _H，H ＝7.6Hz，2H，Fluoren.-H)，7.29(t， ³J _H，H＝5.8Hz，1H，4-NH)，4.27(d， ³J _H，H＝6.4Hz， 2H，CH ₂-O)，4.18(t， ³J _H，H＝6.4Hz，1H，Fluoren.-H)，4.04(dd， ³J _H，H＝8.0Hz， ³J _H，H＝ 4.0Hz，1H，2-H)，3.62(s，3H，1-OCH ₃)，3.07(m，2H，4-H)，1.80&1.62(m，2H， 3-H)。ESI Mass：C ₂₀H ₂₁NO ₅Calculated value m/z, 355.14; Measured value, 356.10[M+H]⁺。

(3) (R)-synthetic (5b) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. Milky oily product. Productive rate: 90.7%.TLC R_f 0.33(CH ₂Cl ₂/ MeOH, 100/1, v/v). analyze HPLC:t_R＝25.9 min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.87(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.66 (d， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.32(t， ³J _H，H＝7.6Hz，2H 2H，Fluoren.-H)，7.25(t， ³J _H，H＝5.6Hz，1H，4-NH)，4.28(d， ³J _H，H＝ 6.8Hz，2H，CH ₂-O)，4.19(t， ³J _H，H＝6.8Hz，1H，Fluoren.-H)，4.09(dd， ³J _H，H＝8.4Hz， ³J _H，H＝4.0Hz，1H，2-H)，3.62(s，3H，1-OCH ₃)，3.04(m，2H，4-H)，1.73-1.61(m，2H， 3-H)，1.09(s，9H，2-OC(CH ₃) ₃)。ESI Mass：C ₂₄H ₂₉NO ₅Calculated value m/z, 411.20; Measured value, 412.27[M+H]⁺。

(4) (R)-synthetic (6b) of 2-tert-butoxy-4-Aminobutanoicacid. White solid. Productive rate: 98.0%.TLC R_f 0.52(CH ₂Cl ₂/MeOH/HOAc，2/2/0.5，v/v). ¹H-NMR(400MHz，DMSO-d ₆)：δ4.19 (dd， ³J _H，H＝8.0Hz， ³J _H，H＝4.0Hz，1H，2-H)，2.89(m，2H，4-H)，1.76 & 1.65(m，2H， 3-H)，1.10(s，9H，2-OC(CH ₃) ₃)。ESI Mass：C ₈H ₁₇NO ₃Calculated value m/z, 175.12; Measured value, 176.16[M+H]⁺；198.20[M+Na] ⁺。

(5) (R)-synthetic (1b) of 2-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid. White solid. Productive rate: 76.0%.R_f0.28 (ethyl acetate/cyclohexane/acetic acid, 2/1/0.01, v/v). Analyze HPLC:t_R＝24.00min。 ¹H-NMR(400MHz，DMSO-d ₆)：δ12.3(br，1H，1-COOH)，7.88(d， ³J _H，H＝7.6Hz， 2H，Fluoren.-H)，7.67(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.40(t， ³J _H，H＝7.4Hz，2H， Fluoren.-H)，7.31(t， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.26(t， ³J _H，H＝4.6Hz，1H， 4-NH)，4.28(m，2H，CH ₂-O)，4.19(t， ³J _H，H＝6.0Hz，1H，Fluoren.-H)，3.95(dd， ³J _H，H ＝8.2Hz， ³J _H，H＝3.6Hz，1H，2-H)，3.05(quartet， ³J _H，H＝6.4Hz，2H，4-H)，1.71 & 1.60(m，2H，3-H)，1.11(s，9H，2-OCMe ₃)。ESI-Mass：C ₂₃H ₂₇NO ₅Calculated value m/z, 397.19; Measured value, 398.23[M+H]⁺，420.16[M+Na] ⁺。

Example 3:(S)-preparation of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1c)

The synthetic route II of employing and synthetic route I all fours can prepare 1c; Each intermediate 3c, 4c, the data characterization of 5c and 6c and final products 1c is as follows:

(1) (S)-synthetic (3c) of 3-hydroxyl-4-Aminobutanoicacid methyl ester hydrochloride. White solid. Productive rate: 98.0%. TLC R_f 0.52(CH ₂Cl ₂/MeOH/HOAc，2/2/1，v/v). ¹H-NMR(400MHz，D ₂O)：δ4.28 (m，1H，3-H)，3.64(s，3H，1-OCH ₃)，3.35(d， ³J _H，H＝1.6Hz，1H，2-OH)，3.10(m，2H， 4-H)，2.28 & 2.11(m，2H，2-H)。ESI Mass：C ₅H ₁₂ClNO ₃Calculated value m/z, 169.15; Measured value, 133.15[M-HCl+H]⁺。

(2) (S)-synthetic (4c) of 3-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. White solid. Productive rate: 75.0%. TLC R_f 0.32(CH ₂Cl ₂/ MeOH, 19/1, v/v). analyze HPLC:t_R＝22.72min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.88(d， ³J _H，H＝7.6Hz，2H，Fluoren.-H)，7.69(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.41(t， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.32(t， ³J _H，H ＝7.2Hz，2H，Fluoren.-H)，7.30(s，1H，4-NH)，4.29(d， ³J _H，H＝7.2Hz，2H，CH ₂-O)， 4.20(t， ³J _H，H＝6.8Hz 1H，Fluoren.-H)，4.26(m，1H，3-H)，3.63(s，3H，1-OCH ₃)， 3.03(m，2H，4-H)，2.35(dd， ²J _H，H＝15.2Hz， ³J _H，H＝3.6Hz，1H，2-H _a)，2.10(dd， ²J _H，H ＝15.2Hz， ³J _H，H＝8.8Hz，1H，2-H _b)。ESI Mass：C ₂₀H ₂₁NO ₅Calculated value m/z, 355.14; Measured value, 356.10[M+H]⁺。

(3) (S)-synthetic (5c) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. Milky oily product. Productive rate: 92.0%. TLC R_f 0.36(CH ₂Cl ₂/ MeOH, 100/1, v/v). analyze HPLC:t_R＝ 26.5min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.89(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)， 7.68(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.41(t， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.33 (m，3H，Fluoren.-H+4-NH)，4.30(d， ³J _H，H＝6.8Hz，2H，CH ₂-O)，4.20(d， ³J _H，H＝6.8 Hz，1H，Fluoren.-H)，3.86(m，1H，3-H)，3.62(s，3H，1-OCH ₃)，3.08 & 2.90(m，2H， 4-H)，2.32(dd， ²J _H，H＝14.8Hz， ³J _H，H＝4.0Hz，1H，2-H _a)，2.10(dd， ²J _H，H＝14.8Hz， ³J _H，H＝6.0Hz，1H，2-H _b)，1.09(s，9H，3-OC(CH ₃) ₃)。ESI Mass：C ₂₄H ₂₉NO ₅Calculated value m/z, 411.20; Measured value, 412.14[M+H]⁺。

(4) (S)-synthetic (6c) of 3-tert-butoxy-4-Aminobutanoicacid. White solid. Productive rate: 97.9%. TLC R_f 0.50(CH ₂Cl ₂/MeOH/HOAc，2/2/0.5，v/v). ¹H-NMR(400MHz，DMSO-d ₆)：δ4.96 (m，1H，3-H)，2.96(dd， ²J _H，H＝13.2Hz， ³J _H，H＝2.8Hz，1H，4-H _a)，2.74(dd， ²J _H，H＝ 13.2Hz， ³J _H，H＝9.6Hz，1H，4-H _b)，2.20(d， ³J _H，H＝6.4Hz，2H，2-H)，1.08(s，9H， 3-OC(CH ₃) ₃)。ESI Mass：C ₈H ₁₇NO ₃Calculated value m/z, 175.12; Measured value, 176.14[M+H]⁺； 198.15[M+Na] ⁺。

(5) (S)-synthetic (1c) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid. White solid. Productive rate: 74.3%. R_f0.30 (ethyl acetate/cyclohexane/acetic acid, 2/1/0.01, v/v). Analyze HPLC:t_R＝23.92min。 ¹H-NMR(400MHz，DMSO-d ₆)：δ12.14(br，1H，1-COOH)，7.90(d， ³J _H，H＝7.2Hz， 2H，Fluoren.-H)，7.70(m，2H，Fluoren.-H)，7.42-7.33(m，5H，Fluoren.-H+4-NH)， 4.29(m，2H，CH ₂-O)，4.21(t， ³J _H，H＝5.6Hz，1H，Fluoren.-H)，3.89(br，1H，3-H)， 3.10 & 2.94(m，2H，4-H)，2.37(dd， ²J _H，H＝14.8Hz， ³J _H，H＝4.0Hz，1H，2-Ha)，2.19 (dd， ²J _H，H＝14.8Hz， ³J _H，H＝6.4Hz，1H，2-H _b)，1.12(s，9H，3-OCMe ₃)。ESI-Mass： C ₂₃H ₂₇NO ₅Calculated value m/z, 397.19; Measured value, 398.25[M+H]⁺，420.18[M+Na] ⁺, anion detects: 396.16[M-H]^-。

Example 4:(R)-preparation of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid (1d)

Adopt synthetic route II, also can prepare 1d; Each intermediate 3d, 4d, the data characterization of 5d and 6d and final products 1d is as follows:

(1) (R)-synthetic (3d) of 3-hydroxyl-4-Aminobutanoicacid methyl ester hydrochloride. White solid. Productive rate: 98.4%. TLC R_f 0.52(CH ₂Cl ₂/MeOH/HOAc，2/2/1，v/v). ¹H-NMR(400MHz，D ₂O)：δ4.27 (m，1H，3-H)，3.64(s，3H，1-OCH ₃)，3.34(d， ³J _H，H＝1.6Hz，1H，2-OH)，3.10(m，2H， 4-H)，2.27 & 2.10(m，2H，2-H)。ESI Mass：C ₅H ₁₂ClNO ₃Calculated value m/z, 169.15; Measured value, 133.17[M-HCl+H]⁺。

(2) (R)-synthetic (4d) of 3-hydroxyl-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. White solid. Productive rate: 74.6%. TLC R_f 0.31(CH ₂Cl ₂/ MeOH, 19/1, v/v). analyze HPLC:t_R＝22.72min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.89(d， ³J _H，H＝7.4Hz，2H，Fluoren.-H)，7.71(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.41(t， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.34(t， ³J _H，H ＝7.2Hz，2H，Fluoren.-H)，7.31(s，1H，4-NH)，4.30(d， ³J _H，H＝6.8Hz，2H，CH ₂-O)， 4.19(t， ³J _H，H＝6.8Hz 1H，Fluoren.-H)，4.24(m，1H，3-H)，3.62(s，3H，1-OCH ₃)， 3.05(m，2H，4-H)，2.34(dd， ²J _H，H＝15.2Hz， ³J _H，H＝4.0Hz，1H，2-H _a)，2.12(dd， ²J _H，H ＝15.2Hz， ³J _H，H＝8.4Hz，1H，2-H _b)。ESI Mass：C ₂₀H ₂₁NO ₅Calculated value m/z, 355.14; Measured value, 356.11[M+H]⁺。

(3) (R)-synthetic (5d) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino methyl butyrate. Milky oily product. Productive rate: 91.2%. TLC R_f 0.36(CH ₂Cl ₂/ MeOH, 100/1, v/v). analyze HPLC:t_R＝ 26.4min. ¹H-NMR(400MHz，DMSO-d ₆)：δ7.89(d， ³J _H，H＝7.6Hz，2H，Fluoren.-H)， 7.69(d， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.41(t， ³J _H，H＝7.2Hz，2H，Fluoren.-H)，7.36 (m，3H，Fluoren.-H+4-NH)，4.31(d， ³J _H，H＝6.4Hz，2H，CH ₂-O)，4.22(d， ³J _H，H＝6.4 Hz，1H，Fluoren.-H)，3.88(m，1H，3-H)，3.63(s，3H，1-OCH ₃)，3.10 & 2.90(m，2H， 4-H)，2.31(dd， ²J _H，H＝15.2Hz， ³J _H，H＝4.0Hz，1H，2-H _a)，2.10(dd， ²J _H，H＝15.2Hz， ³J _H，H＝5.6Hz，1H，2-H _b)，1.07(s，9H，3-OC(CH ₃) ₃)。ESI Mass：C ₂₄H ₂₉NO ₅Calculated value m/z, 411.20; Measured value, 412.14[M+H]⁺。

(4) (R)-synthetic (6d) of 3-tert-butoxy-4-Aminobutanoicacid. White solid. Productive rate: 98.4%. TLC R_f 0.50(CH ₂Cl ₂/MeOH/HOAc，2/2/0.5，v/v). ¹H-NMR(400MHz，DMSO-d ₆)：δ4.98 (m，1H，3-H)，2.95(dd， ²J _H，H＝14.4Hz， ³J _H，H＝3.2Hz，1H，4-H _a)，2.73(dd， ²J _H，H＝ 14.4Hz， ³J _H，H＝8.8Hz，1H，4-H _b)，2.21(d， ³J _H，H＝6.8Hz，2H，2-H)，1.08(s，9H， 3-OC(CH ₃) ₃)。ESI Mass：C ₈H ₁₇NO ₃Calculated value m/z, 175.12; Measured value, 176.10[M+H]⁺； 198.18[M+Na] ⁺。

(5) (R)-synthetic (1d) of 3-tert-butoxy-4-(9-fluorenes methoxyl group) carbonyl amino butyric acid. White solid. Productive rate: 74.4%. R_f0.30 (ethyl acetate/cyclohexane/acetic acid, 2/1/0.01, v/v). Analyze HPLC:t_R＝23.94min。 ¹H-NMR(400MHz，DMSO-d ₆)：δ12.14(br，1H，1-COOH)，7.90(d， ³J _H，H＝7.2Hz， 2H，Fluoren.-H)，7.71(m，2H，Fluoren.-H)，7.43(t， ³J _H，H＝7.4Hz，2H，Fluoren.-H)， 7.32(m，3H，Fluoren.-H+4-NH)，4.29(m，2H，CH ₂-O)，4.20(m，1H，Fluoren.-H)， 3.90(br，1H，3-H)，3.13 & 2.94(m，2H，4-H)，2.38 & 2.20(qq， ²J _H，H＝14.8Hz， ³J _H，H＝ 4.4Hz， ³J _H，H＝6.4Hz，2H，2-H)，1.13(s，9H，3-OCMe ₃)。ESI-Mass：C ₂₃H ₂₇NO ₅Calculated value m/z, 397.19; Measured value, 398.23[M+H]⁺，420.15[M+Na] ⁺, anion detects: 396.27 [M-H]^-。

Example 5: with optimized reaction condition synthesising target compound and their intermediate, following condition is also investigated except above-mentioned:

Table 2 is produced 3a, 3b, 3c, and the investigation of the reaction condition of 3d

No.	Raw material 2a 2b 2c 2d	SOCl 2   (95％，mmol)	CH3OH   (mL)	Other is anti-bAnswer condition	Productive rate (%) 3a 3b 3c 3d	Shape of product	Remarks
								1         2         3         4         5         6         7	1         1         1         1         1         1         1	2.8         1.8         1.8         3.5         3.5         2.5         2.8	1.8         1.2         3.0         1.2         3.0         1.5-1.8         1.8	Example 1 (1) with 1 with 1 with 1 with 1 with 1 with 1	99.2   97.8   98.0   98.4   63.0   66.4   68.0   69.1   60.0   57.4   63.1   64.3   74.0   73.5   75.0   74.7   85.4   84.0   86.7   88.6   95-97.2   94.2-96.8   93.6-98   94-98.5   24.5-40   22.7-44.8   30.0-50.0   32.6-48.9	White solid white solid white solid white solid white solid white solid white solid	The optimizing reaction conditions reaction time prolongs, and productive rate reduces the reaction time prolongation, and productive rate reduces SOCl2When concentration was high, the reaction time was short, but the post processing trouble is polluted large. Excessive SOCl2Make the post processing difficulty, pollute the large reaction condition reaction temperature of optimizing: in the time of 0 ℃-10 ℃, the reaction time is long, the more difficult complete reaction post processing difficulty of raw material

^aTake 1mmol as the calculating standard;^bComprise the reaction time the condition of in remarks, mentioning, temperature, reinforced and post-processing approach etc.

Table 3 is produced 4a, 4b, 4c, and the investigation of the reaction condition of 4d

No.	Raw materiala   3a   3b   3c   3d	FmocONSu   (mmol)	Solvent (mL)	Alkali (mmol)	Other is anti-bAnswer condition	Productive rate (%) 4a 4b 4c 4d	Shape of product	Remarks
									1         2         3         4         5         6         7         8         9         10	1         1         1         1         1         1         1         1         1         1	1.02         1.02         1.02         1.02         1.02         1.02         1.02         1.02         2.0         1.02	1,4-dioxane (3) N, dinethylformamide (3) acetonitrile (3) glycol dimethyl ether (3) 1,4-dioxane (3) 1,4-dioxane (3) Isosorbide-5-Nitrae-dioxane (3) Isosorbide-5-Nitrae-dioxane (6) 1,4-dioxane (3) Isosorbide-5-Nitrae-dioxane (3)	Sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium carbonate (1.2) NaOH (1.2) sodium acid carbonate (3) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2)	Example 1 (2) with 1 with 1 with 1 with 1 with 1 with 1 with 1 with 1 with 1	75.2   78.1   75.0   74.6   68.0   67.6   66.9   67.2   67.2   67.5   66.9   66.5   74.0   74.5   73.0   73.2   55.5   52.0   56.0   58.7   25.7   24.8   27.2   28.5   74.8   76.3   72.1   70.7   75.0   73.6   69.6   66.8   75.0   76.6   74.6   73.8   23-35.0   21-36.5   20-40.5   21-40	White solid white solid white solid white solid white solid white solid white solid white solid white solid white solid	Optimizing reaction conditions post processing difficulty, long preferably reaction condition post processing difficulty of poor and reaction time of productive rate low solubility and strong basicity make the low post processing difficulty of productive rate and too strong basicity make productive rate hang down preferably reaction condition, but the reaction condition that waste alkali is optimized, but the reaction time is long, the reaction condition of the large optimization of solvent load, but consume the FmocONSu reaction temperature: in the time of 0 ℃-10 ℃, the reaction time is long, the more difficult complete reaction post processing difficulty of raw material

^aTake 1mmol as the calculating standard;^bComprise the reaction time the condition of in remarks, mentioning, temperature, reinforced and post-processing approach etc.

Table 4 is produced 5a, 5b, 5c, and the investigation of the reaction condition of 5d

No.	Raw materiala   4a   4b   4c   4d	Isobuteneb	Solvent (mL)	The concentrated sulfuric acid (μ L)	Other is anti-cAnswer condition	Productive rate (%) 5a 5b 5c 5d	Shape of product	Remarks
									1         2         3         4         5         6         7         10	1         1         1         1         1         1         1         1	2         2         2         2         2         0.5         10         1.02	Carrene (50) Isosorbide-5-Nitrae-dioxane (50) glycol dimethyl ether (50) carrene (50) carrene (50) carrene (50) carrene (50) carrene (50)	125         125         125         20         300         125         125         125	Example 1 (3) with 1 with 1 with 1 with 1 with 1 with 1 with 1	88.3   90.7   92.0   91.2   80.0   84.6   82.9   80.0   78.0   75.9   77.5   79.2   78.8   79.0   83.0   81.3   46.8   43.8   51.0   49.5   65.9   64.5   66.1   62.5   85.0   81.6   89.6   87.3   43-68.3   46-65.5   50.7-66.0   53.0-63.4	White oily brown oil shape brown oil solid white oily burgundy oil burgundy oil white look oily white solid	The more difficult desolventizing reaction time of optimizing reaction conditions post processing difficulty is long, the low side reaction of productive rate is many, purification difficult, the productive rate low reaction time is long, the low reaction condition of optimizing of productive rate except excessive isobutene difficulty, consumes the isobutene reaction temperature: in the time of 0 ℃-15 ℃, reaction time is long, the more difficult complete reaction post processing difficulty of raw material

^aTake 1mmol as the calculating standard;^bMultiple with quantity of solvent represents;^cComprise the reaction time the condition of in remarks, mentioning, temperature, reinforced and post-processing approach etc.

Table 5 is produced 6a, 6b, 6c, and the investigation of the reaction condition of 6d

No.	Raw materiala   5a   5b   5c   5d	Alkali (mol/L, mL)	Solvent (mL)	Other is anti-bAnswer condition	Productive rate (%) 6a 6b 6c 6d	Shape of product	Remarks
								1         2         3         4         5         6         7         1	1         1         1         1         1         1         1         1	NaOH (2,2.9) sodium carbonate (2,2.9) NaOH (2,2.9) NaOH (2,2.9) NaOH (1,2.9) NaOH (5,2.9) NaOH (2,20) NaOH (2,2.9)	Isosorbide-5-Nitrae-dioxane (7) Isosorbide-5-Nitrae-dioxane (7) glycol dimethyl ether (7) N, dinethylformamide (7) Isosorbide-5-Nitrae-dioxane (7) Isosorbide-5-Nitrae-dioxane (7) 1,4-dioxane (7) Isosorbide-5-Nitrae-dioxane (7)	Example 1 (4) with 1 with 1 with 1 with 1 with 1 with 1 with 1	98.0   98.0   97.9   98.4   91.0   92.6   90.6   93.0   96.0   95.9   97.3   96.2   92.0   90.1   91.4   93.5   96.0   97.2   96.1   96.2   98.0   97.4   97.1   98.2   96.8   97.4   97.5   95.8   80.3-94.0   83.2-93.0   82-92.9   82.7-90.6	White solid white solid white solid white solid white solid white solid white solid white solid	It is complicated that the slightly difficult solvent difficulty of long post processing of optimizing reaction conditions reaction time is removed slightly long post processing of reaction time, wastes excessive alkali post processing complexity, and waste excessive alkali reaction temperature: in the time of 15 ℃-30 ℃, the reaction time is long

^aTake 1mmol as the calculating standard;^bComprise the reaction time the condition of in remarks, mentioning, temperature, reinforced and post-processing approach etc.

Table 6 is produced 1a, 1b, 1c, and the investigation of the reaction condition of 1d

No.	Raw materiala   6a   6b   6c   6d	FmocONSu   (mmol)	Solvent (mL)	Alkali (mmol)	Other is anti-bAnswer condition	Productive rate (%) 1a 1b 1c 1d	Shape of product	Remarks
									1         2         3         4         5         6         7         8         9         10	1         1         1         1         1         1         1         1         1         1	1.2         1.2         1.2         1.2         1.2         1.2         1.2         1.2         1.2         2.0	Glycol dimethyl ether (5.6) N, dinethylformamide (5.6) acetonitrile (5.6) Isosorbide-5-Nitrae-dioxane (5.6) glycol dimethyl ether (5.6) glycol dimethyl ether (5.6) glycol dimethyl ether (5.6) glycol dimethyl ether (15) glycol dimethyl ether (1) glycol dimethyl ether (5.6)	Sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium carbonate (1.2) NaOH (1.2) sodium acid carbonate (3) sodium acid carbonate (1.2) sodium acid carbonate (1.2) sodium acid carbonate (1.2)	Example 1 (5) with 1 with 1 with 1 with 1 with 1 with 1 with 1 with 1 with 1	80   76.0   74.3   74.4   64.0   63.3   61.2   60.5   65.7   62.4   64.0   64.7   75.8   78.1   73.6   72.0   50.1   49.3   45.2   47.7   30.5   35.0   36.8   38.4   77.2   78.0   73.6   74.0   76.8   75.6   66.6   68.1   72.3   73.0   64.5   65.0   78.0   79.0   73.6   73.9	White solid white solid white solid white solid white solid white solid white solid white solid white solid white solid	Optimizing reaction conditions post processing difficulty, the solvent difficulty remove long preferably reaction condition post processing difficulty of poor solubility and reaction time and strong basicity make the low post processing difficulty of productive rate and too strong basicity make productive rate hang down preferably reaction condition, but the reaction condition that waste alkali is optimized, but the reaction time is long, the large reaction of solvent load begins, and reactant mixture is muddy, can not dissolve fully, affect the reaction condition that productive rate is optimized, but consume FmocONSu

^aTake 1mmol as the calculating standard;^bComprise the reaction time the condition of in remarks, mentioning, temperature, reinforced and post-processing approach etc.

Claims (10)

1. hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that the preparation for the compound of following structure:

Wherein: the carbon of asterisk is asymmetric carbon atom, i.e. S or R configuration; N represents natural number, n=1～10; The hydrogen atom of the carbon atom combination on the main chain can be replaced with D atom; Fmoc is that 9-fluorenylmethyloxycarbonyl (9-fluorenylmethoxycarbonyl) and t-Bu are the tert-butyl group (tert-butyl); Be the precursor compound with optically active S or R configuration be raw material, synthesize by following synthetic route:

Synthesizing of synthetic route I[general formula (1) compound]:

Synthesizing of synthetic route II[general formula (2) compound]:

Its concrete steps are:

(1) compound 3 (a, b, c, d) is synthetic:

The raw material consumption

Compound 2 (a, b, c, d);

Thionyl chloride molar equivalent: 1.8～3.5

Methyl alcohol 1.2～3.0mL/mmol raw material

Reactions steps: 1. above raw material ends in 0～25 ℃ of hybrid reaction to compound 2 (a, b, c, d) complete reaction; 2. rotary evaporation is removed methyl alcohol; 3. use 1: 1 methyl alcohol and carbon tetrachloride solution dissolving, the rotary evaporation desolventizing gets compound 3 (a, b, c, d);

(2) compound 4 (a, b, c, d) is synthetic:

The raw material consumption

Compound 3 (a, b, c, d)

Solvent: DMF, acetonitrile, glycol dimethyl ether

Or Isosorbide-5-Nitrae-dioxane 1.5～6.0mL/mmol raw material

NaOH, sodium carbonate or sodium acid carbonate molar equivalent: 1.0～3.0

FmocONSu molar equivalent: 1.0～2.0

Reactions steps: 1. above raw material ends in 0～45 ℃ of hybrid reaction to compound 3 (a, b, c, d) complete reaction; 2. add frozen water, filter behind 2～5h, reclaim filter cake, separate to get compound 4 (a, b, c, d) by silica gel column chromatography;

(3) compound 5 (a, b, c, d) is synthetic:

The raw material consumption

Compound 4 (a, b, c, d)

The concentrated sulfuric acid 20～300 μ L/mmol raw materials

Solvent: Isosorbide-5-Nitrae-dioxane, glycol dimethyl ether or carrene 4～80mL/mmol raw material

0.5～10 times of quantity of solvent of isobutene

Reactions steps: 1. above raw material only is mixed to compound 4 (a, b, c, d) complete reaction at 0～25 ℃; 2. remove isobutene, 3. through 10% sodium bicarbonate solution, saturated brine washing, anhydrous sodium sulfate drying separates to get compound 5 (a, b, c, d) by silica gel column chromatography;

(4) compound 6 (a, b, c, d) is synthetic:

The raw material consumption

Compound 5 (a, b, c, d)

Dilute sulfuric acid, phosphoric acid or hydrochloric acid conditioned reaction liquid pH=7

Alkali (NaOH or sodium carbonate) solution, concentration is 1～5mol/L, 0.2～20mL/mmol raw material

Solvent: DMF, glycol dimethyl ether

Or Isosorbide-5-Nitrae-dioxane 5～10mL/mmol raw material

Reactions steps: 1. above raw material is at 15～60 ℃ of hybrid reaction 1～3h; 2. transfer pH to neutral with dilute sulfuric acid, phosphoric acid or hydrochloric acid; 3. steaming desolventizes, and dissolves residue with methyl alcohol; 4. the filtering insoluble matter separates to get compound 6 (a, b, c, d) by silica gel column chromatography;

(5) target product 1 (a, b, c, d) is synthetic:

The raw material consumption

Compound 6 (a, b, c, d)

NaOH, sodium carbonate or sodium acid carbonate molar equivalent: 1.0～3.0

Solvent: DMF, acetonitrile, glycol dimethyl ether

Or Isosorbide-5-Nitrae-dioxane 1～15mL/mmol raw material

FmocONSu molar equivalent: 1.0～2.0

Reactions steps: 1. 0～45 ℃ of hybrid reaction 10～15h; 2. make its pH=5 through acidifying; 3. steaming desolventizes, the solid acetic acid ethyl dissolution, and 4. the filtering insoluble matter separates to get target product 1 (a, b, c, d) by silica gel column chromatography.

2. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps:

(1) compound 3 (a, b, c, d) is synthetic:

The raw material consumption

Compound 2 (a, b, c, d);

Thionyl chloride molar equivalent: 2.5～2.8

Methyl alcohol 1.5～1.8mL/mmol raw material.

3. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps:

(2) compound 4 (a, b, c, d) is synthetic:

The raw material consumption

Compound 3 (a, b, c, d)

Solvent: Isosorbide-5-Nitrae-dioxane 1.5～6.0mL/mmol raw material

Sodium acid carbonate molar equivalent: 1.2

FmocONSu molar equivalent: 1.0～1.1.

4. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps: (three) compound 5 (a, b, c, d) synthetic:

The raw material consumption

Compound 4 (a, b, c, d)

The concentrated sulfuric acid 120～125 μ L/mmol raw materials

Solvent: carrene 50mL/mmol raw material

1.5～2.0 times of quantity of solvent of isobutene.

5. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps: (four) compound 6 (a, b, c, d) synthetic:

The raw material consumption

Compound 5 (a, b, c, d)

10% hydrochloric acid conditioned reaction liquid pH=7

Sodium hydroxide solution, concentration are 2mol/L 0.5～3mL/mmol raw material

Solvent: Isosorbide-5-Nitrae-dioxane 5～10mL/mmol raw material.

6. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps: (five) target product 1 (a, b, c, d) synthetic:

The raw material consumption

Compound 6 (a, b, c, d)

Sodium acid carbonate molar equivalent: 1.2

Solvent: glycol dimethyl ether 4～6mL/mmol raw material

FmocONSu molar equivalent: 1.2.

7. each described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection according to claim 1～6 is characterized in that in its concrete steps:

(1) compound 3 (a, b, c, d) is synthetic:

In the reactions steps: 1. raw material 0 ℃ mix 15min after, at room temperature react to compound 2 (a, b, c, d) complete reaction and end;

(2) compound 4 (a, b, c, d) is synthetic:

In the reactions steps: 1. above raw material at room temperature reacts 30h and ends to compound 3 (a, b, c, d) complete reaction behind 0 ℃ of mix and blend 30min;

(3) compound 5 (a, b, c, d) is synthetic:

Reactions steps: 1. above raw material mixes in subzero, behind the stirring 30min, at room temperature continues to stir until till compound 4 (a, b, c, the d) complete reaction;

(4) compound 6 (a, b, c, d) is synthetic:

Reactions steps: 1. above raw material is till 40～45 ℃ of hybrid reaction 1h;

(5) target product 1 (a, b, c, d) is synthetic:

Reactions steps: 1. above raw material mixes at zero degree, behind the stirring 1h, at room temperature continues to stir till the 10h.

8. according to claim 1 described hydroxyl and the amino simultaneously preparation method of the chiral non alpha-amino acid derivativges of protection is characterized in that in its concrete steps:

(1) compound 3 (a, b, c, d) is synthetic:

The raw material consumption

Compound 2 (a, b, c, d);

Thionyl chloride molar equivalent: 2.5～2.8

Methyl alcohol 1.5～1.8ml/mmol raw material

In the reactions steps: 1. above raw material mixes 15min at 0 ℃, after at room temperature react to compound 2 (a, b, c, d) complete reaction and end;

(2) compound 4 (a, b, c, d) is synthetic:

The raw material consumption

Compound 3 (a, b, c, d)

Solvent: Isosorbide-5-Nitrae-dioxane 1.5～6.0ml/mmol raw material

NaOH molar equivalent: 1.2

FmocONSu molar equivalent: 1.0～1.1

In the reactions steps: 1. above raw material is at 0 ℃ of mix and blend 30min, after at room temperature react 30h and end to compound 3 (a, b, c, d) complete reaction;

(3) compound 5 (a, b, c, d) is synthetic:

The raw material consumption

Compound 4 (a, b, c, d)

The concentrated sulfuric acid 120～125 μ l/mmol raw materials

Solvent: carrene 50ml/mmol raw material

1.5～2.0 times of quantity of solvent of isobutene

Reactions steps: 1. above raw material mixes in subzero, behind the stirring 30min, at 22 ℃ of lower reaction 25～70h;

(4) compound 6 (a, b, c, d) is synthetic:

The raw material consumption

Compound 5 (a, b, c, d)

10% hydrochloric acid conditioned reaction liquid pH=7

Sodium hydroxide solution, concentration are 2mol/L 0.5～3mL/mmol raw material

Solvent: Isosorbide-5-Nitrae-dioxane 5～10mL/mmol raw material

Reactions steps: 1. above raw material is till 40～45 ℃ of hybrid reaction 1h;

(5) target product 1 (a, b, c, d) is synthetic:

The raw material consumption

Compound 6 (a, b, c, d)

Sodium acid carbonate molar equivalent: 1.2

Solvent: glycol dimethyl ether 4～6mL/mmol raw material

FmocONSu molar equivalent: 1.2

Reactions steps: 1. above raw material mixes at zero degree, behind the stirring 1h, at room temperature continues to stir till the 10h.