Polysubstituted 4-hydroxy-2-thiabicyclo [3.1. 1] heptane as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of organic synthesis, and discloses a method for preparing polysubstituted 4-hydroxy-2-thiabicyclo [3.1. 1] heptane through cycloaddition reaction catalyzed by lewis acid. The method comprises the following steps: by taking polysubstituted bicyclo [1.1. 0] butane and 2, 5-dihydroxy-1, 4-dithiane as raw materials, in the presence of a Lewis acid catalyst and an organic solvent, obtaining a cyclization product as shown in a formula I; and removing R1 from the cyclization product as shown in the formula I to obtain a molecule as shown in a formula II. In addition, an asymmetric catalysis method is developed to realize enantioselective synthesis of the cyclization product as shown in the formula I. Compared with a traditional method, the preparation method has the advantages that the raw materials and the catalyst are simple and easy to obtain, the synthesis route is short, the atom economy is high, and the substrate universality is wide; the reaction product can be used as a potential bioisostere of drug active molecules containing ortho-position, meta-position or 1, 2, 4-trisubstituted aromatic hydrocarbon, such as diflunisal; the 4-hydroxy-2-thiabicyclo [3.1. 1] heptane product is subjected to pharmacokinetics and other activity tests, and shows a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of organic synthesis technology, and in particular to a polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane, its preparation method and application. Background Technology

[0002] With the rise of the "escape the plane" drug development concept (ref 1. J. Med. Chem. 2009, 52, 6752–675), a surge of research has emerged on isosteric substitution of planar aromatic hydrocarbons using three-dimensional bicyclic [n.1.1]alkane skeletons. Recent studies have shown that bicyclic [3.1.1]heptane can be used as a bioelectron isostere of meta-benzene rings (ref 2. Nature 2022, 611, 721–726), thereby improving the physicochemical properties and activity of drugs. Studies by Mykhaiiuk et al. have shown that introducing heteroatoms such as nitrogen or oxygen into the bicyclic [3.1.1]heptane molecular skeleton can make it an isosteric compound of aromatic heterocycles, while improving the drug's metabolic kinetics (ref 3. Nat. chem. 2023, 15, 1155–1163; ref. 4. Angew. Chem. Int. Ed. 2024, 63, e202318476; ref 5. Angew. Chem. Int. Ed. 2024, 63, e202402730; ref. 6. Angew. Chem. Int. Ed. 2023, 62, e202304246). However, there are currently only two methods for synthesizing 2-thiabicyclo[3.1.1]heptane derivatives. As early as 1981, the Hamanaka group reported a method for synthesizing 2-thiabicyclo[3.1.1]heptane using intramolecular Michael addition reaction as a cyclization strategy. However, the synthetic route is long (more than 10 synthetic steps are required using 3-alkenylcyclobutyl-1-one as the starting material) and cumbersome (ref 7. Tetrahedron Lett. 1981, 22, 1345–1348), which has limited the application of 2-thiabicyclo[3.1.1]heptane derivatives. Our research group recently used pyridinium 1,4-zwitterion thiolate and polysubstituted bicyclo[1.1.0]butane as raw materials, and under the catalysis of scandium trifluoromethanesulfonate (Sc(OTf)3) and with acetonitrile as solvent, at 80 °C, to obtain a polysubstituted 2-thiabicyclo[3.1.1]heptene derivative (ref 8. Angew. Chem. Int. Ed. 2024, e202408578, published).

[0003] Hamanaka's work:

[0004]

[0005] Previous research papers published by our research group:

[0006]

[0007] However, the above reaction requires a temperature of 80°C to obtain a satisfactory yield, which is quite demanding. Furthermore, the pyridine in the reaction is a leaving group, which leads to low atom economy. In addition, the pyridinium 1,4-zwitterion thiolate needs to be prepared in advance and is not commercially available. Summary of the Invention

[0008] Based on the shortcomings of the existing technology, this invention innovatively proposes a (3+3) cycloaddition reaction of multi-substituted bicyclic [1.1.0]butane with 2,5-dihydroxy-1,4-dithiane as the key cyclization strategy, and for the first time achieves the synthesis of hydroxyl-functionalized 2-thiabicyclic [3.1.1]heptane. Using a Lewis acid as a catalyst, this invention constructs a hydroxyl-functionalized 2-thiabicyclic [3.1.1]heptane skeleton by cycloaddition of commercially available 2,5-dihydroxy-1,4-dithiane with bicyclic butane at room temperature using a Lewis acid as a catalyst. The cyclized product containing multiple functional groups, as shown in Formula I, can undergo a series of derivatizations (as shown in Examples 20 to 41). Furthermore, the pharmacokinetics (Example 42) and drug activity tests (Example 43) of the related derivatized products were conducted, demonstrating good application prospects and providing a material basis and theoretical foundation for related medicinal chemistry and new drug development.

[0009] The difficulties to be overcome by this invention include: (1) In the presence of Lewis acids, polysubstituted bicyclic [1.1.0]butane is prone to isomerization reaction and decomposes into cyclobutene. How to find a suitable catalytic system to achieve its chemically selective cyclization reaction is the first difficulty to be overcome by this invention; (2) The hydroxyl groups in 2,5-dihydroxy-1,4-dithiaane and the cyclization product will undergo a ring-opening reaction with polysubstituted bicyclic [1.1.0]butane. How to suppress this ring-opening side reaction and selectively obtain the cyclization product is the second difficulty to be overcome by this invention; (3) This cycloaddition reaction is a novel reaction. How to design this novel (3+3) cycloaddition reaction and find efficient catalytic reaction conditions through condition screening and optimization to achieve efficient and highly selective (3+3) cycloaddition reaction of polysubstituted bicyclic [1.1.0]butane with 2,5-dihydroxy-1,4-dithiaane, and prepare 2-thiabicyclic [3.1.1]heptane derivatives in an atom-economical manner is the third difficulty to be overcome by this invention.

[0010] In this invention, the raw materials and catalysts are inexpensive and readily available, the reaction conditions are mild (room temperature reaction), and the synthetic steps are concise, yielding derivatives of 2-thiabicyclo[3.1.1]heptane as shown in Formula I in high yield. Simultaneously, the asymmetric catalytic system proposed in this invention enables the first enantioselective synthesis of chiral 2-thiabicyclo[3.1.1]heptane derivatives (Examples 15-19). The obtained 2-thiabicyclo[3.1.1]heptane product as shown in Formula I possesses multiple functional groups such as hydroxyl, carbonyl, and sulfur groups, providing conditions for various transformations of the product. This invention also demonstrates various derivatization transformations of the 2-thiabicyclo[3.1.1]heptane product as shown in Formula I, providing an efficient method for the diverse synthesis of a molecular library of 2-thiabicyclo[3.1.1]heptane (Examples 20-41). In addition, this invention also synthesized a 2-thiabicyclo[3.1.1]heptane analogue of diflunisal (an aspirin alternative; a nonsteroidal anti-inflammatory analgesic), and compared the solubility (sol), lipophilicity (logD), and hepatic microsomal enzyme stability (CL) of diflunisal with its bridged ring analogues: I-3o, II-3b, II-3c, and II-3d. int ) and half-life (T) 1 / 2 Pharmacokinetic and other pharmaceutical properties were also assessed (Example 42). Compounds XV-3a-3f containing a 2-thiabicyclic [3.1.1]heptane structure were synthesized, and their effects on osteoclast growth were tested. Among them, compounds XV-3a and XV-3c synthesized in this invention exhibited significant inhibitory activity against osteoclast growth (Example 43).

[0011] The purpose of this invention is to provide an atom-economical method for synthesizing multi-substituted 4-hydroxy-2-thiabicyclo[3,1,1]heptane, addressing the shortcomings of existing technologies. The synthetic method utilizes readily available and inexpensive reactants, constructing the 2-thiabicyclo[3.1.1]heptane skeleton via a (3+3) cycloaddition reaction, synthesizing various cyclized products in yields ranging from 60% to 98%. This method offers advantages such as readily available and simple reactants, 100% atom economy, short steps, environmental friendliness, and high efficiency. Furthermore, the solubility, lipophilicity, hepatic microsomal enzyme stability, and half-life of the derivatized products were tested (Example 42). The effects of these compounds containing the 2-thiabicyclo[3.1.1]heptane structure on osteoclast growth were also investigated, demonstrating promising application prospects (Example 43). In addition, this invention develops an asymmetric catalytic method to achieve the enantioselective synthesis of chiral 4-hydroxy-2-thiabicyclo[3.1.1]heptane.

[0012] This invention proposes a polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane, the structure of which is shown in Formulas I and II below:

[0013]

[0014] R 1 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, alkoxy, substituted amino, C1-C 12 Alkyl or C3-C 12 cycloalkyl, alkenyl, ynyl Where R 4 Arbitrarily selected from alkyl, cycloalkyl, aryl, and heteroaryl groups; R 5 Arbitrarily selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and halogen;

[0015] R 2 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, C1-C 12 Alkyl or C3-C 12 Cycloalkyl, alkenyl, alkynyl, silyl, and boron-containing groups;

[0016] R 3 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, C1-C 12 Alkyl or C3-C 12 Cycloalkyl, alkenyl, alkynyl, silyl, and boron-containing groups;

[0017] Preferably, R 1 For substituted phenyl, heteroaryl, C 1- C 12 alkyl, Where R 4 The derivatives are methyl, ethyl, isopropyl, n-propyl, and cyclopentyl; R 5 It is hydrogen or methyl;

[0018] Preferably, R 2 Hydrogen, substituted phenyl, heteroaryl, C1-C 12 Alkyl, alkenyl, and ynyl groups;

[0019] Preferably, R 3 It can be hydrogen, phenyl, heteroaryl, or boron-containing groups.

[0020] This invention also proposes a method for preparing polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane, comprising the following steps: using polysubstituted bicyclo[1.1.0]butane and 2,5-dihydroxy-1,4-dithiane as raw materials, stirring in the presence of a Lewis acid catalyst and an organic solvent to obtain the cyclized product shown in Formula I; then, the cyclized product shown in Formula I is subjected to R removal... 1The 4-hydroxy-5-carboxy-2-thiabicyclo[3.1.1]heptane derivative shown in Formula II was obtained.

[0021] The specific reaction route is as follows.

[0022]

[0023] Among them, R 1 R 2 R 3 The definitions are the same as those in formulas I and II.

[0024] In this invention, the ratio of the amount of the polysubstituted bicyclic [1.1.0]butane 2 to 2,5-dihydroxy-1,4-dithiaane 1 is 1:0.5 to 5:1; preferably, it is 3:1.

[0025] Based on the amount of compound 1, the amount of the catalyst is 1 mol% to 10 mol%; preferably, it is 10 mol%.

[0026] When the structure of Formula I is obtained in this invention, separation and purification are performed. The separation and purification are performed using a mixed solvent of petroleum ether / ethyl acetate as the eluent, with a volume ratio of 5:1 to 2:1.

[0027] In this invention, the first organic solvent is one or more of dichloromethane, 1,2-dichloroethane, chloroform, toluene, tetrahydrofuran, ethyl acetate, 1,4-dioxane, and acetonitrile; preferably, it is 1,2-dichloroethane, acetonitrile, or dichloromethane.

[0028] In this invention, the second organic solvent is tetrahydrofuran and water; preferably, it is a mixed solvent in which the volume ratio of tetrahydrofuran to water is 1:1.

[0029] In this invention, the alkali is selected from one or more of LiOH, DBU, NaOH, etc.; preferably, it is LiOH.

[0030] Based on the amount of compound of formula I, the amount of base is 1-4 equivalents; preferably, it is 4 equivalents.

[0031] In this invention, the catalyst is one or more of scandium trifluoromethanesulfonate, nickel trifluoromethanesulfonate, gallium trifluoromethanesulfonate, europium trifluoromethanesulfonate, zinc trifluoromethanesulfonate, boron trifluoride ether, silver trifluoromethanesulfonate, and nickel perchlorate; preferably, it is zinc trifluoromethanesulfonate.

[0032] In this invention, the reaction is preferably carried out under a nitrogen atmosphere.

[0033] In this invention, the stirring temperature is 20℃-60℃; preferably, it is room temperature.

[0034] In this invention, the stirring time is 10-20 hours; preferably, it is 12 hours.

[0035] In this invention, the removal of R 1 The temperature is 20℃-50℃; preferably, it is room temperature.

[0036] In this invention, the removal of R 1 The time is 10-20 hours; preferably, it is 12 hours.

[0037] In one specific embodiment, the method includes the following steps:

[0038] S1: Under a nitrogen atmosphere, polysubstituted bicyclic [1.1.0]butane and 2,5-dihydroxy-1,4-dithiane were added to a preheated 25 mL reaction tube;

[0039] S2: Add Lewis acid catalyst to the above reaction tube in the glove box;

[0040] S3: Under a nitrogen atmosphere, a solvent was added and the mixture was stirred overnight at room temperature to obtain the cyclized product shown in Formula I;

[0041] S4: The cyclized product shown in Formula I is subjected to R removal... 1 The 4-hydroxy-5-carboxy-2-thiabicyclo[3.1.1]heptane derivative of formula II was obtained.

[0042] The term "bio-electron isosteres" in this invention refers to a substance that has similar, related, or opposite biological activities produced by groups or substituents with similar physical and chemical properties in a biological sense.

[0043] This invention utilizes the principle of bioisosteric substitution and, through pharmacokinetic testing, shows that, compared with the lipophilic and water-soluble properties of diflunisal, the water-soluble and lipophilic properties of the multi-substituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane described in this invention are significantly improved.

[0044] This invention also proposes the application of the polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane in the preparation of bioelectron isosteres containing, for example, diflunisal, salicylic acid, and other active pharmaceutical ingredients containing 1,2-disubstituted aromatics, 1,3-disubstituted aromatics, and 1,2,4-trisubstituted aromatics. Furthermore, this invention proposes the application of the polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane or its bioelectron isosteres in the preparation of drugs for treating bone cell-related diseases (osteoporosis, osteoarthritis, osteoma, etc.), treating overactive bladder with urinary incontinence, urgency, and frequency, chronic obstructive pulmonary disease and asthma, transthyretin amyloidosis (ATTR), and various mild to moderate pain and anticancer drugs.

[0045] The technical solution of the present invention has the following advantages compared with the prior art:

[0046] (1). The preparation method described in this invention uses the (3+3) cycloaddition reaction as the key cyclization step, which has 100% atom economy and meets the requirements of green chemistry;

[0047] (2). The preparation method described in this invention has the advantages of simple and readily available raw materials, simple operation, short synthesis route, green and environmentally friendly, and high synthesis efficiency. It can synthesize the cyclized product shown in Formula I with medium to excellent yields (60%-98%).

[0048] (3). The cyclized product obtained by the preparation method described in this invention, as shown in Formula I, has multiple functional groups such as hydroxyl, carbonyl, and sulfur groups, which is beneficial to the subsequent functional group transformation diversity of the product for the synthesis of other 2-thiabicyclo[3.1.1]heptane derivatives;

[0049] (4). The asymmetric catalytic method described in this invention can be used for the enantioselective synthesis of chiral cyclized products as shown in Formula I, with an ee value as high as 96%;

[0050] (5) The preparation method described in this invention requires no additional additives and has excellent atom economy; the method completes the preparation of the compound with formula II in two steps. In addition, the product represented by formula II is subjected to pharmacokinetic activities such as solubility, lipid solubility, liver microsomal enzyme stability, and half-life, showing good application prospects. Attached Figure Description

[0051] Figure 1 The effect of the compound on osteoclast growth of RAW264.7 cells induced by NF-κB ligand receptor activator (RANKL) was detected using tartrate-resistant acid phosphatase (TRAP) staining.

[0052] Note: Control: Cell culture in the absence of RANKL and other test compounds; RANKL: NF-κB ligand receptor activator. Detailed Implementation

[0053] The present invention will be further described below with reference to specific embodiments, so that those skilled in the art can better understand and implement the present invention, but the embodiments are not intended to limit the present invention.

[0054] The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit the invention or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0055] In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values.

[0056] This invention belongs to the field of organic synthesis technology and provides a method for the atom-economical synthesis of polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane using Lewis acid catalysis. The method comprises the following steps: using polysubstituted bicyclo[1.1.0]butane and 2,5-dihydroxy-1,4-dithiane as raw materials, stirring in the presence of a Lewis acid catalyst and an organic solvent to obtain the cyclized product shown in Formula I; then, the cyclized product shown in Formula I is subjected to R removal... 1 The 4-hydroxy-5-carboxy-2-thiabicyclo[3.1.1]heptane derivative of Formula II was obtained. The synthetic method of this invention, through condition optimization, yields the target product in moderate to excellent yields. It features low catalyst usage, mild conditions, atom economy, step economy, and environmental friendliness. It can rapidly and efficiently construct multi-substituted 4-hydroxy-2-thiabicyclo[3,1,1]heptane skeletons, which can serve as potential bioisosteres for active pharmaceutical molecules containing 1,2-disubstituted, 1,3-disubstituted, and 1,2,4-trisubstituted aromatics, such as diflunisal and salicylic acid. Simultaneously, the compound was subjected to pharmacokinetic tests, including solubility, lipophilicity, hepatic microsomal enzyme stability, and half-life. Its osteoclast growth inhibition activity was also investigated, demonstrating promising application prospects. Furthermore, an asymmetric catalytic method was developed to achieve the enantioselective synthesis of 4-hydroxy-2-thiabicyclo[3.1.1]heptane.

[0057] In this invention, unless otherwise stated, the abbreviations have the following meanings as shown in Table 1:

[0058] Table 1

[0059]

[0060]

[0061] Example 1: Synthesis of (3,5-dimethyl-1H-pyrazol-1-yl)(4-hydroxy-1-phenyl-2-thiabicyclo[3.1.1]heptane-5-yl) methyl ketone (I-3a)

[0062]

[0063] Take a pre-dried 25 mL reaction tube and accurately weigh 1 (15.2 mg, 0.1 mmol) and 2a (75.7 mg, 0.3 mmol) into it under a nitrogen atmosphere. Then, weigh zinc trifluoromethanesulfonate (7.2 mg, 10 mol%) in a glove box. Add 1,2-dichloroethane solvent and stir overnight at room temperature. Observe by TLC until the starting material reacts completely. Purify by column chromatography to give compound I-3a in 82% yield. 1 H NMR (400MHz, CDCl3) δ7.31(m,2H),7.26-7.22(m,2H),5.98(s,1H),5.27(m,1H),4.75(s,1H),3.50(dd,J=13.2,5.2Hz,1H), 3.29(dd,J=10.4,5.2Hz,1H),3.06(dd,J=13.2,4.4Hz,1H),2.93(m,2H),2.63(d,J=11.2Hz,1H),2.52(s,3H),2.25(s,3H). 13 C NMR (100MHz, CDCl3) δ173.8,153.1,145.8,143.9,128.4,127.1,125.5,110.9,70.1,51.7,47.4,43.7,40.2,31.9,14.2,13.7.

[0064] Example 2

[0065]

[0066] The experimental method used in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3-(4-chlorophenyl)bicyclo[1.1.0]but-1-yl)(3,5-dimethyl-1H-pyrazol-1-yl) methyl ketone, and the product obtained is shown in structural formula (I-3b), with a yield of 83%. 1 HNMR (400MHz, CDCl3) δ7.29-7.26(m,2H),7.20-7.17(m,2H),5.99(s,1H),5.23(s,1H),4.75(s,1H),3.47(dd,J=13.2,4.8Hz,1H) ,3.26(dd,J=10.8,6.4Hz,1H),3.06(dd,J=13.2,2.4Hz,1H),2.94-2.86(m,2H),2.60(d,J=11.2Hz,1H),2.52(s,3H),2.25(s,3H); 13C NMR (100MHz, CDCl3) δ173.6,153.2,145.1,142.4,132.9,128.5,127.0,111.0,70.0,51.6,46.9,43.7,40.1,31.9,14.2,13.7.

[0067] Example 3

[0068]

[0069] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is phenyl (3-phenylbicyclo[1.1.0]but-1-yl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3c), with a yield of 94%. 1 HNMR (400MHz, CDCl3): δ7.86(d,J=6.0Hz,2H),7.55(t,J=7.6Hz,1H),7.46(t,J=7.6Hz,2H),7.33-7.20(m,5H),4.87(s,1 H),3.42-3.46(m,1H),3.25-3.19(m,1H),3.11-3.08(m,1H),2.96(s,2H),2.77(d,J=10.8Hz,1H),2.59(d,J=48.8Hz,1H). 13 C NMR (100MHz, CDCl3): δ202.8,143.4,135.5,133.0,128.7,128.6,128.5,127.3,125.5,72.0,53.8,47.9,47.7,37.1,31.4.

[0070] Example 4

[0071]

[0072] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is phenyl(3-(4-(trifluoromethoxy)phenyl)bicyclo[1.1.0]but-1-yl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3d), with a yield of 94%. 1H NMR (400MHz, CDCl3): δ7.77(d,J=7.6Hz,2H),7.49-7.45(m,1H),7.38-7.34(m,2H),7.18-7.16(m,2H),7.06(d,J=7.6Hz,2H),4.78(d,J=4 .4Hz,1H),3.29(dd,J=12.8,5.6Hz,1H),3.14(dd,J=12.8,6.4Hz,1H),2.99(d,J=10.8Hz,1H),2.85-2.79(m,3H),2.65(d,J=10.8Hz,1H). 13 C NMR (100MHz, CDCl3): δ202.7,148.2,148.2,142.1,135.3,133.1,128.7,128 .6,127.1,121.0,120.4(q,J=255.7Hz),71.9,53.7,47.7,47.2,37.1,31.4.

[0073] Example 5

[0074]

[0075] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3-phenylbicyclo[1.1.0]but-1-yl)(m-tolyl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3e), with a yield of 97%. H NMR (400MHz, CDCl3): δ7.57(d,J=8.4Hz,2H),7.26-7.20(m,4H),7.16-7.13(m,3H),4.78(q,J=6.4Hz,1H),3.28(dd,J=13.2,5.6Hz ,1H),3.13(dd,J=13.2,6.8Hz,1H),2.99(d,J=9.6Hz,1H),2.86(s,2H),2.67(d,J=11.2Hz,1H),2.63(d,J=6.0Hz,1H),2.31(s,3H). 13 C NMR (100MHz, CDCl3): δ202.9,143.4,138.4,135.5,133.8,129.2,128.4,128.4,127.2,125.9,125.5,72.1,53.9,47.9,47.8,36.9,31.4,21.4.

[0076] Example 6

[0077]

[0078] The experimental method used in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is 7-((tert-butyldimethylsiloxy)-1-(3-phenylbicyclo[1.1.0]butan-1-yl)heptan-2-yn-1-one, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3f), with a yield of 82%. 1 HNMR (400MHz, CDCl3): δ7.49-7.45(m,2H),7.40-7.38(m,3H),5.00(s,1H),3.67(t,J=4.8Hz,2H),3.54(dd,J=13.2,6.0Hz,1H),3.1 7(dd,J=13.2,4.8Hz,1H),3.09-2.97(m,4H),2.73-2.68(m,1H),2.54(t,J=6.4Hz,2H),1.78-1.66(m,4H),1.00(s,9H),0.15(s,6H). 13 C NMR (100MHz, CDCl3): δ189.6,143.7,128.5,127.3,125.4,98.5,78.6,69. 5,62.2,54.1,48.7,43.8,37.2,31.7,31.0,25.9,24.3,18.9,18.3,-5.4.

[0079] Example 7

[0080]

[0081] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3-methylbicyclo[1.1.0]but-1-yl)(phenyl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in the structural formula (I-3g), with a yield of 43%. 1 HNMR (400MHz, CDCl3): δ8.65(d,J=4.8Hz,1H),8.02(d,J=7.6Hz,1H),7.90(t,J=7.6Hz, 1H),7.53-7.50(m,1H),7.31-7.27(m,2H),7.22-7.18(m,2H),6.21(s,1H),4.75(d,J=2. 8Hz,1H),3.56(dd,J=13.2,4.8Hz,1H),3.26(dd,J=10.8,7.6Hz,1H),3.10(dd,J=13.2,4 .0Hz,1H),2.95(dd,J=11.2,7.6Hz,1H),2.83(d,J=11.2Hz,1H),2.44(d,J=10.4Hz,1H). 13C NMR (100MHz, CDCl3): δ201.1,152.2,147.9,144.2,138.1,128.4,127.2,127.0,125.5,123.7,69.4,54.3,47.8,42.6,39.6,32.0.

[0082] Example 8

[0083]

[0084] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is 1-(3-phenylbicyclo[1.1.0]but-1-yl)pent-1-one, and the product obtained is shown in the structural formula (I-3h), with a yield of 87%. 1 H NMR (400MHz, CDCl3): δ7.35-3.31(m,2H),7.25-7.23(m,3H),4.69(q,J=6.4H z,1H),3.33(dd,J=13.2,6.0Hz,1H),3.14-3.10(m,2H),2.88(t,J=8.8Hz,1H) ,2.71(d,J=10.4Hz,1H),2.65(d,J=10.8Hz,1H),2.59(t,J=8.4Hz,1H),2.54- 2.45(m,2H),1.57-1.49(m,2H),1.34-1.25(m,2H),0.90(t,J=7.2Hz,3H)ppm. 13 C NMR (100MHz, CDCl3): δ212.5,143.6,128.5,127.2,125.4,70.8,53.7,47.7,45.3,37.5,35.3,31.2,25.2,22.2,13.8.

[0085] Example 9

[0086]

[0087] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3-methylbicyclo[1.1.0]but-1-yl)(phenyl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3i), with a yield of 87%. 1H NMR (400MHz, CDCl3): δ7.84(d,J=7.6Hz,2H),7.54(d,J=7.2Hz,1H),7.45(t,J=7.6Hz,2H),4.76(q,J=6.0Hz,1H),3.29(dd,J=13.2,6.0Hz,1H ),3.07(dd,J=12.8,6.8Hz,1H),2.69(dd,J=11.2,6.8Hz,1H),2.60-2.52(m,2H),2.37(d,J=11.2Hz,1H),2.27(d,J=6.4Hz,1H),1.34(s,3H). 13 C NMR (100MHz, CDCl3): δ202.8,135.6,132.9,128.7,128.5,72.1,54.08,49.3,43.3,38.5,30.6,25.9.

[0088] Example 10

[0089]

[0090] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is bicyclic [1.1.0]but-1-yl(naphth-2-yl)methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3j), with a yield of 60%. 1 HNMR (400MHz, CDCl3): δ8.34(s,1H),7.95-7.85(m,4H),7.63-7.53(m,2H),4.88(s,1H),3.34-3.28(m, 1H),3.16-3.10(m,2H),3.02-2.98(m,1H),2.78-2.73(m,1H),2.66-2.62(m,1H),2.53-2.48(m,1H)ppm. 13 C NMR (100MHz, CDCl3): δ202.8,135.4,132.9,132.3,130.4,129.6,128.57,128.5,127.8,126.8,124.4,72.8,57.1,44.6,34.0,32.3,28.6.

[0091] Example 11

[0092]

[0093] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is naphth-2-yl(3-phenylbicyclo[1.1.0]but-1-yl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in the structural formula (I-3k), with a yield of 96%. 1 HNMR (400MHz, CDCl3): δ8.38(s,1H),7.96-7.85(m,4H),7.62-7.54(m,2H),7.33-7.22(m,5H),4.97(d,J=4.4Hz,1H),3.45(dd,J=13.2,6 .0Hz,1H),3.26(dd,J=13.2,6.8Hz,1H),3.18(dd,J=10.8,5.6Hz,1H),3.09-3.03(m,2H),2.85(d,J=11.2Hz,1H),2.41(d,J=3.6Hz,1H). 13 C NMR (100MHz, CDCl3): δ202.5,143.4,135.4,132.8,132.3,130.4,129.6,128.6,12 8.5,128.5,127.8,127.3,126.9,125.5,124.4,72.2,54.0,48.0,47.9,37.3,31.6.

[0094] Example 12

[0095]

[0096] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is phenyl(3-(o-tolyl)bicyclo[1.1.0]but-1-yl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in structural formula (I-3l), with a yield of 76%. 1 H NMR (400MHz, CDCl3): δ7.77(d,J=7.2Hz,2H),7.47-7.43(m,1H),7.38-7.34(m,2H),7.06-6.97(m,4H),4.77-4.73( m,1H),3.28-3.22(m,1H),3.16-3.08(m,2H),2.99-2.89(m,2H),2.81(s,1H),2.70(d,J=11.2Hz,1H),2.34(s,3H). 13C NMR (100MHz, CDCl3): δ202.9,140.8,136.3,135.5,133.0,131.1,128.7,128.5,127.6,126.4,125.9,72.0,53.8,48.8,48.5,37.6,31.1,20.3.

[0097] Example 13

[0098]

[0099] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3,5-dimethyl-1H-pyrazole-1-yl)(3-(m-tolyl)bicyclo[1.1.0]but-1-yl) methyl ketone, and the product obtained is shown in the structural formula (I-3m), with a yield of 71%. 1 HNMR (400MHz, CDCl3): δ7.19(t,J=7.2Hz,1H),7.07-7.02(m,3H),5.98(s,1H),5.24(s,1H),4.76-4.73(m,1H),3.51-3.45(m ,1H),3.29-3.25(m,1H),3.09-3.03(m,1H),2.96-2.88(m,2H),2.62(d,J=11.Hz,1H),2.51(s,3H),2.32(s,3H),2.25(s,3H). 13 C NMR (100MHz, CDCl3): δ173.8,153.0,145.0,143.8,138.0,128.3,127.9,1 26.2,122.5,110.9,70.1,51.7,47.4,43.8,40.2,31.9,21.3,14.2,13.7.

[0100] Example 14

[0101]

[0102] The experimental method in this embodiment is basically the same as that in Example 1. The BCB used in this embodiment is (3,5-dimethylphenyl)(3-phenylbicyclo[1.1.0]but-1-yl) methyl ketone, the solvent is acetonitrile, and the product obtained is shown in the structural formula (I-3n), with a yield of 94%. 1HNMR (400MHz, CDCl3): δ7.45(s,2H),7.30(t,J=7.2Hz,2H),7.25-7.18(m,4H),4.87(q,J=6.4Hz,1H),3.36(dd,J=13.2,6.0Hz,1 H),3.22(dd,J=13.2,6.8Hz,1H),3.09-3.04(m,1H),2.98-2.94(m,2H),2.76(d,J=10.8Hz,1H),2.70-2.63(m,1H),2.36(s,6H). 13 C NMR (100MHz, CDCl3): δ203.0,143.5,138.2,135.6,134.7,128.4,127.2,126.4,125.5,72.1,53.9,48.0,47.9,36.9,31.4,21.3.

[0103] Example 15

[0104]

[0105] Take a pre-dried 25 mL reaction tube, accurately weigh Zn(OTf)₂ (3.6 mg, 10 mol%) and chiral oxazoline ligand (12 mol%) in a glove box, then add 2b (23.8 mg, 0.1 mmol) and 1 (9.1 mg, 0.6 mmol) under a nitrogen atmosphere, followed by the addition of dichloroethane solvent. Stir overnight at room temperature. Detect by TLC until the reactants are completely reacted. Purify by column chromatography to give compound I-3o in 78% yield with an ee value of 93%. 1 H NMR (400MHz, CDCl3): δ7.32-7.29(m,2H),7.25-7.15(m,2H),7.15(s,1H),7.05(s,1H),6.64(s,1H),4.66(t,J=4.0Hz,1H),3.97(s,3H ),3.55(dd,J=13.2,4.4Hz,1H),3.34(dd,J=10.4,6.0Hz,1H),3.34(dd,J=13.6,2.0Hz,1H),2.89-2.82(m,2H),2.49(d,J=10.8Hz,1H). 13 C NMR (100MHz, CDCl3): δ192.2,144.5,141.6,128.7,128.4,127.0,126.9,125.5,68.5,54.6,47.8,40.6,40.1,36.1,32.2.

[0106] Example 16

[0107]

[0108] The experimental method in this embodiment is basically the same as that in Example 15. The BCB used in this embodiment is 3-(4-chlorophenyl)bicyclo[1.1.0]but-1-yl)(1-methyl-1H-imidazol-2-yl) methyl ketone. The product obtained is shown in the structural formula (I-3p), with a yield of 80% and an ee value of 91%. 1 H NMR (400MHz, CDCl3): δ7.29-7.26(m,2H),7.20-7.16(m,3H),7.06(s,1H),6.65(s,1H),4.65(s,1H),3.99(s,3H ),3.58-3.53(m,1H),3.33(d,J=10.0Hz,1H),2.99(d,J=13.2Hz,1H),2.83-2.79(m,2H),2.45(d,J=10.4Hz,1H). 13 C NMR (100MHz, CDCl3): δ192.0,143.0,141.6,132.8,128.8,128.6,127.0,127.0,68.3,54.5,47.3,40.6,40.0,36.2,32.2.

[0109] Example 17

[0110]

[0111] The experimental method in this embodiment is basically the same as that in Example 15. The BCB used in this embodiment is (1-methyl-1H-imidazol-2-yl)(3-methylbicyclo[1.1.0]but-1-yl) methyl ketone. The product obtained is shown in the structural formula (I-3q), with a yield of 50% and an ee value of 80%. 1 H NMR (400MHz, CDCl3): δ7.13(s,1H),7.04(s,1H),6.53(s,1H),4.55(s,1H),4.00(s,3H),3.66(d d,J=13.2,6.4Hz,1H),2.96-2.6(m,2H),2.47-2.42(m,2H),2.06(d,J=10.4Hz,1H),1.33(s,3H). 13 C NMR (100MHz, CDCl3): δ192.7,141.7,128.7,126.9,68.5,54.8,43.1,42.4,41.6,36.2,31.3,26.6.

[0112] Example 18

[0113]

[0114] The experimental method in this embodiment is basically the same as that in Example 15. The BCB used in this embodiment is (1-methyl-1H-imidazol-2-yl)(3-(o-tolyl)bicyclo[1.1.0]but-1-yl) methyl ketone. The product obtained is shown in the structural formula (I-3r), with a yield of 70% and an ee value of 90%. 1 H NMR (400MHz, CDCl3): δ7.16-7.04(m,6H),6.59(s,1H),4.65(s,1H),3.96(s,3H),3.54(dd,J=13.2 ,4.4Hz,1H),3.45(dd,J=10.4,6.8Hz,1H),3.01-2.89(m,3H),2.54(d,J=10.4Hz,1H),2.45(s,3H). 13 C NMR (100MHz, CDCl3): δ192.3,141.8,141.7,136.3,131.0,128.7,127.3,126.9,126.4,125.8,68.5,54.4,48.4,41.3,41.1,36.1,31.9,20.3.

[0115] Example 19

[0116]

[0117] The experimental method in this embodiment is basically the same as that in Example 15. The BCB used in this embodiment is (1-methyl-1H-imidazol-2-yl)(3-(naphthyl-2-yl)bicyclo[1.1.0]but-1-yl) methyl ketone. The product obtained is shown in the structural formula (I-3s), with a yield of 61% and an ee value of 93%. 1 H NMR (400MHz, CDCl3): δ7.79-7.76(m,3H),7.65(s,1H),7.46-7.41(m,3H),7.15(s,1H),7.02(s,1H),6.65(s,1H),4.70(s,1H),3.96(s,3 H),3.59(dd,J=13.2,4.4Hz,1H),3.44(dd,J=10.4,5.2Hz,1H),3.03(dd,J=13.2,2.8Hz,1H),2.97-2.91(m,2H),2.59(d,J=10.4Hz,1H). 13C NMR (100MHz, CDCl3): δ191.9,146.4,141.5,134.2,129.7,128.7,127.1,127.0,125.9,123.7,68.3,54.5,47.4,40.5,40.1,36.1,32.1.

[0118] Example 204 Synthesis of 4-hydroxy-1-phenyl-2-thiabicyclo[3.1.1]heptane-5-carboxylic acid (II-3a)

[0119]

[0120] Take a pre-dried 25 mL reaction tube, accurately weigh I-3a (32.8 mg, 0.1 mmol) into it under a nitrogen atmosphere, add 2 mL of a mixed solution of THF and water, then add lithium hydroxide (9.6 mg, 0.4 mmol), stir at room temperature for 16 h, and detect by TLC until the starting materials react completely. Extract the reaction solution once with ethyl acetate, separate the layers, adjust the pH of the aqueous phase to 1 with hydrochloric acid, and then extract the aqueous phase three more times with ethyl acetate. Combine the organic phases, dry with anhydrous magnesium sulfate, filter, and concentrate under reduced pressure to obtain compound II-3a in 90% yield. 1 H NMR (400MHz, CDCl3) δ7.335-7.31(m,3H),7.26-7.25(m,2H),6.23-6.13(m,1H),4.76(t,J=5.6Hz,1H),3.38(dd,J=13 .6,6.4Hz,1H),3.08(dd,J=13.6,5.6Hz,1H),2.98-2.92(m,2H),2.80(d,J=10.8Hz,1H),2.60(dd,J=10.0,7.2Hz,1H); 13 C NMR (100MHz, CDCl3) δ178.8,143.5,128.5,127.4,125.4,69.9,48.5,47.8,44.4,36.9,30.6.

[0121] Example 21

[0122]

[0123] The experimental method used in this embodiment is basically the same as that in Example 20. The raw material used in this embodiment is (3,5-dimethyl-1H-pyrazol-1-yl)(1-(4-fluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-yl) methyl ketone, and the product obtained is shown in structural formula (II-3b), with a yield of 88%. 1HNMR (400MHz, DMSO-d6): δ7.33-7.29(m,2H),7.21-7.16(m,2H),5.51(s,1H),4.57(s,1H),3.3 3(dd,J=13.2,6.0Hz,1H),2.96(dd,J=12.8,5.2Hz,1H),2.82-2.78(m,1H),2.65-2.56(m,3H); 13 C NMR(100MHz,DMSO-d6)δ174.6,161.1(d,J=241.8Hz),140.6(d,J=2.9Hz),12 7.6(d,J=8.2Hz),115.2(d,J=21.2Hz),69.2,47.7,46.95,45.0,36.2,31.0. 19 F NMR (376MHz, DMSO-d6): δ115.3.

[0124] Example 22

[0125]

[0126] The experimental method used in this embodiment is basically the same as that in Example 20. The raw material used in this embodiment is (3,5-dimethyl-1H-pyrazol-1-yl)(4-hydroxy-2,2-dioxo-1-phenyl-2-thiabicyclo[3.1.1]heptane-5-yl) methyl ketone, and the product obtained is shown in structural formula (II-3c), with a yield of 91%. 1 H NMR (400MHz, DMSO-d6): δ7.26(d,J=7.2Hz,4H), 4.76(d,J=7.2Hz,1H), 4.14(dd,J=14. 4,7.2Hz,1H),3.37(d,J=14.4Hz,1H),3.10(dd,J=11.6,5.2Hz,1H),2.84-2.76(m,3H); 13 C NMR (100MHz, DMSO-d6): δ173.3, 162.3 (d, J = 243.7Hz), 130.4 (d, J = 8.5Hz), 1 29.8(d,J=3.0Hz),115.1(d,J=21.6Hz),66.4,64.8,57.2,47.3,37.0,33.5. 19 F NMR (376MHz, DMSO-d6): δ116.5.

[0127] Example 23

[0128]

[0129] The experimental method in this embodiment is basically the same as that in Example 20. The raw material used in this embodiment is (1-(2,4-difluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-yl)(3,5-dimethyl-1H-pyrazol-1-yl) methyl ketone, and the product obtained is shown in structural formula (II-3d), with a yield of 84%. 1 H NMR (400MHz, DMSO-d6): δ7.70-7.65(m,1H),7.39-7.01(m,2H),4.52(t,J=5.6Hz,1H), 3.34(dd,J=12.8,6.4Hz,1H),2.97-2.92(m,1H),2.87-2.70(m,3H),2.62-2.57(m,1H); 13 C NMR(100MHz,DMSO-d6)δ174.9(d,J=12.8Hz),160.7(dd,J=247.5,8.6Hz),156.6( dd,J=247.7,8.9Hz),131.7(dd,J=280.3,9.8Hz),120.0(dt,J=169.3,17.8Hz),1 13.3(dd,J=166.1,26.6Hz),104.3(dd,J=21.6,3.8Hz),69.3(d,J=6.3Hz),49.7( d, J=11.3Hz), 46.6 (d, J=26.8Hz), 42.6 (d, J=23.6Hz), 37.9 (d, J=14.6Hz), 31.2. 19 F NMR (376MHz, DMSO-d6): δ-106.3,-113.6.

[0130] Example 24

[0131]

[0132] Take a pre-dried 25 mL reaction tube, accurately weigh I-3a (32.8 mg, 0.1 mmol) into it under a nitrogen atmosphere, add 2 mL of MeOH solution, then add DBU (33.5 mg, 0.11 mmol), stir at room temperature for 16 h, and detect by TLC until the starting material reacts completely. After removing the solvent under reduced pressure, purify by silica gel column chromatography to obtain product III in 92% yield. 1H NMR (400MHz, CDCl3): δ7.36-7.32(m,2H),7.24(d,J=7.8Hz,3H),4.75(t,J=6.0Hz,1H),3.72(s,3H),3.39(dd,J =13.5,6.2Hz,1H),3.13–3.03(m,2H),2.99–2.89(m,2H),2.80(d,J=10.7Hz,1H),2.59(dd,J=10.3,7.6Hz,1H). 13 C NMR (100MHz, CDCl3): δ178.8,143.5,128.5,127.4,125.4,69.9,48.5,47.8,44.4,36.9,30.6.

[0133] Example 25

[0134]

[0135] Take a pre-dried 25 mL reaction tube, accurately weigh I-3a (32.8 mg, 0.1 mmol) into it under a nitrogen atmosphere, add 2 mL of THF solution, then add 0.1 mL (0.3 mmol) of 3 M MeMgBr in diethyl ether. Stir for 1 h at room temperature, then quench with saturated NH4Cl (3 mL). Extract the reaction solution with EtOAc (3 x 10 mL). Dry the combined organic layers on anhydrous MgSO4. Filter, evaporate the solvent under reduced pressure, and purify the mixture by silica gel column chromatography to give product IV in 92% yield. 1 HNMR (400MHz, CDCl3): δ7.35-7.31(m,2H),7.27-7.23(m,3H),4.78(s,1H),3.51(dd,J=13.2,4.4Hz,1H),2.91(d, J=13.6Hz,1H),2.79(dd,J=10.0,5.6Hz,1H),2.53(d,J=10.8Hz,1H),2.40-2.33(m,2H),1.25(s,3H),1.22(s,3H). 13 C NMR (100MHz, CDCl3): δ145.1,128.4,126.9,125.5,74.4,70.9,48.9,47.0,39.5,36.1,32.4,26.5,25.5.

[0136] Example 26

[0137]

[0138] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, at 0 °C, Dess-Martin oxidant (DMP; 0.3 mmol, 127.2 mg) was added to a solution of compound I-3o (62.9 mg, 0.2 mmol) in CH2Cl2 (4 mL), and the mixture was stirred for 5 hours. After the reaction was complete, Na2S2O3 (1 n aq, 10 mL) and a saturated NaHCO3 (10 mL) solution were added, and the mixture was stirred at room temperature for 0.5 hours. The organic layer was washed with brine, dried over anhydrous Mg2SO4, and concentrated under vacuum. The crude product was purified by silica gel column chromatography to give product V in 71% yield. 1 H NMR (400MHz, CDCl3): δ7.31-7.25(m,4H),7.22-7.19(m,1H),7.03(s,1H),6.92(s,1H),3.91(s,3H),3.88(s,2H),3.19(s,4H)ppm. 13 C NMR (100MHz, CDCl3): δ204.2,187.9,142.1,141.1,129.8,128.6,127.7,126.9,125.5,60.6,49.0,44.7,35.7,34.6.

[0139] Example 27

[0140]

[0141] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, Oxone oxidant (0.06 mmol, 34.63 mg, 60%) was added to a solution of compound I-3a (32.84 mg, 0.1 mmol) in EtOH (2 mL) at 0 °C. The mixture was refluxed at 90 °C for 12 hours. After the reaction was complete, ethyl acetate was added, and the mixture was extracted three times with water. The organic layer was washed with brine, dried over anhydrous Mg2SO4, and concentrated under vacuum. The crude product was purified by silica gel column chromatography to give product VI in 70% yield. 1H NMR (400MHz, CDCl3): δ7.39-7.35(m,2H),7.33-7.29(m,1H),7.20(d,J=7.2H z,2H),6.02(s,1H),4.92(t,J=3.2Hz,1H),3.82(d,J=14.0Hz,1H),3.25(dd, J=12.0,6.0Hz,1H),3.11(dd,J=14.0,5.6Hz,1H),2.94(dd,J=12.4,6.0Hz,1 H),2.87(d,J=12.4Hz,1H),2.78(d,J=12.8Hz,1H),2.53(s,3H),2.25(s,3H). 13 C NMR (100MHz, CDCl3): δ172.9,153.7,145.4,138.4,128.7,128.3,126.8,111.3,69.6,60.7,56.2,51.9,34.5,33.9,14.2,13.7.

[0142] Example 28

[0143]

[0144] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, at 0 °C, m-CPBA oxidant (0.3 mmol, 60.9 mg, 85%) was added to a solution of compound I-3a (32.8 mg, 0.1 mmol) in CH2Cl2 (2 mL), and the mixture was stirred for 4 hours. After the reaction was complete, the solution was directly concentrated under vacuum. The crude product was purified by silica gel column chromatography to give product VII in 87% yield. 1 HNMR (400MHz, CDCl3): δ7.40-7.37(m,3H),7.31-7.29(m,2H),6.03(s,1H),4.97-4.93(m,1H),3.97(dd,J= 14.4, 6.8Hz, 1H), 3.73-3.65 (m, 2H), 3.17-3.08 (m, 2H), 2.92 (d, J = 12.4Hz, 1H), 2.52 (s, 3H), 2.27 (s, 3H). 13 C NMR (100MHz, CDCl3): δ172.2,154.1,145.4,132.5,128.9,128.5,128.0,111.6,67.9,66.1,57.3,51.5,38.8,36.9,14.1,13.8.

[0145] Example 29

[0146]

[0147] Take a pre-dried 25 mL reaction tube and, under a nitrogen atmosphere at 0 °C, add m-CPBA oxidant (0.6 mmol, 121.8 mg, 85%) to a solution of compound I-3c (62.1 mg, 0.2 mmol) in CH2Cl2 (4 mL) and stir for 4 hours. Add Et3N (4 eq, 40.48 mg) and CH2Cl2 (2 mL) to the crude product. Place the apparatus in an ice-water bath and slowly add MsCl (2 eq, 22.9 mg, 16 μL). Remove the ice-water bath and react at room temperature for 12 hours. After the reaction is complete, quench with water, extract three times with ethyl acetate, combine the organic phases, dry over anhydrous Mg2SO4, and concentrate under vacuum. Purify the crude product by silica gel column chromatography to obtain product VIII in 71% yield. 1 HNMR (400MHz, CDCl3): δ7.84(d,J=7.2Hz,2H),7.64(t,J=6.8Hz,1H),7.52(t,J=7.2Hz,2H),7.42(d,J=5.6Hz,3H) ,7.36-7.34(m,2H),7.14(d,J=10.0Hz,1H),6.64(d,J=10.0Hz,1H),3.48(d,J=8.8Hz,2H),3.29(d,J=8.8Hz,2H). 13 C NMR (100MHz, CDCl3): δ196.8,142.7,134.1,129.2,129.2,129.1,128.5,128.3,126.4,66.9,48.5,46.3

[0148] Example 30

[0149]

[0150] Take a pre-dried 25 mL reaction tube, and under a nitrogen atmosphere, add pyridine (0.38 mmol, 30.0 mg) and DMAP (0.01 mmol, 2.5 mg) to a solution of compound I-3o (78.6 mg, 0.25 mmol) in CH2Cl2 (10 mL). Place the reaction tube in an ice-water bath, and slowly add acetic anhydride (0.5 mmol, 51.0 mg). React at room temperature for 12 h, and purify by column chromatography to obtain the intermediate.

[0151] Take a pre-dried 25 mL reaction tube, add Ylide reagent (0.3 mmol, 107.2 mg) and THF (2 mL) under a nitrogen atmosphere, place it at 0°C, add potassium tert-butoxide (0.3 mmol, 33.7 mg, 3.0 eq), stir at 0°C for 40 minutes, add the purified intermediate (35.6 mg, 0.1 mmol, 1.0 eq), stir at room temperature until the starting material reacts completely. After the reaction is complete, quench with water, extract three times with ethyl acetate, combine the organic phases, dry over anhydrous Mg2SO4, and concentrate under vacuum. The crude product is purified by silica gel column chromatography to obtain product IX, with a yield of 67%. 1 H NMR (400MHz, CDCl3): δ7.35-7.21(m,5H),7.05(s,1H),6.86(s,1H),5.63(s,1H),5.31(s,1H),5.18(s,1H),3.68(dd ,J=14.0,4.8Hz,1H),3.59(s,3H),3.10-3.07(m,2H),2.83(d,J=14.4Hz,1H),2.74(t,J=9.6Hz,2H),2.03(s,3H)ppm. 13 C NMR (100MHz, CDCl3): δ170.2,145.6,144.7,142.0,128.4,127.9,127.0,125.7,121.9,116.8,73.0,47.7,47.5,44.1,41.1,34.3,29.3,21.0

[0152] Example 31

[0153]

[0154] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, NaBH4 (0.2 mmol, 7.57 mg) was added three times to a solution of compound I-3a (32.84 mg, 0.1 mmol) in MeOH (2 mL). The mixture was stirred at 50 °C for 30 minutes. After the reaction was complete, the mixture was quenched with water, extracted three times with DCM, and the organic phases were combined, dried over anhydrous Mg2SO4, and concentrated under vacuum. The crude product was purified by silica gel column chromatography to give product X in 92% yield. 1H NMR (400MHz, Acetone-d6): δ7.27(m,5H),4.45(s,1H),4.34(s,1H),3.84(s,1H),3.57(s,2H) ,3.40-3.35(m,1H),2.72(m,1H),2.45(d,J=10.4Hz,1H),2.31(m,1H),2.20(d,J=10.4Hz,1H). 13 C NMR (100MHz, Acetone-d6): δ146.6,129.1,127.5,126.5,72.1,67.7,49.3,45.5,42.7,37.3,32.6.

[0155] Example 32

[0156]

[0157] Take a pre-dried 25 mL reaction tube, accurately weigh I-3a (65.6 mg, 0.2 mmol) into it under a nitrogen atmosphere, add 4 mL of MeOH solution, then add DBU (67.0 mg, 0.22 mmol), stir at room temperature for 16 h, and detect by TLC until the starting material reacts completely. After removing the solvent under reduced pressure, purify by silica gel column chromatography to obtain product III.

[0158] Take a pre-dried 25 mL reaction tube, accurately weigh III (26.4 mg, 0.1 mmol) into it under a nitrogen atmosphere, add 2 mL of dried DCM solution, then add DAST (24.2 mg, 0.15 mmol), stir at room temperature for 12 h, and detect by TLC until the starting material reacts completely. Quench with water, extract three times with DCM, combine the organic phases, dry with anhydrous Mg2SO4, and concentrate under vacuum. The crude product is purified by silica gel column chromatography to obtain product XI in 76% yield. 1 H NMR (400MHz, CDCl3): δ7.36-7.23(m,5H),5.70-5.50(m,1H),3.73(s,3H),3.54-3.44(m,1 H),3.29-3.19(m,1H),2.96-2.89(m,2H),2.84(dd,J=10.8,4.4Hz,1H),2.65-2.61(m,1H). 13C NMR (100MHz, CDCl3): δ172.3, 143.1, 128.6, 127.5, 125.4, 90.1 (d, J = 182.3Hz), 52.3 ,48.5,47.2(d,J=21.5Hz),43.6(d,J=8.0Hz),37.2(d,J=6.9Hz),28.8(d,J=24.7Hz). 19 F NMR (376MHz, CDCl3): δ-172.0.

[0159] Example 33

[0160]

[0161] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, at 0 °C, martin sulfurane (154.7 mg, 0.23 mmol, 2.3 eq) was added to a solution of compound I-3c (31.0 mg, 0.1 mmol) in CH2Cl2 (4 mL). The mixture was stirred at room temperature for 24 hours. The reaction was monitored for completeness by TLC. After completion, the mixture was concentrated under vacuum, and the crude product was purified by silica gel column chromatography to obtain product XII in 54% yield. 1 H NMR (400MHz, CDCl3): δ7.85 (d, J = 7.2Hz, 2H), 7.50-7.47 (m, 1H), 7.40-7.36 (m, 2H), 7.31-7.18 (m ,5H),6.44(d,J=8.8Hz,1H),6.22(d,J=8.4Hz,1H),2.86(d,J=6.8Hz,2H),2.53(d,J=6.8Hz,2H). 13 CNMR (100MHz, CDCl3): δ200.1,142.8,134.0,133.3,129.0,128.8,128.7,127.7,126.8,125.6,122.8,50.7,48.8,38.9.

[0162] Example 34

[0163]

[0164] A pre-dried 25 mL reaction tube was used. Under a nitrogen atmosphere, at room temperature, LiF (15.6 mg, 6 eq) and trimethylsilyl reagent (71.2 mg, 2.4 eq) were added to a solution of compound VIII (32.4 mg, 0.1 mmol) in MeCN (2 mL). The mixture was stirred at 80°C for 12 hours. After the reaction was complete, ethyl acetate (10 mL) was added, and the organic phase was extracted three times with K₂CO₃. The organic phase was dried over anhydrous Mg₂SO₄ and concentrated under vacuum. The crude product was purified by silica gel column chromatography to give product XIII in 87% yield. 1 H NMR (400MHz, CDCl3): δ7.73(d,J=7.6Hz,2H),7.52-7.48(m,1H),7.40-7.36(m,2H),7.32-7.18(m,10H),4.14-4.07( m,1H),3.65-3.52(m,3H),3.34(dd,J=11.6,6.0Hz,1H),3.28-3.18(m,2H),3.03-2.95(m,3H),2.43(d,J=6.4Hz,2H). 13 C NMR (100MHz, CDCl3): δ200.1,137.7,133.8,133.6,132.7,128.9,128.8,128.6,1 28.5,128.4,128.3,127.3,64.7,62.2,59.2,55.9,55.3,50.4,45.5,41.2,34.9.

[0165] Example 35

[0166]

[0167] Take a pre-dried 25 mL reaction tube, and under a nitrogen atmosphere at room temperature, add I-3O (31.4 mg, 1.0 eq), 4-(N,N-dipropylaminosulfonyl)benzoic acid (34.2 mg, 1.2 eq), DCC (31.0 mg, 1.5 eq), and DMAP (1.83 mg, 15 mol%). Add 2 mL of CH2Cl2 solution, stir at room temperature for 12 h, and after the reaction is complete, wash with 1N HCl (3 x 10 mL), saturated NaHCO3 (10 mL), and brine (10 mL), respectively. Dry the organic layer with Mg2SO4, filter, concentrate, and purify by flash chromatography on silica gel to obtain product XIV in 87% yield. 1H NMR (400MHz, CDCl3): δ8.04(d,J=8.4Hz,2H),7.84(d,J=8.0Hz,2H),7.36-7.30(m,4H),7. 27-7.23(m,1H),7.14(s,1H),6.96(s,1H),6.51(t,J=5.2Hz,1H),3.90-3.85(m,1H),3.89 (s,3H),3.30(dd,J=10.8,7.2Hz,1H),3.19(dd,J=13.6,4.8Hz,1H),3.09(t,J=7.6Hz,5H) ,2.99(d,J=11.2Hz,1H),2.91(d,J=10.8Hz,1H),1.59-1.50(m,4H),0.87(t,J=7.2Hz,6H). 13 C NMR (100MHz, CDCl3): δ190.9,164.1,144.4,143.7,141.1,133.0,130.2,129.4,128.5, 127.2,126.9,126.7,125.6,75.3,52.6,49.9,48.0,46.2,36.9,35.9,28.6,21.9,11.1.

[0168] Example 361 Synthesis of 1-(2,4-difluorophenyl)-N-(2,5-difluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxamide (XV-3a)

[0169]

[0170] Take a pre-dried 25 mL reaction tube, accurately weigh difluoroaniline (258.2 mg, 10 eq) into it under a nitrogen atmosphere, add THF solution (4 mL), slowly add n-BuLi (1.25 mL, 1.6 mol / L, 10 eq), place the apparatus at -30°C, add a toluene solution of III-1a (60.1 mg, 0.2 mmol), stir at -30°C for 30 min, monitor the reaction progress by TLC, after the reaction is complete, quench with water, add ethyl acetate to the mixture, wash once with HCl, NaHCO3, and water respectively, dry the organic phase with anhydrous magnesium sulfate, filter, concentrate under reduced pressure to obtain compound XV-3a, with a yield of 81%. 1H NMR(400MHz, Acetone-d6)δ9.32(s,1H),8.17-8.11(m,1H),7.66-7.32(m,1H),7.23-7.16(m,1H),7.04-6.96(m,2H),6.86-6.80(m, 1H),5.67-5.63(m,1H),4.87(s,1H),3.78-3.71(m,1H),3.32-3.27(m,1H),3.23-3.17(m,1H),3.03-2.97(m,1H),2.83-2.72(m,2H). 13 C NMR (100MHz, Acetone-d6) δ172.9(d,J=16.6Hz), 160.9(dd,J=249.2,7.9Hz), 160.5(d,J=1.6Hz), 158.1(d,J=2.0 Hz), 150.8 (d, J = 2.7Hz), 133.6 (d, J = 11.1Hz), 130.6 (t, J = 10.4Hz), 121.0 (dt, J = 167.1, 18.7Hz), 116.4 (q, J = 10.1 Hz), 114.3 (dd, J=23.6, 3.9Hz), 110.4 (dd, J=24.7, 13.4Hz), 109.2 (d, J=30.0Hz), 104.8 (dd, J=21.2, 4.1Hz), 69. 5(d,J=9.1Hz), 51.1(d,J=9.8Hz), 43.7(d,J=23.5Hz), 42.1(d,J=18.2Hz), 41.1(d,J=13.0Hz), 32.5(d,J=1.4Hz). 19 F NMR (376MHz, Acetone-d6): δ-106.6,-114.5,-118.2,-136.7.

[0171] Example 37

[0172]

[0173] The experimental method in this embodiment is basically the same as that in Embodiment 36 of the present invention. The raw material used in this embodiment is methyl 1-(4-fluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxylate, and the product obtained is shown in structural formula (XV-3b), with a yield of 85%. 1H NMR(400MHz, Acetone-d6)δ9.38(s,1H),8.18-8.13(m,1H),7.37-7.33(m,2H),7.24-7.18(m,1H),7.14-7.09(m,2H),6.86-6.81(m,1H), 5.67(s,1H),4.87(s,1H),3.70(dd,J=14.0,5.2Hz,1H),3.17-3.12(m,1H),3.01(d,J=11.6Hz,1H),2.67-2.63(m,1H),2.57(d,J=10.0lll Hz,1H). 13 C NMR (100MHz, Acetone-d6) δ173.2, 162.6 (d, J = 242.4Hz), 160.5 (d, J = 11.6Hz), 158. 1(d,J=1.7Hz),150.7(d,J=2.5Hz),148.4(d,J=2.7Hz),141.9(d,J=3.0Hz),129.1(t ,J=12.3Hz),128.4(d,J=8.3Hz),116.4(q,J=10.0Hz),116.0(d,J=21.5Hz),110.3( dd,J=24.5,7.9Hz),109.1(d,J=30.1Hz),69.78,49.53,48.23,40.73,39.77,32.83. 19 F NMR (376MHz, Acetone-d6): δ-116.8,-118.2,-136.8.

[0174] Example 38

[0175]

[0176] The experimental method in this embodiment is basically the same as that in Embodiment 36 of the present invention. The raw material used in this embodiment is methyl 1-(2,4-difluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxylate 2,2-dioxide, and the product obtained is shown in the structural formula (XV-3c), with a yield of 80%. 1HNMR(400MHz, Acetone-d6)δ9.31(s,1H),8.14-8.08(m,1H),7.79-7.46(m,1H),7.25-7.18(m,1H),7.11-7.03(m, 1H),6.91-6.84(m,1H),5.12(s,1H),4.24-4.16(m,1H),3.72-3.61(m,2H),3.50-3.42(m,1H),3.24-3.00(m,3H). 13 C NMR (100MHz, Acetone-d6) δ171.4(d,J=14.5Hz), 162.6(dd,J=251.6,7.1Hz), 159.0(dd,J=250.7,8.1Hz), 160.4(d ,J=11.9Hz),158.0(d,J=1.5Hz),148.6(d,J=2.9Hz),135.4(d,J=9.9Hz),132.5(t,J=10.6Hz),116.5(dd,J=22.0, 9.9Hz), 114.2 (dd, J=23.5, 3.7Hz), 112.7 (dd, J=22.1, 3.1Hz), 109.5 (d, J=29.9Hz), 104.9 (dd, J=21.8, 4.1Hz), 67 .2(d,J=5.3Hz), 63.4(d,J=18.8Hz), 58.3(d,J=2.7Hz), 50.9(d,J=5.8Hz), 37.8(d,J=14.8Hz), 37.2(d,J=8.1Hz). 19 F NMR (376MHz, Acetone-d6): δ-103.9,-112.1,-118.1,-136.1.

[0177] Example 39

[0178]

[0179] The experimental method in this embodiment is basically the same as that in Embodiment 36 of the present invention. The raw material used in this embodiment is methyl 1-(4-fluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxylate 2,2-dioxide, and the product obtained is shown in the structural formula (XV-3d), with a yield of 74%. 1HNMR(400MHz, Acetone-d6)δ9.35(s,1H),8.13-8.09(m,1H),7.38-7.34(m,2H),7.35-7.16(m,J=8.6Hz,3H),6.89-6.84(m,1H),6 .15(s,1H),5.12(m,1H),4.14(dd,J=14.8,6.8Hz,1H),3.60-3.55(m,2H),3.25(d,J=12.4Hz,1H),2.92-2.87(m,1H),2.08(s,1H). 13 C NMR (100MHz, Acetone-d6) δ171.6, 163.8 (d, J = 244.2Hz), 160.4 (d, J = 2.0Hz), 158.1 (d,J=1.4Hz),150.9(d,J=2.4Hz),148.5(d,J=2.5Hz),131.2(d,J=8.5Hz),130.9(d ,J=3.1Hz),128.6(t,J=12.4Hz),116.5(q,J=10.0Hz),115.9(d,J=21.8Hz),110.9( dd,J=24.6,8.1Hz),109.5(d,J=30.1Hz),67.56,66.27,58.43,49.27,36.64,35.71. 19 F NMR (376MHz, Acetone-d6): δ-114.7,-118.1,-136.3.

[0180] Example 40

[0181]

[0182] The experimental method in this embodiment is basically the same as that in Embodiment 36 of the present invention. The raw material used in this embodiment is methyl 1-(4-fluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxylate 2-oxide, and the product obtained is shown in the structural formula (XV-3e), with a yield of 63% (dr = 1.6:1). 1H NMR(600MHz,DMSO-d6)δ9.62(s,1H),7.79-7.76(m,1H),7.33-7.29(m,1H),7.24-7.19(m,4H),7.00-6.97(m,1H),6.22(s,1H),4.86-4.84(m,1 H),3.65(dd,J=14.4,3.0Hz,1H),3.05(dd,J=14.4,6.0Hz,1H),2.90(dd,J=11.4,6.0Hz,1H),2.67-2.64(m,2H),2.55(dd,J=12.0,6.0Hz,1H). 13 C NMR(151MHz,DMSO-d6)δ171.5,161.8(d,J=242.9Hz),158.4(d,J=1.5Hz),156.8 (d,J=1.7Hz),150.5(d,J=2.3Hz),148.9(d,J=2.4Hz),135.9(d,J=2.7Hz),129.0 (d,J=8.3Hz),127.2(t,J=12.0Hz),116.4(q,J=9.8Hz),115.3(d,J=21.5Hz),111 .1(dd,J=24.3,8.1Hz),110.4(d,J=29.7Hz),68.1,59.7,56.0,48.8,32.1,31.6. 19 FNMR (565MHz, DMSO-d6): δ-114.3,-117.3,-131.2.

[0183] Example 41

[0184]

[0185] The experimental method in this embodiment is basically the same as that in Embodiment 36 of the present invention. The raw material used in this embodiment is methyl 1-(4-fluorophenyl)-4-hydroxy-2-thiabicyclo[3.1.1]heptane-5-carboxylate 2-oxide, and the product obtained is shown in structural formula (XV-3f), with a yield of 63% (dr = 1.6:1). 1H NMR(400MHz, Acetone-d6)δ9.52(s,1H),8.17-8.12(m,1H),7.30-7.14(m,5 H),6.88-6.83(m,1H),6.02(d,J=4.0Hz,1H),4.83(s,1H),4.01(dd,J=14.8 ,6.8Hz,1H),3.54(dd,J=11.6,6.4Hz,1H),3.10(d,J=12.8Hz,1H),2.87(dd ,J=15.2,3.2Hz,1H),2.48(d,J=11.6Hz,1H),2.31(dd,J=12.4,6.0Hz,1H). 13 CNMR(100MHz,Acetone-d6)δ172.3,163.3(d,J=243.5Hz),160.5(d,J=2.1Hz),158.1(d ,J=2.2Hz),150.9(d,J=2.7Hz),148.5(d,J=2.9Hz),137.7(d,J=3.1Hz),129.6(d,J=8. 3Hz),128.8(t,J=12.5Hz),116.5(d,J=9.9Hz),116.4(d,J=2.2Hz),116.1(d,J=21.7Hz ), 110.6 (dd, J = 24.6, 7.9Hz), 109.4 (d, J = 30.0Hz), 69.6, 61.0, 54.2, 49.6, 33.8, 28.8. 19 F NMR (376MHz, Acetone-d6): δ-115.7,-118.2,-136.3.

[0186] Example 42: Drug metabolism kinetic test methods and results

[0187]

[0188] Table 2

[0189]

[0190] Notes: Pharmacokinetic properties of the drug Diflunisal and the 4-hydroxy-2-thiabicyclo[3.1.1]heptane II-3d, II-3b, II-3c and I-3o prepared in this invention. Sol (solubility, μg / mL): Experimental kinetic solubility in phosphate buffered saline at pH 7.4. logD (lipophilicity, pH = 7.4): Experimental distribution coefficient in n-octanol / phosphate buffered saline at pH 7.4. CL int Experimental metabolic stability in human liver microsomes (μLmin)-1 mg -1 ). T 1 / 2 (min): Experimental half-life of metabolic breakdown. Due to the high stability of the compound, this parameter should be considered an approximation.

[0191] The drug diflunisal is suitable for rheumatoid arthritis, osteoarthritis, and various mild to moderate pain.

[0192] This invention utilizes the principle of bioisosteric substitution to synthesize compounds II-3d, II-3b, II-3c, and I-3o, and tests their pharmacokinetics. It was found that they can improve water solubility and maintain lipid solubility. Therefore, it is inferred that compounds II-3d, II-3b, II-3c, and I-3o have similar biological functions to the drug diflunisal and can be used to treat rheumatoid arthritis, osteoarthritis, and various mild to moderate pain.

[0193] After preparing II-3d, II-3b, II-3c and I-3o using the method of this invention, the water solubility, lipophilicity and metabolic stability of these drugs were investigated and compared with those of the drug diflunisal.

[0194] Analysis results:

[0195] 1. Solubility: The solubility of diflunisal analogs I-3o, II-3b, II-3c, and II-3d synthesized by isostatic substitution of the benzene ring structure in diflunisal with the 2-thiabicyclo[3.1.1]heptane structural unit was improved. The solubility trend, from smallest to largest, is as follows: 45.8 (diflunisal) < 49.4 (II-3b) < 52.6 (II-3d) < 54.4 (I-3o) < 60.3 (II-3c).

[0196] 2. logD (lipophilicity, pH = 7.4): The lipophilicity of diflunisal analogs II-3b, II-3c, and II-3d, synthesized by isochorically replacing the benzene ring structure in diflunisal with the 2-thiabicyclic [3.1.1]heptane structural unit, was reduced. Compared with diflunisal, analog I-3o had a lipophilicity (logD: 2.53) that was 2.8 times that of diflunisal (logD: 0.89).

[0197] 3. Liver microsomal enzyme stability and half-life: The metabolic stability of diflunisal analogs II-3b, II-3c, and II-3d synthesized by isochorically replacing the benzene ring structure in diflunisal with a 2-thiabicyclo[3.1.1]heptane structural unit was increased to some extent compared with diflunisal (CL). int The values ​​are all less than diflunisal; T 1 / 2All greater than 145 minutes). However, the metabolic stability of compound I-3o (CL) int =60.5μLmin -1 mg -1 ;T 1 / 2 =22.9min) is lower than that of diflunisal.

[0198] In summary, replacing the corresponding benzene ring in the active drug molecule with 2-thiabicyclo[3.1.1]heptane or 2-sulfonylbicyclo[3.1.1]heptane using the bioisosteric principle improves the water solubility of the drug and promotes the reduction of lipophilicity and the enhancement of metabolic stability.

[0199] Example 43: Drug Activity Testing Methods and Results

[0200]

[0201] Table 3

[0202]

[0203] Notes: %inhibition of MNCs: percentage of osteoclasts inhibited; %cell viability: cell survival rate. The 5-(2',4'-difluorophenyl)-salicylic acid derivative XVI is a known osteoclastogenesis inhibitor. Its cell viability (%cell viability: 102.46±1.10) and percentage of osteoclasts inhibited (%inhibition of MNCs: 67.32±4.38) data indicate that this compound has low cytotoxicity and significant inhibitory activity against osteoclast growth. Using the principle of bioisosteric electrons, the analogues XV-3a, XV-3b, XV-3c, XV-3d, XV-3e, and XV-3f synthesized in this invention were analyzed using tartrate-resistant acid phosphatase (TRAP) staining to detect the effects of these compounds on osteoclast growth induced by the NF-κB ligand receptor activator (RANKL) in RAW264.7 cells.

[0204] Analysis results:

[0205] like Figure 1As shown in Table 3, compounds XV-3a, XV-3c, and XV-3d effectively inhibited osteoclast differentiation at a concentration of 5 μM, with inhibition rates (% inhibition of MNCs) exceeding 40%, specifically 42.19 ± 5.98%, 46.18 ± 15.57%, and 68.77 ± 2.08%, respectively. However, compound XV-3d exhibited strong cytotoxicity against RAW264.7 cells, with a cell viability (% cellviability: 54.56 ± 8.02). Compounds XV-3b, XV-3e, and XV-3f showed poor inhibitory activity against osteoclast differentiation, but also exhibited low cytotoxicity.

[0206] In summary, the compounds XV-3a and XV-3c synthesized in this invention exhibit significant inhibitory activity against osteoclast growth. Although their inhibition rate is lower than that of the known osteoclastogenesis inhibitor compound XVI, this example demonstrates the potential application value of the polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane compounds synthesized in this invention in the design and synthesis of drug molecules.

[0207] Solubility test (Sol) method

[0208] 1. Materials

[0209] A: Prepare a 50mm phosphate buffer solution with a pH of 7.4.

[0210] B: Prepare 50ml Na2HPO4: Dissolve 3.549g of Na2HPO4 in 500mL of water. 4， The pH is 9.4

[0211] C: Prepare 50ml NaH2PO4: Dissolve 3.000g of NaH2PO4 in 500mL of water. 4， pH is 4.5

[0212] D: Prepare a 50 mL phosphate buffer solution with a pH of 7.4: Add 15 mL of 50 mL Na₂HPO₄ to a 50 mL centrifuge tube and adjust the pH to 7.4 using the 50 mL NaH₂PO₄ solution.

[0213] 2. Equipment

[0214] Millipore Milli-Q Gradient Water Purification System

[0215] Matrix Multichannel Electronic Pipettes

[0216] SpectraMax Plus microplate scanner

[0217] perforated plate

[0218] vacuum pump

[0219] 3. Method:

[0220] A: Add 10 μL of the DMSO stock solution of the test compound and the control compound to each well of the 96-well plate.

[0221] B: Add 490 μL of culture medium to each well of the 96-well plate.

[0222] C: Vortex dissolve the sample for at least 2 minutes.

[0223] D: Place the 96-well plate at room temperature and shake it at 800 rpm for 24 hours.

[0224] E: Centrifuge at 25°C for 10 minutes.

[0225] F: Transfer the supernatant to a filter plate, then collect the filtrate into a new 96-well plate and centrifuge for at least 5 minutes.

[0226] G: The concentration of the filtrate was quantified using an LC-UV system.

[0227] Test method for lipid-soluble Log D

[0228] 1. Materials

[0229] 1.1 Prepare 100 mM phosphate buffer (PB) with pH 7.4.

[0230] A: Preparation of 100mM NaH2PO4: Dissolve 5.999g of NaH2PO4 in 500mL of water, and the pH value is measured to be approximately 4.5.

[0231] B: Preparation of 100 mmol Na2HPO4: Dissolve 7.098 g of Na2HPO4 in 500 mL of water and measure the pH value to be approximately 9.4.

[0232] C: Preparation of 100mM PB (pH 7.4): Add 15mL of 100mM Na2HPO4 to a 50mL test tube, and then adjust the pH to 7.4±0.05 with 100mM NaH2PO4.

[0233] 1.2 Saturate PB with 1-octanol (pH 7.4): Add 10 mL of 1-octanol to 100 mL of 100 mM PB (pH 7.4), shake the mixture vigorously and let it stand overnight at room temperature before use.

[0234] 1.31-Octanol-saturated PB (pH 7.4): Add 10 mL of 100 mM PB (pH 7.4) to 100 mL of 1-octanol, shake the mixture vigorously, and let it stand overnight at room temperature before use.

[0235] 2. Equipment

[0236] Gradient high performance liquid chromatography system

[0237] API 3000 triple quadrupole mass detector equipped with TurboIonSpray ion source

[0238] VWR membrane nitrogen generator, nitrogen purity 99%+

[0239] Multifunctional Hybrid Rotary Instrument

[0240] laboratory centrifuge

[0241] Water purification system

[0242] Multichannel electronic pipettes: 0.5-12.5μL, 2-125μL, 5-250μL, 15-1250μL

[0243] 3. Method:

[0244] A: Take 2 μL of 10 mM DMSO stock solution and put it into test tubes for the test compound and the control compound, in duplicate.

[0245] B: Add 149 μL of 1-octanol-saturated PB (pH 7.4) to each test tube.

[0246] C: Add 149 μL of PB (pH 7.4) saturated with 1-octanol to the corresponding test tubes.

[0247] D: Mix each test tube vigorously for 2 minutes, then shake at 800 rpm for 1 hour at room temperature.

[0248] E: Then centrifuge at 4000 rpm for 5 minutes at room temperature.

[0249] F: Take an appropriate amount of buffer layer sample and 1-octanol layer sample.

[0250] G: These samples were diluted and then detected using the LC-MS / MS method without the need to run a calibration curve.

[0251] H: Data Analysis: The logarithm D value for each compound is calculated using the following formula:

[0252]

[0253] Human liver microsomal metabolic stability (CL) int ) and half-life (T 1 / 2 Test methods

[0254] method:

[0255] 1. Preparation of test compounds and control working solutions

[0256] 1.1 Working solution: Dilute 5 μL of the compound and the reference stock solution (10 mM, dimethyl sulfoxide (DMSO)) with 495 μL of acetonitrile (CAN) (intermediate solution concentration: 100 μM, 99% CAN).

[0257] 2. Preparation of NADPH cofactor

[0258] 2.1 Materials

[0259] NADPH powder: β-nicotinamide adenine dinucleotide phosphate reduced form, tetrasodium salt; NADPH-4Na;

[0260] 2.2 Preparation Procedure

[0261] Weigh an appropriate amount of NADPH powder and dilute it into a 10 mM MgCl2 solution (working concentration of NADPH: 10 mM; final concentration in the reaction system: 1 mM).

[0262] 3. Preparation of liver microsomes

[0263] 3.1 Materials

[0264] Table 4.1: Liver microsomal information

[0265]

[0266] 3.2 Preparation

[0267] Prepare a microsomal working solution of appropriate concentration in 100 mM potassium dihydrogen phosphate buffer.

[0268] 4. Stop the preparation of the solution.

[0269] Cold (4°C) acetonitrile (CAN) containing 250 nM thiobutazone and 250 nM labetalol internal standard (IS) was used as the stop solution.

[0270] 1. Experimental steps:

[0271] A: Preheat empty “culture” plates T60 and NCF60 for about 10 minutes.

[0272] B: Dilute liver microsomes to 0.56 mg / mL in 100 mM phosphate buffer.

[0273] C: Transfer 445 μL of the microsomal working solution (0.56 mg / mL) to preheated "Incubation" plates T60 and NCF60, and then incubate the "Incubation" plates T60 and NCF60 with continuous shaking at 37°C for 10 minutes. Transfer 54 μL of liver microsomes to a blank plate, then add 6 μL of NADPH cofactor to the blank plate, followed by 180 μL of quenching solution.

[0274] D: Add 5 μL of the compound working solution (100 μM) to the “incubation” plates (T60 and NCF6) containing microparticles and mix thoroughly 3 times.

[0275] E: For NCF60 plates, add 50 μL of buffer and mix thoroughly three times. Start timing; incubate at 37°C with shaking for 60 minutes.

[0276] F: In the "quenching" plate T0, add 180 μL of quenching solution and 6 μL of NADPH cofactor, and ensure the plate is cooled to prevent evaporation.

[0277] G: For T60 plates, mix thoroughly three times and immediately transfer 54 μL of the mixture at time 0 to a "quench" plate. Then add 44 μL of NADPH cofactor to the incubation plate (T60). Start timing; incubate at 37°C with shaking for 60 minutes.

[0278] Table 4.2: Final concentrations of each component in the culture medium

[0279]

[0280] At H: 5, 15, 30, 45 and 60 minutes, 180 μL of quenching solution was added to the "quenching" plate, mixed once, and then 60 μL of sample from each well in the T60 plate was continuously transferred to the "quenching" plate.

[0281] Table 4.3: Reaction Plate Culture

[0282]

[0283] I: For NCF60: Mix once, and then transfer 60 μL of the NCF60 culture medium sample to a "quench" plate containing quench solution at the 60-minute time point.

[0284] Table 4.4: NCF60 cultivation

[0285]

[0286] J: Shake all the sample plates for 10 minutes, then centrifuge at 4000 rpm for 20 minutes at 4°C.

[0287] K: Transfer 60 μL of supernatant to 180 μL of HPLC water and mix with a plate shaker for 10 minutes.

[0288] L: Before performing LC-MS / MS analysis, seal each bioanalytical plate and shake for 10 minutes.

[0289] 5. Calculate T using the first-order dynamic equation. 1 / 2 and C Lint (mic)(µL / min / mg):

[0290]

[0291] Assay for drug inhibition of RANKL-induced osteoclast differentiation

[0292] A: Tartrate-resistant phosphatase (TRAP) staining test

[0293] RAW264.7 cells were seeded at a density of 1 × 10³ cells per well in 96-well plates. Each well was filled with α-MEM (α-MEM, Gibco, USA) medium containing 10% FBS, 2 mM L-glutamate, and 50 ng / mL recombinant soluble mouse RANKL (Peprotech, UK), with or without the added compounds. Cells were cultured for 5 days, with the medium changed every other day. After 5 days, TRAP activity was detected using a TRAP / ALP staining kit (Guangzhou Heguang, China) according to the manufacturer's instructions.

[0294] B: Cell viability assay

[0295] Cell counting kit-8 (Selleck, USA) was used to determine cell viability. RAW264.7 cells were seeded into 96-well plates at a density of 1 × 10⁴ cells per well and treated with a series of compounds for 24 hours. The optical density (OD) at 450 nm was measured using a microplate reader (FLUOstar Omega-ACU, USA).

[0296] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention.

[0297] As used in this invention, the terms "comprising" and "including" are open-ended expressions, meaning they include the contents specified in this invention but do not exclude other aspects.

[0298] As used in this invention, the term "and / or" includes any one or more of the related listed items and all combinations thereof.

[0299] The scope of protection of this invention is not limited to the above embodiments. Any variations and advantages that can be conceived by those skilled in the art without departing from the spirit and scope of the inventive concept are included in this invention and are protected by the appended claims.

Claims

1. A polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane, characterized in that, Its structure is shown in equations I and II below: R 1 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, alkoxy, substituted amino, C1-C 12 Alkyl or C3-C 12 cycloalkyl, alkenyl, ynyl Where R 4 Arbitrarily selected from alkyl, cycloalkyl, aryl, and heteroaryl groups; R 5 Arbitrarily selected from hydrogen, alkyl, cycloalkyl, aryl, heteroaryl, and halogen; R 2 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, C1-C 12 Alkyl or C3-C 12 Cycloalkyl, alkenyl, alkynyl, silyl, and boron-containing groups; R 3 Arbitrarily selected from hydrogen, aryl, heteroaryl, substituted aryl, C1-C 12 Alkyl or C3-C 12 Cycloalkyl, alkenyl, alkynyl, silyl, and boron-containing groups.

2. The polysubstituted 4-hydroxy-2-thiabicyclo[] according to claim 1 3.1.1] Heptane, characterized in that, R 1 Any substituted phenyl, heteroaryl, C1-C 12 alkyl, Where R 4 Arbitrarily selected from methyl, ethyl, isopropyl, n-propyl, cyclopentyl; R 5 Arbitrarily selected from hydrogen and methyl; R 2 Arbitrarily selected from hydrogen, heteroaryl, substituted phenyl, C1-C 12 Alkyl, alkenyl, and ynyl groups; R 3 It can be arbitrarily selected from hydrogen, phenyl, heteroaryl, and boron-containing groups.

3. A method for preparing multi-substituted 4-hydroxy-2-thiabicyclo[ 3.1.1] The method using heptane, characterized in that, Includes the following steps, Using 2,5-dihydroxy-1,4-dithiane 1 and polysubstituted bicyclic [1.1.0]butane 2 as raw materials, the cyclized product shown in Formula I was obtained by stirring in the presence of a Lewis acid catalyst and a first organic solvent; then, in the presence of a second solvent and a base, the cyclized product shown in Formula I was subjected to R removal. 1 The 4-hydroxy-5-carboxy-2-thiabicyclic [3.1.1]heptane derivative of formula II was obtained; the reaction route is as follows: Among them, R 1 R 2 R 3 The definition is as described in formulas I and II of claim 1.

4. The method according to claim 3, characterized in that, The ratio of the amount of the raw material polysubstituted bicyclo[1.1.0]butane 2 to 2,5-dihydroxy-1,4-dithiaane 1 is 1:0.5-5:

1.

5. The method according to claim 3, characterized in that, The first organic solvent is one or more of dichloromethane, 1,2-dichloroethane, chloroform, toluene, tetrahydrofuran, ethyl acetate, 1,4-dioxane, and acetonitrile; and / or, the second organic solvent is one or two of tetrahydrofuran and water.

6. The method according to claim 3, characterized in that, The catalyst is one or more of scandium trifluoromethanesulfonate, nickel trifluoromethanesulfonate, gallium trifluoromethanesulfonate, europium trifluoromethanesulfonate, zinc trifluoromethanesulfonate, boron trifluoride ether, silver trifluoromethanesulfonate, and nickel perchlorate; and / or the base is selected from one or more of LiOH, DBU, and NaOH.

7. The method according to claim 3, characterized in that, The stirring temperature is 20-60℃; and / or the stirring time is 10-20 hours; and / or the R removal... 1 The temperature is 20-50℃; and / or, the removal of R 1 The time is 10-20 hours.

8. The method according to claim 3, characterized in that, Based on the amount of compound 1, the amount of catalyst is 1 mol% to 10 mol%; and / or, based on the amount of compound I, the amount of base is 1 to 4 equivalents.

9. The use of the polysubstituted 4-hydroxy-2-thiabicyclo[3.1.1]heptane according to claim 1 or 2 in the preparation of bioisosteres of pharmaceutical active molecules containing 1,2-disubstituted aromatics, 1,3-disubstituted aromatics and 1,2,4-trisubstituted aromatics.

10. The polysubstituted 4-hydroxy-2-thiabicyclo[] according to claim 1 or 2 3.1.1] Use of heptane, or the bioelectron isosteres according to claim 9, in the preparation of medicaments for treating osteocyte-related diseases, overactive bladder with urinary incontinence, urgency and frequency, chronic obstructive pulmonary disease and asthma, transthyretin amyloidosis (ATTR), and mild to moderate pain, and for anticancer purposes.

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