JPH05247043A - 2-bis(alkoxycarbonyl)methylidenepyrrolidine derivative - Google Patents

Abstract

PURPOSE:To obtain a new compound useful as a carcinostatic agent having strong cell toxicity. CONSTITUTION:A compound of formula I (R<1> and R<2> are 1-5C alkyl or aralkyl) such as (S)-5-(t-butyldiphenylsilyloxy)methylpyrrolidine-2-thione. The compound of formula I is obtained by silylating (S) or (R)-5-hydroxymethylpyrrolidin-2-one of formula II as a starting substance, successively carrying out conversion of carbonyl group, reaction with a halogenomalonic diester and elimination of silyl group to give a compound of formula III, then sulfonylating the hydroxyl group of the compound of formula III and treating with a base (e.g. potassium hydroxide).

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel 2-bis (alkoxycarbonyl) methylidenepyrrolidine derivative having strong cytotoxicity and expected to be used as an anticancer agent, and 5-silyl which is an intermediate for its production. It relates to an oxymethylpyrrolidine-2-thione derivative.

[0002]

BACKGROUND OF THE INVENTION There is a strong demand from society for the development of excellent anticancer agents, and the creation of new compounds having strong cytotoxicity occupies a very important position in the development of excellent anticancer agents. Generally, the antitumor activity and antitumor spectrum of a compound largely depend on its chemical structure, and therefore, from a cytotoxic compound having a novel structure different from the known one,
There is a great possibility that an anticancer drug having characteristics superior to those currently put into practical use will be developed.

[0003]

An object of the present invention is to create a compound having a novel chemical structure and having strong cytotoxicity.

[0004]

Means for Solving the Problems As a result of the inventor's ingenious examination, the present inventors have found that a compound represented by the following general formula is a compound having a novel chemical structure and having strong cytotoxicity.

[0005]

[Chemical 4]

(In the formula, R ¹ and R ² are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aralkyl group.)
And the following general formula

[0007]

[Chemical 5]

(In the formula, R ¹ and R ² are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aralkyl group,
X represents a hydroxyl group, a linear or branched alkanesulfonyloxy group having 1 to 5 carbon atoms, a substituted or unsubstituted allenesulfonyloxy group, a chlorine atom, a bromine atom, or an iodine atom) 2-bis Found an (alkoxycarbonyl) methylidenepyrrolidine derivative,
As a production intermediate of the above general formulas [I] and [II],
The following general formula

[0009]

[Chemical 6]

(Wherein R ³ , R ⁴ and R ⁵ are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aryl group). The 5-silyloxymethylpyrrolidine derivative represented was created and the present invention was completed.

A 2-bis (alkoxycarbonyl) methylidenepyrrolidine derivative represented by the above general formulas [I] and [II], and 5 represented by the above general formula [III]
The -silyloxymethylpyrrolidine-2-thione derivative can be produced by the following synthetic steps.

[0012]

[Chemical 7]

[Chemical 8]

(In the formula, R ¹ and R ² are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aralkyl group,
R ³ , R ⁴ and R ⁵ are the same or different and each represent a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aryl group, and R ⁶ is 1 to 5 carbon atoms.
Represents a linear or branched alkyl group or a substituted or unsubstituted aryl group, and Y represents a chlorine atom, a bromine atom, or an iodine atom.)

First Step This step is based on the literature [S. Saijo et al., Chem. Pharm. Bul.
l., 28, 1449 (1980).]
(S)-or (R) -5-hydroxymethylpyrrolidine-2
Protects the -one hydroxyl group by silylating (S)-or (R)
It is for producing -5-silyloxymethylpyrrolidin-2-one.

As the silylating agent used, chlorotrimethylsilane, bromotrimethylsilane, trimethylsilyltrifluoromethanesulfonate, chlorotriethylsilane, bromotriethylsilane, triethylsilyltrifluoromethanesulfonate, chlorodimethylisopropylsilane, bromodimethylisopropylsilane, dimethylisopropyl Silyltrifluoromethanesulfonate, t-butylchlorodimethylsilane, bromo-t-butyldimethylsilane, t-butyldimethylsilyltrifluoromethanesulfonate, t-butylchlorodiphenylsilane, bromo-t-butyldiphenylsilane, t-butyldiphenyl Silyltrifluoromethanesulfonate, chloromethyldiisopropylsilane, tribenzylchlorosilane, chloroto p- Kishirirushiran, chlorotriisopropylsilane, bromo triphenyl silane,
Etc. are used, but t-butylchlorodiphenylsilane is preferably used. Silylation is performed under normal silylation conditions [TW Green, "Protective Groups in Organic Sy
nthesis ", A Wiley Interscience Publication, John-W
iley & Sons, New York (1981), pp 39-50].

Step 2 In this step, the 2-position carbonyl group of (S)-or (R) -5-silyloxymethylpyrrolidin-2-one is converted to a thiocarbonyl group to give the compound of the present invention (S ) − Or (R) −
It is intended to produce 5-silyloxymethylpyrrolidine-2-thione.

The conversion of this carbonyl group to a thiocarbonyl group is carried out by diphosphorus pentasulfide or 2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphetane-2,4-disulfide (Lawesson). Reagent), and the Lawesson reagent is particularly preferably used. The reaction is carried out in a solvent, and any solvent can be used as long as it does not participate in the reaction, but pentane,
Hydrocarbon-based solvents such as hexane, cyclohexane, benzene, toluene and xylene are used, and toluene is preferably used. The reaction proceeds smoothly from room temperature to 150 ° C.

The third step is the step of (S)-or (R) -silyloxypyrrolidine-
2-thione is reacted with a halogenomalonic acid diester,
Furthermore, the reaction product is treated under weakly basic conditions to produce (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-silyloxymethylpyrrolidine. Examples of the halogenomalonic acid diester used in this step include dimethyl chloromalonate, dimethyl bromomalonate, diethyl chloromalonate, diethyl bromomalonate, dipropyl chloromalonate, dipropyl bromomalonate, diisopropyl chloromalonate, diisopropyl bromomalonate, chloromalonate. Acid di-t-butyl, di-t-butyl bromomalonate, dibenzyl chloromalonate, dibenzyl bromomalonate, dibenzhydryl chloromalonate, dibenzhydryl bromomalonate, ditrityl chloromalonate, ditrityl bromomalonate, di- (1--chloromalonate) Phenylethyl),
Examples include bromomalonate di- (1-phenylethyl), and the like. The reaction with the halogenomamalonic acid diester is carried out in a solvent, and any solvent can be used as long as it does not participate in the reaction, but halogenated hydrocarbon solvents such as dichloromethane, chloroform and carbon tetrachloride are used. Is preferably used. Reaction is from -20 ℃ to 5
It proceeds smoothly at 0 ° C.

Treatment of the reaction product with a halogenomamalonic acid diester with a weak base gives (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-silyloxymethylpyrrolidine. As the base used, alkali metal acid carbonates such as lithium hydrogen carbonate, sodium hydrogen carbonate and potassium hydrogen carbonate are preferably used, and potassium hydrogen carbonate is more preferably used.
Treatment with a weak base is performed with dichloromethane, chloroform,
The reaction is completed in a mixed solvent of a halogenated hydrocarbon solvent such as carbon tetrachloride and water, and the reaction is completed at 5 ° C to 50 ° C.

Fourth Step In this step, the silyl group of (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-silyloxymethylpyrrolidine is removed to obtain the compound of the present invention (S). -Or (R) -2-bis (alkoxycarbonyl) methylidene-5-hydroxymethylpyrrolidine.

Removal of the silyl group is carried out under the reaction conditions usually applied to the silyl group used [TW Green, "Prote.
ctive Groups in Organic Synthesis ", A Wiley Inters
cience Publication, John-Wiley & Sons, New York (1
981), pp 39-50.].

Fifth Step In this step, the hydroxyl group of (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-hydroxymethylpyrrolidine is sulfonylated to give the compound of the present invention (S).
-Or (R) -2-bis (alkoxycarbonyl) methylidene-5-sulfonyloxypyrrolidine.

The sulfonylation includes methanesulfonyl chloride, methanesulfonyl bromide, ethanesulfonyl chloride, ethanesulfonyl bromide, propanesulfonyl chloride, propanesulfonyl bromide, 1-methylethanesulfonyl chloride and 1-bromide.
-Methylethanesulfonyl, butanesulfonyl chloride, butanesulfonyl bromide, benzenesulfonyl chloride, benzenesulfonyl bromide, p-toluenesulfonyl chloride, p-toluene bromide
-Toluenesulfonyl, p-chlorobenzenesulfonyl chloride, p-chlorobenzenesulfonyl bromide, p-nitrobenzenesulfonyl chloride, p-nitrobenzenesulfonyl bromide, 2,4-dimethylbenzenesulfonyl chloride, 2,4-dimethylbenzenesulfonyl bromide, chloride 2, 4, 6
-With sulfonylating agents such as trimethylbenzenesulfonyl, 2,4,6-trimethylbenzenesulfonyl bromide, and the like.

The reaction is carried out in the presence of a base, and examples of the base used include pyridine, 4-N, N-dimethylaminopyridine, triethylamine, 1,5-diazabicyclo [4.3.0] non-5-ene. , Tertiary amines such as 1,8-diazabicyclo [5.4.0] undec-7-ene and 1,4-diazabicyclo [2.2.2] octane are used, and preferably pyridine is used. As the solvent used, any solvent can be used as long as it does not participate in the reaction, but ether solvents such as diethyl ether, tetrahydrofuran and 1,4-dioxane, halogenated hydrocarbons such as dichloromethane, chloroform and carbon tetrachloride. A system solvent is preferably used. When pyridine is used as the base, pyridine can be used also as the reaction solvent. The reaction is from -20 ° C to 5
It proceeds smoothly at 0 ° C.

Step 6 In this step, the sulfonyloxy group of (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-sulfonyloxymethylpyrrolidine is replaced with a chlorine atom, a bromine atom or an iodine atom. And is a compound of the invention,
It is intended to produce (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-halogenomethylpyrrolidine.

For substitution with halogen atoms, alkali metal chlorides such as lithium chloride, sodium chloride and potassium chloride, alkali metal bromides such as lithium bromide, sodium bromide and potassium bromide, lithium iodide, sodium iodide and iodine. Alkali metal iodides such as potassium bromide, quaternary ammonium chloride such as tetraethylammonium chloride, tetrabutylammonium chloride and tetrabenzylammonium chloride, tetraethylammonium bromide, tetrabutylammonium bromide, tetrabenzylammonium bromide and the like Quaternary ammonium iodide such as quaternary ammonium, tetraethylammonium iodide, tetrabutylammonium iodide, tetrabenzylammonium iodide and the like are used.

The reaction is carried out in a solvent, and any solvent that does not participate in the reaction can be used, but acetonitrile, acetone, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, N, N -Dimethylformamide, N, N-dimethylacetamide and the like are used, and preferably acetonitrile and acetone are used. The reaction proceeds smoothly at 0 ° C to 80 ° C.

Step 7 In this step, (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-sulfonyloxymethylpyrrolidine is treated with a base to give the compound of the present invention (S)-. Alternatively, it produces (R) -1-aza-2-bis (alkoxycarbonyl) methylidenebicyclo [3.1.0] hexane.

As the base used in this step, sodium methoxide, sodium ethoxide, potassium t-
Alkali metal alkoxides such as butoxide, lithium hydride, sodium hydride, alkali metal hydrides such as potassium hydride, methyllithium, butyllithium,
Alkyl lithium such as sec-butyl lithium and t-butyl lithium, alkyl magnesium halide and aryl magnesium halide such as methyl magnesium chloride, methyl magnesium bromide, methyl magnesium iodide, butyl magnesium bromide and phenyl magnesium bromide, lithium magnesium, lithium Alkali metal amides such as diisopropylamide, lithium hexamethyldisilazide, sodium hexamethyldisilazide, and sodium amide are used, preferably alkali metal hydrides are used, and more preferably potassium hydride is used. Used.

The reaction is carried out in a solvent, and any solvent can be used as long as it does not participate in the reaction, but diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, 1,3-dioxane, etc. The ether-based solvent is used, and tetrahydrofuran is preferably used. The reaction proceeds smoothly at -50 ° C to 50 ° C.

Eighth Step In this step, (S)-or (R) -2-bis (alkoxycarbonyl) methylidene-5-halogenomethylpyrrolidine is treated with a base to give the compound of the present invention (S)-or (R)-
1-aza-2-bis (alkoxycarbonyl) methylidene bicyclo [3.1.0] hexane. This step proceeds smoothly under the reaction conditions exactly the same as those in the seventh step.

The general formula [I] obtained as described above and
The 2-bis (alkoxycarbonyl) methylidenepyrrolidine derivative represented by [II] was tested for malignant tumor cell growth inhibitory activity, and it was confirmed that these compounds exhibited strong cytotoxicity and had a use as an anticancer agent. ..

The present invention will be described in detail below with reference to Reference Examples, Examples and Test Examples, but it goes without saying that the present invention is not limited thereto.

[0034]

【Example】

 Reference example 1

[Chemical 9]

Reference [S. Saijo et al., Chem. Pharm. Bu
ll., 28, 1449 (1980).] (S) -5.
-Hydroxymethylpyrrolidin-2-one [mp. 86-87
℃, [α] _D ²⁰ + 28.9 ° (c 1.71, EtOH)] (6.10 g, 53.0 mmo
l) was stirred with imidazole (12.0 g, 173 mmol) and t-butylchlorodiphenylsilane (20 mL) in N, N-dimethylformamide (30 mL) at room temperature for 12 hours. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Fractions eluting with acetone-chloroform (5:95) were collected and collected (S) -5-
(T-Butyldimethylsilyloxy) methylpyrrolidin-2-one (19.0 g, 53.0 mmol, yield 100%) was obtained.

Mp. 81-83 ° C (recrystallized from hexane). [Α] _D ²⁰ + 15.3 ° (c 1.66, CHCl ₃ ). IR (nujor) 3200, 3100, 3060, 1690, 1460, 1105, 700, ^{505 cm -1. 1 H-NMR} (200 MHz, CDCl 3) δ = 1.05 (9H, s, C (CH 3) 3), 1.72 (1H, m, C 4 H), 2.18 (1
H, m, C ₄ H), 2.32 (2H, m, C ₃ H ₂ ), 3.51 (1H, dd, J = 7.6
_{and 10.2 Hz, C 5 CH 2} 0 -), 3.62 (1H, dd, J = 4.0 and 10.2
Hz, C ₅ CH ₂ 0-), 3.79 (1H, m, C ₅ H), 5.84 (1H, br s, N
H), 7.45 (6H, m, aromatic protons), 7.69 (4H, m, aro
matic protons). EI-MS (rel int.%) m / z = 354 (0.35, MH ⁺ ), 296 (100, Mt-Bu ⁺ ). Elemental analysis: (C ₂₁ H ₂₇ NO ₂ Si) Calculated value C = 71.35%, H = 7.70%, N = 3.96%. Analytical value C = 71.21%, H = 7.71%, N = 3.92%.

Reference Example 2

[Chemical 10]

As in Reference Example 1, the literature [S. Saijo et al.,
Chem. Pharm. Bull., 28, 1449 (1980).] (R) -5-hydroxymethylpyrrolidine-2
-. On _{[mp 86-88 ℃, [α] D} 20 -31.8 ° (c 1.75, EtO
H)) (4.00 g, 34.8 mmol), imidazole (5.00 g, 85.2
mmol), N, N-dimethylformamide (30 mL), t-butylchlorodiphenylsilane (10.0 g, 36.5 mmol),
(R) -5- (t-butyldimethylsilyloxy) methylpyrrolidin-2-one (12.2 g, 34.6 mmol, yield 99%) was obtained.

Mp. 81-82 ° C (recrystallized from hexane). [Α] _D ²⁰ -15.7 ° (c 1.71, CHCl ₃ ). The ¹ H-NMR spectrum was completely in agreement with that of the (S) -form. .. Elemental analysis: (C ₂₁ H ₂₇ NO ₂ Si) Calculated value C = 71.35%, H = 7.70%, N = 3.96%. Analytical value C = 71.21%, H = 7.84%, N = 4.03%.

Example 1

[Chemical 11]

(S) -5- (t-Butyldiphenylsilyloxy) methylpyrrolidin-2-one (5.00 g, 14.2 mmo
l) was heated to reflux with 2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphetan-2,4-disulfide (Lawesson reagent) in toluene (10 mL) for 20 minutes. After cooling to 0 ° C., the precipitated solid was filtered by suction filtration. The filtrate was concentrated under reduced pressure, and the obtained residue was purified by silica gel column chromatography. Fractions eluting with ethyl acetate-benzene (4:96) were collected and combined with (S) -5- (t-butyldiphenylsilyloxy) methylpyrrolidine-2-thione (5.03 g, 13.7 mmo).
l, yield 97%) was obtained.

Mp. 117-118 ° C. (recrystallized from ether-hexane). [Α] _D ²⁰ + 16.9 ° (c 1.06, CHCl ₃ ). IR (nujor) 3200, 2870, 2830, 1535, 1455, 1370, 1300, 1100, 10
^{15,695, 610, 500 cm -1.} 1 H-NMR (200 MHz, CDCl 3) δ = 1.07 (9H, s, C (CH 3) 3), 1.82 (1H, dddd, J = 6.1, 7.
9, 9.1, and 12.8 Hz, C ₄ H), 2.22 (1H, ddt, J = 6.4, 1
2.8, 8.3 Hz, C ₄ H), 2.94 (2H, m, C ₃ H ₂ ), 3.58 (1H, dd,
J = 7.7 and 10.6 Hz, C ₅ CH ₂ O-), 3.68 (1H, dd, J = 3.9 a
nd 10.6 Hz, C ₅ CH ₂ O-), 4.05 (1H, m, C ₅ H), 7.40-7.70
(10H, m, aromatic protons), 7.75 (1H, br s, NH) .EI-MS (rel int.%) M / z = 369 (7.0, M ⁺ ), 312 (100, Mt-Bu ⁺ ), 234 (99, M-
t-Bu-PhH ⁺ ). Elemental analysis: (C ₂₁ H ₂₇ NOSSi) Calculated value C = 68.24%, H = 7.36%, N = 3.79%, S = 8.67%. Analytical value C = 68.08%, H = 7.41 %, N = 3.75%, S = 8.60%.

Example 2

[Chemical 12]

As in Example 1, (R) -5- (t-butyldiphenylsilyloxy) methylpyrrolidin-2-one
(10.2 g, 28.9 mmol), Lawesson's reagent (6.40 g, 15.9 mm
ol) and toluene (40 mL), (R) -5- (t-butyldiphenylsilyloxy) methylpyrrolidine-2-thione (1
0.2 g, 27.6 mmol, yield 96%) were obtained.

Mp. 116-117 ° C. (recrystallized from ether-hexane). [Α] _D ²⁰ -17.4 ° (c 1.12, CHCl ₃ ). The ¹ H-NMR spectrum shows that it is completely the same as that of the (S) -form. Matched Elemental analysis: (C ₂₁ H ₂₇ NOSSi) Calculated value C = 68.24%, H = 7.36%, N = 3.79%, S = 8.67%. Analytical value C = 68.28%, H = 7.37%, N = 3.91%, S = 8.68%.

Reference Example 3

[Chemical 13]

(S) -5- (t-Butyldiphenylsilyloxy) methylpyrrolidine-2-thione (3.00 g, 8.13 mm
ol) and diethyl bromomalonate (2.0 mL) were stirred in dichloromethane (2.0 mL) at room temperature for 12 hours. An aqueous solution of potassium hydrogen carbonate (4.00 g, 40.0 mmol) (7.0 mL) was added, and the mixture was further stirred at room temperature for 4 hours. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Fractions eluting with ethyl acetate-benzene (4:96) were collected and collected (S) -2- (t-butyldiphenylsilyloxy) methyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (3.89 g, 7.85 mmol, yield). Rate 97%).

[Α] _D ²⁰ -29.4 ° (c 2.47, CHCl ₃ ). IR (film) 3300, 2930, 1685, 1645, 1565, 1243, 1105, 1085, 79
^{5,700, 500 cm -1. 1 H} -NMR (200 MHz, CDCl 3) δ = 1.05 (9H, s, C (CH 3) 3), 1.29, 1.31 (each 3H, t, J
= 7.3 Hz, CH ₃ × 2), 1.77 (1H, m, C ₃ H), 2.08 (1H, m, C ₃
H), 3.36 (2H, m, C ₄ H ₂ ), 3.58 (2H, m, C ₂ CH ₂ O-), 3.97
(1H, m, C ₂ H), 4.20, 4.21 (each 2H, q, J = 7.3 Hz, MeC
H ₂ O- × 2), 7.30-7.70 (10H, m, aromatic protons), 9.60
(1H, br s, NH). ¹³ C-NMR (100 MHz, CDCl3, deuterated chloroform chemical shift was 77.0 ppm.) Δ = 14.40, 14.45, 19.09, 23.81, 26.70, 33.77, 59.4
0, 59.53, 61.23, 66.55, 87.34, 127.70, 129.73, 132.
87, 135.54, 167.62, 169.56, 172.62 ppm.EI-MS (rel int.%) M / z = 495 (0.80, M ⁺ ), 450 (5.4, M-EtO ⁺ ), 438 (6.1, M
-t-Bu ⁺ ), 392 (100, Mt-Bu-PhH ⁺ ). EI-HIMS: (C ₂₈ H ₃₇ NO ₅ Si) Calculated value 495.2442 Analytical value m / z 495.2422

Reference Example 4

[Chemical 14]

As in Reference Example 3, (R) -5- (t-butyldiphenylsilyloxy) methylpyrrolidine-2-thione (3.00 g, 8.13 mmol), diethyl bromomalonate (2.0
mL), dichloromethane (2.0 mL), potassium hydrogen carbonate (4.0
0 g, 40.0 mmol) aqueous solution (7.0 mL), (R) -2- (t-
Butyldiphenylsilyloxy) methyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (3.90 g, 7.8
8 mmol, yield 97%) was obtained.

The [α] _D ²⁰ + 27.9 ° (c 1.76, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Reference Example 5

[Chemical 15]

In the same manner as in Reference Example 3, (S) -5- (t-butyldiphenylsilyloxy) methylpyrrolidine-2-thione (1.01 g 2.75 mmol), dibenzyl bromomalonate (1.0
g, 2.75 mmol) in dichloromethane (2.0 mL) at room temperature for 12 hours, and then potassium hydrogen carbonate (1.00 g, 10.0 mmo)
l) An aqueous solution (3.0 mL) was added, and the mixture was further stirred at room temperature for 3 hours. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Collect the fractions eluting with ether-hexane (40:60).
(S) -2-bis (benzyloxycarbonyl) methylidene-5- (t-butyldiphenylsilyloxy) methylpyrrolidine (1.50 g, 2.42 mmol, yield 88%) was obtained.

[Α] _D ²⁰ -19.4 ° (c 1.89, CHCl ₃ ). IR (film) 3300, 2980, 2950, 2930, 2860, 1685, 1645, 1570, 12
85,1245, 1110, 1085, 700, 500 cm -1. 1 H-NMR (400 MHz, CDCl 3) δ = 1.04 (9H, s, C (CH 3) 3), 1.75 (1H, ddt, J = 9.3, 13.
0, and 6.0 Hz, C ₄ H), 2.05 (1H, dddd, J = 7.0, 8.4, 9.
5, and 13.0 Hz, C ₄ H), 3.12 (1H, ddd, J = 6.0, 9.5, an
d 18.6 Hz, C ₃ H), 3.17 (1H, ddd, J = 7.0, 9.3, and 18.
6 Hz, C ₃ H), 3.52 (1H, dd, J = 6.4 and 10.3 Hz, C ₅ CH ₂ O
-), 3.62 (1H, dd, J = 4.3 and 10.3 Hz, C ₅ CH ₂ O-), 4.01
(1H, m, C ₅ H), 5.18 (2H, s, PhCH ₂ O-), 5.23, 5.26 (eac
h 1H, d, J = 12.8 Hz, PhCH ₂ O-), 7.23-7.43 (20H, m, ar
omatic protons), 9.83 (1H, br s, NH) .EI-MS (rel int.%) m / z = 619 (0.65, M ⁺ ), 562 (4.6, Mt-Bu ⁺ ), 454 (6.3, M)
-PhCH ₂ OH-t-Bu ⁺ ), 91 (100, PhCH ₂ ⁺ ). EI-HIMS: (C ₃₈ H ₄₁ NO ₅ Si) Calculated value 619.2755 Analytical value m / z 619.2736

Reference Example 6

[Chemical 16]

In the same manner as in Reference Example 3, (R) -5- (t-butyldiphenylsilyloxy) methylpyrrolidine-2-thione (2.94 g 7.96 mmol) and dibenzyl bromomalonate (2.
90 g, 7.99 mmol) from dichloromethane (2.0 mL) and potassium hydrogen carbonate (4.00 g, 40.0 mmol) aqueous solution (1.0 mL) were added to (R) -2-bis (benzyloxycarbonyl) methylidene- 5- (t-Butyldiphenylsilyloxy) methylpyrrolidine (4.87 g, 7.88 mmol, yield 99%) was obtained.

The [α] _D ²⁰ + 19.2 ° (c 2.01, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 3

[Chemical 17]

(S) -2- (t-butyldiphenyloxy)
A tetrabutylammonium fluoride-tetrahydrofuran solution (1.0 M, 9.0 mL) was added to a tetrahydrofuran solution (20 mL) of methyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (3.80 g, 7.67 mmol), and the mixture was stirred at room temperature for 2 hours. .. After removing the solvent under reduced pressure, the obtained residue was purified by silica gel column chromatography. Collect the fractions eluting with acetone-chloroform (20:80) (S)-
2-Bis (ethoxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (1.80 g, 7.00 mmol, yield 91%)
Got

[Α] _D ²⁰ + 22.4 ° (c 1.06, CHCl ₃ ). IR (film) 3430, 3320, 2980, 1685, 1650, 1575, 1440, 1250, 10
^{80, 800 cm -1. 1 H} -NMR (400 MHz, CDCl 3) δ = 1.29, 1.30 (each 3H, t, J = 7.1 Hz, CH 3 × 2), 1.78
(1H dddd, J = 6.0, 6.7, 9.6, and 13.0 Hz, C ₄ H), 2.13
(1H dddd, J = 6.0, 8.2, 9.6, and 13.0 Hz, C ₄ H), 3.0
8 (1H, ddd, J = 6.7, 9.6, and 18.3 Hz, C ₃ H), 3.18 (1
H, ddd, J = 6.0, 9.6, and 18.3 Hz, C ₃ H), 3.55 (1H, br
dd, J = 6.8 and 11.0 Hz, C ₅ CH ₂ O-), 3.78 (1H, brdd, J =
4.0 and 11.0 Hz, C ₅ CH ₂ O-), 4.01 (1H, m, C ₅ H), 4.18
(4H, m, MeCH ₂ O- × 2), 9.59 (1H, br s, NH) .EI-MS (rel int.%) M / z = 257 (20, M ⁺ ), 226 (36, M- CH ₂ OH ⁺ ), 212 (36, ME
tO ⁺ ), 180 (M-CH ₂ OH-EtO ⁺ ). EI-HIMS: (C ₁₂ H ₁₉ NO ₅ ) calculated 257.1263 analytical m / z 257.1282

Example 4

[Chemical 18]

In the same manner as in Example 3, (R) -2- (t-butyldiphenyloxy) methyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (3.90 g, 7.88 mmol), tetrahydrofuran (20 mL), fluorine was added. Tetrabutylammonium chloride-tetrahydrofuran solution (1.0 M, 8.5 mL)
(R) -2-bis (ethoxycarbonyl) methylidene-5
-Hydroxymethylpyrrolidine (1.94 g, 7.54 mmol, yield 96%) was obtained.

The [α] _D ²⁰ -22.4 ° (c 1.21, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 5

[Chemical 19]

As in Example 3, (S) -2-bis (benzyloxycarbonyl) methylidene-5- (t-butyldiphenylsilyloxy) methylpyrrolidine (1.50 g,
2.42 mmol) in tetrahydrofuran solution (10 mL) in tetrabutylammonium fluoride-tetrahydrofuran solution
(1.0 M, 3.0 mL) was added, and the mixture was stirred at room temperature for 1 hr. After removing the solvent under reduced pressure, the obtained residue was purified by silica gel column chromatography. Acetone-chloroform (10:
The fractions eluted at 90) were collected to obtain (S) -2-bis (benzyloxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (890 mg, 2.34 mmol, yield 97%).

[Α] _D ²⁰ + 17.5 ° (c 1.71, CHCl ₃ ). IR (film) 3450, 3300, 2940, 1680, 1640, 1560, 1435, 1245, 10
^{70, 690 cm -1. 1 H} -NMR (400 MHz, CDCl 3) δ = 1.75 (1H, dddd, J = 6.1, 6.9, 9.3, and 13.0 Hz, C
₄ H), 2.10 (1H, dddd, J = 6.4, 8.4, 9.1, and 13.0 Hz,
C ₄ H), 3.08 (1H, ddd, J = 6.1, 9.1, and 18.7 Hz, C ₃ H),
3.22 (1H, ddd, J = 6.4, 9.3, and 18.7 Hz, C ₃ H), 3.51
(1H, dd, J = 6.6 and 11.1 Hz, C ₅ CH ₂ O-), 3.72 (1H, dd,
J = 3.7 and 11.1 Hz, C ₅ CH ₂ O-), 4.00 (1H, m, C ₅ H), 5.1
8, 5.20 (each 2H, s, PhCH ₂ O- × 2), 7.20-7.40 (10H,
m, aromatic protons), 9.70 (1H, br s, NH) .EI-MS (rel int.%) m / z = 381 (1.7, M ⁺ ), 350 (0.89, M-CH ₂ OH ⁺ ), 274 (1.9,
M-PhCH ₂ O ⁺ ), 247 (2.4, MH-PhCH ₂ O ₂ C ⁺ ), 242 (2.2, MC
H ₂ OH-PhCH ₂ O ⁺ ), 216 (4.0, M-CH ₂ OH-PhCH ₂ O ₂ C ⁺ ), 91 (10
0. PhCH ₂ ⁺ ). EI-HIMS: (C ₂₂ H ₂₃ NO ₅ ) calculated value 381.1577 analytical value m / z 381.1603

Example 6

[Chemical 20]

As in Example 3, (R) -2-bis (benzyloxycarbonyl) methylidene-5- (t-butyldiphenylsilyloxy) methylpyrrolidine (4.88 g,
7.86 mmol), tetrahydrofuran (20 mL), tetrabutylammonium fluoride-tetrahydrofuran solution (1.0
M, 8.5 mL), (R) -2-bis (benzyloxycarbonyl) methylidene-5-hydroxymethylpyrrolidine
(2.80 mg, 7.35 mmol, yield 93%) was obtained.

The [α] _D ²⁰ -17.3 ° (c 1.83, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 7

[Chemical 21]

(S) -2-bis (ethoxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (1.80 g,
7.00 mmol), p-toluenesulfonyl chloride (2.00 g, 10.5
mmol) and pyridine (2.0 mL) in dichloromethane (20 mL)
The mixture was stirred at room temperature for 12 hours. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Fractions eluting with ethyl acetate-benzene (20:80) were collected and combined with (S) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (2.66 g, 6.48 mmol, yield 92
%).

Mp. 61-63 ° C (recrystallized from ether-hexane). [Α] _D ²⁰ -21.4 ° (c 1.18, CHCl ₃ ). IR (nujor) 3320, 1680, 1630, 1650, 1460 1360, 1270. , 1245, 117
0, 1080, 940,870, 665, 550 cm -1. 1 H-NMR (400 MHz, C 6 D 6) δ = 0.85 (1H, m, C 4 H), 1.09 (1H, m, C 4 H), 1.39 (6H,
t, J = 7.1 Hz, CH ₃ × 2), 1.84 (3H, s, CH ₃ Ar), 2.71 (1H,
ddd, J = 6.2, 9.6, and 18.5 Hz, C ₃ H), 2.81 (1H, ddd,
J = 6.5, 9.4, and 18.5 Hz, C ₃ H), 3.13 (1H, m, C ₅ H),
3.30 (1H, dd, J = 6.7 and 10.2 Hz, C ₅ CH ₂ O-), 3.47 (1
H, dd, J = 4.5 and 10.2 Hz, C ₅ CH ₂ O-), 4.15-4.26 (4H,
m, MeCH ₂ O- × 2), 6.75 (2H, brd, J = 7.7 Hz, aromatic
protons), 7.71 (2H, brd, J = 7.7 Hz, aromatic proton
s), 9.47 (1H, br s, NH). Elemental analysis: (C ₁₉ H ₂₅ NO ₇ S) Calculated value C = 55.46%, H = 6.12%, N = 3.40%, S = 7.79%. = 55.17%, H = 5.96%, N = 3.35%, S = 7.96%.

Example 8

[Chemical formula 22]

As in Example 7, (R) -2-bis (ethoxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (1.00 g, 3.89 mmol), p-toluenesulfonyl chloride (1.50 g, 7.89 mmol), pyridine (1.0 mL) and dichloromethane (7.0 mL) gave (R) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (1.60 g, 3.89 mmol, yield 100%). ..

[Α] _D ²⁰ + 21.3 ° (c 1.11, CHCl ₃ ). Mp. 62-64 ° C. (recrystallized from ether-hexane). ¹ H-NMR spectrum shows that of (S) -form Matched. Elemental analysis: (C ₁₉ H ₂₅ N0 ₇ S) Calculated value C = 55.46%, H = 6.12%, N = 3.40%, S = 7.79%. Analytical value C = 55.31%, H = 6.02%, N = 3.41% , S = 7.94%.

Example 9

[Chemical formula 23]

In the same manner as in Example 7, (S) -2-bis (benzyloxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (600 mg, 1.57 mmol) and p-toluenesulfonyl chloride (600 mg, 3.16 mmol) were added. Pyridine (300 μL) was stirred in dichloromethane (3.0 mL) at room temperature for 24 hours. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Collect the fractions eluting with ethyl acetate-benzene (7:93) to obtain (S) -2
-Bis (benzyloxycarbonyl) methylidene-5-
(P-Toluenesulfonyloxy) methylpyrrolidine (78
6 mg, 1.47 mmol, yield 94%) were obtained.

[Α] _D ²⁰ -18.5 ° (c 1.08, CHCl ₃ ). IR (film) 3300, 2950, 1685, 1640, 1560, 1360, 1270, 1245, 11
90, 1170,1070, 960, 810, 790, 695, 660, 550 cm -1. 1 H-NMR (200 MHz, CDCl 3) δ = 1.72 (1H, m, C 4 H), 2.13 (1H, m , C ₄ H), 2.41 (3H,
m, CH ₃ Ar), 3.19 (2H, br t, J = 8.2 Hz, C ₃ H ₂ ), 3.80-4.2
0 (3H, m, C ₅ , C ₅ CH ₂ O-), 5.16, 5.21 (each 2H, s, PhC
H ₂ O- × 2), 7.20-7.40 (12H, m, aromatic protons), 7.
76 (2H, dt, J = 1.0 and 8.0 Hz, aromatic protons),
9.60 (1H, br s, NH) .EI-MS (rel int.%) M / z = 535 (1.6, M ⁺ ), 488 (0.89, M-PhCH ₂ O ⁺ ), 401 (2.1,
MH-PhCH ₂ O ₂ C ⁺ ), 91 (100. PhCH ₂ ⁺ ). EI-HIMS: (C ₂₉ H ₂₉ NO ₇ S) Calculated value 535.1666 Analytical value m / z 535.1677

Example 10

[Chemical formula 24]

In the same manner as in Example 7, (R) -2-bis (benzyloxycarbonyl) methylidene-5-hydroxymethylpyrrolidine (1.00 g, 2.63 mmol), p-toluenesulfonyl chloride (1.00 g, 5.26 mmol), Pyridine (1.0 mL),
From dichloromethane (7.0 mL) (R) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (1.37 g, 2.56 mmo
l, yield 98%) was obtained.

[Α] _D ²⁰ + 17.6 ° (c 1.08, CHCl ₃ ). ¹ H-NMR spectrum was completely in agreement with that of the (S) -form.

Example 11

[Chemical 25]

(S) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (200 mg, 487 μmol) was added to acetonitrile.
(1.5 mL) in tetrabutylammonium bromide (300 mg, 9
After refluxing with 30 μmol) for 3 hours, the mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Fractions eluting with ethyl acetate-benzene (10:90) were collected and (S) -2-bromomethyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (150 mg,
469 μmol, yield 96%) was obtained.

Mp. 85-86 ° (recrystallized from hexane). [Α] _D ²⁰ -17.4 ° (c 1.06, CHCl ₃ ). IR (nujor) 3240, 1665, 1560, 1435, 1360, 1315, 1280, 1240, 80
^{0 cm -1. 1 H-NMR} (200 MHz, CDCl 3) δ = 1.29, 1.30 (each 3H, t, J = 7.1 Hz, CH 3 × 2), 1.83
(1H dddd, J = 5.5, 6.7, 9.3, and 12.2 Hz, C ₃ H), 2.24
(1H dddd, J = 6.5, 8.0, 9.2, and 12.2 Hz, C ₃ H), 3.1
0 (1H, ddd, J = 6.6, 9.1, and 18.5 Hz, C ₄ H), 3.18 (1
H, ddd, J = 6.5, 9.3, and 18.5 Hz, C ₄ H), 3.37 (1H, d
d, J = 6.8 and 10.3 Hz, C ₂ CH ₂ Br), 3.42 (1H, dd, J = 5.4
and 10.3 Hz, C ₂ CH ₂ Br), 4.17 (1H, m, C ₂ H), 4.18, 4.1
2 (each 2H, q, J = 7.1 HZ, MeCH ₂ O- × 2), 9.70 (1H, br
s, NH). Elemental analysis: (C ₁₂ H ₁₈ NO ₄ Br) Calculated value C = 45.02%, H = 5.67%, N = 4.37%, Br = 24.96%. Analytical value C = 44.95%, H = 5.43% , N = 4.29%, Br = 24.80%.

Example 12

[Chemical formula 26]

As in Example 11, (R) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (200 mg, 487 μmo)
l), acetonitrile (1.5 mL), and tetrabutylammonium bromide (300 mg, 930 μmol) to give (R) -2-bromomethyl-5-bis (ethoxycarbonyl) methylidenepyrrolidine (145 mg, 454 μmol, yield). 93%).

Mp. 85-86 ° (recrystallized from hexane). [Α] _D ²⁰ + 17.6 ° (c 1.03, CHCl ₃ ). ¹ H-NMR spectrum was completely in agreement with that of the (S) -form. .. Elemental analysis: (C ₁₂ H ₁₈ NO ₄ Br) Calculated value C = 45.02%, H = 5.67%, N = 4.37%, Br = 24.96%. Analytical value C = 45.00%, H = 5.56%, N = 4.29% , Br = 24.88%.

Example 13

[Chemical 27]

(S) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (172 mg, 418 μmol) was added to acetone (3.0 m).
L) with sodium iodide (144 mg, 961 μmol) in
After refluxing for an hour, the mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Collect the fractions eluting with ethyl acetate-benzene (10:90).
(S) -2-Bis (ethoxycarbonyl) methylidene-5
-Iodomethylpyrrolidine (151 mg, 411 μmol, yield 98
%).

Mp. 119-120 ° (recrystallized from ether-hexane). [Α] _D ²⁰ -58.7 ° (c 1.19, CHCl ₃ ). IR (nujor) 3240, 1665, 1625, 1560, 1435, 1280, 1240, 1090, 80
^{0 cm -1. 1 H-NMR} (200 MHz, CDCl 3) δ = 1.28 (3H, t, J = 7.1 Hz, CH 3), 1.30 (3H, t, J = 7.2
Hz, CH ₃ ), 1.77 (1H, dddd, J = 5.6, 6.8, 9.4, and 13.2
Hz, C ₄ H), 2.26 (1H, dddd, J = 6.1, 7.9, 9.3, and 13.
2 Hz, C ₄ H), 3.00-3.38 (4H, m, C ₃ H ₂ , C ₅ CH ₂ I), 4.09 (1
H, br quint, J = 6.0 Hz, C ₅ H), 4.17 (2H, q, J = 7.1 Hz,
MeCH ₂ O-), 4.20 (2H, q, J = 7.2 Hz, MeCH ₂ O-), 9.70 (1
H, br s, NH). Elemental analysis: (C ₁₂ H ₁₈ NO ₄ I) Calculated value C = 39.25%, H = 4.94%, N = 3.81%. Analytical value C = 39.09%, H = 5.00%, N = 3.74%.

Example 14

[Chemical 28]

As in Example 13, (R) -2-bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (162 mg, 394 μmo)
l), acetone (2.5 mL), sodium iodide (240 mg, 1.6
(R) -2-bis (ethoxycarbonyl) methylidene-5-iodomethylpyrrolidine (133 mg, 362 μ)
mol, yield 92%) was obtained.

Mp. 117-118 ° (recrystallized from ether-hexane). [Α] _D ²⁰ + 57.7 ° (c 1.11, CHCl ₃ ). ¹ H-NMR spectrum shows that it is completely that of the (S) -form. Matched Elemental analysis: (C ₁₂ H ₁₈ NO ₄ I) calculated C = 39.25%, H = 4.94%, N = 3.81%. Analytical C = 39.30%, H = 4.78%, N = 3.68%.

Example 15

[Chemical 29]

In the same manner as in Example 11, (S) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (149 mg, 279
μmol) was refluxed with tetrabutylammonium bromide (193 mg, 599 μmol) in acetonitrile (1.5 mL) for 4 hours, poured into water and extracted with ether three times. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. Fractions eluting with ethyl acetate-benzene (2:98) were collected and collected (S) -2-
Bis (benzyloxycarbonyl) methylidene-5-bromomethylpyrrolidine (117 mg, 264 μmol, yield 95%) was obtained.

[Α] _D ²⁰ -12.9 ° (c 0.933, CHCl ₃ ). IR (film) 3300, 3030, 2950, 1685, 1640, 1575, 1430, 1275, 12
40, 1070,1020, 795, 725 cm -1. 1 H-NMR (200 MHz, CDCl 3) δ = 1.81 (1H, dddd, J = 5.6, 6.6, 9.3, and 13.2 Hz, C
₄ H), 2.24 (1H, dddd, J = 6.5, 8.1, 9.2, and 13.2 Hz,
C ₄ H), 3.14 (1H, ddd, J = 6.6, 9.2, and 18.7 Hz, C ₃ H),
3.26 (1H, ddd, J = 6.5, 9.4, and 18.7 Hz, C ₃ H), 3.33
(1H, dd, J = 6.7 and 10.4 Hz, C ₅ CH ₂ Br), 3.44 (1H, dd,
J = 5.4 and 10.4 Hz, C ₅ CH ₂ Br), 4.19 (1H, br quint, J
= 6.0 Hz, C ₅ H), 5.18, 5.23 (each 2H, s, PhCH ₂ O- × 2),
7.20-7.40 (10H, m, aromatic protons), 9.75 (1H, br
s, NH) .EI-MS (rel int.%) m / z = 445 (0.75, M ⁺ ), 443 (0.60, M ⁺ ), 338 (0.51, MP
hCH ₂ O ⁺ ), 336 (0.53, M-PhCH ₂ O ⁺ ), 311 (0.66, MH-PhCH ₂
O ₂ C ⁺ ), 309 (0.68, MH-PhCH ₂ O ₂ C ⁺ ), 248 (0.91, M-PhCH ₂
O-PhCH ₂ ⁺ ), 246 (0.93, M-PhCH ₂ O-PhCH ₂ ⁺ ), 91 (100. Ph
CH ₂ ⁺ ) .EI-HI-MS: (C ₂₂ H ₂₂ NO ₄ ⁸¹ Br): (C ₂₂ H ₂₂ NO ₄ ⁷⁹ Br) Calculated value 445.0713: Calculated value 443.0732 Analytical value m / z 445.0710: Analytical value m / z 443.0726

Example 16

[Chemical 30]

In the same manner as in Example 15, (R) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (178 mg, 333
μR), acetonitrile (2.0 mL), tetrabutylammonium bromide (200 mg, 620 μmol), and then (R) -2-bis (benzyloxycarbonyl) methylidene-5-bromomethylpyrrolidine (137 mg, 307 μmol, yield) 92%).

The [α] _D ²⁰ + 12.1 ° (c 1.08, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 17

[Chemical 31]

In the same manner as in Example 13, (S) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (160 mg, 299
μmol) in acetone (3.0 mL) with sodium iodide (103
After refluxing with (mg, 687 μmol) for 18 hours, the mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure. The crude product was purified using silica gel column chromatography. The fractions eluted with ethyl acetate-benzene (5:95) were collected to give (S) -2-bis (benzyloxycarbonyl) methylidene-5-iodomethylpyrrolidine (143 mg, 290 μmol, 97% yield). ..

[Α] _D ²⁰ -43.4 ° (c 1.04, CHCl ₃ ). IR (film) 3270, 2950, 1685, 1640, 1560, 1270, 1240, 1070, 10
^{10,795, 730, 695 cm -1.} 1 H-NMR (200 MHz, CDCl 3) δ = 1.79 (1H, dddd, J = 5.7, 6.6, 9.5, and 12.4 Hz, C
₄ H), 2.26 (1H, dddd, J = 6.1, 7.9, 9.3, and 12.4 Hz,
C ₄ H), 3.00-3.40 (4H, m, C ₃ H ₂ , C ₅ CH ₂ I), 4.19 (1H, br
quint, J = 6.0 Hz, C ₅ H), 5.21, 5.23 (each 2H, s, PhCH
₂ O- × 2), 7.20-7.40 (10H, m, aromatic protons), 9.73
(1H, br s, NH) .EI-MS (rel int.%) M / z = 491 (0.39, M ⁺ ), 384 (0.79, M-PhCH ₂ O ⁺ ), 357 (1.
2, MH-PhCH ₂ O ₂ C ⁺ ), 294 (1.1, MH-PhCH ₂ O-PhCH ₂ ⁺ ), 254
(0.94, MH-PhCH ₂ O-PhCH ₂ O ₂ C ⁺ ), 91 (100, PhCH ₂ ⁺ ) .EI-HI-MS: (C ₂₂ H ₂₂ NO ₄ I) Calculated value 491.0595 Analytical value m / z 491.0568

Example 18

[Chemical 32]

In the same manner as in Example 17, (R) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (158 mg, 295
μmol), acetone (2.5 mL), sodium iodide (176 m
g, 1.17 mmol), (R) -2-bis (benzyloxycarbonyl) methylidene-5-iodomethylpyrrolidine (1
33 mg, 270 μmol, yield 92%) were obtained.

The [α] _D ²⁰ + 44.0 ° (c 1.13, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 19

[Chemical 33]

[0107] Potassium hydride (10.0 mg, 250 µmol) degreased with hexane was placed in a dried 25 mL round-bottomed flask, and the mixture was placed under an argon atmosphere, and then tetrahydrofuran (10 m
L), and the mixture was stirred and suspended, and a tetrahydrofuran solution (1.0 mL) of (S) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (103 mg, 250 μmol) was added. Stir for 15 minutes. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were washed together with saturated saline, dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a crude product (65.0 mg, 200 MHz).
^{In the 1} H-NMR spectrum, almost no signals other than the raw material and the target substance were detected. The yield of the crude product was calculated to be 80% from the integrated ratio). The crude product was quickly purified using Florisil column chromatography. Fractions eluting with ethyl acetate-benzene (3:97) were collected and the desired (S) -1-aza-2-bis (ethoxycarbonyl) methylidenebicyclo [3.1.0] hexane (20.7 mg, 85.6 μmol, yield 34%), ethyl acetate-
Fractions eluting with benzene (10:90) were collected and further subjected to silica gel column chromatography to collect fractions eluting with ethyl acetate-benzene (10:90) to collect (S) -2-bis (ethoxycarbonyl) methylidene as a raw material. -5- (p-Toluenesulfonyloxy) methylpyrrolidine (11.3 mg, 47.3 μmol, recovery rate 18%) was obtained. The isolation yield was 38% when the recovery of the raw materials was taken into consideration.

[Α] _D ²⁰ + 104 ° (c 0.750, CHCl ₃ ). IR (film) 2980, 1730, 1710, 1640, 1290, 1260, 1230, 1070, 10
^{30 cm -1. 1 H-NMR} (400 MHz, CDCl 3) δ = 1.20, 1.28 (each 3H, t, J = 7.1 Hz, CH 3 × 2), 1.61
(1H, d, J = 5.1 Hz, C ₆ H α), 2.14 (2H, m, C ₄ H ₂ ), 2.40
(1H, dd, J = 1.0 and 4.3 Hz, C ₆ Hβ), 2.61 (1H, ddt, J
= 1.0, 17.9, and 9.1 Hz, C ₃ H α), 2.88 (1H, m, C ₅ H),
3.25 (1H, ddd, 6.4, 8.7, and 17.9 Hz, C ₃ Hβ), 4.11
(2H, q, J = 7.1 Hz, MeCH ₂ O-), 4.22, 4.31 (each 1H, d
q, J = 10.8 and 7.2 Hz, MeCH ₂ O-). ¹³ C-NMR (100 MHz, CDCl3, deuterated chloroform chemical shift was 77.0 ppm) δ = 14.07, 14.09, 24.34, 29.79, 39.76, 44.77 , 60.5
4, 61.16 115.76, 164.26, 166.29, 174.64 ppm.EI-MS (rel int.%) M / z = 239 (2.3, M ⁺ ), 210 (2.6, M-Et ⁺ ), 194 (4.3, ME
tO ⁺ ), 180 (100, M-EtO-CH ₂ ⁺ ) .EI-HI-MS: (C ₁₂ H ₁₇ NO ₄ ) calculated value 239.1158 analytical value m / z 239.1158

Example 20

[Chemical 34]

As in Example 19, potassium hydride (13.
5 mg, 388 μmol), tetrahydrofuran (10 mL), (R)-
2-bis (benzyloxycarbonyl) methylidene-5
-(P-Toluenesulfonyloxy) methylpyrrolidine
(139 mg, 388 μmol) in tetrahydrofuran (1.0 m
(R) -1-aza-2-bis (ethoxycarbonyl) methylidenebicyclo [3.1.0] hexane (15.7 mg, 65.7 μmol, yield 19%), the target (R) ) -2-
Bis (ethoxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (28.3 mg,
68.9 μmol, recovery rate 20%) was obtained. 200 MHz ¹ H- of crude product
The yield of the crude product was calculated to be 70% from the integration ratio of the NMR spectrum. The isolation yield is 26 considering the recovery of raw materials.
%Met.

The [α] _D ²⁰ -93.0 ° (c 0.800, CHCl ₃ ). ¹ H-NMR spectrum was in perfect agreement with that of the (S) -form.

Example 21

[Chemical 35]

[0113] Potassium hydride (3.60 mg, 90.0 µmol) degreased with hexane was placed in a dried 25 mL round-bottomed flask, and the mixture was placed under an argon atmosphere, and then tetrahydrofuran (1.5
mL) and stirred to suspend, and a solution of (S) -2-bis (benzyloxycarbonyl) methylidene-5-bromomethylpyrrolidine (40.0 mg, 90.1 μmol) in tetrahydrofuran is added.
(1.0 mL) was added and stirred for 15 minutes. Pour the reaction mixture into water,
Extract 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The crude product was quickly purified using Florisil column chromatography. Fractions eluting with ethyl acetate-benzene (2:98) were collected and the raw material (S) -2-bis (benzyloxycarbonyl) methylidene-5-bromomethylpyrrolidine (16.0 mg, 36.0 μmol, recovery rate 40%),
Fractions eluting with ethyl acetate-benzene (5:95) were collected to obtain the desired product, (S) -1-aza-2-bis (benzyloxycarbonyl) methylidenebicyclo [3.1.
0] hexane (5.7 mg, 15.7 μmol, yield 17%) was obtained. The isolation yield was 29% in consideration of the recovery of the raw materials.

IR (film) 3030, 2950, 1720, 1640, 1560, 1290, 1260, 1220, 10
^{70,1015, 750, 695 cm -1.} 1 H-NMR (200 MHz, CDCl 3) δ = 1.67 (1H, d, J = 4.3 Hz, C 6 Hα), 2.19 (2H, m, C
₄ H ₂ ), 2.42 (1H, dd, J = 0.9 and 5.0 Hz, C ₆ H β), 2.70
(1H, ddt, J = 0.9, 20.1, and 8.0 Hz, C ₃ H α), 2.98 (1
H, m, C ₅ H), 3.33 (1H, ddd, J = 6.9, 7.1, and 20.1 Hz,
C ₃ H β), 5.15 (2H, s, PhCH ₂ O-), 5.21, 5.35 (each 1H,
d, J = 12.3 Hz, PhCH ₂ O-), 7.20-7.40 (10H, m, aromati
EI-MS (rel int.%) m / z = 363 (0.25, M ⁺ ), 350 (0.20, M-CH2-H ⁺ ), 272 (5.
4, M-PhCH ₂ ⁺ ), 256 (0.48, M-PhCH ₂ O ⁺ ), 242 (4.6, M-Ph
CH ₂ O-CH2-H ⁺ ), 91 (100, PhCH ₂ ⁺ ). EI-HI-MS: (C ₂₂ H ₂₁ NO ₄ ) calculated value 363.1471 analytical value m / z 363.1459

Example 22

[Chemical 36]

As in Example 21, potassium hydride (3.7
mg, 92.5 μmol), tetrahydrofuran (1.5 mL), (R)
-2-bis (benzyloxycarbonyl) methylidene-
Using a solution of 5-bromomethylpyrrolidine (41.0 mg, 92.3 μmol) in tetrahydrofuran (1.0 mL), the starting material (R)-
2-bis (benzyloxycarbonyl) methylidene-5
-Bromomethylpyrrolidine (6.4 mg, 14.0 μmol, recovery rate
16%), the desired product (R) -1-aza-2-bis (benzyloxycarbonyl) methylidenebicyclo [3.1.
0] hexane (6.2 mg, 17.0 μmol, yield 18%) was obtained.
The isolation yield was 22% in consideration of the recovery of the raw materials.

¹ H-NMR was in perfect agreement with that of the (S) -form.

Test Example Mouse lymphocytic leukemia cells (P388) were added to RPMI-1940 culture medium containing 10% fetal calf serum, and the cultured cells were added to 5 cells.
It was adjusted to × 10 ⁴ cells / mL. The 2-bis (alkoxycarbonyl) methylidenepyrrolidine derivative obtained in Examples 3 to 22, which is a compound of the present invention, was added thereto to a predetermined concentration, and the mixture was cultured at 37 ° C for 2 days. The number of floating cells was counted using a Coulter counter, and IC ₅₀ was calculated from the growth inhibition rate relative to the control group. 20 of the results obtained
Table 1 summarizes the compounds showing IC ₅₀ values of μg / mL or less.

[0119]

[Table 1]

[0120]

[Table 2]

─────────────────────────────────────────────────── ───

[Procedure amendment]

[Submission date] January 22, 1993

[Procedure Amendment 1]

[Document name to be amended] Statement

[Correction target item name] 0015

[Correction method] Change

[Correction content]

As the silylating agent used, chlorotrimethylsilane, bromotrimethylsilane, trimethylsilyltrifluoromethanesulfonate, chlorotriethylsilane, bromotriethylsilane, triethylsilyltrifluoromethanesulfonate, chlorodimethylisopropylsilane, bromodimethylisopropylsilane, dimethylisopropyl Silyltrifluoromethanesulfonate, t-butylchlorodimethylsilane, bromo-t-butyldimethylsilane, t-butyldimethylsilyltrifluoromethanesulfonate, t-butylchlorodiphenylsilane, bromo-t-butyldiphenylsilane, t-butyldiphenyl Silyltrifluoromethanesulfonate, chloromethyldiisopropylsilane, tribenzylchlorosilane, chloroto - p-Kishirirushiran, chlorotriisopropylsilane, bromo triphenyl silane, but is used like, preferably, t- butyl butylchlorodiphenylsilane is used. Silylation is performed under normal silylation conditions [TW Green, "Protective Groups in Organi
c Synthesis ", A Wiley Interscience Publication, Jo
hn-Wiley & Sons, New York (1981), pp39-50].

[Procedure Amendment 2]

[Document name to be amended] Statement

[Name of item to be corrected] 0027

[Correction method] Change

[Correction content]

The reaction is carried out in a solvent, and any solvent that does not participate in the reaction can be used, but acetonitrile, acetone, diethyl ether, tetrahydrofuran, 1,2-dimethoxyethane, N, N -Dimethylformamide, N, N-dimethylacetamide and the like are used, and preferably acetonitrile and acetone are used. The reaction proceeds smoothly at 0 ° C to 82 ° C.

[Procedure 3]

[Document name to be amended] Statement

[Correction target item name] 0084

[Correction method] Change

[Correction content]

Mp. 85-86 ° (recrystallized from hexane). [Α] _D ²⁰ -17.4 ° (c 1.06, CHCl ₃ ). IR (nujor) 3240, 1665, 1560, 1435, 1360, 1315, 1280, 1240, 80
^{0 cm -1. 1 H-NMR} (200 MHz, CDCl 3) δ = 1.29, 1.30 (each 3H, t, J = 7.1 Hz, CH 3 × 2), 1.83
(1H dddd, J = 5.5, 6.7, 9.3, and 12.2 Hz, C ₃ H), 2.24
(1H dddd, J = 6.5, 8.0, 9.2, and 12.2 Hz, C ₃ H), 3.1
0 (1H, ddd, J = 6.6, 9.1, and 18.5 Hz, C ₄ H), 3.18 (1
H, ddd, J = 6.5, 9.3, and 18.5 Hz, C ₄ H), 3.37 (1H, d
d, J = 6.8 and 10.3 Hz, C ₂ CH ₂ Br), 3.42 (1H, dd, J = 5.4
and 10.3 Hz, C ₂ CH ₂ Br), 4.17 (1H, m, C ₂ H), 4.18, 4.2
1 (each 2H, q, J = 7.1 HZ, MeCH ₂ O- × 2), 9.70 (1H, br
s, NH). Elemental analysis: (C ₁₂ H ₁₈ NO ₄ Br) Calculated value C = 45.02%, H = 5.67%, N = 4.37%, Br = 24.96%. Analytical value C = 44.95%, H = 5.43% , N = 4.29%, Br = 24.80%.

[Procedure amendment 4]

[Document name to be amended] Statement

[Name of item to be corrected] 0107

[Correction method] Change

[Correction content]

[0107] Potassium hydride (10.0 mg, 250 µmol) degreased with hexane was placed in a dried 25 mL round-bottomed flask, and the mixture was placed under an argon atmosphere, and then tetrahydrofuran (10 m
L), and the mixture was stirred and suspended, and a tetrahydrofuran solution (1.0 mL) of (S) -2-bis (benzyloxycarbonyl) methylidene-5- (p-toluenesulfonyloxy) methylpyrrolidine (103 mg, 250 μmol) was added. Stir for 15 minutes. The reaction mixture was poured into water and extracted 3 times with ether. The ether layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give the crude product (65.0 mg, 200 MHz
^{In the 1} H-NMR spectrum, almost no signals other than the raw material and the target substance were detected. The crude product yield was calculated to be 80% from the integral ratio). The crude product was quickly purified using Florisil column chromatography. Fractions eluting with ethyl acetate-benzene (3:97) were collected and the desired (S) -1-aza-2-bis (ethoxycarbonyl) methylidenebicyclo [3.1.0] hexane (20.7 mg, 85.6 μmol, yield 34%), ethyl acetate-
Fractions eluting with benzene ( 5:95 ) were collected and further subjected to silica gel column chromatography to collect fractions eluting with ethyl acetate-benzene (10:90) to collect (S) -2-bis (ethoxycarbonyl) methylidene as a raw material. -5- (p-Toluenesulfonyloxy) methylpyrrolidine (11.3 mg, 47.3 μmol, recovery rate 18%) was obtained. The isolation yield was 38% in consideration of the recovery of the raw materials.

Claims (3)

[Claims] 1. The following general formula: (In the formula, R ¹ and R ² are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms, or a substituted or unsubstituted aralkyl group)
Bis (alkoxycarbonyl) methylidenepyrrolidine derivative. 2. The following general formula: (In the formula, R ¹ and R ² are the same or different and each represents a linear or branched alkyl group having 1 to 5 carbon atoms or a substituted or unsubstituted aralkyl group, and X is a hydroxyl group or 1 carbon atom. ~ Represents a linear or branched alkanesulfonyloxy group, a substituted or unsubstituted allenesulfonyloxy group, a chlorine atom, a bromine atom, or an iodine atom.
A 2-bis (alkoxycarbonyl) methylidenepyrrolidine derivative represented by: 3. The following general formula: (In the formula, R ³ , R ⁴ , and R ⁵ are the same or different and are linear or branched alkyl groups having 1 to 5 carbon atoms;
Or represents a substituted or unsubstituted aryl group), a 5-silyloxymethylpyrrolidine-2-thione derivative.