Synthesis method and application of pinenyl thiazolidone derivative

Abstract

The invention relates to the technical field of biological medicine, in particular to a synthesis method and application of pinenyl thiazolidone derivatives, and the synthesis method comprises the step of synthesizing thiosemicarbazide A at the yield of 90% under the promotion of hydrochloric acid through condensation reaction between myrtenal and 4-methyl-3-thiosemicarbazide. Cyclizing the compound A and chloroacetic acid to obtain a key intermediate thiazolidone B; then, the intermediate reacts with substituted benzaldehyde, and the target compound C1-C13 is obtained. The adopted substrate myrtenal has a better anti-proliferation effect on malignant cells, strong water solubility is enhanced by introducing thiazolidines, and remarkably enhanced anti-cancer activity can be shown by introducing the thiazolidine derivative modified by Schiff base. The series of products are novel in structure, high in yield, simple in separation and purification operation and suitable for mass production, and part of products such as C5 have good biological activity.

Description

Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to a method for synthesizing and applying a pinene-based thiazolidinone derivative. Background Art

[0002] It is well known that medicinal plants are a valuable source of potential anticancer compounds, and essential oils containing a pinane core have various biological activities, including the treatment of cancer, arthritis, and pain. Oxygenated derivatives of α-pinene are natural product components with a pinane core and have various pharmacological properties such as antioxidant, antibacterial, antifungal, antidiabetic, anxiolytic, and gastroprotective activities, which are mainly derived from the separation and extraction or semi-synthesis of essential oils from medicinal plants. According to the "Chinese Herbal Medicine Atlas", Myrtus communis has the effects of nourishing blood and stopping bleeding, astringing the intestine and strengthening menstruation, and can be used to treat symptoms such as weakness due to blood deficiency, hematemesis, epistaxis, hemoptysis due to overwork, hematochezia, spermatorrhea, scalds, and traumatic bleeding.

[0003] In medicinal chemistry, nitrogen-containing heterocycles are considered privileged motifs due to their extensive applications in medicinal chemistry. Thiazolidinone derivatives, as heterocyclic compounds, play a crucial role in various bioactive natural products. Thiazolidine compounds have functional fluids such as C=O, -NH-, and -S-, and can interact with biomolecules at multiple sites through non-covalent bonds such as p-p stacking and hydrogen bonding, thereby enhancing their water solubility. Several studies have shown that the thiazolidine fragment, combining an electron-accepting group and electron-donating (-NH- and -S-) groups, can easily interact with enzymes and other receptors, including protein tyrosine phosphatase 1B, monoamine oxidase, tyrosine phosphatase SHP-2, α-glucosidase, and cyclooxygenase-2, through weak bonds such as p-p stacking, hydrogen bonding, hydrophobic interaction, and van der Waals forces. Recently, some studies have reported that the thiazolthione skeleton has inhibitory activity against CDK2. Therefore, the incorporation of the thiazolidinone moiety can significantly improve the binding affinity with biomolecules on various targets. Schiff base is a classical pharmacophore, which usually helps to enhance the properties of the whole molecule. Therefore, thiazolidine derivatives modified with Schiff base may exhibit significantly enhanced anticancer activity.

[0004] Considering the above factors, through calculations such as CADD (computer-aided drug design), the goal of this application is to synthesize pinane derivatives containing the thiazolidinone moiety, hoping to screen out lead compounds with good anti-tumor activity from them. Summary of the Invention

[0005] The purpose of the present invention is to solve the deficiencies in the prior art, and to propose a method for synthesizing and applying a pinene-based thiazolidinone derivative, the substrate myrtenal has good anti-proliferation effects on malignant cells, the introduction of thiazolidine enhances water solubility, and the introduction of thiazolidine derivatives modified with Schiff base may exhibit significantly enhanced anticancer activity.

[0006] To achieve the above object, the present invention adopts the following technical solutions:

[0007] A method for synthesizing a pinene-based thiazolidinone derivative, comprising the following steps:

[0008] Through the condensation reaction between myrtenal and 4-methyl-3-thiosemicarbazide, promoted by hydrochloric acid, thiosemicarbazide A is synthesized with a yield of 90%;

[0009] Cyclize thiosemicarbazide A with chloroacetic acid to obtain the key intermediate thiazolidinone B;

[0010] Subsequently, this intermediate thiazolidinone B reacts with a substituted benzaldehyde to obtain the target compound, a pinene-based thiazolidinone derivative.

[0011] Preferably, the specific steps are as follows:

[0012] Synthesis of compound thiosemicarbazide A: Charge 1 mmol of myrtenal (150.2 mg, 1.0 mmol), 1.2 mmol of thiosemicarbazide (109.4 mg, 1.2 mmol), 20 mL of ethanol and three drops of concentrated hydrochloric acid (0.15 mL) into a three-necked flask; React the mixture at 50 °C for 2 hours; Monitor the reaction by TLC until it ends, add 30 mL of water to quench the reaction, cool the reaction solution to room temperature, a white solid forms in the flask, then separate the pure product, and obtain thiosemicarbazide compound A by recrystallization from methanol;

[0013] Synthesis of compound thiazolidinone B: Dissolve an equimolar mixture of chloroacetic acid and compound thiosemicarbazide A (474.8 mg, 2.0 mmol) in 20 mL of absolute ethanol, stir the mixed solution at 80 °C for 10 hours; Monitor the reaction by TLC until it ends, add 20 mL of water to quench the reaction, let the solution stand overnight, filter the white solid and wash it with water to obtain compound thiazolidinone B;

[0014] Synthesis of pinanyl thiazolidinone derivatives C1-C13: Dissolve compound thiazolidinone B (139.7 mg, 0.5 mmol) and a substituted benzaldehyde (0.6 mmol) in 20 mL of ethanol, add a catalytic amount of piperidine (0.1 mL); React the mixture at 80 °C for 2 hours, monitor the reaction by TLC until it ends, quench the reaction and cool the solution to room temperature, filter the precipitated crystalline powder, wash it with a small amount of ethanol (2-10 mL), and dry it to obtain the target compound, pinanyl thiazolidinone derivatives C1-C13.

[0015] The present invention also provides an application of a pinene-based thiazolidinone derivative obtained by the above synthesis method in the preparation of an anti-glioma drug.

[0016] Compared with the prior art, the present invention has the following beneficial effects:

[0017] The present invention uses myrtenal as a substrate, which has good anti-proliferative effects on malignant cells. By introducing thiazolidines, the water solubility is enhanced, and the introduction of Schiff base-modified thiazolidine derivatives can exhibit significantly enhanced anti-cancer activities. This series of products has novel structures, high yields, and simple separation and purification operations, making them suitable for large-scale production. Some products, such as C5, have already shown good biological activities. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 Schematic diagram of the synthetic method of the present invention; wherein, a. 4-methylthiosemicarbazone, HCl, EtOH, 50 °C, 2 h; b. chloroacetic acid, sodium acetate, EtOH, 80 °C, overnight; c. benzaldehyde derivatives, morpholine, EtOH, 80 °C, 2 h;

[0019] Figure 2 1H NMR spectrum of pinanyl thiazolidinone derivative C1 in Example 1 of the present invention;

[0020] Figure 3 13C NMR spectrum of pinanyl thiazolidinone derivative C1 in Example 1 of the present invention;

[0021] Figure 4 1H NMR spectrum of pinanyl thiazolidinone derivative C2 in Example 2 of the present invention;

[0022] Figure 5 13C NMR spectrum of pinanyl thiazolidinone derivative C2 in Example 2 of the present invention;

[0023] Figure 6 1H NMR spectrum of pinanyl thiazolidinone derivative C3 in Example 3 of the present invention;

[0024] Figure 7 13C NMR spectrum of pinanyl thiazolidinone derivative C3 in Example 3 of the present invention;

[0025] Figure 8 1H NMR spectrum of pinanyl thiazolidinone derivative C4 in Example 4 of the present invention;

[0026] Figure 9 13C NMR spectrum of pinanyl thiazolidinone derivative C4 in Example 4 of the present invention;

[0027] Figure 10 1H NMR spectrum of pinanyl thiazolidinone derivative C5 in Example 5 of the present invention;

[0028] Figure 11 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C5 in Example 5 of the present invention;

[0029] Figure 12 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C6 in Example 6 of the present invention;

[0030] Figure 13 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C6 in Example 6 of the present invention;

[0031] Figure 14 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C7 in Example 7 of the present invention;

[0032] Figure 15 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C7 in Example 7 of the present invention;

[0033] Figure 16 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C8 in Example 8 of the present invention;

[0034] Figure 17 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C8 in Example 8 of the present invention;

[0035] Figure 18 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C9 in Example 9 of the present invention;

[0036] Figure 19 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C9 in Example 9 of the present invention;

[0037] Figure 20 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C10 in Example 10 of the present invention;

[0038] Figure 21 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C10 in Example 10 of the present invention;

[0039] Figure 22 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C11 in Example 11 of the present invention;

[0040] Figure 23 It is the carbon nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C11 in Example 11 of the present invention;

[0041] Figure 24 It is the proton nuclear magnetic resonance spectrum of the pinanyl thiazolidinone derivative C12 in Example 12 of the present invention;

[0042] Figure 2513C NMR spectrum of pinanylthiazolidinone derivative C12 of Example 1 of the present invention;

[0043] Figure 26 1H NMR spectrum of pinanylthiazolidinone derivative C13 of Example 13 of the present invention;

[0044] Figure 27 13C NMR spectrum of pinanylthiazolidinone derivative C13 of Example 13 of the present invention;

[0045] Figure 28 Representative image of anti-glioma staining of pinanylthiazolidinone derivative C5 of the present invention. Detailed implementation manners

[0046] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings, so that those skilled in the art can better understand the advantages and features of the present invention, and thus make a clearer definition of the protection scope of the present invention. The embodiments described in the present invention are only a part of the embodiments of the present invention, rather than all the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0047] Unless otherwise specified, all reagents and solvents were commercially purchased and used without further purification. All experiments were carried out using distilled water. Thin layer chromatography (TLC, UV 254 nm) on silica gel plates (0.25 mm, China Co., Ltd.) was used to verify the subsequent reactions. The melting point was determined using an Optimelt-MPA100 melting point apparatus. 1H NMR (400 MHz) and 13C NMR (100 MHz) spectra were recorded on a Bruker AV-500 MHz spectrometer, with the solutions and tetramethylsilane as the internal standard. Chemical shifts were expressed in ppm and coupling constants (J) in hertz (Hz). High resolution mass spectrometry data was obtained on a QSTAR Elite high resolution liquid chromatography tandem mass spectrometer. 1 1H NMR(400MHz) and 13 13C NMR(100MHz) spectra, with the solutions and tetramethylsilane as the internal standard. Chemical shifts were expressed in ppm and coupling constants (J) in hertz (Hz). High resolution mass spectrometry data was obtained on a QSTAR Elite high resolution liquid chromatography tandem mass spectrometer.

[0048] A synthesis method of a pinenylthiazolidinone derivative, as Figure 1 shown. According to the synthetic route, pinanylthiazolidinone derivatives C1-C13 were synthesized in three steps, including the following steps:

[0049] By the condensation reaction between myrtenal and 4-methyl-3-thiosemicarbazide, promoted by hydrochloric acid, thiosemicarbazide A was synthesized with a yield of 90%;

[0050] Cyclize thiosemicarbazide A with chloroacetic acid to obtain the key intermediate thiazolidinone B;

[0051] Subsequently, this intermediate thiazolidinone B reacts with the substituted benzaldehyde to obtain the target compounds pinanyl thiazolidinone derivatives C1-C13.

[0052] The experimental part details the synthesis of all the new compounds. Using 1 1H NMR and 13 13C NMR spectra to thoroughly characterize each compound.

[0053] Example

[0054] Synthesis of Pinanyl Thiazolidinone Derivatives

[0055] Synthesis of compound thiosemicarbazide A: Charge 1 mmol of myrtenal (150.2 mg, 1.0 mmol), 1.2 mmol of thiosemicarbazide (109.4 mg, 1.2 mmol), 20 mL of ethanol and three drops of concentrated hydrochloric acid (0.15 mL) into a three-necked flask. React the mixture at 50 °C for 2 hours. Monitor the end of the reaction by TLC, add 30 mL of water to quench the reaction, cool the reaction solution to room temperature, a white solid forms in the flask, then isolate the pure product and obtain thiosemicarbazide compound A (213.7 mg, 0.90 mmol) by recrystallization from methanol, with a yield of 90%. 1 1H NMR (400 MHz, CDCl3) δ 10.37 (s, 1H), 7.58 (s, 1H), 7.27 (s, 1H), 5.97 (s, 1H), 3.16 (d, J = 4.9 Hz, 3H), 2.82 (t, J = 5.6 Hz, 1H), 2.47–2.33 (m, 3H), 2.10 (s, 1H), 1.29 (s, 3H), 1.07 (d, J = 9.0 Hz, 1H), 0.73 (s, 3H). 13 13C NMR (101 MHz, CDCl3) δ 177.67, 144.67, 144.57, 134.05, 40.71, 39.94, 37.69, 32.51, 31.07, 30.93, 26.04, 20.89.

[0056] Synthesis of compound thiazolidinone B: Dissolve an equimolar mixture of chloroacetic acid and compound thiosemicarbazide A (474.8 mg, 2.0 mmol) in 20 mL of absolute ethanol, and stir the mixed solution at 80 °C for 10 hours. Monitor the end of the reaction by TLC, add 20 mL of water to quench the reaction, let the solution stand overnight, filter the white solid and wash it with water to obtain compound thiazolidinone B (279.4 mg, 1.0 mmol), with a yield of 50%. 11H NMR (400 MHz, CDCl3) = δ 8.01 (s, 1H), 6.10 (s, 1H), 3.74 (s, 2H), 3.26 (s, 3H), 3.02 (t, J = 6.2 Hz, 1H), 2.51 - 2.44 (m, 3H), 2.16 (s, 1H), 1.36 (s, 3H), 1.17 (d, J = 9.0 Hz, 1H), 0.81 (s, 3H). 13 13C NMR (101 MHz, CDCl3) = δ 172.10, 162.88, 159.47, 146.23, 135.26, 40.72, 40.25, 37.65, 32.72, 32.39, 31.17, 29.70, 26.08, 20.92.

[0057] Synthesis of pinanyl thiazolidinone derivatives C1 - C13: Dissolve compound thiazolidinone B (139.7 mg, 0.5 mmol) and substituted benzaldehyde (0.6 mmol) in 20 mL of ethanol, and add a catalytic amount of piperidine (0.1 mL). React the mixture at 80 °C for 2 hours. Monitor the end of the reaction by TLC. Quench the reaction by cooling the solution to room temperature. Filter the precipitated crystalline powder, wash it with a small amount of ethanol (2 - 10 mL), and dry it to obtain the target compounds C1 - C13.

[0058] Example 1: Pinanyl thiazolidinone derivative C1

[0059] 2 - (((6,6 - dimethylbicyclo[3.1.1]hept - 2 - en - 2 - yl)methylene)hydrazineylidene)-5-(4 - methoxybenzylidene)-3 - methylthiazolidin - 4 - one (C1): Yellow solid, yield 91%. 1 1H NMR (400 MHz, CDCl3) = δ 8.08 (s, 1H), 7.66 (s, 1H), 7.54 (d, J = 8.7 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 6.14 (s, 1H), 3.86 (s, 3H), 3.41 (s, 3H), 3.11 (t, J = 5.6 Hz, 1H), 2.58 - 2.46 (m, 3H), 2.19 (s, 1H), 1.67 (s, 1H), 1.42 (s, 3H), 1.21 (d, J = 9.0 Hz, 1H), 0.84 (s, 3H). ( Figure 2 as shown) 1313C NMR (101 MHz, CDCl3) = δ 167.09, 160.11, 158.54, 146.27, 141.54, 136.05, 130.44, 129.87, 126.02, 121.20, 40.78, 40.30, 37.77, 32.84, 31.27, 29.82, 26.14, 20.96, 15.10.( Figure 3 as shown

[0060] Example 2: Pinanylthiazolidinone Derivative C2

[0061] 2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-5-(4-methoxybenzylidene)-3-methylthiazolidin-4-one (C2): Yellow solid, yield 93%. 1 1H NMR (400 MHz, CDCl3) = δ 8.08 (s, 1H), 7.66 (s, 1H), 7.54 (d, J = 8.8 Hz, 2H), 7.01 (d, J = 8.8 Hz, 2H), 6.14 (s, 1H), 3.86 (s, 3H), 3.41 (s, 3H), 3.11 (td, J = 5.7, 1.5 Hz, 1H), 2.60 - 2.39 (m, 3H), 2.19 (s, 1H), 1.42 (s, 3H), 1.21 (d, J = 9.0 Hz, 1H), 0.84 (s, 3H).( Figure 4 as shown 13 13C NMR (101 MHz, CDCl3) = δ 167.26, 160.76, 159.95, 158.85, 146.30, 135.86, 131.98, 130.30, 126.79, 119.52, 114.60, 55.44, 40.80, 40.30, 37.77, 32.82, 31.27, 29.76, 26.14, 20.96.( Figure 5 as shown

[0062] Example 3: Pinanylthiazolidinone Derivative C3

[0063] 2-(((6,6-Dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-3-methyl-5-(4-(methylthio)benzylidene)thiazolidin-4-one (C3): Yellow solid, yield 92%. 1 1H NMR (400 MHz, CDCl3) δ 8.08 (s, 1H), 7.64 (s, 1H), 7.50 (d, J = 8.5 Hz, 2H), 7.32 (d, J = 8.5 Hz, 2H), 6.15 (s, 1H), 3.41 (s, 3H), 3.09 (t, J = 5.6 Hz, 1H), 2.59 - 2.46 (m, 6H), 2.19 (s, 1H), 1.64 (s, 1H), 1.42 (s, 3H), 1.21 (d, J = 9.1 Hz, 1H), 0.84 (s, 3H).( Figure 6 as shown 13 13C NMR (101 MHz, CDCl3) δ 167.09, 160.11, 158.54, 146.27, 141.53, 136.05, 130.47, 130.44, 129.87, 126.02, 121.20, 40.78, 40.30, 37.77, 32.84, 31.26, 29.82, 26.14, 20.96, 15.10.( Figure 7 as shown

[0064] Example 4: Pinanylthiazolidinone derivative C4

[0065] 5-(4-(Dimethylamino)benzylidene)-2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-3-methylthiazolidin-4-one (C4): Yellow solid, yield 92%. 11H NMR (400 MHz, CDCl3) = δ 8.08 (s, 1H), 7.64 (s, 1H), 7.49 (d, J = 8.9 Hz, 2H), 6.76 (d, J = 9.0 Hz, 2H), 6.13 (s, 1H), 3.40 (s, 3H), 3.16–3.10 (m, 1H), 3.05 (s, 6H), 2.93 (d, J = 2.9 Hz, 1H), 2.58–2.46 (m, 3H), 2.19 (tdd, J = 6.7, 3.0, 1.6 Hz, 1H), 1.64 (s, 2H), 1.42 (s, 3H), 1.22 (d, J = 9.0 Hz, 1H), 0.84 (s, 3H).( Figure 8 as shown 13 13C NMR (101 MHz, CDCl3) = δ 167.63, 159.46, 151.05, 146.36, 135.36, 132.17, 131.42, 121.71, 115.83, 111.97, 40.81, 40.31, 40.10, 37.78, 32.79, 31.30, 29.65, 26.16, 20.96.( Figure 9 as shown

[0066] Example 5: Pinanylthiazolidinone Derivative C5

[0067] 2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-5-(4-hydroxybenzylidene)-3-methylthiazolidin-4-one (C5): Yellow solid, yield 94%. 1 1H NMR (400 MHz, CDCl3) = δ 8.09 (s, 1H), 7.65 (s, 1H), 7.48 (d, J = 8.7 Hz, 2H), 6.97 (d, J = 8.7 Hz, 2H), 6.16 (s, 1H), 3.42 (s, 3H), 3.10 (t, J = 6.3 Hz, 1H), 2.58–2.41 (m, 3H), 2.18 (s, 1H), 1.39 (s, 3H), 1.21 (d, J = 9.0 Hz, 1H), 0.83 (s, 3H).( Figure 10 as shown 1313C NMR (101 MHz, CDCl3) = δ 167.51, 160.19, 158.87, 157.49, 146.21, 136.27, 132.26, 130.70, 126.60, 119.19, 116.24, 116.03, 40.77, 40.31, 37.77, 32.84, 31.27, 29.82, 26.11, 20.94.( Figure 11 as shown

[0068] Example 6: Pinanylthiazolidinone Derivative C6

[0069] 2 - (((6,6 - dimethylbicyclo[3.1.1]hept - 2 - en - 2 - yl)methylene)hydrazineylidene)-5-(4 - hydroxy - 3 - methoxybenzylidene)-3 - methylthiazolidin - 4 - one (C6): Yellow solid, yield 92%. 1 1H NMR (400 MHz, DMSO - d6) = δ 9.90 (s, 1H), 8.13 (s, 1H), 7.60 (s, 1H), 7.23 (d, J = 2.1 Hz, 1H), 7.10 (dd, J = 8.3, 2.0 Hz, 1H), 6.95 (d, J = 8.2 Hz, 1H), 6.29 (s, 1H), 3.84 (s, 3H), 3.38 (s, 1H), 3.28 (s, 3H), 3.04 - 2.92 (m, 1H), 2.49 - 2.37 (m, 2H), 2.16 (s, 1H), 1.35 (s, 3H), 1.11 (d, J = 8.9 Hz, 1H), 0.78 (s, 3H).( Figure 12 as shown 13 13C NMR (101 MHz, DMSO - d6) δ 166.64, 159.49, 159.07, 149.31, 148.29, 145.78, 136.40, 130.86, 125.47, 124.02, 118.23, 116.61, 114.80, 55.85, 37.67, 32.73, 31.11, 30.04, 26.36, 21.30.( Figure 13 as shown

[0070] Example 7: Pinanylthiazolidinone Derivative C7

[0071] 5-(4-chlorobenzylidene)-2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-3-methylthiazolidin-4-one (C7): Yellow solid, yield 93%. 1 H NMR (400 MHz, CDCl3) δ 8.09 (s, 1H), 7.65 (s, 1H), 7.48 (d, J = 8.7 Hz, 2H), 6.97 (d, J = 8.7 Hz, 2H), 6.16 (s, 1H), 3.42 (s, 3H), 3.09 (t, J = 5.5 Hz, 1H), 2.58 - 2.46 (m, 3H), 2.18 (s, 1H), 1.39 (s, 3H), 1.21 (d, J = 9.0 Hz, 1H), 0.83 (s, 3H). ( Figure 14 as shown) 13 C NMR (101 MHz, CDCl3) = δ 167.51, 160.19, 158.87, 157.49, 146.21, 136.27, 132.27, 130.70, 126.60, 119.19, 116.24, 40.77, 40.31, 37.77, 32.85, 31.27, 29.83, 26.11, 20.95. ( Figure 15 as shown)

[0072] Example 8: Pinanylthiazolidinone derivative C8

[0073] 5-(4-(diethylamino)benzylidene)-2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)

[0074] methylene)hydrazineylidene)-3-methylthiazolidin-4-one (C8): Yellow solid, yield 91%. 11H NMR (400 MHz, Pyridine-d5) = δ 10.19 (s, 2H), 9.85 (s, 1H), 9.43 (s, 1H), 9.09 - 8.98 (m, 3H), 8.03 (d, J = 9.0 Hz, 2H), 4.92 (s, 4H), 4.67 (q, J = 7.1 Hz, 4H), 3.81 (dt, J = 6.1, 3.6 Hz, 3H), 3.45 (s, 1H), 2.62 (s, 3H), 2.60 (d, J = 8.9 Hz, 1H), 2.47 (t, J = 7.0 Hz, 6H), 2.26 (s, 3H).( Figure 16 as shown 13 13C NMR (101 MHz, Pyridine-d5) = δ 169.25, 161.79, 161.46, 151.07, 148.44, 134.74, 133.48, 123.00, 117.29, 113.86, 46.44, 42.96, 42.60, 39.72, 34.81, 33.38, 31.68, 27.92, 22.97, 14.56.( Figure 17 as shown

[0075] Example 9: Pinanylthiazolidinone Derivative C9

[0076] 2 - (((6,6 - dimethylbicyclo[3.1.1]hept - 2 - en - 2 - yl)methylene)hydrazineylidene)-5-(2 - hydroxy - 5 - methylbenzylidene)-3 - methylthiazolidin - 4 - one (C9): Yellow solid, yield 91%. 1 1H NMR (400 MHz, DMSO - d6) = δ 10.19 (s, 1H), 8.14 (s, 1H), 7.89 (s, 1H), 7.22 (s, 1H), 7.11 (d, J = 8.4 Hz, 1H), 6.87 (d, J = 8.2 Hz, 1H), 6.30 (s, 1H), 3.38 (s, 1H), 3.28 (s, 3H), 2.97 (t, J = 5.6 Hz, 1H), 2.44 (t, J = 21.0 Hz, 2H), 2.27 (s, 3H), 2.16 (s, 1H), 1.35 (s, 3H), 1.11 (d, J = 8.9 Hz, 1H), 0.78 (s, 3H).( Figure 18 as shown 13¹³C NMR (101 MHz, DMSO-d₆) = δ 166.64, 159.71, 158.97, 155.33, 145.76, 136.53, 132.79, 128.90, 128.45, 125.45, 120.73, 120.52, 116.41, 37.68, 32.74, 31.11, 30.07, 26.34, 21.29, 20.77.( Figure 19 as shown

[0077] Example 10: Pinanylthiazolidinone Derivative C10

[0078] 5-(3-chloro-4-hydroxybenzylidene)-2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)met hylene)hydrazineylidene)-3-methylthiazolidin-4-one (C10): Yellow solid, yield 93%. 1 ¹H NMR (400 MHz, DMSO-d₆) = δ 11.12 (s, 1H), 8.14 (s, 1H), 7.66 (s, 1H), 7.58 (s, 1H), 7.46 (d, J = 8.6 Hz, 1H), 7.16 (d, J = 8.5 Hz, 1H), 6.31 (s, 1H), 3.28 (s, 3H), 2.98 (t, J = 5.6 Hz, 1H), 2.50 - 2.37 (m, 2H), 2.16 (s, 1H), 1.37 (s, 3H), 1.22 (s, 1H), 1.11 (d, J = 8.8 Hz, 1H), 0.78 (s, 3H).( Figure 20 as shown 13 ¹³C NMR (101 MHz, DMSO-d₆) = δ 166.43, 160.07, 158.33, 155.29, 145.68, 136.88, 132.65, 130.15, 129.19, 126.19, 120.95, 119.68, 117.75, 37.73, 32.75, 31.11, 30.15, 26.38, 21.30.( Figure 21 as shown

[0079] Example 11: Pinanylthiazolidinone Derivative C11

[0080] 2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-5-(3-hydroxy-4-methoxybenzylidene)-3-methylthiazolidin-4-one (C11): Yellow solid, yield 93%. 1 H NMR (400 MHz, DMSO-d6) = δ 9.47 (s, 1H), 8.15 (s, 1H), 7.54 (s, 1H), 7.10 (d, J = 5.2 Hz, 3H), 6.32 (s, 1H), 3.84 (s, 3H), 3.28 (s, 3H), 3.06 - 2.94 (m, 1H), 2.45 (t, J = 18.8 Hz, 2H), 2.17 (s, 1H), 1.38 (s, 3H), 1.12 (d, J = 8.9 Hz, 1H), 0.79 (s, 3H).( Figure 22 as shown 13 C NMR (101 MHz, DMSO-d6) = δ 166.57, 160.05, 158.62, 150.02, 147.29, 145.68, 136.85, 130.58, 126.70, 123.63, 118.92, 116.30, 112.87, 56.09, 37.77, 32.74, 31.12, 30.10, 26.42, 21.32.( Figure 23 as shown

[0081] Example 12: Pinanylthiazolidinone derivative C12

[0082] 2-(((6,6-dimethylbicyclo[3.1.1]hept-2-en-2-yl)methylene)hydrazineylidene)-5-(3-ethoxy-4-hydroxybenzylidene)-3-methylthiazolidin-4-one (C12): Yellow solid, yield 92%. 11H NMR (400 MHz, DMSO-d6) = δ 9.84 (s, 1H), 8.14 (s, 1H), 7.58 (s, 1H), 7.25 - 7.04 (m, 2H), 6.95 (d, J = 8.4 Hz, 1H), 6.30 (s, 1H), 4.10 (d, J = 7.1 Hz, 2H), 3.37 (s, 2H), 3.28 (s, 3H), 2.98 (s, 1H), 2.41 (d, J = 17.5 Hz, 2H), 2.17 (s, 1H), 1.37 (d, J = 11.4 Hz, 6H), 1.11 (d, J = 8.8 Hz, 1H), 0.78 (s, 3H).( Figure 24 as shown 13 13C NMR (101 MHz, DMSO-d6) = δ 166.67, 159.40, 159.25, 149.54, 147.41, 145.79, 136.41, 130.89, 124.57, 118.22, 116.69, 115.27, 64.15, 37.68, 32.73, 31.11, 30.04, 26.39, 21.30, 15.09.( Figure 25 as shown

[0083] Example 13: Pinanylthiazolidinone Derivative C13

[0084] 2 - (((6,6 - dimethylbicyclo[3.1.1]hept - 2 - en - 2 - yl)methylene)hydrazineylidene)-5-(2 - hydroxy - 4 - methoxybenzylidene)-3 - methylthiazolidin - 4 - one (C13): Yellow solid, yield 90%. 1 1H NMR (400 MHz, DMSO-d6) = δ 10.53 (s, 1H), 8.14 (s, 1H), 7.89 (s, 1H), 7.38 (d, J = 8.7 Hz, 1H), 6.64 (d, J = 8.9 Hz, 1H), 6.51 (d, J = 2.6 Hz, 1H), 6.30 (s, 1H), 3.77 (s, 3H), 3.28 (s, 3H), 2.97 (t, J = 5.6 Hz, 1H), 2.42 (d, J = 19.9 Hz, 1H), 2.16 (s, 1H), 1.36 (s, 3H), 1.11 (d, J = 8.5 Hz, 1H), 0.78 (s, 3H).( Figure 26 as shown 1313C NMR (101 MHz, DMSO-d6) = δ 166.67, 159.40, 159.25, 149.54, 147.41, 145.79, 136.41, 130.89, 124.57, 118.22, 116.69, 115.27, 64.15, 37.68, 32.73, 31.11, 30.04, 26.39, 21.30, 15.09.( Figure 27 as shown

[0085] The development of traditional anti-cancer drugs was carried out using a monolayer cell-based 2D method. However, due to the defects of 2D models in simulating cancer cell development, it is difficult to accurately evaluate the anti-cancer effects of candidate drugs. On this basis, a three-dimensional cell culture method was used to evaluate the therapeutic effect of C5 on U251 cancer cells. After treatment with C5 for 24 hours, U251 cells were stained with PI and calcein AM in DMEM for 1.5 hours, and then observed under a laser scanning confocal microscope to analyze the live / dead U251 (human glioblastoma cells) MCTSs (multicellular tumor spheroids,).

[0086] as Figure 28 shown, representative images of C5-treated U251 cells stained with calcein AM (green live) and PI (red dead). Most cells in untreated spheroids were alive (green fluorescence), and only a few dead cells (red fluorescence) were found in the center of the spheroids, which may be due to overgrowth and lack of nutrients and oxygen, ultimately leading to apoptosis. In contrast, C5-treated MCTS showed a streamlined shape, and the necrotic core in C5-treated spheroids expanded with increasing C5 concentration. In summary, compound C5 inhibited the MCTS of U251 cells in a dose-dependent manner.

[0087] In summary, the present invention synthesized a series of pinanylthiazolidinone derivatives with different structures. Compound C5 inhibited the migration and cell cycle of U251 cells, and further confirmed its anti-tumor efficacy using 3D cell culture. Generally speaking, compound C5 represents a novel thiazolidinone CDK2 inhibitor with potent anti-glioma effects.

[0088] The descriptions and practices disclosed in the present invention are easy to think about and understand for those of ordinary skill in the art. Without departing from the principle of the present invention, several improvements and refinements can be made. Therefore, the modifications or improvements made without deviating from the spirit of the present invention should also be regarded as within the protection scope of the present invention.

Claims

1. A method for synthesizing a pinene-based thiazolidinone derivative, characterized in that, It includes the following steps: Through the condensation reaction between myrtenal and 4-methyl-3-thiosemicarbazide, promoted by hydrochloric acid, thiosemicarbazide A is synthesized with a yield of 90%; Cyclize thiosemicarbazide A with chloroacetic acid to obtain the key intermediate thiazolidinone B; Subsequently, this intermediate thiazolidinone B reacts with substituted benzaldehyde to obtain the target compound pinene-based thiazolidinone derivative.

2. The synthesis method of a pinene-based thiazolidinone derivative according to claim 1, characterized in that, The specific steps are as follows: Synthesis of compound thiosemicarbazide A: Charge 1 mmol of myrtenal, 1.2 mmol of thiosemicarbazide, 20 mL of ethanol and three drops of concentrated hydrochloric acid into a three-necked flask; React the mixture at 50 °C for 2 hours; Monitor the end of the reaction by TLC, add 30 mL of water to quench the reaction, cool the reaction solution to room temperature, a white solid forms in the flask, then separate the pure product and obtain thiosemicarbazide compound A by recrystallization from methanol; Synthesis of compound thiazolidinone B: Dissolve an equimolar mixture of chloroacetic acid and compound thiosemicarbazide A in 20 mL of anhydrous ethanol, stir the mixed solution at 80 °C for 10 hours; Monitor the end of the reaction by TLC, add 20 mL of water to quench the reaction, let the solution stand overnight, filter the white solid and wash it with water to obtain compound thiazolidinone B; Synthesis of pinanyl thiazolidinone derivatives C1-C13: Dissolve compound thiazolidinone B and substituted benzaldehyde in 20 mL of ethanol, add a catalytic amount of piperidine; React the mixture at 80 °C for 2 hours, monitor the end of the reaction by TLC, quench the reaction and cool the solution to room temperature, filter the precipitated crystalline powder, wash it with ethanol, dry it to obtain the target compound pinanyl thiazolidinone derivatives C1-C13.

3. Use of a pinene-based thiazolidinone derivative obtained by the synthesis method according to any one of claims 1-2 in the preparation of an anti-glioma drug.

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