Chiral spiro compounds, processes for their preparation and uses thereof

Abstract

The application provides a chiral spiro compound and a preparation method and application thereof, and belongs to the technical field of organic synthesis. The chiral spiro compound is obtained by mixing spiro olefin, alkyl iodide, cobalt salt, a chiral ligand, an organic solvent, a reducing agent and an alkaline substance to perform a hydrogen alkylization reaction. A cobalt-catalyzed stereodivergent asymmetric synthesis method is disclosed, high-efficiency preparation of a spiro compound containing 1,3-non-adjacent stereogenic centers is realized by ligand regulation, and the product has excellent biological activity. The method has wide substrate compatibility and high selectivity, and provides a new way for drug development.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of organic synthesis, and particularly relates to a chiral spiro compound and a preparation method and application thereof. BACKGROUND

[0002] In the field of natural and pharmacologically active compounds, chiral spiro compounds with multiple stereogenic centers are a unique and important structure, exhibiting a wide range of biological activities (such as anti-tumor and anti-inflammatory properties) and pharmacological effects (including the treatment of hypertension and nephrotic syndrome). It is worth noting that the introduction of a spiro skeleton into a drug candidate molecule can improve its pharmacokinetic properties, such as potency, selectivity, protein binding affinity, and metabolic stability. Due to the unique interaction between the chiral environment formed by the molecule and the biological macromolecule, different stereoisomers have significantly different effects on human physiological activities, however, the diastereomeric divergent asymmetric synthesis strategy for such skeletons with 1,3-non-adjacent stereogenic centers is still a great challenge, and the main problems include:

[0003] 1. Difficulty in stereochemical control: the rigid spiro skeleton leads to a highly restricted spatial environment of the substrate, making it difficult to achieve diastereomeric and enantioselective control;

[0004] 2. Limitations of existing methods: related technologies mostly rely on cyclization or cycloaddition reactions, which are only suitable for the construction of consecutive stereogenic centers, and have narrow substrate range and insufficient selectivity;

[0005] 3. Catalytic system is not developed: metal hydride catalyzed hydrogen alkylation reaction has not been successfully used for diastereomeric divergent synthesis of non-adjacent stereogenic centers. SUMMARY

[0006] Therefore, the purpose of the present application is to provide a chiral spiro compound and a preparation method and application thereof. The preparation method of the present application realizes the efficient stereodivergent synthesis of spiro compounds containing 1,3-non-adjacent stereogenic centers.

[0007] In order to achieve the above-mentioned purpose of the application, the present application provides the following technical solutions:

[0008] The present application provides a preparation method of a chiral spiro compound, comprising the following steps:

[0009] mixing a spiro olefin, an alkyl iodide, a cobalt salt, a chiral ligand, an organic solvent, a reducing agent, and a basic substance to perform a hydrogen alkylation reaction, to obtain the chiral spiro compound;

[0010] When the chiral ligand is a first ligand, the chiral spiro compound is a cis product, and the first ligand has a structure shown in formula a:

[0011] Formula a,

[0012] R is -H, -Me, -Et or -tBuC6H4 in formula a;

[0013] when the chiral ligand is a second ligand, the chiral spiro compound is a trans product, the second ligand having a structure shown in formula b:

[0014] formula b,

[0015] R is -Me, R' is -tBu or R is -iPr, R' is -tBu or R is -tBu, R' is -tBu or R is -Me, R' is -phenyl in formula b;

[0016] when the chiral ligand is a third ligand, the chiral spiro compound is an enantiomeric cis product, the third ligand being an enantiomer of the first ligand;

[0017] when the chiral ligand is a fourth ligand, the chiral spiro compound is an enantiomeric trans product, the fourth ligand being an enantiomer of the second ligand.

[0018] Preferably, the spiroalkene includes one of indolinone, benzofuranone, 3-isochromanone, pyrrolidinone and piperidinone and derivatives thereof.

[0019] Preferably, the spiroalkene has a structure shown in any one of formulas 1, 6-29:

[0020] .

[0021] Preferably, the alkyl iodide is methyl iodide, ethyl iodide, propyl iodide, propyl sulfide derived alkyl iodide or has a structure shown in any one of formulas I-X, the propyl sulfide derived alkyl iodide having a structure shown in formula A:

[0022] 、 formula A.

[0023] Preferably, the molar equivalent ratio of the spiroalkene to the alkyl iodide is 1:1.2-4.

[0024] Preferably, the cobalt salt includes one or more of cobalt halide, cobalt (II) bromide ethylene glycol dimethyl ether adduct and cobalt tetrafluoroborate; the molar amount of the cobalt salt is 5-40% of the molar amount of the spiroalkene.

[0025] Preferably, the organic solvent includes methyl tert-butyl ether and / or ethylene glycol dimethyl ether; the reducing agent includes dimethoxymethylsilane; the basic substance includes potassium phosphate hydrate.

[0026] Preferably, the hydrogen alkylating reaction is carried out at a temperature of -20 to 25°C for 48 to 96 hours.

[0027] The present application also provides a chiral spiro compound, including a cis product, a trans product, an enantiomeric cis product and an enantiomeric trans product.

[0028] The cis product has a structure shown in formula M:

[0029] Formula M;

[0030] In formula M, X is C, O or NR, R is methyl, ethyl, t-butyl, benzyl, aryl, substituted aryl or t-butyloxycarbonyl; alkyl is alkyl or substituted alkyl; n is 0 or 1; the dotted line represents a fused aromatic ring, a fused substituted aromatic ring, a fused heteroaromatic ring, a fused substituted heteroaromatic ring or no fused ring;

[0031] The trans product has a structure shown in formula N:

[0032] Formula N;

[0033] In formula N, X is C, O or NR, R is methyl, ethyl, t-butyl, benzyl, aryl, substituted aryl or t-butyloxycarbonyl; alkyl is alkyl or substituted alkyl; n is 0 or 1; the dotted line represents a fused aromatic ring, a fused substituted aromatic ring, a fused heteroaromatic ring, a fused substituted heteroaromatic ring or no fused ring;

[0034] The enantiomeric cis product has a structure shown in formula P:

[0035] Formula P;

[0036] In formula P, X is C, O or NR, R is methyl, ethyl, t-butyl, benzyl, aryl, substituted aryl or t-butyloxycarbonyl; alkyl is alkyl or substituted alkyl; n is 0 or 1; the dotted line represents a fused aromatic ring, a fused substituted aromatic ring, a fused heteroaromatic ring, a fused substituted heteroaromatic ring or no fused ring;

[0037] The enantiomeric trans product has a structure shown in formula Q:

[0038] Formula Q;

[0039] In formula Q, X is C, O or NR, R is methyl, ethyl, t-butyl, benzyl, aryl, substituted aryl or t-butyloxycarbonyl; alkyl is alkyl or substituted alkyl; n is 0 or 1; the dotted line represents a fused aromatic ring, a fused substituted aromatic ring, a fused heteroaromatic ring, a fused substituted heteroaromatic ring or no fused ring.

[0040] The application further provides application of the chiral spiro compound in preparation of anti-inflammatory drugs.

[0041] Compared with the prior art, the application has the following beneficial effects:

[0042] The application discloses a cobalt-catalyzed stereodivergent asymmetric synthesis method, which is a cobalt-catalyzed ligand-regulated C(sp3)-C(sp3) coupling reaction.

[0043] The application further provides the chiral spiro compound prepared by the preparation method, which has excellent biological activity. BRIEF DESCRIPTION OF DRAWINGS

[0044] Figure 1 The compound inhibits the expression level of an inflammatory factor IL-6 in a BV2 cell model induced by LPS;

[0045] Figure 2 The compound inhibits the expression level of an inflammatory factor TNF-alpha in a BV2 cell model induced by LPS;

[0046] Figure 3 The compound inhibits the expression level of an inflammatory factor IL-1beta in a BV2 cell model induced by LPS. DETAILED DESCRIPTION

[0047] The application provides a preparation method of a chiral spiro compound, which comprises the following steps:

[0048] The chiral spiro compound is obtained by mixing a spiro olefin, an alkyl iodide, a cobalt salt, a chiral ligand, an organic solvent, a reducing agent and an alkaline substance for a hydrogen alkylation reaction.

[0049] When the chiral ligand is a first ligand, the chiral spiro compound is a cis product, and the first ligand has a structure shown in formula a.

[0050] When the chiral ligand is a second ligand, the chiral spiro compound is a trans product, and the second ligand has a structure shown in formula b.

[0051] When the chiral ligand is a third ligand, the chiral spiro compound is an enantiomeric cis product, and the third ligand is an enantiomer of the first ligand.

[0052] When the chiral ligand is a fourth ligand, the chiral spiro compound is an enantiomeric trans product, and the fourth ligand is an enantiomer of the second ligand.

[0053] In the present application, the raw materials used are all commercially available products or products obtained by conventional methods in the art, unless otherwise specified.

[0054] In the present application, the spirocyclic olefin preferably includes indolinone, benzofuranone, 3-isochromone, pyrrolidinone and piperidinone, and derivatives thereof, and more preferably has a structure represented by any one of formulas 1, 6-29.

[0055] In the present application, the alkyl iodide is preferably methyl iodide, ethyl iodide, n-propyl iodide, or has a structure represented by any one of formulas I-X.

[0056] In the present application, the molar equivalent ratio of the spirocyclic olefin to the alkyl iodide is preferably 1:1.2-4, and more preferably 1:2.

[0057] In the present application, the cobalt salt preferably includes one or more of halogenated cobalt, cobalt (II) bromide ethylene glycol dimethyl ether adduct, and cobalt tetrafluoroborate, the halogenated cobalt more preferably includes one or more of cobalt chloride, cobalt bromide, and cobalt iodide, the cobalt tetrafluoroborate is preferably used in the form of cobalt tetrafluoroborate hydrate; the molar amount of the cobalt salt is preferably 4-40% of the molar amount of the spirocyclic olefin, more preferably 10-18%, and further preferably 12-15%, and the molar amount of the chiral ligand is preferably 6-60% of the molar amount of the spirocyclic olefin, more preferably 12-18%.

[0058] In the present application, the organic solvent preferably includes methyl tert-butyl ether and / or ethylene glycol dimethyl ether; the reducing agent preferably includes dimethoxymethylsilane; and the basic substance preferably includes potassium phosphate hydrate.

[0059] In the present application, the molar equivalent ratio of the spirocyclic olefin to the reducing agent is preferably 1:2-6, and more preferably 1:3-5.

[0060] In the present application, the molar equivalent ratio of the spirocyclic olefin to the basic substance is preferably 1:2-6, and more preferably 1:2.5-5.

[0061] In the present application, the first ligand preferably has a structure represented by formula L1:

[0062] Formula L1 (i.e., is ( S, S, R, R ) L1).

[0063] In the present application, the second ligand preferably has a structure represented by formula L2:

[0064] Formula L2 (i.e., is ( S, S ) L2).

[0065] In the present application, the third ligand is preferably (R, R, S, S, ) L1.

[0066] In the present application, the fourth ligand is (L2). R, R ) L2.

[0067] In the present application, the temperature of the hydrogen alkylation reaction is preferably -20-25°C, more preferably 0°C, and the time is preferably 48-96h.

[0068] In the present application, the hydrogen alkylation reaction is preferably carried out under stirring, and the stirring speed is preferably 650rpm.

[0069] After the hydrogen alkylation reaction is completed, the present application preferably dilutes the obtained reaction mixture with saturated aqueous ammonium chloride solution and ethyl acetate, extracts the obtained aqueous phase with ethyl acetate, combines the organic phases, and concentrates under reduced pressure to obtain a crude mixture, which is purified by silica gel column chromatography with petroleum ether / ethyl acetate (volume ratio, 10:1) as the eluent to obtain the chiral spiro compound.

[0070] The present application realizes diastereomeric divergent synthesis (dr>20:1, ee>99%) by switching the chirality of the first ligand and the second ligand.

[0071] In the present application, the dr of the cis product is >20:1, and the ee is >99%; the dr of the trans product is >20:1, and the ee is 90:10-99:1; and the dr of the stereodivergent isomers (including the enantiomeric cis product and the enantiomeric trans product) is >10:1.

[0072] The present application is suitable for a variety of spirocyclic olefins (such as indolinones, 3-isochromane ketone derivatives) and alkyl iodides (including drug molecule derivatives such as ibuprofen, naproxen); and has the potential for scale-up: still maintains high yield (55%) and stereoselectivity (dr>20:1) at a scale of 5.0 mmol.

[0073] The present application also provides a chiral spiro compound, including a cis product, a trans product, an enantiomeric cis product, and an enantiomeric trans product.

[0074] In the present application, the alkyl in the alkyl group in the cis product, the trans product, the enantiomeric cis product and the enantiomeric trans product preferably includes methyl, ethyl or propyl, and the substituted alkyl group preferably includes phenethyl, 3-phenylpropyl, 3-(4-methoxyphenyl)propyl, 3-(4-bromophenyl)propyl, 3-methoxypropyl, 4-(benzyloxy)butyl, 4-(ethyl butyrate)butyl, 4-(4-methoxyphenoxy)butyl, 3-phenoxypropyl, 3-(tert-butyldimethylsiloxy)propyl, thien-2-carboxylic acid propyl ester, furan-2-carboxylic acid propyl ester, 3-(benzoyloxy)propyl or p-[(dipropylamino)sulfonyl]benzoic acid propyl ester.

[0075] In the present application, the chiral spiro compound preferably has a structure represented by formula trans -3af、 cis -3ab、 cis -3f、 cis- 3y、 cis- 3i'、 trans -3ax、 cis -3ao、 cis -3j、 trans -3az、 ent-cis -3ar、 trans -3ah、 cis -3o、 trans -3an、 cis -3s、 trans -3v、 trans -3ao、 trans -3ah'、 trans- 3pp、 cis- 3al、cis-3e、 cis -3v、 trans -3at、 cis -3a、 cis -3b、 cis -3c、 cis -3d、 cis -3g、 cis -3h、 cis -3i、 trans -3a、 trans -3c、 trans -3ai、 trans -3aj、 trans -3ak、 trans -3d、 trans -3u or trans-3ar, and specific structures are shown in the examples, which are not described herein again.

[0076] The present application also provides a use of the chiral spiro compound in the above technical solution in the preparation of an anti-inflammatory drug.

[0077] The technical solutions in the present application will be described clearly and completely below in connection with the embodiments in the present application. Obviously, the described embodiments are only some of the embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all the other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0078] Example 1

[0079] (1R,3R)-1'-Benzyl-3-(3-methoxypropyl)spiro[cyclopentane-1,3'-indolin]-2'-one

[0080] cis -3a

[0081] Under a nitrogen atmosphere, using glovebox techniques, an oven-dried Teflon screw-cap test tube was charged with a magnetic stir bar, followed by cobalt chloride (0.015 mmol, 15 mol%) and the first ligand L1 (0.018 mol, 18 mol%). Anhydrous methyl-tert-butyl ether was then added and stirred at room temperature for 15 min. Potassium phosphate (hydrate) (0.20 mmol, 2 equiv), spiroalkene 1 (0.10 mmol, 1 equiv), 1-iodo-3-methoxypropane (0.12 mmol, 1.2 equiv) and dimethoxymethylsilane (0.20 mmol, 2 equiv) were then added sequentially. The test tube was then sealed with airtight insulating tape, removed from the glovebox and stirred at 0 °C at 650 rpm for 48 h. Upon completion of the reaction, the reaction mixture was diluted with saturated aqueous ammonium chloride solution and ethyl acetate. The aqueous phase was extracted with ethyl acetate and the combined organic phases were concentrated under reduced pressure. The crude mixture was purified by flash column chromatography on silica gel using petroleum ether / ethyl acetate (10:1 by volume) as eluent to give the target cis product 22.6 mg, 65% yield.

[0082] 1H NMR (600 MHz, CDCl3) δ 7.33 – 7.28 (m, 2H), 7.28 – 7.22 (m, 3H),7.20 (dd, J = 7.4, 1.3 Hz, 1H), 7.10 (td, J = 7.7, 1.3 Hz, 1H), 7.00 (td, J =7.5, 1.0 Hz, 1H), 6.67 (d, J = 7.8 Hz, 1H), 4.89 (d, J = 3.0 Hz, 2H), 3.41(td, J = 6.5, 1.3 Hz, 2H), 3.34 (s, 3H), 2.50 (m, J = 10.4, 7.1 Hz, 1H), 2.35(dd, J = 13.1, 7.5 Hz, 1H), 2.28 – 2.18 (m, 2H), 1.99 – 1.93 (m, 1H), 1.72 –1.48 (m, 6H).

[0083] 13 C NMR (151 MHz, CDCl3) δ 182.4, 142.0, 136.7, 136.2, 128.7, 127.5,127.2, 127.2, 122.6, 122.2, 108.6, 77.2, 77.0, 76.8, 72.9, 58.5, 53.5, 45.0,43.6, 40.7, 38.1, 33.8, 32.0, 28.7.

[0084] HRMS (ESI) m / z: [M + H] + Calcd for C 23 H 28 NO2 + 350.2115; Found 350.2117.

[0085] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 90:10, 1.0 mL / min, 254 nm, t minor = 17.9 min,t major = 23.6 min.

[0086] [α]D 20= -133.5 (c = 0.60, CHCl3).

[0087] Example 2

[0088] (1R,3S)-5'-chloro-3-methyl-1'-phenylspiro[cyclopentane-1,3'-indolin]-2'-one

[0089] cis -3b

[0090] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by iodomethane and spirocyclic alkene 1 was replaced by spirocyclic alkene 7.

[0091] 1 H NMR (500 MHz, CDCl3) δ 7.67 (d, J = 8.0 Hz, 2H), 7.61 – 7.49 (m,2H), 7.30 (d, J = 8.0 Hz, 1H), 7.22 (d, J = 7.5 Hz, 2H), 7.09 -6.95 (m, 2H),6.54 (d, J = 3.0 Hz, 1H), 5.83 (q, J = 7.0 Hz, 1H), 1.93 (d, J = 7.0 Hz, 3H),1.30 (s, 12H).

[0092] 13 C NMR (126 MHz, CDCl3) δ 147.4, 137.0, 135.7 (2C), 129.8, 126.1,125.8, 121.9, 121.3, 120.0, 111.0, 102.0, 84.4, 55.5, 25.1, 21.8.

[0093] HRMS (ESI) m / z: [M + H] + Calcd for C 19 H 19 ClNO + 312.1150; Found 312.1155.

[0094] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, tminor = 10.2 min, t major = 14.5 min.

[0095] [α]D 20 = -81.1 (c = 1.08, CHCl3).

[0096] Example 3

[0097] (1R,3S)-3-ethyl-1'-phenylspiro[cyclopentane-1,3'-indolin]-2'-one

[0098] cis -3c

[0099] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by iodoethane, and spirocyclic olefin 1 was replaced by spirocyclic olefin 13.

[0100] 1 H NMR (500 MHz, CDCl3) δ 7.87 (s, 1H), 7.82 (d, J = 8.0 Hz, 1H), 7.55(d, J = 7.5 Hz, 1H), 7.26 – 7.15 (m, 2H), 7.11 (d, J = 6.5 Hz, 2H), 7.01 (t,J = 8.5 Hz, 2H), 6.49 (d, J = 3.5 Hz, 1H), 5.60 (q, J = 7.0 Hz, 1H), 3.79 (s,3H), 1.83 (d, J = 7.0 Hz, 3H).

[0101] 13 C NMR (126 MHz, CDCl3) δ 166.8, 143.2, 136.0, 130.6, 130.4, 128.9,128.8, 128.7, 127.1, 124.6, 121.6, 121.0, 119.6, 109.9, 101.8, 54.6, 52.1,21.6.

[0102] HRMS (ESI) m / z: [M + H] + Calcd for C 20 H 22 NO + 292.1696; Found 292.1695.

[0103] HPLC: >99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 14.1 min,t major = 20.3 min.

[0104] [α]D 20 = -221.4 (c = 0.80, CHCl3).

[0105] Example 4

[0106] (1R,3S)-1'-Benzyl-3-propylspiro[cyclopentane-1,3'-indolin]-2'-one

[0107] cis -3d

[0108] Preparation method was the same as example 1, except that 1-iodo-3- methoxypropane was replaced by iodopropane.

[0109] 1 H NMR (500 MHz, CDCl3) δ 7.35 – 7.18 (m, 6H), 7.11 (td, J = 7.7, 1.3Hz, 1H), 7.05 – 6.96 (m, 1H), 6.69 (d, J = 7.6 Hz, 1H), 4.90 (d, J = 1.8 Hz,2H), 2.37 (m, J = 10.1, 8.7, 5.2 Hz, 1H), 2.29 (m, J = 13.0, 8.8, 4.1 Hz,1H), 2.13 (m, J = 12.1, 7.4, 3.8 Hz, 1H), 1.99 (dd, J = 12.7, 7.1 Hz, 1H),1.93 – 1.82 (m, 2H), 1.73 (dt, J = 12.4, 9.1 Hz, 1H), 1.50 (m, J = 14.6,11.0, 6.1 Hz, 2H), 1.43 – 1.32 (m, 2H), 0.94 (t, J = 7.3 Hz, 3H).

[0110] 13C NMR (126 MHz, CDCl3) δ 182.0, 141.8, 137.3, 136.1, 128.7, 127.4,127.2, 127.2, 122.5, 122.4, 108.7, 77.3, 77.0, 76.7, 53.9, 44.9, 43.7, 41.3,37.8, 37.7, 33.0, 21.9, 14.3.

[0111] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 26 NO2 + 320.2009; Found 320.2010.

[0112] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 9.8 min, t major = 11.7 min.

[0113] [α]D 20 = -58 (c = 1.24, CHCl3).

[0114] Example 5

[0115] (1R,3R)-1'-Benzyl-3-(3-phenoxypropyl)spiro[cyclopentane-1,3'-indolin]-2'-one

[0116] cis -3e

[0117] Preparation method was the same as example 1, except that 1-iodo-3-methoxypropane was replaced by (3-iodopropoxy)benzene to give the cis target product 20.8 mg, yield 50%.

[0118] 1H NMR (500 MHz, CDC13) δ 7.34 - 7.18 (m, 8H), 7.12 (td, J = 7.7, 1.2 Hz, 1H), 7.00 (t, J = 7.5 Hz, 1H), 6.96 - 6.87 (m, 3H), 6.69 (d, J = 7.7 Hz, 1H), 4.91 (s, 2H), 3.99 (t, J = 6.4 Hz, 2H), 2.41 (m, J = 11.8, 8.3, 7.7 Hz, 1H), 2.31 (m, J = 12.9, 8.7, 4.1 Hz, 1H), 2.17 (m, J = 11.9, 9.4, 5.6 Hz, 1H), 2.04 (dd, J = 12.7, 7.1 Hz, 1H), 1.96 - 1.81 (m, 4H), 1.82 - 1.63 (m, 3H).

[0119] 13 C NMR (126 MHz, CDC13) δ 181.9, 159.0, 141.8, 137.0, 136.1, 129.4, 128.7, 127.5, 127.3, 127.2, 127.2, 122.6, 122.4, 120.5, 114.5, 108.8, 77.3, 77.0, 76.7, 67.8, 53.9, 44.8, 43.7, 41.2, 37.8, 33.0, 31.8, 28.5.

[0120] HRMS (ESI) m / z: [M + H] + Calcd for C 28 H 30 NO2 + 412.2171; Found 412.2173.

[0121] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 80:20, 1.0 mL / min, 254 nm, t minor = 11.7 min,t major = 14.5 min.

[0122] [α]D20 = -164.0 (c = 0.56, CHCl3).

[0123] Example 6

[0124] (1R,3S)-1'-Benzyl-3-(4-(4-methoxyphenoxy)butyl)spiro[cyclopentane-1,3'- indolin]-2'-one

[0125] cis -3f

[0126] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by 1-(3-iodopropoxy)-4-methoxybenzene to obtain the cis target product 23.7 mg, yield 52%.

[0127] 1 H NMR (500 MHz, CDCl3) δ 7.35 – 7.18 (m, 6H), 7.11 (m, J = 7.7, 1.3Hz, 1H), 7.00 (m, J = 7.5, 1.1 Hz, 1H), 6.83 (d, J = 1.2 Hz, 4H), 6.71 – 6.65(m, 1H), 4.94 – 4.84 (m, 2H), 3.92 (t, J = 6.5 Hz, 2H), 3.76 (s, 3H), 2.42 –2.33 (m, 1H), 2.29 (m, J = 13.0, 8.7, 4.2 Hz, 1H), 2.19 – 2.11 (m, 1H), 2.01(dd, J = 12.7, 7.1 Hz, 1H), 1.93 – 1.84 (m, 2H), 1.83 – 1.72 (m, 3H), 1.63 –1.49 (m, 4H).

[0128] 13 C NMR (126 MHz, CDCl3) δ 182.0, 153.6, 153.2, 141.8, 137.1, 136.1,128.7, 127.5, 127.2, 127.2, 127.1, 122.5, 122.4, 115.4, 114.6, 108.7, 77.3,77.0, 76.7, 68.5, 55.7, 53.9, 44.8, 43.7, 41.4, 37.8, 35.2, 33.0, 29.5, 25.3.

[0129] HRMS (ESI) m / z: [M + H] + Calcd for C 30 H 34 NO3 + 456.2533; Found 456.2530.

[0130] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 ℃, Hexane:i-PrOH = 80:20, 1.0 mL / min, 254 nm, t minor = 23.7 min,t major = 43.6 min.

[0131] [α]D 20 = -95.0 (c = 1.00, CHCl3).

[0132] Example 7

[0133] (1R,3S)-1'-Benzyl-3-(3-phenylpropyl)spiro[cyclopentane-l,3'-indolin]-2'-one

[0134] cis -3g

[0135] The preparation method is the same as Example 1, except that 1-iodo-3- methoxypropane is replaced by 3-iodopropylbenzene.

[0136] 1H NMR (500 MHz, CDC13) δ 7.38 - 7.14 (m, 11H), 7.14 - 7.04 (m, 1H), 6.99 (t, J = 7.4 Hz, 1H), 6.68 (d, J = 7.7 Hz, 1H), 4.89 (d, J = 2.4 Hz, 2H), 2.64 (t, J = 7.6 Hz, 2H), 2.36 (m, J = 10.1, 7.1 Hz, 1H), 2.28 (m, J = 13.0, 8.8, 4.2 Hz, 1H), 2.13 (m, J = 8.2, 3.5 Hz, 1H), 1.99 (dd, J = 12.6, 7.0 Hz, 1H), 1.93 - 1.81 (m, 2H), 1.78 - 1.62 (m, 3H), 1.62 - 1.47 (m, 2H).

[0137] 13 C NMR (126 MHz, CDC13) δ 181.9, 142.6, 141.8, 137.1, 136.1, 128.7, 128.4, 128.3, 128.2, 127.4, 127.2, 127.1, 125.6, 122.5, 122.4, 122.3, 108.7, 108.7, 77.3, 77.0, 76.7, 53.9, 44.8, 43.7, 43.6, 41.4, 38.5, 37.8, 36.2, 35.1, 33.0, 30.7, 26.7.

[0138] HRMS (ESI) m / z: [M + H] + Calcd for C 28 H 30 NO + 396.2322; Found 396.2320.

[0139] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® AD-Hcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 11.1 min,t major = 18.6 min.

[0140] [α]D 20 = -124 (c = 0.60, CHCl3).

[0141] Example 8

[0142] (1R,3S)-1'-Benzyl-3-(3-(4-methoxyphenyl)propyl)spiro[cyclopentane-1,3'- indolin]-2'-one

[0143] cis -3h

[0144] The preparation method was the same as in Example 1, except that 1-iodo-3- methoxypropane was replaced by 1-(3-iodopropyl)-4-methoxybenzene.

[0145] 1 H NMR (500 MHz, CDCl3) δ 7.32 – 7.28 (m, 2H), 7.27 – 7.23 (m, 3H),7.21 (dd, J = 7.4, 1.3 Hz, 1H), 7.11 (td, J = 7.8, 1.3 Hz, 3H), 6.99 (td, J =7.5, 1.0 Hz, 1H), 6.85 – 6.80 (m, 2H), 6.68 (d, J = 7.5 Hz, 1H), 4.90 (d, J =2.3 Hz, 2H), 3.78 (s, 3H), 2.58 (t, J = 7.5 Hz, 2H), 2.36 (m, J = 10.0, 8.6,5.4 Hz, 1H), 2.28 (m, J = 13.0, 8.8, 4.1 Hz, 1H), 2.17 – 2.08 (m, 1H), 1.99(dd, J = 12.7, 7.0 Hz, 1H), 1.92 – 1.82 (m, 2H), 1.73 (m, J = 12.4, 9.1 Hz,1H), 1.68 – 1.62 (m, 2H), 1.59 – 1.49 (m, 2H).

[0146] 13C NMR (126 MHz, CDCl3) δ 181.9, 157.6, 141.8, 137.2, 136.1, 134.7,129.2, 129.2, 128.7, 127.4, 127.2, 127.2, 127.1, 122.5, 122.4, 113.7, 108.7,77.3, 77.0, 76.7, 55.2, 53.9, 44.9, 43.7, 41.4, 37.8, 35.2, 35.0, 33.0, 30.9.

[0147] HRMS (ESI) m / z: [M + H] + Calcd for C 29 H 32 NO2 + 426.2428; Found 426.2430.

[0148] HPLC:>99:1er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 17.7 min,t major = 27.6 min.

[0149] [α]D 20 = -134.5 (c = 0.60, CHCl3).

[0150] (1R,3R)-1'-Benzyl-3-(3-((tert-butyldimethylsilyl)oxy)propyl)spiro[cyclopentane-1,3'-indolin]-2'-one

[0151] Example 9

[0152] (1R,3R)-1'-Benzyl-3-(3-((tert-butyldimethylsilyl)oxy)propyl)spiro[cyclopentane-1,3'-indolin]-2'-one

[0153] cis -3i

[0154] The preparation method was the same as that in Example 1, except that 1-iodo-3-methoxypropane was replaced with tert-butyl (3-iodopropyl) dimethylsilane to obtain 38.7 mg of the cis-target product in a yield of 86%.

[0155] 1 H NMR (600 MHz, CDCl3) δ 7.34 – 7.28 (m, 2H), 7.28 – 7.22 (m, 4H),7.12 (td, J = 7.7, 1.3 Hz, 1H), 7.01 (td, J = 7.5, 1.0 Hz, 1H), 6.69 (d, J =7.7 Hz, 1H), 4.91 (d, J = 3.7 Hz, 2H), 3.64 (t, J = 6.3 Hz, 2H), 2.36 (m, J =16.9, 9.8, 8.4, 5.0 Hz, 1H), 2.30 (m, J = 13.0, 8.9, 4.1 Hz, 1H), 2.18 – 2.11(m, 1H), 2.00 (dd, J = 12.7, 7.0 Hz, 1H), 1.92 – 1.85 (m, 2H), 1.73 (m, J =12.5, 9.1 Hz, 1H), 1.61 – 1.58 (m, 2H), 1.58 – 1.50 (m, 2H), 0.90 (s, 9H),0.06 (s, 6H).

[0156] 13 C NMR (151 MHz, CDCl3) δ 182.0, 141.8, 137.2, 136.1, 128.7, 127.5,127.2, 127.2, 122.5, 122.4, 108.7, 77.2, 77.0, 76.8, 63.4, 53.9, 45.0, 43.7,41.4, 37.8, 33.1, 32.1, 31.6, 26.0, 18.4, -5.3.

[0157] HRMS (ESI) m / z: [M + H] + Calcd for C 28 H 40 NO2Si + 450.2823; Found 450.2823.

[0158] [α]D 20 = -8.0 (c = 0.60, CH2Cl2).

[0159] Example 10

[0160] cis -3i’

[0161] The preparation method is treating cis-3i with t-butyl ammonium fluoride (TBAF) to give.

[0162] 1 H NMR (500 MHz, CDCl3) δ 7.29 – 7.11 (m, 6H), 7.05 (m, J = 7.7, 1.2Hz, 1H), 6.93 (m, J = 7.5, 1.1 Hz, 1H), 6.62 (d, J = 7.7 Hz, 1H), 4.83 (s,2H), 3.61 (t, J = 6.2 Hz, 2H), 2.30 (m, J = 10.0, 7.0 Hz, 1H), 2.22 (m, J =12.9, 8.7, 4.3 Hz, 1H), 2.12 – 2.04 (m, 1H), 1.95 (dd, J = 12.8, 7.1 Hz, 1H),1.87 – 1.77 (m, 2H), 1.69 (m, J = 12.5, 8.9 Hz, 1H), 1.62 – 1.55 (m, 3H),1.52 (m, J = 12.5, 5.6, 3.5 Hz, 2H).

[0163] 13 C NMR (126 MHz, CDCl3) δ 182.0, 141.8, 137.1, 136.1, 128.7, 127.5,127.3, 127.2, 122.6, 122.4, 108.8, 77.3, 77.0, 76.7, 63.1, 53.9, 44.7, 43.7,41.2, 37.8, 33.1, 31.9, 31.4.

[0164] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 80:20, 1.0 mL / min, 254 nm, t minor = 14.4 min,t major = 19.1 min.

[0165] [α]D 20 = -1.5 (c = 0.84, CH2Cl2).

[0166] Example 11

[0167] cis -3j

[0168] Preparation method is the same as example 1, the only difference is that 1-iodo-3- methoxypropane is replaced by 4-iodobutyric acid ethyl ester, to get the cis target product 24.1 mg, yield 62%.

[0169] 1 H NMR (500 MHz, CDCl3) δ 7.34 – 7.17 (m, 6H), 7.11 (m, J = 7.7, 1.3Hz, 1H), 7.00 (t, J = 7.5 Hz, 1H), 6.69 (d, J = 7.7 Hz, 1H), 4.90 (d, J = 2.5Hz, 2H), 4.13 (q, J = 7.1 Hz, 2H), 2.42 – 2.25 (m, 4H), 2.19 – 2.10 (m, 1H),2.01 (dd, J = 12.8, 7.1 Hz, 1H), 1.93 – 1.83 (m, 2H), 1.80 – 1.65 (m, 3H),1.56 (qt, J = 9.7, 6.7 Hz, 2H), 1.26 (t, J = 7.1 Hz, 3H).

[0170] 13 C NMR (126 MHz, CDCl3) δ 181.9, 173.6, 141.8, 137.0, 136.1, 128.7,127.4, 127.3, 127.1, 122.5, 122.3, 108.7, 77.3, 77.0, 76.7, 60.2, 53.8, 44.7,43.7, 41.2, 37.7, 34.9, 34.5, 32.9, 24.2, 14.2.

[0171] HRMS (ESI) m / z: [M + H] + Calcd for C 25 H 30 NO3 + 392.2220; Found 392.2223.

[0172] HPLC: >99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 °C, Hexane:i-PrOH = 96:4, 1.0 mL / min, 254 nm, t minor = 58.1 min, t major = 52.4 min.

[0173] [α]D 20 = -55.0 (c = 1.00, CHCl3).

[0174] Example 12

[0175] cis -3o

[0176] Preparation method was the same as example 1, except that 1-iodo-3- methoxypropane was replaced by iodoethane, and spiro olefin 1 was replaced by spiro olefin 17, to give the cis target product 21.9 mg, yield 93%.

[0177] 1 H NMR (500 MHz, CDCl3) δ 7.28 – 7.21 (m, 2H), 7.04 (td, J = 7.5, 1.1Hz, 1H), 6.81 (dd, J = 8.0, 1.0 Hz, 1H), 3.20 (s, 3H), 2.30 – 2.18 (m, 2H),2.12 (m, J = 12.2, 7.5, 4.5, 1.2 Hz, 1H), 1.97 – 1.87 (m, 1H), 1.85 – 1.76(m, 2H), 1.68 (m, J = 12.5, 8.9 Hz, 1H), 1.51 (m, J = 13.5, 6.7 Hz, 2H), 0.95(t, J = 7.4 Hz, 3H).

[0178] 13 C NMR (126 MHz, CDCl3) δ 181.8, 142.7, 137.3, 127.3, 122.4, 122.3,107.6, 77.3, 77.0, 76.7, 54.0, 44.5, 43.2, 37.4, 32.6, 28.3, 26.2, 13.1.

[0179] HRMS (ESI) m / z: [M + H] + Calcd for C 15 H 20 NO + 230.1539; Found 230.1541.

[0180] HPLC: 98:2 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 ℃, Hexane:i-PrOH = 98:2, 0.6 mL / min, 250 nm, t minor = 12.8 min,t major = 14.1 min.

[0181] [α]D 20 = -392.5 (c = 0.44, CHCl3).

[0182] Example 13

[0183] cis -3s

[0184] Preparation method is the same as example 1, except that 1-iodo-3-methoxypropane is replaced by iodomethane, spiro olefin 1 is replaced by spiro olefin 18, to get the cis target product 17.2 mg, yield 75%.

[0185] 1H NMR (500 MHz, CDC13) δ 7.23 (td, J = 7.6, 1.2 Hz, 2H), 7.03 (td, J= 7.5, 1.0 Hz, 1H), 6.86 - 6.79 (m, 1H), 3.76 (q, J = 7.2 Hz, 2H), 2.44 (m, J= 13.5, 10.3, 6.7 Hz, 1H), 2.23 (m, J = 13.0, 8.7, 4.1 Hz, 1H), 2.09 (m, J= 11.3, 7.3, 4.1, 1.3 Hz, 1H), 1.91 (m, J = 12.6, 7.0, 1.3 Hz, 1H), 1.86 - 1.76 (m, 2H), 1.67 (m, J = 12.5, 9.9, 8.9 Hz, 1H), 1.26 (t, J = 7.2 Hz, 3H), 1.16 (d, J = 6.5 Hz, 3H).

[0186] 13 C NMR (126 MHz, CDC13) δ 181.5, 141.8, 137.7, 127.2, 122.5, 122.2, 107.8, 77.3, 77.0, 76.7, 54.2, 46.6, 38.0, 36.0, 35.1, 34.6, 19.7, 12.7.

[0187] HRMS (ESI) m / z: [M + H] + Calcd for C 15 H 20 NO + 230.1539; Found 230.1541.

[0188] HPLC: 98:2 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 °C, Hexane:i-PrOH = 98:2, 0.6 mL / min, 250 nm, t minor = 11.3 min,t major = 12.2 min.

[0189] [α]D 20 = -227.5 (c = 0.44, CHCl3).

[0190] Example 14

[0191] cis -3v

[0192] The preparation method was the same as Example 1, except that spirocycloalkene 1 was replaced by spirocycloalkene 7 to give the cis target product 30.9 mg in 84% yield.

[0193] 1 H NMR (500 MHz, CDCl3) δ 7.51 (dd, J = 8.7, 7.0 Hz, 2H), 7.42 – 7.37(m, 3H), 7.26 (s, 1H), 7.14 (dd, J = 8.4, 2.1 Hz, 1H), 6.75 (d, J = 8.4 Hz,1H), 3.41 (t, J = 6.3 Hz, 2H), 3.35 (s, 3H), 2.36 (m, J = 17.2, 9.1, 4.0 Hz,2H), 2.21 – 2.12 (m, 1H), 2.08 (dd, J = 12.8, 7.1 Hz, 1H), 1.93 (m, J = 13.1,9.1, 7.1 Hz, 2H), 1.76 (dt, J = 12.6, 9.2 Hz, 1H), 1.67 (m, J = 14.7, 12.7,6.8 Hz, 2H), 1.58 (m, J = 10.5, 7.5, 5.2 Hz, 2H).

[0194] 13 C NMR (126 MHz, CDCl3) δ 180.7, 141.1, 138.5, 134.4, 129.6, 128.2,128.0, 127.1, 126.3, 123.1, 110.0, 77.3, 77.0, 76.7, 72.9, 58.6, 54.1, 45.0,41.4, 38.2, 32.9, 31.8, 28.8.

[0195] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 25 ClNO2 + 370.1568; Found 370.1570.

[0196] HPLC: >99:1 er determined by analytical HPLC, Daicel CHIRALPAK® IC column, 25 °C, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 26.0 min, t major = 33.6 min.

[0197] [α]D 20 = -162.0 (c = 0.56, CHCl3).

[0198] Example 15

[0199] cis -3y

[0200] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by iodomethane, and spiro olefin 1 was replaced by spiro olefin 7, to give the cis target product 27.8 mg in 89% yield.

[0201] 1 H NMR (500 MHz, CDCl3) δ 7.56 – 7.50 (m, 2H), 7.44 – 7.37 (m, 3H),7.20 (d, J = 7.9 Hz, 1H), 7.04 (dd, J = 8.0, 1.9 Hz, 1H), 6.80 (d, J = 1.9Hz, 1H), 2.47 (m, J = 10.3, 6.8 Hz, 1H), 2.36 (m, J = 13.0, 8.8, 4.1 Hz, 1H),2.12 (m, J = 12.6, 7.3, 4.1, 1.3 Hz, 1H), 2.03 (m, J = 12.8, 7.1, 1.3 Hz,1H), 1.96 – 1.87 (m, 2H), 1.72 (m, J = 12.5, 9.1 Hz, 1H), 1.18 (d, J = 6.5Hz, 3H).

[0202] 13C NMR (126 MHz, CDCl3) δ 181.2, 143.8, 135.5, 134.3, 133.0, 129.6,128.1, 126.5, 123.6, 122.8, 109.6, 77.3, 77.0, 76.7, 54.1, 47.0, 38.6, 36.1,35.0, 19.6.

[0203] HRMS (ESI) m / z: [M + H] + Calcd for C 19 H 19 ClNO + 312.1150; Found 312.1153.

[0204] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALCEL ® ICcolumn, 25 ℃, Hexane:i-PrOH = 97:3, 1.0 mL / min, 254 nm, t minor = 5.7 min, t major = 6.5 min.

[0205] [α]D 20 = -88.0 (c = 0.64, CHCl3).

[0206] Example 16

[0207] cis -3ab

[0208] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by iodomethane, and spiroolefin 1 was replaced by spiroolefin 11, to obtain the cis target product 18.2 mg, yield 63%.

[0209] 1H NMR (500 MHz, CDC13) δ 7.55 - 7.48 (m, 2H), 7.40 (m, J = 15.0, 6.9, 1.4 Hz, 3H), 7.18 (d, J = 7.5 Hz, 1H), 6.89 (dd, J = 7.4, 1.4 Hz, 1H), 6.64 (d, J = 1.5 Hz, 1H), 2.48 (m, J = 10.2, 6.7 Hz, 1H), 2.35 (m, J = 13.0, 8.9, 4.2 Hz, 1H), 2.30 (s, 3H), 2.12 (m, J = 16.0, 7.3, 4.7, 1.3 Hz, 1H), 2.06 - 2.00 (m, 1H), 1.96 - 1.86 (m, 2H), 1.70 (m, J = 12.4, 9.1 Hz, 1H), 1.18 (d, J = 6.6 Hz, 3H).

[0210] 13 C NMR (126 MHz, CDC13) δ 181.7, 142.6, 137.3, 134.9, 134.3, 129.4, 127.7, 126.6, 123.4, 122.4, 109.8, 77.3, 77.0, 76.7, 54.2, 47.1, 38.6, 36.0, 35.1, 21.6, 19.7.

[0211] HRMS (ESI) m / z: [M + H] + Calcd for C 20 H 22 NO + 292.1696; Found 292.1697.

[0212] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 °C, Hexane:i-PrOH = 99:1, 1.0 mL / min, 254 nm, t minor = 35.4 min,t major = 56.9 min.

[0213] [α]D 20 = -3.8 (c = 0.44, CH2Cl2).

[0214] Example 17

[0215] cis -3ao

[0216] Preparation method was the same as example 1, except that 1-iodo-3- methoxypropane was replaced by iodopropane, and spiro olefin 1 was replaced by spiro olefin 17, to give the cis target product 20.0 mg, yield 81%.

[0217] 1 H NMR (500 MHz, CDCl3) δ 7.28 – 7.20 (m, 2H), 7.04 (td, J = 7.5, 1.0Hz, 1H), 6.81 (dd, J = 8.1, 1.1 Hz, 1H), 3.20 (s, 3H), 2.38 – 2.29 (m, 1H),2.22 (m, J = 13.1, 8.9, 4.4 Hz, 1H), 2.15 – 2.05 (m, 1H), 1.92 (dd, J = 12.7,7.0 Hz, 1H), 1.85 – 1.76 (m, 2H), 1.68 (m, J = 12.5, 9.0 Hz, 2H), 1.53 – 1.41(m, 2H), 1.40 – 1.30 (m, 2H), 0.92 (t, J = 7.3 Hz, 3H).

[0218] 13 C NMR (126 MHz, CDCl3) δ 181.8, 142.8, 137.3, 127.3, 122.5, 122.3,107.7, 77.3, 77.0, 76.7, 54.0, 44.8, 41.1, 37.7, 37.5, 32.9, 26.3, 21.9,14.3.

[0219] HRMS (ESI) m / z: [M + H] + Calcd for C 16 H 22 NO + 244.1696; Found 244.1695.

[0220] HPLC: >99:1 er determined by analytical HPLC, Daicel CHIRALPAK® AZ-3 column, 25 °C, Hexane:i-PrOH = 99:1, 1.0 mL / min, 254 nm, t minor = 14.7 min,t major = 16.8 min.

[0221] [α]D 20 = -1.8 (c = 0.25, CH2Cl2).

[0222] Example 18

[0223] cis -3al

[0224] The preparation method was the same as Example 1, except that 1-iodo-3- methoxypropane was replaced by 4-iodobutyric acid ethyl ester, and spirocyclic olefin 1 was replaced by spirocyclic olefin 13, to obtain the cis target product 23. 4 mg, yield 62%.

[0225] 1 H NMR (500 MHz, CDCl3) δ 7.46 – 7.41 (m, 2H), 7.36 – 7.29 (m, 3H),7.22 (dd, J = 7.5, 1.3 Hz, 1H), 7.09 (td, J = 7.7, 1.3 Hz, 1H), 7.01 (td, J =7.5, 1.1 Hz, 1H), 6.75 (dd, J = 7.9, 1.1 Hz, 1H), 4.06 (q, J = 7.2 Hz, 2H),2.38 – 2.23 (m, 4H), 2.14 – 2.05 (m, 1H), 2.02 (dd, J = 12.8, 7.1 Hz, 1H),1.91 – 1.82 (m, 2H), 1.71 – 1.58 (m, 3H), 1.48 (m, J = 13.9, 12.3, 7.3, 3.9Hz, 2H), 1.19 (t, J = 7.1 Hz, 3H).

[0226] 13C NMR (126 MHz, CDCl3) δ 181.2, 173.6, 142.6, 136.8, 134.8, 129.4,127.7, 127.2, 126.5, 123.0, 122.6, 109.0, 77.3, 77.0, 76.7, 60.2, 54.0, 45.0,41.2, 38.2, 34.9, 34.5, 32.9, 24.2, 14.2.

[0227] HRMS (ESI) m / z: [M + H] + Calcd for C 24 H 28 NO3 + 378.2064; Found 378.2065.

[0228] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 80:20, 1.0 mL / min, 254 nm, t minor = 20.8 min,t major = 27.5 min.

[0229] [α]D 20 = -0.7 (c = 1.04, CH2Cl2).

[0230] Example 19

[0231] (1R,3S)-1'-Benzyl-3-(3-methoxypropyl)spiro[cyclopentane-1,3'-indolin]-2'-one

[0232] trans -3a

[0233] Under a nitrogen atmosphere, using a glovebox technique, an oven-dried Teflon screw cap test tube was charged with a magnetic stir bar, followed by cobalt chloride (0.015 mmol, 15 mol%) and the second ligand (0.018 mol, 18 mol%). Anhydrous methyl-tert-butyl ether was then added and stirred at room temperature for 15 min. Potassium phosphate (hydrate) (0.20 mmol, 2 equiv), spirocyclic olefin 1 (0.10 mmol, 1 equiv), 1-iodo-3-methoxypropane (0.12 mmol, 1.2 equiv) and dimethoxymethylsilane (0.20 mmol, 2 equiv) were then added sequentially. The test tube was then sealed with a gas-tight insulating tape, removed from the glovebox and stirred at 0 °C at 650 rpm for 48 h. Upon completion of the reaction, the reaction mixture was diluted with saturated aqueous ammonium chloride and ethyl acetate. The aqueous phase was extracted with ethyl acetate and the combined organic phases were concentrated under reduced pressure. The crude mixture was purified by flash column chromatography on silica gel with petroleum ether / ethyl acetate (10:1 by volume) as eluent to give the target trans product 30.4 mg, 87% yield.

[0234] 1 H NMR (600 MHz, CDCl3) δ 7.32 – 7.28 (m, 2H), 7.28 – 7.22 (m, 3H),7.20 (dd, J = 7.4, 1.3 Hz, 1H), 7.10 (td, J = 7.7, 1.3 Hz, 1H), 7.00 (td, J =7.5, 1.0 Hz, 1H), 6.67 (d, J = 7.8 Hz, 1H), 4.89 (d, J = 3.0 Hz, 2H), 3.41(td, J = 6.5, 1.3 Hz, 2H), 3.34 (s, 3H), 2.50 (m, J = 10.2, 6.9 Hz, 1H), 2.35(dd, J = 13.1, 7.5 Hz, 1H), 2.26 – 2.18 (m, 2H), 1.98 – 1.93 (m, 1H), 1.70 –1.62 (m, 2H), 1.62 – 1.56 (m, 2H), 1.55 – 1.49 (m, 2H).

[0235] 13C NMR (151 MHz, CDCl3) δ 182.4, 142.0, 136.7, 136.2, 128.7, 127.5,127.2, 127.2, 122.6, 122.2, 108.6, 77.2, 77.0, 76.8, 72.9, 58.5, 53.5, 45.0,43.6, 40.7, 38.1, 33.8, 32.0, 28.7.

[0236] HRMS (ESI) m / z: [M + H] + Calcd for C 23 H 28 NO2 + 350.2115; Found 350.2112.

[0237] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 90:10, 1.0 mL / min, 254 nm, t minor = 16.3 min,t major = 20.8 min.

[0238] [α]D 20 = -2.6 (c = 0.34, CH2Cl2).

[0239] Example 20

[0240] (1R,3S)-3-(3-((tert-Butyldimethylsilyl)oxy)propyl)-6'-chloro-1'-methylspiro[cyclopentane-1,3'-indolin]-2'-one

[0241] trans -3ah

[0242] The preparation method was the same as that in Example 19, except that 1-iodo-3-methoxypropane was replaced by tert-butyl (3-iodopropyl) dimethylsilane, and spiroolefin 1 was replaced by spiroolefin 20, to obtain 24.5 mg of the trans target product in a yield of 60%.

[0243] 1H NMR (500 MHz, CDCl3) δ 7.05 (d, J = 7.9 Hz, 1H), 6.96 (dd, J = 7.9,1.9 Hz, 1H), 6.74 (d, J = 1.8 Hz, 1H), 3.57 (t, J = 6.3 Hz, 2H), 3.12 (s,3H), 2.42 – 2.32 (m, 1H), 2.24 – 2.18 (m, 1H), 2.10 (m, J = 32.6, 13.0, 9.8,6.8 Hz, 2H), 1.81 (m, J = 13.0, 7.9, 2.6 Hz, 1H), 1.53 – 1.36 (m, 6H), 0.84(s, 9H).

[0244] 13 C NMR (126 MHz, CDCl3) δ 182.2, 144.2, 135.2, 133.1, 123.1, 122.3,108.4, 77.3, 77.2, 77.0, 76.7, 63.3, 53.3, 44.8, 40.6, 38.0, 33.9, 32.0,31.7, 26.3, 26.0, 18.4, -5.3.

[0245] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 35 ClNO2Si + 408.2120; Found408.2125.

[0246] [α]D 20 = -0.3 (c = 0.68, CH2Cl2).

[0247] Example 21

[0248] (1R,3R)-3-Ethyl-1'-phenylspiro[cyclopentane-1,3'-indolin]-2'-one

[0249] trans -3c

[0250] The preparation method is the same as that of Example 19, except that 1-iodo-3-methoxypropane is replaced by iodoethane, and spirocyclic olefin 1 is replaced by spirocyclic olefin 13.

[0251] 1H NMR (600 MHz, CDCl3) δ 7.53 – 7.48 (m, 2H), 7.46 – 7.40 (m, 2H),7.38 (td, J = 7.3, 1.3 Hz, 1H), 7.28 (dd, J = 7.4, 1.3 Hz, 1H), 7.16 (td, J =7.7, 1.3 Hz, 1H), 7.08 (td, J = 7.5, 1.1 Hz, 1H), 6.81 (dd, J = 7.8, 1.0 Hz,1H), 2.47 – 2.37 (m, 2H), 2.31 – 2.19 (m, 2H), 2.02 (m, J = 12.2, 8.2, 2.7Hz, 1H), 1.65 – 1.56 (m, 2H), 1.51 (m, J = 13.4, 10.5, 6.6 Hz, 2H), 0.97 (t,J = 7.4 Hz, 3H).

[0252] 13 C NMR (151 MHz, CDCl3) δ 181.8, 142.8, 136.8, 134.8, 129.4, 127.7,127.2, 126.5, 123.0, 122.5, 109.0, 77.2, 77.0, 76.8, 53.7, 45.2, 42.6, 38.4,33.6, 28.4, 13.1.

[0253] HRMS (ESI) m / z: [M + H] + Calcd for C 20 H 22 NO + 292.1696; Found 292.1699.

[0254] HPLC: 88:12 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 14.5 min,t major = 16.2 min.

[0255] [α]D 20= -110.0 (c = 0.92, CHCl3).

[0256] Example 22

[0257] (1R,3S)-1'-Methyl-3-(3-phenoxypropyl)spiro[cyclopentane-1,3'- indolin]-2'-one

[0258] trans -3ai

[0259] The preparation method was the same as in Example 19, except that 1-iodo-3- methoxypropane was replaced by iodoethane and spirocyclic alkene 1 was replaced by spirocyclic alkene 17.

[0260] 1 H NMR (600 MHz, CDCl3) δ 7.27 (dd, J = 8.7, 7.2 Hz, 3H), 7.22 (m, J = 11.1, 7.5, 1.2 Hz, 2H), 7.06 (td, J = 7.5, 1.0 Hz, 1H), 6.96 – 6.88 (m, 3H), 6.80 (d, J = 7.8 Hz, 1H), 3.98 (td, J = 6.4, 1.5 Hz, 2H), 3.19 (s, 3H), 2.50 (m, J = 10.2, 7.1 Hz, 1H), 2.31 (m, J = 13.2, 7.6, 1.1 Hz, 1H), 2.26 – 2.19 (m, 1H), 2.16 (m, J = 13.0, 10.2, 7.4 Hz, 1H), 1.92 (m, J = 12.9, 8.0, 2.8 Hz, 1H), 1.89 – 1.80 (m, 2H), 1.67 – 1.59 (m, 3H), 1.54 (dd, J = 13.3, 10.2 Hz, 1H).

[0261] 13 C NMR (151 MHz, CDCl3) δ 182.3, 159.0, 143.0, 136.8, 129.4, 127.4, 122.6, 122.1, 120.5, 114.5, 107.7, 77.2, 77.0, 76.8, 67.9, 53.5, 44.8, 40.6, 37.9, 33.9, 31.9, 28.5, 26.2.

[0262] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 26 NO2 + 336.1958; Found 336.1960.

[0263] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALCEL ® OD-3column, 25 ℃, Hexane:i-PrOH = 95:5, 0.8 mL / min, 254 nm, t minor = 26.9 min,t major = 15.2 min.

[0264] [α]D 20 = -3.6 (c = 0.28, CH2Cl2).

[0265] Example 23

[0266] Ethyl 4-((lR,3R)-l'-methyl-2'-oxospiro[cyclopentane-l,3'-indolin]-3-yl)butanoate

[0267] trans -3aj

[0268] The preparation method is the same as Example 19, except that 1-iodo-3- methoxypropane is replaced by ethyl 4-iodobutyrate and spiroolefin 1 is replaced by spiroolefin 17.

[0269] 1H NMR (600 MHz, CDCl3) δ 7.24 (td, J = 7.7, 1.2 Hz, 1H), 7.21 – 7.18(m, 1H), 7.05 (td, J = 7.5, 1.0 Hz, 1H), 6.80 (dt, J = 7.7, 0.8 Hz, 1H), 4.13(q, J = 7.2 Hz, 2H), 3.19 (s, 3H), 2.44 (tq, J = 10.3, 7.2 Hz, 1H), 2.33 (t,J = 7.4 Hz, 2H), 2.28 (m, J = 13.2, 7.6, 1.2 Hz, 1H), 2.23 – 2.10 (m, 2H),1.90 (m, J = 13.0, 8.0, 2.7 Hz, 1H), 1.74 – 1.63 (m, 2H), 1.60 – 1.53 (m,1H), 1.53 – 1.46 (m, 3H), 1.26 (t, J = 7.1 Hz, 3H).

[0270] 13 C NMR (151 MHz, CDCl3) δ 182.2, 173.6, 142.9, 136.8, 127.3, 122.6,122.1, 107.6, 77.2, 77.0, 76.8, 60.2, 53.5, 44.7, 40.5, 37.9, 35.0, 34.5,33.8, 26.2, 24.0, 14.2.

[0271] HRMS (ESI) m / z: [M + H] + Calcd for C 19 H 26 NO3 + 316.1907; Found 316.1912.

[0272] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALCEL ® OD-Hcolumn, 25 ℃, Hexane:i-PrOH = 98:2, 1.0 mL / min, 254 nm, t minor = 15.7 min,t major = 11.6 min.

[0273] [α]D 20 = -3.9 (c = 0.31, CH2Cl2).

[0274] Example 24

[0275] 3-((1R,3S)-1'-Methyl-2'-oxospiro[cyclopentane-1,3'-indolin]-3-yl)propyl furan-2- carboxylate

[0276] trans -3ak

[0277] The preparation was the same as Example 19, except that 3-iodopropyl furan-2- carboxylate and spiroalkene 17 were used.

[0278] 1 H NMR (500 MHz, CDCl3) δ 7.58 (dd, J = 1.8, 0.9 Hz, 1H), 7.24 (td, J= 7.7, 1.3 Hz, 2H), 7.23 – 7.16 (m, 2H), 7.05 (td, J = 7.5, 1.0 Hz, 1H), 6.82– 6.78 (m, 1H), 6.51 (dd, J = 3.5, 1.7 Hz, 1H), 4.33 (t, J = 6.7 Hz, 2H),3.19 (s, 3H), 2.54 – 2.44 (m, 1H), 2.30 (m, J = 13.3, 7.6, 1.2 Hz, 1H), 2.25– 2.19 (m, 1H), 2.15 (m, J = 13.0, 10.0, 7.3 Hz, 1H), 1.91 (m, J = 13.0, 8.0,2.6 Hz, 1H), 1.82 (m, J = 15.7, 13.7, 7.6, 4.6 Hz, 2H), 1.61 – 1.49 (m, 4H).

[0279] 13 C NMR (126 MHz, CDCl3) δ 182.2, 158.8, 146.2, 144.8, 143.0, 136.7,127.4, 122.6, 122.1, 117.8, 111.8, 107.7, 77.3, 77.2, 77.0, 76.7, 65.1, 53.5,44.7, 40.5, 37.8, 33.8, 31.8, 27.9, 26.2.

[0280] HRMS (ESI) m / z: [M + H] + Calcd for C 21 H 24 NO4 + 354.1700; Found 354.1703.

[0281] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK ® AZ-3column, 25 ℃, Hexane:i-PrOH = 90:10, 1.0 mL / min, 254 nm, t minor = 48.7 min,t major = 43.9 min.

[0282] [α]D 20 = -1.2 (c = 0.48, CH2Cl2).

[0283] Example 25

[0284] (1R,3R)-1'-Benzyl-3-propylspiro[cyclopentane-1,3'-indolin]-2'-one

[0285] trans -3d

[0286] The preparation method was the same as Example 19, except that iodoethane was used instead.

[0287] 1 H NMR (500 MHz, CDCl3) δ 7.34 – 7.18 (m, 6H), 7.11 (td, J = 7.7, 1.3Hz, 1H), 7.01 (td, J = 7.5, 1.1 Hz, 1H), 6.67 (dd, J = 7.8, 0.9 Hz, 1H), 4.89(d, J = 2.6 Hz, 2H), 2.48 (m, J = 10.4, 6.7, 3.3 Hz, 1H), 2.34 (m, J = 13.2,7.7, 1.1 Hz, 1H), 2.26 – 2.16 (m, 2H), 1.99 – 1.91 (m, 1H), 1.62 – 1.49 (m,2H), 1.49 – 1.33 (m, 4H), 0.94 (t, J = 7.1 Hz, 3H).

[0288] 13 C NMR (126 MHz, CDCl3) δ 182.5, 142.0, 137.0, 136.2, 128.7, 127.5,127.2, 127.2, 122.6, 122.2, 108.6, 77.3, 77.0, 76.7, 53.5, 45.1, 43.6, 40.7,38.1, 37.9, 33.9, 21.8, 14.4.

[0289] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 26 NO + 320.2009; Found 320.2012.

[0290] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 ℃, Hexane:i-PrOH = 97:3, 1.0 mL / min, 254 nm, t minor = 18.5 min,t major = 23.2 min.

[0291] [α]D 20 = -0.7 (c = 0.36, CH2Cl2).

[0292] Example 26

[0293] (1R,3S)-3-(3-Methoxypropyl)-4'-methyl-1'-phenylspiro[cyclopentane-1,3'-indolin]-2'-one

[0294] trans -3u

[0295] The preparation method is the same as that of Example 19, except that spirocyclic olefin 6 is used.

[0296] 1H NMR (600 MHz, CDCl3) δ 7.50 (m, J = 8.2, 2.3 Hz, 2H), 7.41 – 7.36(m, 3H), 7.06 (t, J = 7.8 Hz, 1H), 6.87 (d, J = 7.7 Hz, 1H), 6.64 (d, J = 7.8Hz, 1H), 3.40 (td, J = 6.5, 2.5 Hz, 2H), 3.34 (d, J = 4.2 Hz, 3H), 2.65 –2.55 (m, 1H), 2.42 (s, 3H), 2.35 – 2.20 (m, 3H), 2.14 (m, J = 13.0, 6.4, 1.4Hz, 1H), 1.85 (dd, J = 13.0, 11.9 Hz, 1H), 1.70 – 1.61 (m, 3H), 1.55 – 1.49(m, 2H).

[0297] 13 C NMR (151 MHz, CDCl3) δ 182.4, 143.5, 134.8, 133.7, 132.0, 129.4,127.8, 127.2, 126.8, 125.5, 106.9, 77.2, 77.0, 76.8, 73.0, 58.5, 53.8, 43.2,40.7, 33.9, 33.9, 31.5, 28.8, 18.3.

[0298] HRMS (ESI) m / z: [M + H] + Calcd for C 23 H 28 NO2 + 350.2115; Found 350.2114.

[0299] HPLC: 91:9 er determined by analytical HPLC, Daicel CHIRALPAK ® IF-3column, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 20.6 min,t major = 27.7 min.

[0300] [α]D 20= -1.8 (c = 0.52, CH2Cl2).

[0301] Example 27

[0302] (1R,3S)-5'-chloro-3-(3-methoxypropyl)-1'-phenylspiro[cyclopentane-1,3'- indolin]-2'-one

[0303] trans -3v

[0304] Preparation method was the same as example 19, except that 1-iodo-3- methoxypropane and spiroalkene 7 were used to give the trans target product 18.6 mg, yield 50%.

[0305] 1 H NMR (500 MHz, CDCl3) δ 7.53 – 7.50 (m, 2H), 7.42 – 7.37 (m, 3H),7.23 (d, J = 2.1 Hz, 1H), 7.13 (dd, J = 8.4, 2.1 Hz, 1H), 6.74 (d, J = 8.4Hz, 1H), 3.41 (t, J = 6.4 Hz, 2H), 3.35 (s, 3H), 2.54 – 2.39 (m, 2H), 2.32 –2.20 (m, 2H), 2.05 – 2.00 (m, 1H), 1.70 – 1.63 (m, 2H), 1.62 (d, J = 1.6 Hz,1H), 1.59 (d, J = 2.9 Hz, 1H), 1.57 – 1.51 (m, 2H).

[0306] 13 C NMR (126 MHz, CDCl3) δ 181.2, 141.3, 138.3, 134.4, 129.6, 128.3,128.0, 127.1, 126.4, 123.0, 110.0, 77.3, 77.0, 76.7, 72.8, 58.6, 53.8, 45.4,40.7, 38.4, 33.9, 31.9, 28.8.

[0307] HRMS (ESI) m / z: [M + H] + Calcd for C 22 H 25 ClNO2 +370.1568; Found 370.1573.

[0308] HPLC: 91:9 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 26.9 min,t major = 22.8 min.

[0309] [α]D 20 = -0.3 (c = 0.28, CH2Cl2).

[0310] Example 28

[0311] trans -3ah’

[0312] The preparation method is to treat trans-3ah with tert-butyl ammonium fluoride.

[0313] 1 H NMR (500 MHz, CDCl3) δ 7.10 (d, J = 7.8 Hz, 1H), 7.01 (dd, J = 7.9,1.9 Hz, 1H), 6.79 (d, J = 1.8 Hz, 1H), 3.67 (t, J = 6.4 Hz, 2H), 3.17 (s,3H), 2.43 (m, J = 10.3, 7.1 Hz, 1H), 2.26 (m, J = 13.2, 7.5, 1.2 Hz, 1H),2.23 – 2.08 (m, 2H), 1.87 (m, J = 13.0, 8.0, 2.5 Hz, 1H), 1.66 – 1.58 (m,2H), 1.56 – 1.50 (m, 2H), 1.49 – 1.43 (m, 1H), 1.40 (s, 1H).

[0314] 13C NMR (126 MHz, CDCl3) δ 182.2, 144.1, 135.1, 133.1, 123.0, 122.3,108.4, 77.3, 77.2, 77.0, 76.7, 63.0, 53.2, 44.8, 40.6, 37.9, 33.9, 31.8,31.6, 26.3.

[0315] HRMS (ESI) m / z: [M + H] + Calcd for C 16 H 21 ClNO2 + 294.1255; Found 294.1256.

[0316] HPLC: 92:8 er determined by analytical HPLC, Daicel CHIRALPAK ® ICcolumn, 25 ℃, Hexane:i-PrOH = 85:15, 1.0 mL / min, 220 nm, tminor = 20.7 min,tmajor = 16.7 min.

[0317] [α]D 20 = -0.4 (c = 0.66, CH2Cl2).

[0318] Example 29

[0319] trans -3af

[0320] Preparation Method is the same as Example 19, except that methyl iodide and spiro olefin 26 are used, to give the trans target product 19. 1 mg, 85% yield.

[0321] 1H NMR (500 MHz, CDC13) δ 7.68 - 7.63 (m, 2H), 7.36 (m, J = 10.9, 5.7, 2.1 Hz, 2H), 7.14 - 7.10 (m, 1H), 3.75 - 3.70 (m, 2H), 2.38 - 2.22 (m, 2H), 2.14 - 1.97 (m, 4H), 1.74 - 1.68 (m, 1H), 1.31 - 1.22 (m, 1H), 1.17 (dd, J = 12.4, 8.0 Hz, 1H), 1.04 (d, J = 6.4 Hz, 3H).

[0322] 13 C NMR (126 MHz, CDC13) δ 179.4, 139.9, 128.7, 124.0, 119.4, 119.4, 77.3, 77.0, 76.7, 52.3, 45.7, 45.2, 36.6, 35.0, 34.8, 34.6, 20.5.

[0323] HRMS (ESI) m / z: [M + H] + Calcd for C 15 H 20 NO + 230.1539; Found 230.1543.

[0324] HPLC: 92:8 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 °C, Hexane:i-PrOH = 97:3, 1.0 mL / min, 254 nm, t minor = 32.8 min, t major = 37.5 min.

[0325] [α]D 20 = -492.0 (c = 0.64, CHCl3).

[0326] Example 30

[0327] trans -3ax

[0328] The preparation method was the same as Example 19, except that spiroolefin 9 was used, to give the trans target product 27.8 mg, 76% yield.

[0329] 1 H NMR (500 MHz, CDCl3) δ 7.50 (t, J = 7.9 Hz, 2H), 7.45 – 7.39 (m,2H), 7.39 – 7.33 (m, 1H), 6.87 (d, J = 2.6 Hz, 1H), 6.75 (d, J = 8.6 Hz, 1H),6.68 (dd, J = 8.6, 2.6 Hz, 1H), 3.81 (s, 3H), 3.41 (t, J = 6.4 Hz, 2H), 3.34(s, 3H), 2.46 (m, J = 28.5, 11.9, 7.3 Hz, 2H), 2.32 – 2.18 (m, 2H), 2.04 –1.98 (m, 1H), 1.69 – 1.48 (m, 6H).

[0330] 13 C NMR (126 MHz, CDCl3) δ 181.4, 156.4, 138.0, 136.2, 134.9, 129.4,127.5, 126.2, 111.2, 109.9, 109.3, 77.3, 77.0, 76.7, 72.9, 58.5, 55.8, 54.0,45.5, 40.6, 38.4, 33.9, 32.0, 28.7.

[0331] HRMS (ESI) m / z: [M + H] + Calcd for C 23 H 28 NO3 + 366.2064; Found 366.2067.

[0332] HPLC: 87:13 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 ℃, Hexane:i-PrOH = 90:10, 1.0 mL / min, 254 nm, t minor = 25.0 min,t major = 32.9 min.

[0333] [α]D 20 = -32.0 (c = 1.16, CHCl3).

[0334] Example 31

[0335] trans -3az

[0336] Preparation method was the same as example 19, except that spiroolefin 16 was used to give the trans target product 19. 2 mg, yield 67%.

[0337] 1 H NMR (500 MHz, CDCl3) δ 7.04 (dd, J = 7.0, 1.8 Hz, 1H), 6.99 – 6.90(m, 2H), 3.47 (s, 3H), 3.39 (td, J = 6.5, 1.2 Hz, 2H), 3.34 (s, 3H), 2.57 (s,3H), 2.44 (m, J = 10.4, 7.1 Hz, 1H), 2.25 (m, J = 13.1, 7.4, 1.2 Hz, 1H),2.21 – 2.08 (m, 2H), 1.87 (m, J = 12.9, 7.9, 2.3 Hz, 1H), 1.68 – 1.58 (m,2H), 1.58 – 1.43 (m, 4H).

[0338] 13 C NMR (126 MHz, CDCl3) δ 183.1, 140.6, 137.5, 131.0, 122.5, 120.1,119.2, 77.3, 77.0, 76.7, 72.9, 58.5, 52.9, 45.4, 40.7, 38.3, 33.9, 31.9,29.5, 28.7, 19.0.

[0339] HRMS (ESI) m / z: [M + H] + Calcd for C 18 H 26 NO2 + 288.1958; Found 288.1954.

[0340] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALCEL® OJ-3 column, 25 °C, Hexane:i-PrOH = 98:2, 1.0 mL / min, 254 nm, t minor = 7.9 min, t major = 11.8 min.

[0341] [α]D 20 = -1.1 (c = 0.48, CH2Cl2).

[0342] Example 32

[0343] trans -3at

[0344] Preparation method was the same as Example 19, except that spiro olefin 11 was used to give the trans target product 21.5 mg, 61% yield.

[0345] 1 H NMR (500 MHz, CDCl3) δ 7.55 – 7.48 (m, 2H), 7.43 – 7.36 (m, 3H),7.15 (d, J = 7.6 Hz, 1H), 6.90 (dt, J = 7.5, 1.2 Hz, 1H), 6.63 (d, J = 1.5Hz, 1H), 3.41 (td, J = 6.4, 1.0 Hz, 2H), 3.34 (s, 3H), 2.53 – 2.37 (m, 2H),2.24 (m, J = 12.5, 6.2, 2.4 Hz, 2H), 2.03 – 1.97 (m, 1H), 1.69 – 1.61 (m,3H), 1.61 – 1.58 (m, 1H), 1.57 – 1.50 (m, 2H).

[0346] 13 C NMR (126 MHz, CDCl3) δ 182.0, 142.8, 137.4, 134.8, 133.6, 129.4,127.7, 126.6, 123.5, 122.3, 109.8, 77.3, 77.0, 76.7, 72.9, 58.5, 53.4, 45.4,40.6, 38.4, 33.8, 32.0, 28.7, 21.6.

[0347] HRMS (ESI) m / z: [M + H] +Caled for C 23 H 28 NO2 + 350.2115; Found 350.2111.

[0348] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK ® IGcolumn, 25 °C, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 21.8 min,t major = 25.9 min.

[0349] [α]D 20 = -1.3 (c = 0.68, CH2Cl2).

[0350] Example 33

[0351] trans -3ao

[0352] Preparation method was the same as example 19, except that iodopropane and spiro olefin 17 were used to give the trans target product 18.5 mg, 76% yield.

[0353] 1 H NMR (500 MHz, CDCl3) δ 7.28 – 7.20 (m, 1H), 7.23 – 7.18 (m, 1H),7.05 (td, J = 7.5, 1.0 Hz, 1H), 6.80 (d, J = 7.7 Hz, 1H), 3.19 (s, 3H), 2.44(m, J = 10.1, 6.8 Hz, 1H), 2.27 (dd, J = 13.2, 7.6 Hz, 1H), 2.23 – 2.07 (m,2H), 1.93 – 1.80 (m, 1H), 1.60 – 1.28 (m, 5H), 1.37 (s, 1H), 0.93 (t, J = 7.1Hz, 3H).

[0354] 13C NMR (126 MHz, CDCl3) δ 182.4, 142.9, 137.1, 127.3, 122.5, 122.1, 107.6, 77.3, 77.0, 76.7, 53.5, 44.8, 40.6, 38.0, 37.9, 34.0, 26.2, 21.8, 14.4.

[0355] HRMS (ESI) m / z: [M + H] + Calcd for C 16 H 22 NO + 244.1696; Found 244.1698.

[0356] HPLC: 91:9 er determined by analytical HPLC, Daicel CHIRALPAK ® AZ-3column, 25 ℃, Hexane:i-PrOH = 99:1, 1.0 mL / min, 254 nm, t minor = 13.8 min,t major = 15.9 min.

[0357] [α]D 20 = -2.8 (c = 0.32, CH2Cl2).

[0358] Example 34

[0359] trans -3an

[0360] Preparation method is the same as example 19, except that iodopropane and spiro olefin 13 are used, to give the trans target product 19.6 mg, yield 74%.

[0361] 1H NMR (500 MHz, CDC13) δ 7.54 - 7.47 (m, 2H), 7.44 - 7.35 (m, 3H), 7.28 (dd, J = 7.3, 1.4 Hz, 1H), 7.16 (td, J = 7.7, 1.4 Hz, 1H), 7.08 (td, J = 7.5, 1.1 Hz, 1H), 6.81 (dd, J = 7.8, 1.1 Hz, 1H), 2.54 - 2.37 (m, 2H), 2.32 - 2.16 (m, 2H), 2.02 (m, J = 12.1, 7.9, 2.4 Hz, 1H), 1.64 - 1.54 (m, 2H), 1.52 - 1.32 (m, 4H), 0.94 (t, J = 7.1 Hz, 3H).

[0362] 13 C NMR (126 MHz, CDC13) δ 181.8, 142.7, 136.8, 134.7, 129.4, 127.7, 127.2, 126.5, 123.0, 122.5, 108.9, 77.3, 77.0, 76.7, 53.7, 45.5, 40.6, 38.4, 37.9, 34.0, 21.8, 14.4.

[0363] HRMS (ESI) m / z: [M + H] + Calcd for C 21 H 24 NO + 306.1852; Found 306.1855.

[0364] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 °C, Hexane:i-PrOH = 95:5, 1.0 mL / min, 254 nm, t minor = 9.2 min, t major = 10.0 min.

[0365] [α]D 20 = -1.7 (c = 0.12, CH2Cl2).

[0366] Example 35

[0367] trans -3pp

[0368] Preparation method was the same as example 19, except that propylthio derived iodoalkane and spiroalkene 17 were used to give the trans target 21.5 mg, 41% yield.

[0369] 1 H NMR (500 MHz, CDCl3) δ 8.19 – 8.11 (m, 2H), 7.92 – 7.84 (m, 2H),7.27 – 7.17 (m, 2H), 7.05 (td, J = 7.5, 1.0 Hz, 1H), 6.81 (d, J = 7.7 Hz,1H), 4.38 (t, J = 6.6 Hz, 2H), 3.20 (s, 3H), 3.14 – 3.04 (m, 4H), 2.52 (m, J= 10.4, 7.2 Hz, 1H), 2.31 (dd, J = 13.1, 7.4 Hz, 1H), 2.27 – 2.12 (m, 2H),1.97 – 1.79 (m, 3H), 1.65 – 1.49 (m, 8H), 0.87 (t, J = 7.4 Hz, 6H).

[0370] 13 C NMR (126 MHz, CDCl3) δ 182.2, 165.3, 144.1, 142.9, 136.6, 133.7,130.2, 127.4, 127.0, 122.6, 122.1, 107.7, 77.3, 77.2, 77.0, 76.7, 65.8, 53.4,49.9, 44.8, 40.5, 37.8, 33.8, 31.8, 27.9, 26.2, 21.9, 11.1.

[0371] HRMS (ESI) m / z: [M + H] + Calcd for C 29 H 39 N2O5S + 527.2574; Found 527.2578.

[0372] HPLC: 90:10 er determined by analytical HPLC, Daicel CHIRALPAK® IA-3 column, 25 °C, Hexane:i-PrOH = 90:10, 1.0 mL / min, 254 nm, t minor = 32.7 min,t major = 23.8 min.

[0373] [α]D 20 = -106.7 (c = 0.67, CHCl3).

[0374] Example 36

[0375] trans -3ar

[0376] Preparation Method was the same as Example 19, except that 4-iodobutyric acid ethyl ester and spiroalkene 21 were used to give the trans target product 24.5 mg, 68% yield.

[0377] 1 H NMR (600 MHz, CDCl3) δ 8.10 (dd, J = 5.2, 1.7 Hz, 1H), 7.33 (dd, J= 7.2, 1.7 Hz, 1H), 6.86 (dd, J = 7.3, 5.2 Hz, 1H), 4.12 (m, J = 7.2, 3.7 Hz,2H), 2.43 – 2.34 (m, 1H), 2.33 – 2.25 (m, 3H), 2.20 – 2.09 (m, 2H), 1.85 –1.79 (m, 1H), 1.76 (d, J = 3.9 Hz, 9H), 1.72 – 1.59 (m, 3H), 1.51 – 1.42 (m,3H), 1.24 (td, J = 7.1, 2.0 Hz, 3H).

[0378] 13 C NMR (151 MHz, CDCl3) δ 182.7, 173.6, 158.1, 145.3, 131.4, 128.7,117.4, 77.2, 77.0, 76.8, 60.2, 58.2, 52.9, 44.8, 40.7, 38.1, 34.9, 34.4,33.9, 28.8, 24.0, 14.2.

[0379] HRMS (ESI) m / z: [M + H] +Calcd for C 21 H 31 N2O3 + 359.2329; Found 359.2325.

[0380] HPLC: 91:9 er determined by analytical HPLC, Daicel CHIRALPAK ® IDcolumn, 25 ℃, Hexane:i-PrOH = 99:1, 1.0 mL / min, 254 nm, t minor = 16.3 min,t major = 21.8 min.

[0381] [α]D 20 = -0.6 (c = 0.64, CH2Cl2).

[0382] Example 37

[0383] ent-cis -3ar

[0384] Preparation method is same as example 1, only change ligand to its enantiomer, use ethyl 4-iodobutyrate and spiroalkene 21 as reaction substrates, get enantiomeric cis target product 19.7 mg, yield 55%.

[0385] 1 H NMR (500 MHz, CDCl3) δ 8.12 (dd, J = 5.2, 1.7 Hz, 1H), 7.36 (dd, J= 7.3, 1.7 Hz, 1H), 6.87 (dd, J = 7.3, 5.2 Hz, 1H), 4.13 (q, J = 7.2 Hz, 2H),2.32 (t, J = 7.5 Hz, 2H), 2.29 – 2.17 (m, 2H), 2.12 – 2.03 (m, 1H), 1.90 (dd,J = 12.7, 7.2 Hz, 1H), 1.77 (m, 10H), 1.72 – 1.63 (m, 3H), 1.51 (m, J = 9.5,7.6, 4.9 Hz, 2H), 1.26 (m, J = 7.1 Hz, 4H).

[0386] 13C NMR (151 MHz, CDCl3) δ 182.1, 173.7, 157.9, 145.3, 131.6, 128.8,117.3, 77.3, 77.0, 76.7, 60.3, 58.3, 53.4, 44.7, 41.2, 37.6, 34.8, 34.5,32.9, 28.8, 24.1, 14.2.

[0387] HRMS (ESI) m / z: [M + H] + Calcd for C 21 H 31 N2O3 + 359.2329; Found 359.2334.

[0388] HPLC:>99:1 er determined by analytical HPLC, Daicel CHIRALPAK® IDcolumn, 25 ℃, Hexane:i-PrOH = 99:1, 1.0 mL / min, 254 nm, t minor = 25.8 min,t major = 17.4 min.

[0389] [α]D 20 = +22.5 (c = 0.12, CH2Cl2).

[0390] Anti-inflammatory activity test

[0391] Mouse microglial cells, i.e. BV2 cell line, were purchased from the Cell Bank of Chinese Academy of Sciences, catalog number: GNM45.

[0392] After being stimulated by lipopolysaccharide (LPS), the gene expression level of pro-inflammatory factors in BV2 cells significantly increases, prompting the cells to secrete a large amount of inflammatory factors such as TNF-a, IL-6 and IL-1b. These inflammatory factors play an important role in the process of neuroinflammation, especially TNF-a and IL-6, which can activate the downstream inflammatory cascade reaction, thereby exacerbating the inflammatory state of the nervous system.

[0393] BV2 cell culture

[0394] The gas phase composition of the culture is: air 95vol%, carbon dioxide 5vol%; temperature 37°C; humidity 70%~80%. The composition of the complete culture medium is: 88wt% DMEM high-sugar culture medium, 10wt% fetal bovine serum, 1wt% L-glutamine, 1wt% penicillin-streptomycin.

[0395] Recovery, the freeze tube containing 1 mL cell suspension was thawed in a 37°C water bath, shaken and transferred to a centrifuge tube containing 4 mL complete medium, centrifuged at 800 rpm for 3 min, the supernatant was discarded, 2 mL medium was added and shaken gently. 1 mL cell suspension was transferred to a T25 culture flask, 5 mL complete medium was added and placed in the incubator for routine culture.

[0396] When the cell density reached 80%, the cells were passaged. The culture supernatant was discarded, washed with 1 mL PBS, 1 mL digestion solution was added to the culture flask, and the flask was incubated in a 37°C incubator for 1 min. The digestion was observed, and if most of the cells were rounded and detached, 2 mL complete medium was added to terminate the digestion. The cells were transferred to a centrifuge tube, centrifuged at 800 rpm for 3 min, the supernatant was discarded, 1 mL medium was added and shaken, and finally the cell suspension was transferred to a T25 flask and complete medium was added to 5 mL, which was shaken crossly.

[0397] Compound inhibition of LPS-induced BV2 cell model experiment

[0398] First, the normally cultured BV2 cells were transferred from a T25 cell culture flask to a 15 mL centrifuge tube after digestion, centrifugation and resuspension of the cells. Then, part of the cells were diluted ten times for counting, and 3.2 x 10 4 cells / mL of cell density were plated into a 6-well plate, and finally the plate was placed in the incubator for 12-24 h of continuous culture. When the cell density reached 50%, 100 ng / mL of LPS was added to establish the BV2 cell inflammation model, and the mixture of compound and 100 ng / mL LPS was treated. After 24 h, the cells were collected to extract RNA for reverse transcription and qPCR analysis, or the cell supernatant was collected for ELISA detection.

[0399] qPCR detection of inflammatory factor expression level

[0400] (1) Extraction of BV2 cell RNA

[0401] After removing the BV2 cell culture medium supernatant, 500 μL PBS was added to each well for washing. Then, 500 μL Buffer RL was added for cell digestion and lysis. The lysed sample was transferred to FastPure gDNA-Filter Columns III, centrifuged at 12,000 rpm for 30 s, and then the FastPure gDNA-Filter Columns III was discarded and the filtrate was collected. 250 μL of absolute ethanol was added to the filtrate and mixed well. The mixture was transferred to FastPure RNA Columns III, centrifuged at 12,000 rpm for 30 s, and the filtrate was discarded.

[0402] Next, 700 μL Buffer RW1 was added to the FastPure RNA Columns III, centrifuged at 12,000 rpm for 30 s, and the filtrate was discarded. Subsequently, 700 μL Buffer RW2 (with the addition of anhydrous ethanol) was added, centrifuged in the same manner, and the filtrate was discarded. Another 500 μL Buffer RW2 (with the addition of anhydrous ethanol) was added, centrifuged at 12,000 rpm for 2 min, and the filtrate was discarded. Finally, the FastPure RNA Columns III were placed back into the collection tube, centrifuged at 12,000 rpm for 1 min to prevent contamination by ethanol.

[0403] The adsorption column was carefully transferred to a new 1.5 mL centrifuge tube, 50 μL RNase-free MQ H2O was added dropwise in the center of the adsorption column, and it was left to stand at room temperature for 3 min, and then centrifuged at 12,000 rpm for 1 min to elute the RNA. The total RNA extracted can be directly used for reverse transcription and qPCR, or stored at -80°C.

[0404] (2) Reverse transcription

[0405] To remove genomic DNA, 1 μg of template RNA, 4 μL of 4×gDNA WiperMix, and RNase-free MQ H2O were added to an RNase-free centrifuge tube, and the mixture was mixed and centrifuged briefly. The mixture was heated at 42°C for 2 min. Then 4 μL of 5×HiScript III qRT SuperMix was added to make the reaction system 20 μL, mixed and centrifuged briefly.

[0406] The reverse transcription reaction was performed at a temperature of 37°C for 15 min, and then the temperature was raised to 85°C for 5 s. The 20 μL of cDNA product generated can be immediately used for qPCR reaction, or stored at 4°C for short-term storage to avoid repeated freeze-thawing.

[0407] (3) Real-time fluorescent quantitative polymerase chain reaction (qPCR)

[0408] Reaction solution preparation: 10 μL of 2×Taq Pro Universal SYBR qPCR Master Mix, 0.4 μL of Primer 1 (5 µM), 0.4 μL of Primer 2 (5 µM), 2 μL of Template DNA / cDNA, and water to 20 μL.

[0409] Reaction program settings: Stage 1, pre-denaturation, Rep: 1, 95℃, 30s; Stage 2, cycle reaction, Rep: 40, 95℃, 10s, 60℃, 30s; Stage 3, melting curve, using the default melting curve collection program of the instrument.

[0410] Data processing: First, record the fluorescence signal intensity (Ct value) of each reaction in the Roche qPCR instrument, subtract the Ct value of the target gene from the reference gene, calculate the ΔCt value (ΔCt = Ct target gene - Ct reference gene), and then calculate the ΔΔCt value (ΔΔCt = ΔCt - ΔCt control group). Next, calculate the relative expression of the target gene using the 2^(-ΔΔCt) method.

[0411] The primers used and their sequences are as follows.

[0412] qPCR primer sequences

[0413]

[0414] In the present application, the FORWARD primer sequence of IL-1β is as shown in SEQ ID NO. 1, the REVERSE primer sequence of IL-1β is as shown in SEQ ID NO. 2, the FORWARD primer sequence of IL-6 is as shown in SEQ ID NO. 3, the REVERSE primer sequence of IL-6 is as shown in SEQ ID NO. 4, the FORWARD primer sequence of TNF-α is as shown in SEQ ID NO. 5, and the REVERSE primer sequence of TNF-α is as shown in SEQ ID NO. 6.

[0415] Inhibition of compound on mRNA level of inflammatory factor IL-6 in BV2 cells

[0416] A blank control group, an LPS modeling group, and an LPS and drug combined treatment group were designed. After 24h of treatment, the cells were collected and RNA was extracted, and after reverse transcription, the expression level of pro-inflammatory genes was detected using qPCR. Figure 1 To inhibit the expression level of inflammatory factor IL-6 in the LPS-induced BV2 cell model by the compound, Table 1 is the inhibition rate of the compound on IL-6, and the results show that LPS stimulation significantly up-regulated the expression of IL-6 in BV2 cells, and most of the compounds had obvious inhibitory effect on IL-6, with an inhibition rate of more than 30%, only cis -3e、 cis -3v and trans -3at had an inhibition rate of less than 30%.

[0417] Table 1 Inhibition rate of compound on IL-6

[0418]

[0419] Inhibition of mRNA level of inflammatory factor TNF-α in BV2 cells by compounds

[0420] The blank control group, LPS modeling group and LPS and drug combined treatment group were designed. After 24h treatment, the cells were collected and RNA was extracted, and after reverse transcription, the expression level of pro-inflammatory genes was detected by qPCR. Figure 2 To inhibit the expression level of inflammatory factor TNF-α in LPS-induced BV2 cell model by compounds, Table 2 is the inhibition rate of TNF-α by compounds, and the results show that LPS stimulation significantly up-regulates the expression of TNF-α in BV2 cells, and 13 compounds have inhibitory effect on TNF-α, of which: cis -3o, trans -3ah', cis -3s, trans -3ao, cis -3j.

[0421] Table 2 Inhibition rate of TNF-α by compounds

[0422]

[0423] Inhibition of mRNA level of inflammatory factor IL-1β in BV2 cells by compounds

[0424] The blank control group, LPS modeling group and LPS and drug combined treatment group were designed. After 24h treatment, the cells were collected and RNA was extracted, and after reverse transcription, the expression level of pro-inflammatory genes was detected by qPCR. Figure 3 To inhibit the expression level of inflammatory factor IL-1β in LPS-induced BV2 cell model by compounds, the results show that LPS stimulation significantly up-regulates the expression of IL-1β in BV2 cells, most of the compounds have no obvious inhibitory effect on IL-1β, and even promote the expression of IL-1β, and only trans -3ah and trans -3ah' have an inhibition rate greater than 30%.

[0425] In summary, in addition to cis -3e, cis -3v and trans -3at, there are 20 compounds with an inhibition rate greater than 30% on IL-6; 15 compounds have inhibitory effect on TNF-α, of which: cis -3o, trans -3ah', cis -3s, trans -3ao,cis -3j; most of the compounds have no significant inhibitory effect on IL-1β, only trans -3ah and trans -3ah' is greater than 30%. Therefore, the compounds with the inhibitory rate greater than 30% on IL-6, TNF-α and IL-1β are: trans -3ah'.

[0426] The above description is only the preferred embodiments of the present application, and does not limit the present application in any form. It should be noted that for those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which should also be considered as the protection scope of the present application.

Claims

1. A chiral spiro compound, characterized by, having trans -3ah' the structure: trans -3ah'.

2. Use of the chiral spiro compound of claim 1 for the manufacture of an anti-inflammatory drug.

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