CN1031508C - Method for synthesizing epi-rape lactone and likes - Google Patents

Abstract

The present invention relates to a method for synthesizing steroid plant hormones, particularly to a method for preparing bioactive epi-brassinolide and analogues thereof from vegetable oil screenings (rape oil or soybean oil). Proved by field experiment, the bioactive epi-brassinolide has obvious yield increase effect on crops. The method of the present invention has the advantages of rich source of raw materials, low cost and easy purification; a one-step ozonization reaction provided by the method enables alkoxyl to be oxidized into carbonyl; the method reduces reaction procedures so that the method of the present invention is possible to be applied in industrial scale.

Description

The method of synthesizing epi-rape lactone and analogue thereof

The present invention relates to the synthetic of a kind of steroidal class plant hormone, especially produce the method for the epi-rape lactone and the analogue thereof of biologically active from vegetables oil leftover bits and pieces.

Epi-rape lactone (2 α, 3 α, 22R, 23R-tetrahydroxy-B-height-7-oxygen-5 α-ergot steroid-6-ketone) and analogue (its chemical formula is shown in I, the II) be a class novel plant hormone, field experiment shows through Shanghai Inst. of Plant Physiology, Chinese Academy of Sciences, and farm crop are had stronger production-increasing function.Epi-rape lactone with 0.01ppm concentration is handled the florescence wheat, and the setting percentage and the fringe grain that can increase the wheat head are heavy, make wheat increase yield 10%, handle corn ear with same concentration, can make corn yield increasing 9-18%; Handle cucumber and watermelon seedling with 0.1ppm concentration, can impel cucumber production promoting 30%, increasing yield of water melons 20% can expect that the application on agricultural of epi-rape lactone and analogue thereof will bring tangible economic benefit.Therefore, people pay much attention to its synthetic.At present, existing several route synthesizing epi-rape lactones and analogue thereof.Thompson etc. are raw material with the ergosterol, make epi-rape lactone [referring to document J.Org.Chem.1979,44 (26), 5002] through ten single step reactions.Ikekawa etc. are raw material with the Brassicasterin, and reaction makes epi-rape lactone [referring to Japanese Patent JP60,139,685] through five steps.These methods all are to make raw material with suitable natural steroid compound, and the structure of modification of process A/B ring and the coplane oxidation of pendant double bonds are finished.But the reactions steps of article one route is long, and preparative-scale is little, and total recovery is low, is difficult to industrialization; Though second route reactions steps is short, raw material sources are difficult for, and total recovery is also low.

The objective of the invention is to obtain a kind of abundant raw material, synthesis step is simple, overall yield is higher and can industrialized method prepare epi-rape lactone and analogue thereof.

Main points of the present invention are to be raw material with vegetables oil leftover bits and pieces, through general saponification, extract and mix sterol III, and without separation and purification, just directly sulfonylation and alcoholysis, the two keys with on the sealing B ring then, make the oxidation of pendant double bonds coplane.Because the polarity of sterol structure and gonane structure differs greatly, thus by column chromatography can be easily oxidation products-diol compound with mix sterol in original contained can not separating by oxidized saturated alkane of having.After making hydroxyl protection with suitable protecting group, carry out ozonization, make alkoxyl group be direct oxidation into carbonyl.Subsequently, open loop, and make the two key coplane oxidations of A ring, and introduce the two hydroxyls of cis, protecting group is gone in the reaction of B cyclic lactone, thereby makes epi-rape lactone and analogue thereof.Use method provided by the present invention,, can produce epi-rape lactone as being raw material with rape oil leftover bits and pieces; With soya-bean oil leftover bits and pieces is raw material, can produce high rape lactone.

Method of the present invention can be represented with following chemical equation:

The present invention is elaborated as follows:

A, get a foothold to extract mix sterol III from vegetables oil:

Rape oil or soya-bean oil are got a foothold with 10% potassium hydroxide aqueous solution, and 100 ℃ were boiled 10-24 hours, carried out saponification reaction, again with ether or ethyl acetate extraction, remove and desolvate, and ethyl alcohol recrystallization makes and mixes sterol III.

Being got a foothold by rape oil, its main component is based on β-Gu Zaichun and Brassicasterin in the mixing sterol extracted, and from structure, their A, B cyclopentanoperhydro-phenanthrene are consistent.Its difference mainly shows on the side chain.Saturated and unsaturated branch is arranged.If will separately be the comparison difficulty with both.[referring to document J.Am.Chem.Soc., 1939,61,142; JP60,215,699; DE3,506,938].

The present invention adopts the two keys of sealing cyclopentanoperhydro-phenanthrene earlier, the way of rear oxidation pendant double bonds, utilize the significant difference on o-dihydroxy structure and the saturated alkane structure polarity, make both separately, that is to say, after the two key sealings of cyclopentanoperhydro-phenanthrene, oxide side chain, make the side chain unsaturated compound be oxidized to diol compound, thereby separate with not oxidized saturated compound.Reached like this and separated the purpose of mixing sterol, and method is simple, separates easily.

B, the present invention seal that the reaction of the two keys of cyclopentanoperhydro-phenanthrene is performed such:

Make mixing sterol III sulfonylation with general sulfonylation agent, sulfonylation agent is a methylsulfonyl chloride preferably, p-toluenesulfonyl chloride.

3 β that make like this-mesyloxy or 3 β-right-tosyloxy compound is the mixture of steroid alkene and gonane, through alcoholysis (reacting with methyl alcohol in the presence of pyridine), makes 6 'beta '-methoxies-3 α, 5-ring-5 α-steroid alkene and gonane mixture.

C, said mixture IV, in polar solvent, use suitable oxidizers, make the side chain coplane oxidation of steroid ene compound, obtain vicinal diol compound V and steric isomer VI thereof, through chromatographic separation, itself and unoxidized gonane compound are separated, and suitable oxygenant is N-methyl-morphine quinoline-N-oxide compound, in the presence of the little amount of catalyst perosmic anhydride, reaction solvent uses mixed solvent, and suitable proportioning is the trimethyl carbinol: tetrahydrofuran (THF): water=10: 8: 1.

D, formula V, VI vicinal diol compound with appropriate protection base protection hydroxyl, make respective compound VII, VIII that hydroxyl is protected, in the formula R be that sec.-propyl, benzyl, the dimethyl tertiary butyl are silica-based, ethanoyl etc.

E, in organic solvent, with suitable oxygenant, in the presence of little amount of catalyst, make alkoxyl group be oxidized to carbonyl, thereby make sterone Compound I X, X.

Oxidizing reaction is carried out between 0～-78 ℃ usually, and optimum temperature range is-50～-78 ℃, and used oxygenant is an ozone, four rubidium oxides.Preferred ozone, available pyridine is made catalyzer.

Utilize method of the present invention can make alkoxyl group be direct oxidation into carbonyl, make key intermediate 22R, 23R-diacetoxy-3 α, 5-encircles-5 α-ergot steroid-6-ketone.

F, formula IX, the open loop of X compound make formula XI, XII compound, wherein RR such as above-mentioned definition.The used A cycloisomerisation reaction of method of the present invention utilizes pyridine hydrobromide salt to carry out, and [referring to document Steroids 1,985 45,561 ,]

G, in the presence of polar solvent and perosmic anhydride, make two key coplane oxidations on the A ring of formula XI, XII compound, make the compound that chemical formula is XIII, XIV.

H, make the B cyclic lactoneization and the pendant hydroxyl group of formula XIII, XIV compound go protecting group, make compound and corresponding steric isomer II thereof that chemical formula is I.

Method raw material sources of the present invention are abundant, and are with low cost.In view of Semen Brassicae campestris is the large oil crops of China, rapeseed germplasm is very abundant, and utilizing rape oil to get a foothold not only turns waste into wealth as the steroidal raw material, and has alleviated vegetable seed and add a large amount of environmental pollutions of getting a foothold and being caused in man-hour.Because the mixing sterol is direct reaction without separation and purification, make and be difficult to relatively separate easily of compound separately originally, simplified purification process greatly, and provided by the present invention one the step ozonization can make alkoxyl group be direct oxidation into carbonyl, shortened reactions steps.Obviously, according to above-mentioned advantage, be possible in industrial scale applications method of the present invention.

The present invention is described in further detail by the following example:

Embodiment 1:

Mix the extraction of sterol (III)

Vegetables oil (rape oil or soya-bean oil) tankage add 10% potassium hydroxide-ethanol solution, boil saponification 10 hours, add two times in water after the cooling, ether extraction, boil off behind the solvent yellow solid.Use ethyl alcohol recrystallization again, make and mix sterol III.

Rape mixing sterol from the rape oil tankage extract, mainly contain:

β-Gu Zaichun (Sitosterol) percentage composition :～55%

Brassicasterol (Campesterol)～25%

Brassicasterin (Brassicasterol) 5-20%

The soybean of extracting from the soya-bean oil tankage mixes sterol, mainly contains:

β-Gu Zaichun percentage composition :～65%

Soyasterol (stigmasterol)～20%

Embodiment 2:

The mixture preparation of 3 β-mesyloxy gonane and steroid alkene.

20 grams mix sterol and are dissolved in 100 milliliters of methylene dichloride, add 20 milliliters of triethylamines, be cooled to below 0 ℃, drip 7 milliliters of methylsulfonyl chlorides and stir and spend the night, add 300 milliliters of ether after boiling off most of solvent, washing, dry (using anhydrous sodium sulphate) removes ether, chromatographic separation, the methanesulfonates 20-24 gram that must mix sterol, yield 83-100%.

H?NMR(100MHz，CDCL ₃)

0.59～1.0(m)，2.88(s，CH?SO)

4.00(m)，5.1-5.3(m，＝CH)

Embodiment 3:

6 'beta '-methoxies-3 α, the preparation of 5-ring-5 α-gonane and steroid alkene mixture.

Mix in the methanesulfonates compound of sterol at 20 grams, add 210 milliliters of dry methyl alcohol, 9 milliliters of pyridines refluxed 5 hours, boil off solvent after the cooling, in the impouring water, use ethyl acetate extraction, drying, remove the rear pillar chromatography that desolvates, get product 13-15 gram, yield 75-85%.

H?NMR(100MHz，CDCL ₃)

0.6～0，9(m) 3.10(s，-OCH) 5.10(m，＝CH)

Embodiment 4:

22 ξ, 23 ξ-dihydroxyl-6 'beta '-methoxy-3 α, the preparation of 5-ring-5 α-ergostane.

At 6 'beta '-methoxies-3 α of 12 grams, in 5-ring-5 α-gonane alkene mixture, add 100 milliliters of trimethyl carbinols by the preparation of rape mixing sterol, 80 milliliters of tetrahydrofuran (THF)s, 10 ml waters stir 3 gram N-methylmorpholine-N-oxide compound and the 300 milligrams of perosmic anhydrides of adding down, stirring at room three days.Add 100 milliliters of saturated sodium bisulfite water solvents, restir half an hour use extracted with diethyl ether, washing, drying boil off column chromatography behind the solvent, product 2 grams, by product (mainly be 6 'beta '-methoxies-3 α, 5-encircle-5 α-stigmastane) 9 restrains.

H-NMR(100MHz，CDCL ₃)

δ?0.64(s，3H)， 0.8～1.0(m)

3.24(3H，s，-OCH)，?3.60(1H，m，CH-OH)，

4.00(1H，m，CH-OH)。

Embodiment 5:

22S, 23S-and 22R, 23R-diacetoxy-6 'beta '-methoxy-3 α, the preparation of 5-ring-5 α-ergostane.

At 2 grams, 22 ξ, 23 ξ-dihydroxyl-6 'beta '-methoxy-3 α is in 5-ring-5 α-ergostane, add 35 milliliters of pyridines, 11 ml acetic anhydride, stirring at room 24 hours, in 150 milliliters of saturated sodium bicarbonate solutions of impouring, extracted with diethyl ether boils off solvent behind the washing and drying, column chromatography gets 0.5 gram 22S, 23S-diacetoxy-6 'beta '-methoxy-3 α, 5-encircles-5 α-ergostane (XV), 1.5 gram 22R, 23R-diacetoxy-6 'beta '-methoxy-3 α, 5-encircles-5 α-ergostane (X VI).

X?V?：

IR?1735，1460，1370cm ^-1

H-NMR(100MHz，CDCL ₃)

δ?0.62(3H，s)， 0.70(3H，d)，

0.80(6H，d)， 0.94(3H，s)，

1.20(3H)， 1.98(3H，s，OAc)

2.02(3H，s，-OAc)， 3.20(3H，s，-OCH)，

4.98(1H，m，CH-OAc) 5.02(1H，m，CH-OAc)，

MS(m/e)：?531(M+1)，516，499，476，

IR?1740，1460，1370cm ^-1

X?VI：

H-NMR(100HMz，CDCL ₃)

δ?0.62(3H，s)， 0.70(3H)，

0.88(9H)， 0.96(3H，s)，

1.98(3H，s，-OAc)， 2.01(3H，s，OAc)

3.21(3H，s，-OCH)， 5.02(1H，m，CH-0Ac)，

5.17(1H，m，CH-OAc).

MS(m/e)：531(M+1)，516，499，476，

Embodiment 6:

22R, 23R-diacetoxy-3 α, the preparation of 5-ring-5 α-ergot steroid-6-ketone.

At 0.4 gram 22R, 23R-diacetoxy-6 'beta '-methoxy-3 α adds 40 milliliters of methylene dichloride in 5-ring-5 α-ergostane, 0.4 the milliliter pyridine is cooled to-78 ℃-50 ℃ down logical ozone 1 hour, boils off column chromatography behind the solvent, get product 0.32-0.35 gram, yield 75%-85%.

IR?1740，1690，1460，1370cm ^-1

H-NMR(400HMz，CDCL ₃)

δ?0.70(3H，s)， 0.8?(3H，d，J＝7)，

0.86(3H，d，J＝6.6)， 0.90(3H，d，J＝7)，

0.98(3H，s)， 0.99(3H，d，J＝6.5)，

2.00(3H，s，-OAc) 2.01(3H，s，-OAc)，

5.05(1H，m，CH-OAc)， 5.23(1H，m，CH-OAc)

MS(m/e)：514(M)，394(M-2×AcOH)

Embodiment 7:

22R, 23R-diacetoxy-6-carbonyl-5 α-ergot steroid-2-alkene.

1.1 gram 22R, 23R-diacetoxy-3 α in 5-ring-5 α-ergot steroid-6-ketone, is dissolved in 40 milliliters of N, and N-dimethylformamide (DMF) adds 1.1 gram pyridine hydrobromide salt.Stirred 3 hours down at 160 ℃, in the cooling hypsokinesis entry, use extracted with diethyl ether, wash, drying boils off behind the solvent column chromatography on silicagel column, with the mixture of sherwood oil and ether (volume ratio is 3: 1) as eluent.Get product 0.85 gram, yield 78%.

IR?3020、1744，1710，1460，cm ^-1

H-NMR(400KHZ，CDCL ₃)；

δ?0.65(3H，s)， 0.70(3H，s)，

0.80(3H，d，J＝7)， 0.86(3H，d，J＝7)，

0.92(3H，d，J＝7)， 0.99(3H，d，J＝7)，

2.00(3H，s，-OAc) 2.02(3?H，s，-OAc)，

5.04(3H，m，CH-OAc)， 5.22(1H，m，CH-OAc)

5.55(1H，m，＝CH) 5.66(1II，m，＝CH)。

MS(m/e)：514(M)，499，486

Embodiment 8:

22R, 23R-diacetoxy-2 α, the preparation of 3 α-dihydroxyl-5 α-ergot steroid-6-ketone.

At 750 milligrams of 22R, in 23R-diacetoxy-6-carbonyl-5 α-ergot steroid-2-alkene, add 20 milliliters of trimethyl carbinols, 16 milliliters of tetrahydrofuran (THF)s, 4 ml waters, stir 800 milligrams of N-methylmorpholine-N-oxide compounds of adding, 100 milligrams of perosmic anhydrides, the gained mixture stirred 16-24 hours at 25 ℃, then, in reaction mixture, add 100 milliliters of saturated sodium sulfite solution, use ethyl acetate extraction after half an hour, anhydrous sodium sulfate drying, distillation removes and desolvates, residue obtained column chromatography is refining, gets 780 milligrams of products, yield 97%.

IR?3450，1740，1710，1460，cm ^-1

H-NMR(400MHz，CDCL ₃)δ：

0.64(3H，s)， 0.73(3H，s)，

0.80(3H，d，J＝7)， 0.84(3H，d，J＝6.7)，

0.90(3H，d，J＝6.8)， 0.97(3H，d，J＝7)，

2.00(3H，s，-OAc)， 2.01(3H，s，-OAc)，

2.64(1H，m)， 3.72(1H，m，CH-OH)，

4.01(1H，-m，CH-OH)， 5.05(1H，m，CH-OAc)，

5.20(1H，m，CH-OAc) MS(m/e)：549(M+1)，531，513.

Embodiment 9:

2 α, 3 α, 22R, the preparation of 23R-tetrahydroxy-B-height-7-oxygen-5 α-ergot steroid-6-ketone (epi-rape lactone).

500 milligrams of 22R, 23R-diacetoxy-2 α, 3 α-dihydroxyl-5 α-ergot steroid-6-ketone is dissolved in 20 milliliters of methylene dichloride, ice bath adds 1.3 milliliters of trifluoroacetic dichloromethane solutions of peroxide (10 milliliters of methylene dichloride, ice bath add 0.75 milliliter of 30% hydrogen peroxide down and make above-mentioned peroxide trifluoroacetic acid solution) down, and 0 ℃ was stirred 1-3 hours down, in the gained reaction mixture impouring saturated solution of sodium bicarbonate, with ethyl acetate extraction, the pressure reducing and steaming solvent gets 550 milligrams of crude products.Add 40 milliliters of 5% potassium hydroxide-methanol solutions subsequently, 70 ℃ were refluxed 1 hour down, and the cooling back adds concentrated hydrochloric acid, regulates PH＜3, stirred 1-3 hours, ethyl acetate extraction is removed the rear pillar chromatography that desolvates, use re-crystallizing in ethyl acetate again, get 250 milligrams of products, yield 60%.

mp?250℃

IR?3420，1730，1715cm ^-1

Embodiment 10:

22S, 23S-diacetoxy-3 α, the preparation of 5-ring-5 α-ergot steroid-6-ketone.

Press the preparation of embodiment 6 conditions.

IR?1730，1590，1460，1370。cm ^-1

H-NMR(400MHz，CDCL ₃)

δ?0.69(3H，s)， 0.71(3H，d，J＝7)

0.73(3H，d，J＝6.4)， 0.89(3H，d，J＝6.4)，

0.90(3H，d，J＝7)， 0.98(3H，s)，

2.06(3H，s，-OAc)， 2.11(3H，s，-OAc)，

5.08(2H，m，CH-OAc) MS(m/e)：514(M)，454，394.

Embodiment 11:

22S, the preparation of 23S-diacetoxy-6-carbonyl-5 α-ergot steroid-2-alkene.

Connect the preparation of embodiment 7 conditions.

IR，3020，1740，1709，1460，1430。cm ^-1

H-NMR (400MHz, CDCL ₃) the δ value:

0.63(3H，s)， 0.68(3H，s)

0.72(3H，d，J＝7)， 0.81(3H，d，J＝7)

0.89(3H，d，J＝6.4)， 0.96(3H，d，J＝7)，

0.05(3H，s，-OAc)， 2.07(3H，s，-OAc)，

5.06(1H，m，CH-OAc) 5.18(1H，m，CH-OAc)，

5.54(1H，m，＝CH)， 5.66(1H，m，＝CH) MS(m/e)：514(M)，499，486.

Embodiment 12:

22S, the preparation of 23S-diacetoxy-2 α, 3 alpha-dihydroxy-s-5 α-ergot steroid-6-ketone.

Press the preparation of embodiment 8 conditions.

IR?3450，1740，1710，1460，1370cm ^-1

H-NMR (400HMz, CDCL ₃) the δ value:

0.60(3H，s)， 0.69(3H)

0.72—0.80(6H)， 0.90—0.94(6H)，

1.99(3H，s，-OAc)， 2.01(3H，s，-OAc)。

3.70(1H，m)， 3.98(1H，m)，

5.00(1H，m) 5.16(1H，m) MS(m/e)：531(M-17)，513，428.

Embodiment 13:

2 α, 3 α, 22S, the preparation of 22S-tetrahydroxy-B-height-7-oxygen-5 α-ergot steroid-6-ketone.

Press the preparation of embodiment 9 conditions.

IR3?420，1730，1715?cm ^-1

mp：193℃。

Embodiment 14:

22 ξ, 22 ξ-dihydroxyl-6 'beta '-methoxy-3 α, the preparation of 5-ring-5 α-big stigmastane.

From mixing 6 'beta '-methoxies-3 α that sterol makes by soybean, 5-ring-5 α-gonane alkene mixture prepares by embodiment 4 conditions.

IR?3420，1730，1715cm ^-1

Embodiment 15:

22S, 23S-and 22R, 23R-diacetoxy-6-methoxyl group-3 α, the preparation of 5-ring-5 α-big stigmastane.

Press the preparation of embodiment 5 conditions.

Embodiment 16:

22S, 23S-diacetoxy-3 α, the preparation of 5-ring-5 α-soybean steroid-6-ketone.

Press the preparation of embodiment 6 conditions.

IR?1730cm ^-1

H-NMR(400HMZ，CDCL ₃) δ?0.70(3H，s，18-CH)， 0.85(3H，d，J＝7，20-CH)

0.92(9H)， 1.02(3H，s，19-CH)，

2.03(3H，s，-OAc)， 2.07(3H，s，-OAc)，

2.45(2H，m)， 5.00(1H，m)，

5.22(1H，m)， MS(m/e)：528(M)，468(M-AcOH)，408(M-2×AcOH)。

Claims (3)

1. the preparation method of a class formation formula I compound and corresponding steric isomer II thereof, wherein R ₁Be methyl or ethyl and their corresponding steric isomers

It is characterized in that this method comprises that its reaction sequence is:

A. be raw material with vegetables oil leftover bits and pieces, extract to such an extent that chemical formula is the mixing sterol of III through saponification R wherein ₂Be side chain alkyl with following formula (i)-(iv), or thiazolinyl and corresponding steric isomer thereof

B. compound III is through the sulfonylation agent sulfonylation, and the two keys on the B ring are sealed in alcoholysis under the alkaline condition, get the mixture of Formula I V, R in the formula ₂Be as above definition, R ₃Be methyl, ethyl, sec.-propyl or benzyl

C. mixture IV is in polar solvent, with oxygenant N-methyl-morphine quinoline-N-oxide compound under perosmic anhydride catalysis, oxidation steroid-22-ene compound wherein, make two keys be oxidized to corresponding adjacent diol structure, get compound V and steric isomer VI thereof, with column chromatography separately, R in the formula with unoxidized gonane compound ₁, R ₃As above-mentioned definition

V 22S，23S-

VI?22R，23R-

D. vicinal diol compound V, VI get respective compound VII, the VI of hydroxyl protection, R in the formula through aceticanhydride pyridine protection hydroxyl ₄Be sec.-propyl, benzyl, silica-based, the ethanoyl of the dimethyl tertiary butyl

R ₁, R ₃As above definition;

E. in organic solvent, get Formula I X, X compound, R through ozone oxidation ₁, R ₄As above-mentioned definition

F. Compound I X, X open loop in the presence of pyridine hydrobromide salt is made XI, XII

G. in polar solvent, in the presence of the perosmic anhydride, make oxygenant, make the two key coplane oxidation on compounds X I, the XIIA ring, make R in XIII, the XIV formula with N-methylmorpholine-N-oxide compound ₁, R ₄As above definition

H.g step reaction gained XIII, XIV product make the B cyclic lactoneization in the presence of peroxide is trifluoroacetic, the deprotection base makes I, II compound again.

2. method according to claim 1 is characterized in that in the presence of basic catalyst, makes alkoxyl group be oxidized to carbonyl, and temperature of reaction is between 0--78 ℃.

3. preparation method according to claim 2 is characterized in that the oxygenant that makes alkoxyl group be oxidized to carbonyl is an ozone.