Compound rhus chinensis furolactone, preparation method and pharmaceutical composition thereof, and application of compound rhus chinensis furolactone in analgesia and anti-inflammation

Abstract

The invention belongs to the field of medicines, and discloses a rhus chinensis furolactone compound, a preparation method and a pharmaceutical composition thereof, and application of the rhus chinensis furolactone compound in analgesia and anti-inflammation. The rhus chinensis furolactone is shown as a formula (I) and has analgesic and anti-inflammatory effects; the reaction time for the photo-thermal induced mouse pain effect is prolonged; the effect of improving croton oil induced mouse ear swelling is achieved. The rhus chinensis furolactone can relieve and / or treat analgesic and anti-inflammatory related diseases such as rheumatoid arthritis, osteoarthritis, rheumatic arthritis, gouty arthritis, lupus erythematosus syndrome and the like. # imgabs0 #

Description

Technical Field

[0001] The invention relates to the field of medicines, and relates to a compound Rhus chinensis furanolide, a preparation method and a pharmaceutical composition thereof, and uses thereof in analgesia and anti-inflammation. Background Art

[0002] Rhus chinensis Mill. is a plant of the genus Rhus in the family Anacardiaceae. There are about 250 species of this genus in the world, of which 6 species and 3 variants are found in my country. They mostly grow in the shrubs and sparse forests of limestone mountains at an altitude of 350-2300 meters, and are widely distributed throughout the country (except Heilongjiang, Jilin, Liaoning, Xinjiang, and Qinghai). As a medicinal plant, Rhus chinensis Mill. was first recorded in the Kaibao Materia Medica of the Song Dynasty. It is currently included in the National Compilation of Chinese Herbal Medicines and the Chinese Medicine Dictionary. Rhus chinensis Mill. has the effects of clearing away heat and detoxifying, dispersing blood stasis and stopping bleeding. It is often used for colds, fever, bronchitis, coughing and coughing up blood, diarrhea, dysentery, and hemorrhoidal bleeding. Modern pharmacological studies have shown that Rhus chinensis Mill. has antiviral, anti-inflammatory, analgesic, antioxidant, cytotoxic and anti-ulcer pharmacological activities.

[0003] Pain is a protective reaction of the body after being subjected to harmful stimulation, often accompanied by emotional activities such as fear, tension, and anxiety. Pain is also a symptom of certain diseases and can make people feel miserable. In addition to the pain in the senses and the emotional anxiety, severe pain can also lead to physiological dysfunction, insomnia, and even induce shock and endanger life. Modern medicine has listed "pain" as the fifth vital sign after breathing, pulse, blood pressure, and body temperature. Therefore, it is necessary to use analgesics appropriately in clinical practice to relieve severe pain and prevent shock, which is of great significance in the treatment of diseases and trauma rescue.

[0004] Inflammation is a reaction caused by damage to the body. It is a means used by higher animals to fight infection and trauma. It can remove harmful factors and damaged tissues, and then repair tissues to restore them to normal. There are many factors that cause inflammation, such as microorganisms, metabolites of the body, harmful chemicals, abnormal immune responses, dead tissues, etc. The general symptoms of inflammatory response can be summarized as: redness, swelling, heat, pain, and dysfunction. On the one hand, the body's inflammatory response can fight against foreign invasion, and on the other hand, it can also cause damage to itself. If the body's inflammatory response exceeds a certain range, it will cause a variety of diseases. Therefore, the appropriate use of anti-inflammatory drugs is crucial to relieve symptoms and eliminate the cause of the disease.

[0005] The compounds involved in this study are new compounds, which have been found to have analgesic and anti-inflammatory activities. So far, the above research results have not been reported in patents or literature.

[0006] The purpose of the present invention is to make full use of my country's unique plant resources, conduct in-depth research and development, and find compounds with unique chemical structures, strong activity, and small toxic side effects, in order to provide new analgesic and anti-inflammatory drugs for clinical research to relieve and / or treat analgesic and anti-inflammatory related diseases such as rheumatoid arthritis, osteoarthritis, rheumatoid arthritis, gouty arthritis, lupus erythematosus syndrome, etc. Summary of the invention

[0007] The problem solved by the present invention is to provide a compound Rhus chinensis furanolide, a preparation method and a pharmaceutical composition thereof, and the use thereof in analgesia and anti-inflammation. To solve the technical problem of the present invention, the present invention provides the following technical solutions:

[0008] The first aspect of the technical solution of the present invention is to provide a compound Rhus chinensis furanolide and a pharmaceutically acceptable salt thereof, characterized in that: the chemical structural formula is

[0009]

[0010] The second aspect of the technical solution of the present invention is to provide a method for preparing the compound Rhus chinensis furanolide described in the first aspect, characterized in that:

[0011] 1) extracting the dried roots of Rhus chinensis with 95% ethanol-water under reflux for 2 times, each time for 2 hours, combining the extracts, recovering the ethanol under reduced pressure, and obtaining an extract;

[0012] 2) The extract was separated using a Diaion HP-20 macroporous resin column (140×15 cm).

[0013] ion, and then washed with water, 10% ethanol-water, 30% ethanol-water, 50% ethanol-water,

[0014] 70% ethanol-water and 95% ethanol-water were used for elution, and the eluents of each fraction were evaporated to dryness;

[0015] 3) The 95%-water elution fraction of the macroporous resin was subjected to silica gel column chromatography and gradient eluted with petroleum ether-ethyl acetate (98:2-50:50). After combining the fractions, 6 fractions were obtained.

[0016] (Fr.1 to Fr.6), the Fr.6 portion was subjected to silica gel column chromatography, eluted with petroleum ether:ethyl acetate, and then to reverse phase C 18 Semi-preparative column (250 × 9.4 mm, 5 μm;

[0017] YMC-C 18 Column; 12mL / min) for purification, acetonitrile: 0.1% formic acid / water

[0018] (85:15) as the mobile phase, at t R= 21 min to obtain the compound according to claim 1.

[0019] The third aspect of the technical solution of the present invention is to provide a pharmaceutical composition, characterized in that: the pharmaceutical composition contains the compound Rhus furanolide and its pharmaceutically acceptable salt described in the first aspect and a pharmaceutically acceptable carrier or excipient.

[0020] The fourth aspect of the technical solution of the present invention provides the use of the compound Rhus chinensis furanolide and its pharmaceutically acceptable salts described in the first aspect in the preparation of anti-inflammatory or analgesic drugs.

[0021] Beneficial technical effects:

[0022] The compound Rhus chinensis furanolide described in the present invention is a brand-new compound, and experiments have proved that the compound of the present invention has anti-inflammatory and analgesic pharmacological activities. Description of the drawings:

[0023] Figure 1 The structure of the compound Rhus chinensis furanolide

[0024] Figure 2 High-resolution mass spectrum of the compound Rhus chinensis furanolide

[0025] Figure 3 UV spectrum of the compound Rhus chinensis furanolide

[0026] Figure 4 IR spectrum of the compound Rhus chinensis furanolide

[0027] Figure 5 Compound Rhus chinensis furanolide 1 H NMR (400MHz,CDCl3) spectrum

[0028] Figure 6 Compound Rhus chinensis furanolide 13 C NMR (100MHz,CDCl3) spectrum

[0029] Figure 7 HSQC spectrum of the compound Rhus chinensis furanolide

[0030] Figure 8 HMBC spectrum of the compound Rhus chinensis furanolide

[0031] Fig. 9 NOE spectrum of the compound Rhus chinensis furanolide

[0032] Fig.10 Three-dimensional simulation diagram of the compound Rhus chinensis furanolide (a): the double bond C3-C10 configuration is E; (b): the double bond C3-C10 configuration is Z DETAILED DESCRIPTION

[0033] Example: Isolation of the compound Rhus chinensis furanolide

[0034] 20 kg of dried roots of Rhus chinensis were extracted twice with 95% ethanol-water under reflux for 2 hours each time, and the extracts were combined and the ethanol was recovered under reduced pressure to obtain 2.1 kg of extract. The extract was separated using a Diaion HP-20 macroporous resin column (140×15 cm), eluted with water, 10% ethanol-water, 30% ethanol-water, 50% ethanol-water, 70% ethanol-water and 95% ethanol-water, and the eluents of each part were evaporated to dryness. The 95% ethanol-water elution part (320 g) of the macroporous resin was subjected to silica gel column chromatography, eluted with petroleum ether-ethyl acetate (98:2–50:50) gradient, and 6 parts (Fr.1 to Fr.6) were obtained after combining the fractions. The Fr.6 part (7 g) was subjected to silica gel column chromatography, eluted with petroleum ether:ethyl acetate, and then subjected to reverse phase C 18 Preparative column (250×20 mm, 5 μm; YMC-C 18 Column; 12mL / min) for purification, acetonitrile: 0.1% formic acid / water (85:15) as the mobile phase, at t R = 21 min to obtain the compound Rhus chinensis furanolide (3 g).

[0035] The compound Rhus chinensis furanolide is an orange amorphous powder. UVλ max 369 (3.91), 262 (3.56), 237 (3.65) and 204 (4.28) nm, indicating the absorption bands of long conjugated aromatic compounds in the structure. IRν max 3454、3250cm -1 is the hydroxyl absorption peak, 1726cm -1 is the carbonyl absorption peak, 1616cm -1 The absorption peak of benzene ring is 1480cm -1 The presence of a double bond is indicated. According to high-resolution mass spectrometry (-) HR-ESI-MS, the quasi-molecular ion peak m / z 529.2960 [MH] - (calcd.forC 34 H 41 O5,529.2959), the molecular formula is determined to be C 34 H 42 O5, unsaturation is 14.

[0036] 1 H-NMR spectrum (in CDCl3, 400MHz, δppm): There are two meta-coupled benzene ring hydrogen signals in the low field region at δ H 7.77 (1H, d, J = 2.8 Hz, H-5) and δ H6.73 (1H, d, J = 2.8 Hz, H-7), 1 double bond singlet hydrogen signal at δ H 7.31 (1H, s, H-10), two ortho-coupled furan ring hydrogen signals at δ H 6.80 (1H, d, J = 3.2 Hz, H-12) and δ H 6.49 (1H, d, J = 3.2 Hz, H-13), 4 non-conjugated double bond multiplet hydrogen signals at δ H 5.32 (1H, m, H-2′) and δ H 5.09 (3H, m, H-6′, H-10′, H-14′); there is a group of singlet methylene signals in the high field region at δ H 4.80 (2H, s, H-15), a double peak of methylene hydrogen signals at δ H 3.35 (2H, d, J = 7.2 Hz, H-1′), 6 groups of methylene multiplet hydrogen signals at δ H 2.11-1.94 (12H, m, H-4′, H-5′, H-8′, H-9′, H-12′, H-13′), 5 groups of methyl broad singlet hydrogen signals in δ H 1.73(3H,br s,H-19′), 1.66(3H,br s,H-16′), 1.59(3H,br s,H-18′), 1.58(6H,br s,H-17′,H-20′).

[0037] 13 The C-NMR spectrum (in CDCl3, 100MHz, δppm) showed 34 carbon signals, including 21 carbon signals in the low field region: 1 ester carbonyl carbon signal (δ C 170.8), 4 furan ring carbon signals (δ C 157.7, 151.5, 121.6, 112.0), 6 benzene ring carbon signals (δ C 152.5, 146.5, 125.7, 122.2, 118.1, 109.1), 10 double bond carbon signals (δ C 137.7, 135.3, 135.0, 131.4, 124.5, 124.3, 124.1, 122.6, 120.7, 118.5); there are 8 methylene carbon signals in the high field region (δ C 57.5, 39.8×3, 27.7, 26.9, 26.8, 26.7,) and 5 methyl carbon signals (δ C 25.8, 17.8, 16.4, 16.2, 16.1).

[0038] The above signals are basically consistent with those of the known compound cannabifolactone A, with the only difference being that an additional geranylgeranyl fragment is attached to C-8 of the compound Rhus chinensis furanolide.

[0039] The corresponding relationship between carbon and hydrogen was determined by correlation analysis of HSQC spectra.

[0040] HMBC spectrum shows that H-1′(δ H 3.35) and C-9 (δ C 146.5), C-3′(δ C 137.7) and C-7 (δ C 118.1), indicating that the geranylgeranyl group is connected to the C-8 position of the B ring; H-10 (δ H 7.31) and C-2(δ C 170.8), C-4(δ C 122.2) and C-12 (δ C 121.6), indicating that the 2-hydroxymethylfuranyl group is connected to the C-3 position of the A ring via a carbon-carbon double bond through C-10.

[0041] NOE spectrum shows that irradiated H-5(δ H 7.77), then H2-15(δ H 4.80) has a gain, and similarly, irradiation of H2-15 (δ H 4.80), then H-5(δ H 7.77) also has a gain. In the ChemDraw 3D simulation diagram, when the double bond configuration is E, the distance between H-5 and H2-15 is There is NOE; when the double bond configuration is Z, the distance between H-5 and H2-15 is No NOE. Therefore, it can be determined that the double bond C3-C10 is of E configuration, which is consistent with the double bond configuration of the known compound cannabifolactone A.

[0042] In summary, the compound was determined to be Rhus chinensis furanolide.

[0043] Table 1 Compound Rhus chinensis furanolide 1 H NMR and 13 C NMR data

[0044]

[0045] Pharmacological experiments

[0046] Experimental Example 1: Effect of the compound Rhus chinensis furanolide on photothermal-induced pain in mice

[0047] Experimental method: Kunming mice, male, weighing 18-23g, compound Rhus furanolide was fully dissolved in physiological saline, and subcutaneous injection was given to mice in the administration group at a dose of 50mg / kg, 100mg / kg, and 200mg / kg, and the blank control group was given an equal dose of physiological saline. Analgesia experiment was performed with a photosensitive analgesic instrument 1 hour after administration, and the pain reaction time of mice was recorded (seconds) according to the tail-flick reaction of mice, and the extension rate of pain reaction time was calculated: extension rate (%) = (average tail-flick time of the administration group-average tail-flick time of the blank control group) / average tail-flick time of the administration group × 100%.

[0048] Experimental results:

[0049] The results showed that the compound Rhus chinensis furanolide had a significant analgesic effect in vivo: the pain time extension rate of the compound Rhus chinensis furanolide administered subcutaneously at 200 mg / kg was 48.55% compared with the blank control group, indicating that the compound can significantly prolong the pain reaction time of mice (p < 0.05) and has significant analgesic activity in vivo (as shown in Table 1).

[0050] Table 1. Effects of the compound Rhus chinensis furanolide on photothermal-induced pain response in mice

[0051]

[0052] Experimental Example 2: Effect of the compound Rhus chinensis furanolide on croton oil-induced otitis in mice

[0053] Experimental method: Kunming mice, male, weighing 18-23g, compound Rhus furanolide was fully dissolved in physiological saline, and subcutaneous injection was given to mice in the administration group at a dose of 50mg / kg and 100mg / kg, and the blank control group was given an equal dose of physiological saline. One hour after administration, 50μL of 4% croton oil solution was prepared and applied to one ear of the animal to model otitis. The mice were killed by dislocation 4 hours later, the two ears were cut along the auricle, and a round ear piece was punched at the same position (along the ear edge) of the left and right ears with a puncher with a diameter of 8mm. The weight difference between the left ear piece and the right ear weight (mg) was weighed and recorded. Calculate the ear swelling inhibition rate: ear swelling inhibition rate (%) = (average ear swelling degree of the model group - average ear swelling degree of the administration group) / average ear swelling degree of the model group × 100%.

[0054] Experimental results:

[0055] The results showed that the compound Rhus chinensis furanolide had a significant effect on improving ear swelling in mice with croton oil-induced otitis model: the ear swelling inhibition rate of 100 mg / kg of the compound Rhus chinensis furanolide subcutaneously injected was 44.03% compared with the blank control group, indicating that the compound can significantly improve the degree of ear swelling in the croton oil-induced otitis model in mice (p < 0.05) and has significant in vivo anti-inflammatory activity (as shown in Table 2).

[0056] Table 2. Inhibition of ear swelling in croton oil-induced mouse otitis model by compound Rhus chinensis furanolide

[0057]

[0058] In summary, the compound Rhus chinensis furanolide can significantly inhibit the photothermal-induced pain response in mice and the croton oil-induced otitis in mice, indicating that it has significant analgesic and anti-inflammatory activity.

Claims

1. A compound as shown in formula (I) and a pharmaceutically acceptable salt thereof, characterized in that:

2. The method for preparing the compound according to claim 1, characterized in that: 1) extracting the dried roots of Rhus chinensis with 95% ethanol-water under reflux for 2 times, each time for 2 hours, combining the extracts, recovering the ethanol under reduced pressure, and obtaining an extract; 2) Separating the extract using a Diaion HP-20 macroporous resin column (140×15 cm), eluting with water, 10% ethanol-water, 30% ethanol-water, 50% ethanol-water, 70% ethanol-water and 95% ethanol-water, and evaporating the eluents from each part to dryness; 3) The 95% ethanol-water elution fraction of the macroporous resin was subjected to silica gel column chromatography and gradient eluted with petroleum ether-ethyl acetate (98:2–50:50). After combining the fractions, 6 fractions (Fr.1 to Fr.6) were obtained. The Fr.6 fraction was subjected to silica gel column chromatography and eluted with petroleum ether:ethyl acetate and then subjected to reverse phase C 18 Semi-preparative column (250×9.4mm, 5μm; YMC-C 18 Column; 12mL / min) for purification, acetonitrile: 0.1% formic acid / water (85:15) as the mobile phase, at t R = 21 min to obtain the compound according to claim 1.

3. A pharmaceutical composition, characterized in that: The pharmaceutical composition contains the compound according to claim 1 and a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

4. Use of the compound according to claim 1 and its pharmaceutically acceptable salt in the preparation of anti-inflammatory or analgesic drugs.

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