Use of a forskolin tetra cyclic analogue in the preparation of an anti-inflammatory medicament
By modifying the structure of forscolin, a tetracyclic analog of forscolin with a unique tetracyclic structure was synthesized, which solved the shortcomings of existing anti-inflammatory drugs in inhibiting pro-inflammatory factors and achieved a significant anti-inflammatory effect.
Patent Information
- Application Number
- CN202311391227.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-10-25
- Publication Date
- 2026-02-10
- Estimated Expiration
- 2043-10-25
AI Technical Summary
Existing anti-inflammatory drugs have limited effectiveness in inhibiting pro-inflammatory factors such as tumor necrosis factor, interleukins, prostaglandins, and nitric oxide, and there is a lack of effective chemical structural modifications to enhance the anti-inflammatory activity of these drugs.
By structurally modifying forscolin, a forscolin tetracyclic analogue with a unique tetracyclic structure was synthesized and prepared into an anti-inflammatory drug. Its unique tetracyclic structure endows it with excellent anti-inflammatory activity and inhibits pro-inflammatory factors.
It effectively inhibited pro-inflammatory factors such as tumor necrosis factor, interleukin, prostaglandin and nitric oxide, showing significant anti-inflammatory effects and no obvious cytotoxicity.
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Figure CN117224529B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the fields of medicinal chemistry and pharmacology, specifically to the application of a tetracyclic analog of foscorine in the preparation of anti-inflammatory drugs. Background Technology
[0002] Forsocolin is a diterpenoid compound with significant medicinal value in anticancer, anti-asthmatic, antihypertensive, and positive inotropic effects. Furthermore, forsocolin can interact with certain other proteins, such as glucose transporters and ion channels. Pharmacodynamic studies have shown that forsocolin can effectively promote neuronal differentiation and neurite spur growth in the central and peripheral nervous systems, thus having a significant impact on the cardiovascular, respiratory, and tumor systems, exhibiting pharmacological effects such as cardiotonic, antiasthmatic, antitumor, antithrombotic, and intraocular pressure-lowering properties. It is currently being used clinically to treat cardiovascular diseases, tumors, and common geriatric diseases, demonstrating significant pharmacological effects and promising clinical application prospects. Developing new compounds by using inexpensive and readily available forsocolin as a raw material and modifying its chemical skeleton through chemical transformation, and then exploring their applications in medicinal chemistry and pharmacology, is of great significance. Summary of the Invention
[0003] In view of this, the purpose of the present invention is to provide an application of a tetracyclic analog of foscorine in the preparation of anti-inflammatory drugs. Such compounds have a unique tetracyclic structure and strong anti-inflammatory activity, and can be used to prepare anti-inflammatory drugs.
[0004] This invention provides a tetracyclic analog of foscorine, which has the structure shown in formula (I):
[0005]
[0006] Wherein, R represents one of phenyl, p-trifluoromethoxyphenyl, p-tert-butylphenyl, p-chlorophenyl, p-methylphenyl, p-methoxyphenyl, p-nitrophenyl, and p-cyanophenyl.
[0007] Furthermore, the fuscolin tetracyclic analogue has a structure as shown in any one of formulas 4a to 4c:
[0008]
[0009] in,
[0010] When R is phenyl, the tetracyclic analog of foscorine is a compound with the structure shown in formula 4a;
[0011] When R is p-trifluoromethoxyphenyl, the tetracyclic analog of foscorine is a compound with the structure shown in formula 4b.
[0012] When R is p-tert-butylphenyl, the tetracyclic analog of foscorine is a compound with the structure shown in formula 4c;
[0013] This invention also provides a method for preparing tetracyclic analogues of foscorine, comprising the following steps:
[0014] Forsocolin was reacted with sodium methoxide in methanol to give 7-deacetylated forsocolin 1. Then, 7-deacetylated forsocolin 1 was reacted with sodium periodate in a DCM-H2O mixed solution to give forsocolin rearrangement product 2. Forsocolin rearrangement product 2 was reacted with DMP and pyridine in acetonitrile to give forsocolin rearrangement product 3. Finally, forsocolin rearrangement product 3 was reacted with aryl iodide in anhydrous dichloroethane under the catalysis of anhydrous silver acetate, anhydrous copper acetate and palladium acetate to give forsocolin tetracyclic analog 4.
[0015] The reaction formula is as follows:
[0016]
[0017] Wherein, R represents one of phenyl, p-trifluoromethoxyphenyl, p-tert-butylphenyl, p-chlorophenyl, p-methylphenyl, p-methoxyphenyl, p-nitrophenyl, and p-cyanophenyl.
[0018] Specifically, the preparation method includes the following steps:
[0019] (1) Dissolve fuscaline in methanol, add sodium methoxide, react at 40°C for 2 hours to obtain the first reaction solution. After cooling the first reaction solution, concentrate it under reduced pressure, dilute it with an organic solvent, wash it with water, wash it with saturated brine, dry it with anhydrous MgSO4, concentrate it under reduced pressure, and then obtain a white solid 7-deacetylated fuscaline 1 by flash column chromatography. The molar ratio of fuscaline to sodium methoxide is 1:2.
[0020] (2) Dissolve 7-deacetylfoscorine 1 in a mixed solution of dichloromethane and water, add silica gel and sodium periodate, react at 40°C for 24 h to obtain a second reaction solution. After cooling the second reaction solution, concentrate under reduced pressure, dilute with organic solvent, filter, wash the organic phase with water and saturated brine, dry with anhydrous MgSO4, concentrate under reduced pressure, and then obtain white solid foscorine rearrangement product 2 by flash column chromatography. The volume ratio of dichloromethane to water is 4:1, the mass ratio of silica gel to 7-deacetylfoscorine 1 is 1:1, and the molar ratio of 7-deacetylfoscorine 1 to sodium periodate is 1:10.
[0021] (3) The forscoline rearrangement product 2 was dissolved in acetonitrile, and the oxidant Dysmartin and pyridine were added. The mixture was reacted at 20°C for 12 h to obtain a third reaction solution. The third reaction solution was diluted with an organic solvent and washed successively with saturated sodium carbonate aqueous solution, saturated sodium thiosulfate aqueous solution, and saturated brine. The mixture was dried over anhydrous MgSO4, concentrated under reduced pressure, and then obtained by flash column chromatography as a white solid forscoline skeleton rearrangement product 3. The molar ratio of forscoline rearrangement product 2, Dysmartin oxidant, and pyridine was 1:3:2.
[0022] (4) The forscoline rearrangement product 3 was dissolved in anhydrous dichloroethane, and anhydrous silver acetate, anhydrous copper acetate, palladium acetate, and aryl iodide were added sequentially. The mixture was reacted at 80°C for 18 hours to obtain a fourth reaction solution. The fourth reaction solution was cooled, diluted with an organic solvent, filtered, and the filtrate was concentrated under reduced pressure. A white solid was obtained by flash column chromatography. The white solid was forscoline tetracyclic analog 4. The molar ratio of forscoline rearrangement product 3, anhydrous silver acetate, anhydrous copper acetate, palladium acetate, and aryl iodide was 1:1.2:2:0.05:2.
[0023] The aryl iodide is one of the following: iodobenzene, p-trifluoromethoxyiodobenzene, p-tert-butyliodobenzene, p-chloroiodobenzene, p-methyliodobenzene, p-methoxyiodobenzene, p-nitroiodobenzene, and p-cyanoiodobenzene.
[0024] Furthermore, in the above preparation method, the organic solvent is at least one selected from methanol, dichloromethane, ethyl acetate, and dichloroethane.
[0025] Furthermore, the anti-inflammatory drug is a drug that inhibits pro-inflammatory factors.
[0026] Furthermore, the pro-inflammatory factors include one or more of tumor necrosis factor, interleukin, prostaglandin, nitric oxide, and reactive oxygen species.
[0027] Furthermore, the pro-inflammatory factor is nitric oxide.
[0028] Furthermore, the dosage form of the drug is an oral formulation.
[0029] Furthermore, the dosage form of the drug is one of tablets, capsules, pills, granules, decoctions, ointments, elixirs, oral liquids, drop pills, and syrups.
[0030] Furthermore, the drug also includes pharmaceutically acceptable excipients.
[0031] Furthermore, the pharmaceutically acceptable excipients include one or more of the following: fruit powder, flavoring, sweetener, acidulant, filler, lubricant, preservative, suspending agent, food coloring, diluent, emulsifier, disintegrant, and plasticizer.
[0032] Compared with the prior art, this application provides the application of a new class of compounds, foscorine tetracyclic analogs, in the preparation of anti-inflammatory drugs. These compounds contain a unique tetracyclic structure, which endows these derivatives with excellent anti-inflammatory activity and can be applied to the preparation of anti-inflammatory drugs. Attached Figure Description
[0033] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below.
[0034] Figure 1 Nuclear magnetic resonance imaging of the forscolin rearrangement product (3) provided in Example 2 of this invention 1 H spectrum;
[0035] Figure 2 Nuclear magnetic resonance imaging of the forscolin rearrangement product (3) provided in Example 2 of this invention 13 C spectrum;
[0036] Figure 3 X-ray single-crystal diffraction structure of the forscolin rearrangement product (3) provided in Example 2 of the present invention;
[0037] Figure 4 Nuclear magnetic resonance imaging of the tetracyclic analogue (4a) of foscorine provided in Embodiment 3 of the present invention 1 H spectrum;
[0038] Figure 5 Nuclear magnetic resonance imaging of the tetracyclic analogue (4a) of foscorine provided in Embodiment 3 of the present invention 13 C spectrum;
[0039] Figure 6 The nuclear magnetic resonance 1H spectrum of the tetracyclic analogue (4b) of foscorine provided in Embodiment 3 of the present invention;
[0040] Figure 7 The nuclear magnetic resonance 13C spectrum of the tetracyclic analogue (4b) of foscorine provided in Embodiment 3 of the present invention. Detailed Implementation
[0041] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.
[0042] Example 1
[0043] 410 mg (1.0 mmol) of foscorline was dissolved in methanol (10 mL), and 360 mg of sodium methoxide (30 wt% inMeOH, 2.0 mmol) was added. The reaction mixture was reacted at 40 °C for 2 hours. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate (10 mL), and then washed successively with water and saturated brine. The solution was dried over anhydrous MgSO4, concentrated under reduced pressure, and obtained by flash column chromatography (petroleum ether:ethyl acetate = 8:1) to give 360 mg of a white solid, namely 7-deacetylated foscorline 1 (yield 98%).
[0044] 1 H NMR(400MHz,Chloroform-d)δ6.60(m,1H,9-OH),6.13(dd,J=17.4,10.7Hz,1H,H-14),5.20(d,J=
[0045] 17.4Hz,1H,H-15),4.99(d,J=10.6Hz,1H,H-15),4.62(s,1H,H-1),4.48(s,1H,H-6),4.13(t,J=3.5Hz,1H,H-7),3.18(d,J=1 7.2Hz,1H,H-12),2.93(dt,J=9.4,3.1Hz,1H,1-OH),2.70(q,J=3.7Hz,1H,7-OH),2.50(d,J=17.2Hz,1H,H-12),2.42–2.36(m, 1H,H-2e),2.09(d,J=2.7Hz,1H,H-5),1.84–1.79(m,1H,6-OH),1.73(td,J=13.9,3.6Hz,1H,H-3a),1.65(s,3H,CH3),1.42(s ,3H,CH3),1.42-1.41(m,1H,H-2a),1.41(s,3H,CH3),1.27(s,3H,CH3),1.12(dt,J=13.4,3.5Hz,1H,H-3e),1.06(s,3H,CH3).
[0046] Example 2
[0047] 368 mg (1.0 mmol) of 7-deacetylated foscorine 1 was dissolved in a dichloromethane-water mixture (10 mL, v:v = 4:1), and 368 mg of silica gel and 2.14 g (10.0 mmol) of sodium periodate were added. The reaction mixture was reacted at 40 °C for 24 hours. The reaction solution was concentrated under reduced pressure, diluted with ethyl acetate (10 mL), filtered, and the organic phase was washed successively with water and saturated brine. The solution was dried over anhydrous MgSO4, concentrated under reduced pressure, and obtained by flash column chromatography (petroleum ether:ethyl acetate = 10:1) to give 340 mg of white solid, which was foscorine rearrangement product 2 (yield 98%). Then, 366 mg (1.0 mmol) of rearrangement product 2 was dissolved in acetonitrile (10 mL), and 158 μL (2.0 mmol) of pyridine and 1.27 g (3.0 mmol) of Des Martin oxidant were added. The reaction mixture was reacted at 20 °C for 12 hours. The reaction solution was concentrated under reduced pressure, then diluted with ethyl acetate (10 ml), washed successively with saturated sodium carbonate aqueous solution, saturated sodium thiosulfate aqueous solution, and saturated brine, dried over anhydrous MgSO4, concentrated under reduced pressure, and obtained by flash column chromatography to yield 122 mg of white solid, namely foscarline rearrangement product 3 (yield 35%).
[0048] 1 H NMR (400MHz, DMSO-d6) δ9.58(s,1H,H-16),5.91(dd,J=17.4,10.6Hz,1H,H-13),5.40(s,1H,H-11),5.20(dd,J=17. 4,1.3Hz,1H,H-14),4.98(dd,J=10.5,1.2Hz,1H,H-14),4.26(dd,J=6.0,3.4Hz,1H,H-1),2.73(s,1H,H-5),2.06(dd t,J=15.6,6.8,5.1Hz,1H),1.75(dddd,J=15.2,8.9,5.0,3.4Hz,1H),1.57(dddd,J=14.5,9.7,4.9Hz,1H),1.37(s,3 H,CH3),1.32(ddd,J=12.6,5.2,2.0Hz,1H),1.28(s,3H,CH3),1.21(s,3H,CH3),1.10(s,3H,CH3),1.08(s,3H,CH3). 13 C NMR (100MHz, DMSO) δ200.8,172.1,151.1,144.0,112.2,106.8,90.3,81.7,81.6,76.7,52.6,51.1,34.8,31.2,30.4,28.1,24.1,23.5,20.1,13.7.
[0049] Example 3
[0050] 346 mg (1.0 mmol) of forsocrine rearrangement product 3 was dissolved in anhydrous dichloroethane (10 mL), followed by the addition of 200 mg (1.2 mmol) of anhydrous silver acetate, 363 mg (2.0 mmol) of anhydrous copper acetate, 12 mg (0.05 mmol) of palladium acetate, and 408 mg (2.0 mmol) of iodobenzene. The reaction mixture was reacted at 80 °C for 18 hours. The reaction solution was diluted with ethyl acetate (10 mL), filtered, and the filtrate was washed successively with water and saturated brine, dried over anhydrous MgSO4, concentrated under reduced pressure, and obtained by flash column chromatography (petroleum ether:ethyl acetate = 30:1) to give 238 mg of a white solid, namely forsocrine tetracyclic analog 4a (yield 69%).
[0051] 1 H NMR(400MHz,Chloroform-d)δ9.67(s,1H,CHO),7.45–7.10(m,5H,ArH),6.48(d,J=16.1Hz,1H,H- 14),6.26(d,J=16.2Hz,1H,H-13),5.25(s,1H,H-11),4.06(dd,J=6.2,3.5Hz,1H,H-1),2.72(s,1 H,H-5),1.98(dq,J=18.1,6.0Hz,1H),1.84–1.73(m,1H),1.53(td,J=9.8,5.0Hz,1H),1.46(s,3H ,CH3),1.37–1.30(m,1H),1.29(s,3H,CH3),1.21(s,3H,CH3),1.11(s,3H,CH3),1.08(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ201.07,171.83,151.30,136.84,134.89,128.51,127.53,127.36,126.67,107.20,90.1 2,81.95,81.73,76.40,52.84,51.28,34.95,31.21,30.33,28.07,24.36,23.40,20.14,13.69.HRMS(ESI):m / z calcd for C 26 H 30 O5Na:445.1991; found:445.1989[M+Na] + .
[0052] Examples 4-5
[0053] According to the method in Example 3 above, iodobenzene was replaced with p-trifluoromethoxyiodobenzene and p-tert-butyliodobenzene, respectively, to prepare compounds 4b and 4c.
[0054] The following is a list of the physicochemical data for compounds 4b-4c:
[0055] 4b: Yield 63%. 1 H NMR(400MHz,Chloroform-d)δ9.67(s,1H,CHO),7.45–7.29(m,2H,ArH),7.14–7.02(m,2H,ArH),6.48(d,J=16. 1Hz,1H,H-14),6.24(d,J=16.1Hz,1H,H-13),5.24(s,1H,H-11),4.08(dd,J=6.1,3.5Hz,1H,H-1),2.71(s,1H,H -5),1.99(dtd,J=15.6,6.6,5.0Hz,1H),1.79(dddd,J=15.5,10.0,5.2,3.5Hz,1H),1.54(tt,J=10.0,5.0Hz,1H ),1.46(s,3H,CH3),1.38–1.31(m,1H),1.29(s,3H,CH3),1.22(s,3H,CH3),1.11(s,3H,CH3),1.09(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ200.8,171.8,151.5,148.4,135.9,135.7,127.9,125.9,121.7,121.0,119.2,10 6.8,90.1,82.0,81.8,76.3,52.8,51.3,34.9,31.2,30.3,28.2,24.4,23.4,20.1,13.7.HRMS(ESI):m / z calcd for C 27 H 29 O6NaF3:529.1814; found:529.1815[M+Na] + .
[0056] 4c: Yield 24%. 1H NMR(400MHz,)δ9.66(d,J=0.8Hz,1H,CHO),7.36–7.21(m,4H,ArH),6.46(d,J=16.1Hz,1H,H-14),6.22( d,J=16.1Hz,1H,H-13),5.24(s,1H,H-11),4.05(dd,J=6.1,3.4Hz,1H,H-1),2.70(s,1H,H-5),2.08-1.9 0(m,1H),1.78(dddd,J=15.7,9.6,5.4,3.7Hz,1H),1.55–1.47(m,1H),1.45(s,3H,CH3),1.40–1.32(m,1 H),1.29–1.26(m,3H,CH3),1.22(s,9H,C(CH3)3),1.21(s,3H,CH3),1.11(s,3H,CH3),1.08(s,3H,CH3). 13 C NMR (100MHz, CDCl3) δ201.1,171.8,151.2,150.6,134.2,134.1,127.1,126.3,125.4,107.3,90.1,81 .9,81.7,76.5,52.8,51.3,35.0,34.6,31.3,31.2,30.3,28.2,24.3,23.4,20.1,13.7.HRMS(ESI):m / z calcd for C 30 H 38 O5Na:501.2617; found:501.2625[M+Na] + .
[0057] To better understand the essence of this invention, the following pharmacological experimental results demonstrating the inhibitory effect of the tetracyclic foscorine analogue provided by this invention on LPS-induced NO release in RAW264.7 cells illustrate its novel application in the field of anti-inflammatory drug research. The pharmacological examples provide partial activity data for representative compounds. It must be noted that the pharmacological examples of this invention are for illustrative purposes only and not for limiting the invention. Simple modifications to this invention based on its essence are all within the scope of protection claimed by this invention.
[0058] Drug Example 1: Inhibitory activity of compounds 4a-4c, forscorin, and dexamethasone on LPS-induced NO release from RAW264.7 macrophages
[0059] RAW264.7 macrophages were used at a rate of 1×10⁻⁶. 5RAW264.7 macrophages were seeded at a density of cells / mL in 96-well plates and incubated at 37°C in a 5% CO2 incubator for 24 h. The experiment included a control group, an LPS-induced group, a positive control group treated with dexamethasone (DEX), and experimental groups. The positive control and experimental groups were treated with 0.2, 1.0, 5.0, and 25.0 μM of the test compound 4a-4c or DEX, respectively. After 4 h of incubation, 10 μL of culture medium was added to the control group, while LPS (1.0 μg / mL) was added to the other groups to induce cell growth. After 24 h of incubation, the cells were centrifuged, and the supernatant was collected. The NO release was determined using a nitric oxide kit, and the IC50 was calculated using Graphpad 5 software. 50 The effect of the compound at 20 μM on the growth of RAW264.7 macrophages was detected using the MTT assay.
[0060] Table 1. Inhibitory activity of compounds 4a-4c, foscorine, and dexamethasone on LPS-induced NO release from RAW264.7 macrophages.
[0061] compound <![CDATA[IC 50 (μM)]]> Cell viability / % 4a 1.7 99.43±2.10 4b 0.8 99.76±3.11 4c 0.3 99.24±1.46 Fuscorin 25.1 100.33±1.88 DEX 8.6 99.27±2.43
[0062] As shown in Table 1, the tetracyclic analogue of forscorin provided by the present invention has important biological activity. The inhibitory activity test of LPS-induced NO release from RAW 264.7 macrophages showed that the tetracyclic analogue of forscorin with the structure shown in formula (1) has good inhibitory activity on LPS-induced NO release from RAW 264.7 macrophages. In the examples, the inhibitory effect of compounds 4a-4c was significantly better than that of the positive control drugs dexamethasone and forscorin, and there was no obvious cytotoxicity. It is possible to develop it into a new anti-inflammatory drug.
[0063] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.
Claims
1. The application of a tetracyclic analogue of fuscaline with the structure shown in Formula I in the preparation of anti-inflammatory drugs. in, R represents one of phenyl, p-trifluoromethoxyphenyl, p-tert-butylphenyl, p-chlorophenyl, p-methylphenyl, p-methoxyphenyl, p-nitrophenyl, and p-cyanophenyl.
2. The application according to claim 1, characterized in that, The fuscaline tetracyclic analogue has a structure as shown in any one of formulas 4a to 4c:
3. The application according to claim 1, characterized in that, The anti-inflammatory drug is a drug that inhibits pro-inflammatory factors.
4. The application according to claim 3, characterized in that, The pro-inflammatory factors include one or more of tumor necrosis factor, interleukin, prostaglandin, nitric oxide, and reactive oxygen species.
5. The application according to claim 3, characterized in that, The pro-inflammatory factor is nitric oxide.
6. The application as described in any one of claims 1-5, characterized in that, The drug is an oral formulation.
7. The application as described in any one of claims 1-5, characterized in that, The dosage form of the drug is one of the following: tablets, capsules, pills, granules, decoctions, ointments, elixirs, oral liquids, drop pills, and syrups.
8. The application as described in any one of claims 1-5, characterized in that, The drug also includes pharmaceutically acceptable excipients.
9. The application as described in claim 8, characterized in that, The pharmaceutically acceptable excipients include one or more of the following: fruit powder, flavoring, sweetener, acidulant, filler, lubricant, preservative, suspending agent, food coloring, emulsifier, disintegrant, and plasticizer.