Phenylpropanoid compound and preparation method and use thereof

Abstract

The present invention provides a phenylpropanoid compound and a preparation method and use thereof. The phenylpropanoid compound has a structural formula shown as formula (I), and the pharmaceutically acceptable salt has a structural formula shown as formula (II), formula (III), formula (IV), or formula (V). The phenylpropanoid compound and the pharmaceutically acceptable salt thereof have an effect on inhibiting a content of an inflammatory cytokine NO and an expression of an inflammatory cytokine TNF-, have an effect on inhibiting a hydroxyl radical (OH), thereby have anti-inflammatory and antioxidant activities, and has a good application prospect in preparing of a medicine for treating an inflammatory disease related to the above factors, such as cervicitis, endometritis, pelvic inflammatory disease, mastitis, sphagitis and/or arthritis. ##STR00001##

Claims

1. A pharmaceutically acceptable salt of a phenylpropanoid compound, wherein the pharmaceutically acceptable salt has a structural formula shown as formula (II), formula (III), formula (IV), or formula (V): ##STR00006## wherein the R is an inorganic acid.

2. The pharmaceutically acceptable salt of the phenylpropanoid compound according to claim 1, wherein the inorganic acid is selected from a group consisting of hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid and phosphoric acid.

3. The pharmaceutically acceptable salt of the phenylpropanoid compound according to claim 1, wherein the pharmaceutically acceptable salt is an ammonium salt.

4. A pharmaceutical composition, wherein the pharmaceutical composition contains the pharmaceutically acceptable salt of the phenylpropanoid compound according to claim 1.

5. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition further contains a pharmaceutically permissible supplementary material and/or carrier.

6. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition further contains one or more of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, or an extract thereof.

7. The pharmaceutical composition according to claim 4, wherein the pharmaceutical composition has a form selected from a group consisting of a tablet, a capsule, a powder, a granule, a pill, a solution, a suspension, a syrup, an injection, an ointment, a suppository and a spray.

8. A preparation method of a phenylpropanoid compound, wherein the preparation method comprises steps as follows: S1, taking a root of Moghania macrophylla (Willd.) O. Kuntze as a raw material, drying, stripping and slicing, extracting with an ethanol solution to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use; S2, dissolving the extractum obtained in the step S1 with water, eluting the dissolved extractum by a macroporous adsorption resin column, with an eluent being an ethanol-water system, collecting first three column volumes of an eluate, naming the eluate as MM-1, for future use; S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use; S4, separating the fraction MM-12 collected in the step S3 by a preparative liquid phase, with a mobile phase being a methanol-water-acetic acid system, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a mobile phase being the methanol-water-acetic acid system, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing, wherein the phenylpropanoid compound has a structural formula shown as formula (I): ##STR00007##

9. The preparation method according to claim 8, wherein in the step S1, a concentration of the ethanol solution is 50 to 80% by volume.

10. The preparation method according to claim 8, wherein in the step S1, extracting with ethanol performs 2 to 4 times, and each time is 1 to 3 hours.

11. The preparation method according to claim 8, wherein in the step S2, a volume ratio of ethanol to water is 0:100 to 15:85.

12. The preparation method according to claim 8, wherein in the step S3, for the eluent, a volume ratio of methanol to water is 20:80 to 30:70.

13. The preparation method according to claim 8, wherein in the step S4, a volume ratio of methanol to water to acetic acid is 10:90:0.01 to 35:65:0.01.

14. The preparation method according to claim 8, wherein in the step S4 and/or the step S5, a chromatographic column of the preparative liquid phase is YMC, 20 mm*250 mm, and a flow rate of the mobile phase is 5 to 10 mL/min.

15. The preparation method according to claim 8, wherein in the step S5, a volume ratio of methanol to water to acetic acid is 15:85:0.01.

16. A method of using a phenylpropanoid compound and a pharmaceutically acceptable salt thereof for treating an inflammatory disease, the method comprises: preparing a medicine by using the phenylpropanoid compound or the pharmaceutically acceptable salt thereof, wherein the phenylpropanoid compound has a structural formula shown as formula (I), and the pharmaceutically acceptable salt has a structural formula shown as formula (II), formula (III), formula (IV), or formula (V): ##STR00008## wherein the R is an inorganic acid; and administering the medicine for treating the inflammatory disease.

17. The method according to claim 16, wherein the medicine is for inhibiting a content of an inflammatory cytokine NO or inhibiting an expression of an inflammatory cytokine TNF- or inhibiting an activity of a hydroxyl radical.

18. The method according to claim 16, wherein the inflammatory disease is cervicitis, endometritis, pelvic inflammatory disease, mastitis, sphagitis and/or arthritis.

19. The method according to claim 16, wherein the medicine contains a pharmaceutically permissible supplementary material and/or carrier.

20. The method according to claim 16, wherein the medicine further contains one or more of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, or an extract thereof.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

(1) FIG. 1 shows an effect diagram of the phenylpropanoid compound of the present invention on cell viability.

(2) FIG. 2 shows an inhibition effect diagram of the phenylpropanoid compound of the present invention on NO.

(3) FIG. 3 shows an inhibition effect diagram of the phenylpropanoid compound of the present invention on TNF-.

(4) FIG. 4 shows an inhibition effect diagram of the phenylpropanoid compound of the present invention on IL-1.

(5) FIG. 5 shows an inhibition effect diagram of the phenylpropanoid compound of the present invention on IL-6.

(6) FIG. 6 shows an inhibition effect diagram of the phenylpropanoid compound of the present invention on OH.

DESCRIPTION OF THE EMBODIMENTS

(7) The present invention is further described below in combination with the accompanied drawings and specific embodiments, but the embodiments do not limit the present invention in any forms. Unless specified, reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment used in the art. Unless specified, raw materials and equipment used in the present invention are commercially available conventional raw materials and equipment used in the art.

(8) The compounds of the present invention are a phenylpropanoid compound shown as formula (I) and a pharmaceutically acceptable salt thereof shown as formula (II) or formula (III). The compounds can be prepared by a method of extracting a raw material Moghania macrophylla (Willd.) O. Kuntze, which the method is provided by the present invention, and also can be prepared by methods such as in combination with chemical synthesis in the art according to a structural formula provided by the present invention.

(9) As the salts of the phenylpropanoid compound of the present invention, they only need to be the pharmaceutically acceptable salts, listed as inorganic salts generated by the phenylpropanoid compound with inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid and the like; sulfonate generated by the phenylpropanoid compound with sulfonic acid; alkali metal salts generated by the phenylpropanoid compound with hydroxides of alkali metals such as potassium, sodium, calcium, magnesium, lithium and the like; and ammonium salts generated by the phenylpropanoid compound with ammoniums.

(10) The phenylpropanoid compound of the present invention can be used for a medicine for treating inflammatory diseases such as cervicitis, endometritis, pelvic inflammatory disease, mastitis, sphagitis and/or arthritis and the like.

(11) The compound of the present invention can be used as a pharmaceutical composition with a pharmaceutically permissible supplementary material and/or carrier, and also can be used as a pharmaceutical composition with a group of one or more of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, or an extract thereof under the condition of adding the pharmaceutically permissible supplementary material and/or carrier. The compound of the present invention can also be used as a pharmaceutical composition with other pharmaceutically permissible medicinal ingredients.

(12) As the pharmaceutical composition, it can have a form such as a tablet, a capsule, a powder, a granule, a pill, a solution, a suspension, a syrup, an injection, an ointment, a suppository, a spray and the like.

(13) Further, the tablet can be prepared to a sugarcoated tablet, a film-coated tablet, an enteric coated tablet or a double-layer tablet or a multi-layer tablet, under the condition of adding the pharmaceutically permissible supplementary material and/or carrier.

(14) The supplementary material and/or carrier of the present invention can be prepared into follows:

(15) a solid formulation, that is, an additive can be used, for example, sucrose, lactose, cellulosic saccharide, maltitol, glucose, starches, agar, alginates, chitin, chitosans, pectins, arabic gums, gelatins, collagens, casein, albumin, calcium phosphate, sorbitol, glycine, glycerol, polyethylene glycol, sodium bicarbonate, talc and the like;

(16) a semisolid formulation, that is, animal and vegetable oils (olive oil, corn oil, castor oil, etc.), mineral oils (Vaseline, white petrolatum, solid paraffin, etc.), waxes (jojoba oil, carnauba wax, beeswax, etc.), partially synthetic or fully synthetic glycerol fatty acid esters (lauric acid, myristic acid, palmitic acid, etc.) and the like can be used;

(17) a liquid formulation, that is, an additive can be used, for example, sodium chloride, glucose, sorbitol, glycerol, olive oil, propylene glycol, ethanol and the like. Especially under the condition of preparing into an injection, that is, sterile aqueous solution can be used, for example, physiological saline, isotonic solution and oily liquid, such as sesame oil and soybean oil. Besides, a suitable suspending agent can be used simultaneously according to needs, for example, sodium carboxymethyl cellulose, nonionic surfactant and cosolvent, such as benzyl benzoate and benzyl alcohol.

(18) A content of active ingredients in these formulations is 0.01 to 80 wt % of the formulation, suitably 1 to 50 wt %, and a dosage varies according to differences in symptom, body weight, age of a patient.

Example 1

(19) Preparation of a Phenylpropanoid Compound

(20) The present example provides a preparation method of the phenylpropanoid compound shown as formula (I), comprising steps as follows:

(21) S1, taking 50 kg of Moghania macrophylla (Willd.) O. Kuntze, with a root as a raw material, drying, stripping and slicing into small pieces, carrying out a reflux extraction with 8 multiples of 60% ethanol in 3 times while 2 hours for each time to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use;

(22) S2, dissolving the extractum after concentration obtained in the step S1 with 10 L of water, eluting the dissolved extractum by D101 macroporous adsorption resin column, with an eluent being water for three column volumes, collecting an eluate, naming the eluate as MM-1, for future use;

(23) S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system in a volume ratio of 25:75, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use; S4, separating the fraction MM-12 collected in the step S3 by a preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being a methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 25:75:0.01, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and

(24) S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being the methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 15:85:0.01, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing.

Example 2

(25) Preparation of a Phenylpropanoid Compound

(26) The present example provides a preparation method of the phenylpropanoid compound shown as formula (I), comprising steps as follows:

(27) S1, taking 40 kg of Moghania macrophylla (Willd.) O. Kuntze, with a root as a raw material, drying, stripping and slicing into small pieces, carrying out a reflux extraction with 6 multiples of 50% ethanol in 2 times while 1 hour for each time to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use;

(28) S2, dissolving the extractum after concentration obtained in the step S1 with 5 L of water, eluting the dissolved extractum by D101 macroporous adsorption resin column, with an eluent being ethanol and water in a volume ratio of 15:85 for three column volumes, collecting an eluate, naming the eluate as MM-1, for future use;

(29) S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system in a volume ratio of 25:80, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use;

(30) S4, separating the fraction MM-12 collected in the step S3 by a preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 10 ml/min, a mobile phase being a methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 35:65:0.01, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and

(31) S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being the methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 15:85:0.01, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing.

Example 3

(32) Preparation of a Phenylpropanoid Compound

(33) The present example provides a preparation method of the phenylpropanoid compound shown as formula (I), comprising steps as follows:

(34) S1, taking 60 kg of Moghania macrophylla (Willd.) O. Kuntze, with a root as a raw material, drying, stripping and slicing into small pieces, carrying out a reflux extraction with 7 multiples of 70% ethanol in 4 times while 3 hours for each time to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use;

(35) S2, dissolving the extractum after concentration obtained in the step S1 with 8 L of water, eluting the dissolved extractum by D101 macroporous adsorption resin column, with an eluent being ethanol and water in a volume ratio of 10:90 for three column volumes, collecting an eluate, naming the eluate as MM-1, for future use;

(36) S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system in a volume ratio of 30:70, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use;

(37) S4, separating the faction MM-12 collected in the step S3 by a preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 10 ml/min, a mobile phase being a methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 30:70:0.01, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and

(38) S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being the methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 15:85:0.01, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing.

Example 4

(39) Preparation of a Phenylpropanoid Compound

(40) The present example provides a preparation method of the phenylpropanoid compound shown as formula (I), comprising steps as follows:

(41) S1, taking 50 kg of Moghania macrophylla (Willd.) O. Kuntze, with a root as a raw material, drying, stripping and slicing into small pieces, carrying out a reflux extraction with 8 multiples of 60% ethanol in 2 times while 1.5 hours for each time to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use;

(42) S2, dissolving the extractum after concentration obtained in the step S1 with 6 L of water, eluting the dissolved extractum by D101 macroporous adsorption resin column, with an eluent being ethanol and water in a volume ratio of 5:95 for three column volumes, collecting an eluate, naming the eluate as MM-1, for future use;

(43) S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system in a volume ratio of 25:75, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use;

(44) S4, separating the fraction MM-12 collected in the step S3 by a preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 10 ml/min, a mobile phase being a methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 25:75:0.01, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and

(45) S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being the methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 15:85:0.01, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing.

Example 5

(46) Preparation of a Phenylpropanoid Compound

(47) The present example provides a preparation method of the phenylpropanoid compound shown as formula (I), comprising steps as follows:

(48) S1, taking 50 kg of Moghania macrophylla (Willd.) O. Kuntze, with a root as a raw material, drying, stripping and slicing into small pieces, carrying out a reflux extraction with 8 multiples of 80% ethanol in 2 times while 1.5 hours for each time to obtain an extracted solution, merging the extracted solution, concentrating till no alcohol taste, obtaining an extractum for future use;

(49) S2, dissolving the extractum after concentration obtained in the step S1 with 6 L of water, eluting the dissolved extractum by D101 macroporous adsorption resin column, with an eluent being ethanol and water in a volume ratio of 10:90 for three column volumes, collecting an eluate, naming the eluate as MM-1, for future use;

(50) S3, eluting a fraction MM-1 collected in the step S2 by a reverse phase material ODS column chromatography, with an eluent being the ethanol-water system in a volume ratio of 28:72, eluting for 18 column volumes, collecting one fraction of eluate per three column volumes, thereby collecting 6 fractions in order, naming the 6 fractions as MM-11, MM-12, MM-13, MM-14, MM-15, MM-16 respectively, for future use;

(51) S4, separating the fraction MM-12 collected in the step S3 by a preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 10 ml/min, a mobile phase being a methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 10:90:0.01, collecting eluates in peak-order, collecting 7 fractions in total, naming the 7 fractions as MM-121, MM-122, MM-123, MM-124, MM-125, MM-126, MM-127 respectively, for future use; and

(52) S5, purifying the fraction MM-125 collected in the step S4 by the preparative liquid phase, with a chromatographic column for the preparative liquid phase being YMC of 20 mm*250 mm with a velocity of 5 ml/min, a mobile phase being the methanol-water-acetic acid system, and a volume ratio of methanol-water-acetic acid being 15:85:0.01, collecting an eluate, obtaining the phenylpropanoid compound after recrystallizing.

(53) The compounds obtained from Examples 1 to 5 were carried out tests of MS, H-NMR and C-NMR. The results verify that the obtained compounds are 3,5-dihydroxy-4-glucosyl-phenyl-3-hydroxypropanoic acid methyl ester, with a structural formula shown as formula (I):

(54) ##STR00004##

(55) Spectrogram data of MS, H-NMR and C-NMR thereof is as follows:

(56) HR-ESIMS shows [M+Na]+ as m/z 399.1889, and in combination with nuclear magnetic characteristics, a molecular formula was obtained as C.sub.15H.sub.20O.sub.11 with an unsaturation degree being 6.

(57) .sup.1H-NMR (600 MHz, CD.sub.3OD): 6.60 (d, 1H), 6.47 (d, 1H), 4.95 (s, 1H), 4.94 (m, 2H), 3.00-4.00 (glc-H), 2.71 (m, 2H).

(58) .sup.13C-NMR (150 MHz, CD.sub.3OD): 170.5 (C-1), 150.7 (C-1), 150.2 (C-4), 135.5 (C-3,5), 114.7 (C-2,6), 101.2 (C-1), 61.5-77.8 (C2-C6), 61.1 (C-2), 38.69 (C-3).

Example 6

(59) Preparation of a Salt of a the Phenylpropanoid Compound

(60) Preparation of Hydrochlorides of the Phenylpropanoid Compound:

(61) Under stirring, a methanol solution of the compound was dripped with saturated hydrochloric acid to reach a pH value of 2 to 3, and then added with acetonitrile. White powder solid was obtained by filtration and drying, that is, hydrochloric acid salt of the compound.

(62) Preparation of Sulfonates of the Phenylpropanoid Compound:

(63) Alkali metal hydroxides, solvent, sulfuric acid, neutral oil, and accelerator were added to a reaction system containing the phenylpropanoid compound. Solvent, lower alcohol, and accelerator, and carbon dioxide were introduced into the reaction system to separate the white powder solid, i.e. sulfonates of the compound.

(64) Preparation of Potassium or Sodium Salts of the Phenylpropanoid Compound:

(65) Under stirring, KOH or NaOH dissolved in ethanol was added to the compound, heated to reflux under stirring, cooled to room temperature, dripped with acetonitrile, and then filtered and dried to white solid, i.e. potassium or sodium salt of the compound.

(66) Preparation of Ammonium Salts of the Phenylpropanoid Compound:

(67) Under stirring, the compound in methanol solution was dripped with saturated ammonia to reach a pH value of 9 to 11, stirred with acetonitrile, and then filtered and dried to white solid, i.e. ammonium salts of the phenylpropanoid compound.

(68) Spectrogram Data of the Salt of the Compound Mentioned Above:

(69) The hydrochlorides of the phenylpropanoid compound: ESIMS displays m/z 412.67, .sup.1H-NMR (600 MHz, CD.sub.3OD): 6.60 (s, 2H), 6.47 (m, 1H), 5.48 (s, 1H), 5.35 (s, 1H), 4.88 to 4.95 (m, 3H), 4.41 (s, 1H), 3.33 to 3.50 (m, 6H), 2.89 (s, 1H), 2.70 (m, 2H), 1.24 (m, 2H).

(70) Potassium or Sodium Salts of the Phenylpropanoid Compound:

(71) The potassium salts of the phenylpropanoid compound: ESIMS displays m/z 452.19, .sup.1H-NMR (600 MHz, CD.sub.3OD): 6.62 (s, 2H), 4.86 to 4.97 (m, 3H), 4.42 (s, 1H), 3.33 to 3.51 (m, 6H), 2.90 (s, 1H), 2.71 (m, 2H), 1.24 (m, 2H).

(72) The sodium salts of the phenylpropanoid compound: ESIMS displays m/z 420.06, .sup.1H-NMR (600 MHz, CD.sub.3OD): 6.62 (s, 2H), 6.47 (m, 1H), 4.86 to 4.97 (m, 3H), 4.42 (s, 1H), 3.33 to 3.51 (m, 6H), 2.90 (s, 1H), 2.71 (m, 2H), 1.24 (m, 2H).

(73) Ammonium salts of the phenylpropanoid compound: ESIMS displays m/z 410.15, .sup.1H-NMR (600 MHz, CD.sub.3OD): 6.60 (s, 2H), 6.47 (m, 1H), 5.48 (s, 1H), 5.35 (s, 1H), 4.88 to 4.95 (m, 3H), 4.41 (s, 1H), 3.33 to 3.50 (m, 6H), 2.89 (s, 1H), 2.70 (m, 2H), 1.24 (m, 2H).

(74) The structural formula of the phenylpropanoid compounds mentioned above is shown in Formulas (III) to (VII).

(75) ##STR00005##

(76) Wherein, Formula (III) is one of the prepared potassium salts of the phenylpropanoid compound, Formula (IV) is one of the prepared sodium salts of the phenylpropanoid compound, Formula (V) is one of the ammonium salts of the phenylpropanoid compound, Formula (VI) is the hydrochloride of the phenylpropanoid compound, and Formula (VII) is one of the sulfonates of the phenylpropanoid compound.

Example 7

(77) Application Experiments

(78) The compound and salts of present invention have effects on LPS-induced oxidative stress and inflammation of RAW 264.7 macrophages. (For convenience of recording during experiment, the phenylpropanoid compound described in present invention is labelled as: MM-125, that is, MM-125 described in present invention refers to the phenylpropanoid compound shown in formula (I) of present invention or their pharmaceutically acceptable salts.)

(79) 1 Materials and Methods

(80) 1.1 Medicine and Devices

(81) Lipopolysaccharide (LPS), MTT purchased from Sigma, Raw 264.7 mouse macrophage purchased from Xiangya cell bank; PBS; DMEM high glucose medium, fetal bovine serum, penicillin and streptomycin, automatic enzyme labelling instrument, constant temperature CO.sub.2 incubator.

(82) Mouse IL-1-beta ELISA kit, batch number: 2014/06 (96T), Mouse IL-6 ELISA kit, batch number: 2014/06 (96T), Mouse tumor necrosis factor-alpha (TNF-) ELISA kit, batch number: 2014/06 (96T), Mouse NO ELISA Kit Batch number: 2014/10 (96T), mouse hydroxyl radical (OH) ELISA detection kit, batch number: 2014/10 (96T).

(83) 1.2 Preparation of Medicine

(84) Medicine was first dissolved in a small amount of DMSO, and then diluted to a certain concentration with DMEM, so that final concentration of DMSO was less than 1%.

(85) 1.3 Cell Culture

(86) Raw 264.7 mouse macrophages were incubated in DMEM medium containing 10% heat-inactivated fetal bovine serum (FBS), 10 U/mL penicillin sodium and 100 g/ml streptomycin, and incubated in 37 C., 5% CO.sub.2 incubator.

(87) 1.4 Determination of Cell Vitality

(88) Cell vitality was determined by MTT method. Cell suspension liquid was inoculated into a 96-well plate (1*104 holes/pore) to incubate for 24 hours and then synchronize for 24 hours. The cells were treated with different concentration of medicine for 2 hours, and then stimulated by LPS (30 g/ml) for 24 hours. Original medium was abandoned, and MTT (0.5 mg/ml) was added into each hole for 4 hours. The medium was then abandoned and DMSO (150 g/ml) was added into each hole. Absorbance was measured at 490 nm after shaking for 10 min.

(89) 1.5 Determination of NO Concentration

(90) Raw 264.7 cells were inoculated on a 96-well plate for 24 hours, then synchronized for 24 hours. The cells were treated with different concentration of medicine for 2 hours, and then stimulated with LPS (30 g/ml) for 24 hours. Lastly, supernatant was collected and centrifuged for 5 minutes at 10,000 rpm. The supernatant was separated and stored at 80 C. for reserve. The concentration of NO was determined by mouse NO kit.

(91) 1.6 Measurement of Inflammatory Factors TNF-, IL-1, IL-6

(92) Sample prepared in at 1.5 was used for subsequent inflammatory factor determination. The amount of TNF-, IL-1, IL-6 produced by cells was measured by mouse TNF-, IL-1, and IL-6 kit.

(93) 1.7 Determination of OH Concentration

(94) Sample prepared at 1.5 was used for OH factor determination. The concentration was determined by OH kit.

(95) 1.8 Statistical Analysis

(96) SPSS17.0 software was used, and experiment data was expressed as xs. The data obtained was analysed by one-way ANOVA. Homogeneity of variance is tested by LSD, and Dunnett T3 was used for variant homogeneity.

(97) 2 Experimental Results

(98) 2.1 Cell Vitality

(99) The effect of medicine on cell vitality was evaluated by MTT method. As shown in FIG. 1, MM-125 had no significant effect on vitality of Raw 264.7 cells in a range of 1.37 to 12.37 g/ml. Therefore, concentration of MM-125 in this range was suitable for subsequent experiments.

(100) 2.2 Medicine Inhibits NO Production

(101) As shown in FIG. 2, NO production (48.661.66 IU/mL) of Raw 264.7 cells stimulated by LPS was significantly higher (p<0.01) than that of normal Raw 264.7 cells (29.050.85 IU/ml). In a range of 6.8812.38 g/ml, MM-125 could significantly inhibit the increase of NO concentration induced by LPS in a dose-dependent manner.

(102) 2.3 Medicine Inhibits Production of TNF-, IL-1, and IL-6

(103) As shown in FIGS. 3 to 5, concentrations of inflammatory cytokines TNF- (127.984.65 pg/ml), IL-1 (347.559.56 pg/ml), and IL-6 (201.6710.32 pg/ml) were significantly higher than those in normal controls (p<0.01): TNF- (67.124.00 pg/ml), IL-1 (160.88113.10 pg/ml), and IL-6 (111.138.06 pg/ml), which indicates that LPS could stimulate Raw 264.7 cells to produce a large number of inflammatory factors.

(104) In a range of 6.8812.38 g/ml, MM-125 could significantly inhibit production of TNF- (p<0.05) in Raw 264.7 cells in a dose-dependent manner, but could not inhibit production of IL-1 or IL-6 (p>0.05) at various concentrations.

(105) 2.4 Medicine Inhibits OH Production

(106) As shown in FIG. 6, Raw 264.7 cells stimulated by LPS produced higher OH concentration (106.003.90 ng/ml) than normal controls (64.622.18 ng/ml). (p<0.01)

(107) In a range of 4.13-12.38 g/ml, MM-125 could significantly inhibit the increase of OH concentration induced by LPS in a dose-dependent manner.

(108) Effects of MM-125 on production of NO, TNF-, IL-1, IL-6, and OH in mouse macrophages were studied in vitro.

(109) MM-125 has significant inhibitory effects on production of cytokine NO and also certain level of inhibiting effects on TNF-alpha, but it has no obvious inhibitory effect on IL-1 or IL-6, indicating that MM-125 has certain anti-inflammatory capability. It has significant inhibitory effects on production of OH, indicating that it has better antioxidant capability.

Example 8

(110) Preparation of tablets: the phenylpropanoid compound shown in Formula (I) was prepared according to the method in Example 1, using salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium. Tablets are pelleted and tableted by adding excipients in a proportion of 1:10 weight ratio (of the compound or any of its salts to the excipient).

Example 9

(111) Preparation of powder: the phenylpropanoid compound shown in Formula (I) was prepared according to the method in Example 1, using salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium. The compound or its salts was made into powder using conventional methods.

Example 10

(112) Preparation of capsules or granules: the phenylpropanoid compound shown in Formula (I) was prepared according to the method in Example 1, using salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium. Excipients were added to the compound or any of its salts in a weight ratio of 1:10 (of the compound or any of its salts to the excipient) to make capsules or granules.

Example 11

(113) Preparation of injections: the phenylpropanoid compound shown in Formula (I) was prepared according to the method in Example 1, using salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium. Injection was made by conventional water injection, fine filtration, encapsulation and sterilization.

Example 12

(114) A pharmaceutical composition, containing a phenylpropanoid compound shown in Formula (I) prepared according to the method in Example 1, and salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium, and powder made of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, and supplementary materials.

Example 13

(115) A pharmaceutical composition, containing a phenylpropanoid compound shown in Formula (I) prepared according to the method in Example 1, and powder made of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, and supplementary materials.

Example 14

(116) A pharmaceutical composition, containing a phenylpropanoid compound shown in Formula (I) prepared according to the method in Example 1, and extracts of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, and supplementary materials. The extract was prepared according to methods described in one or more patent documents among CN1078079C, CN1170549C, CN1158087C, CN1330335C, CN1296071C, CN1321631C, CN1296072C, CN1296073C.

Example 15

(117) A pharmaceutical composition, containing a phenylpropanoid compound shown in Formula (I) prepared according to the method in Example 1, and salts made of the compound with inorganic acids (e.g. hydrochloric acid, hydrobromic acid, hydrofluoric acid, hydroiodic acid, sulfuric acid, nitric acid, carboxylic acid, phosphoric acid, lactic acid) or sulfonic acid or hydroxide salts of alkali metals (e.g. potassium hydroxide, sodium hydroxide, calcium hydroxide, magnesium hydroxide, lithium hydroxide) or ammonium, and extracts of Radix Rosa Laevigata, Zanthoxylum dissitum Hemsl., Caulis Spatholobi, Caulis Mahoniae, Herba Andrographis, Radix Angelicae Sinensis, Radix Codonopsis, and supplementary materials. The extract was prepared according to methods described in one or more patent documents among CN1078079C, CN1170549C, CN1158087C, CN1330335C, CN1296071C, CN1321631C, CN1296072C, CN1296073C.

(118) The above shows and describes basic principles and main features and advantages of present invention. It should be understood by those skilled in the art that present invention is not limited by embodiments and specifications mentioned above, which describe only the principles of present invention, and that present invention will undergo various changes and improvements without departing from the spirit and scope of present invention, which is obvious to those skilled in the art. These changes and improvements fall into the scope of the requirement to protect the invention. The invention requires that the scope of protection be defined by appended claims and their equivalents.