CN115403658B - Ursolic acid-Tyr-Gly-Phe-Gly-Gly, synthesis, activity and application thereof - Google Patents

Ursolic acid-Tyr-Gly-Phe-Gly-Gly, synthesis, activity and application thereof Download PDF

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CN115403658B
CN115403658B CN202211069067.1A CN202211069067A CN115403658B CN 115403658 B CN115403658 B CN 115403658B CN 202211069067 A CN202211069067 A CN 202211069067A CN 115403658 B CN115403658 B CN 115403658B
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赵明
张筱宜
冯琦琦
原笑笑
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Abstract

The invention discloses an ursolic acid-Tyr-Gly-Phe-Gly-Gly and preparation and application thereof, wherein the ursolic acid-Tyr-Gly-Phe-Gly-Gly has a structure shown in a general formula I. In addition, the invention also discloses a preparation method of the compound, and an anti-osteoporosis activity and application of the compound in preparation of anti-osteoporosis medicines.

Description

Ursolic acid-Tyr-Gly-Phe-Gly-Gly, synthesis, activity and application thereof
Technical Field
The invention relates to an ursolic acid-Tyr-Gly-Phe-Gly-Gly and a preparation method thereof, and also relates to an anti-osteoporosis activity and application thereof in preparation of anti-osteoporosis drugs. The invention belongs to the field of biological medicine.
Background
Ursolic acid is a triterpene existing in leaves, fruits and pericarps of medicinal plants such as fructus Ligustri Lucidi and Prunellae Spica. The main interest in ursolic acid research in the published literature is anti-tumor. Some ursolic acid has recently been studied for treating osteoporosis. Based on the association of tumor growth and bone loss and the structure of ursolic acid being like estradiol, the inventor connects ursolic acid and ursolic acid-Tyr-Gly-Phe-Gly-Gly with the active pockets of tumor-related cytokines Tth-1, IL-6, IL-8 and TNF-alpha. Virtual screening based on molecular docking shows that neither ursolic acid nor ursolic acid-Tyr-Gly-Phe-Gly-Gly can enter into active pockets of Tth-1, IL-6 and IL-8. The virtual screening based on molecular docking further shows that, although ursolic acid and ursolic acid-Tyr-Gly-Phe-Gly-Gly can enter the active pocket of TNF-alpha, the docking score of the ursolic acid is obviously lower than that of the ursolic acid-Tyr-Gly-Phe-Gly-Gly. Figure 2 of the specification is a graph showing the butt joint morphology and score of ursolic acid-Tyr-Gly-Phe-Gly-Gly and ursolic acid in a TNF-alpha active pocket.
The virtual screening completed by the inventor shows that the anti-osteoporosis activity of the ursolic acid-Tyr-Gly-Phe-Gly-Gly is superior to that of the ursolic acid in theory. The inventor expands the experimental study of ursolic acid-Tyr-Gly-Phe-Gly-Gly. Experimental study shows that the anti-osteoporosis activity of the ursolic acid-Tyr-Gly-Phe-Gly-Gly is indeed superior to that of the ursolic acid. Based on these findings, the inventors have proposed the present invention.
Disclosure of Invention
The first object of the present invention is to provide an ursolic acid-Tyr-Gly-Phe-Gly having the following formula,
The second object of the present invention is to provide a preparation method of ursolic acid-Tyr-Gly-Phe-Gly of the structure, which comprises:
1) Synthesizing Boc-Tyr-Gly-Phe-Gly-Gly-OBzl;
2) Synthesizing HCl.Tyr-Gly-Phe-Gly-Gly-OBzl;
3) 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate is used as a condensing agent, and ursolic acid is condensed with HCl.Tyr-Gly-Phe-Gly-Gly-OBzl to prepare ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl;
4) And (3) adopting H 2/Pb to deprotect the OBzl of the ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl to prepare the ursolic acid-Tyr-Gly-Phe-Gly-Gly.
The synthetic route diagram of the ursolic acid-Tyr-Gly-Phe-Gly-Gly is shown in figure 1.
The third object of the present invention is to evaluate the anti-osteoporosis activity of ursolic acid-Tyr-Gly-Phe-Gly having the structure as described, and the use thereof in the preparation of anti-osteoporosis drugs.
Drawings
FIG. 1 shows the synthesis route of ursolic acid-Tyr-Gly-Phe-Gly-Gly. i) N, N' -Dicyclohexylcarbodiimide (DCC), 1-hydroxybenzotriazole (HOBt), N-methylmorpholine, anhydrous tetrahydrofuran; ii) 10% palladium on carbon, hydrogen; iii) Ethyl acetate solution of hydrogen chloride (4M); iv) 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate, anhydrous N, N-dimethylformamide, anhydrous N, N-diisopropylethylamine.
FIG. 2 is a graph showing the docking morphology and score of ursolic acid-Tyr-Gly-Phe-Gly-Gly and ursolic acid in TNF-alpha active pocket.
Detailed Description
To further illustrate the invention, a series of examples are given below. These examples are given solely for the purpose of illustration and are not to be construed as limitations of the present invention.
EXAMPLE 1 preparation of Boc-Gly-Gly-OBzl
1.75G (10 mmol) of Boc-Gly,1.35g (10 mmol) of N-hydroxybenzotriazole (HOBt), 80mL of anhydrous tetrahydrofuran and 2.50g (12 mmol) of N, N' -Dicyclohexylcarbodiimide (DCC) were stirred at 0℃for 0.5 hours to give a reaction solution A. To the reaction solution A, 3.40g (10 mmol) of Tos Gly-OBzl was added, and the pH was adjusted to 9 with N-methylmorpholine (NMM) at 0 ℃. After that, the reaction mixture was stirred at room temperature for 4h, and TLC showed Tos.Gly-OBzl disappeared, and the stirring was stopped. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved with 100mL of ethyl acetate. The resulting solution was filtered and the filtrate was washed three times with 30mL of saturated NaHCO 3 solution, three times with 30mL of saturated NaCl solution, three times with 30mL of 5% KHSO 4 solution, and three times with 30mL of saturated NaCl solution. Thereafter, the mixture was dried over anhydrous Na 2SO4 for 12 hours. Na 2SO4 was filtered off and the filtrate was concentrated under reduced pressure to give 3.15g (98%) of Boc-Gly-Gly-OBzl as a pale yellow solid. TLC (Petroleum ether/ethyl acetate, 4/1; UV-coloration; rf=0.25), ESI-MS (m/e): 323[ M+H ] +.
EXAMPLE 2 preparation of HCl.Gly-Gly-OBzl
3.00G (9.3 mmol) of Boc-Gly-Gly-OBzl was dissolved in 30mL of ethyl acetate solution of hydrogen chloride (4M), stirred at 0deg.C for 4 hours, and TLC showed disappearance of Boc-Gly-Gly-OBzl, and stirring was stopped. The reaction mixture was concentrated under reduced pressure, and the residue was dissolved with 10mL of anhydrous ethyl acetate and concentrated under reduced pressure. This operation was repeated 3 times. The residue was suspended in 10mL petroleum ether and the suspension was concentrated under reduced pressure. This operation was repeated 3 times. The obtained HCl.Gly-Gly-OBzl was pale yellow solid, and was directly used for the subsequent reaction. ESI-MS (m/e): 223[ M+H ] +.
EXAMPLE 3 preparation of Boc-Gly-Phe-OBzl
From 1.75g (10 mmol) of Boc-Gly and 2.90g (10 mmol) of HCl Phe-OBzl, 4.08g (99%) of Boc-Gly-Phe-OBzl were obtained as colorless solid by the method of example 1. TLC (CH 2Cl2/MeOH, 40/1; UV-developed; rf=0.35), ESI-MS (m/e): 413[ M+H ] +.
EXAMPLE 4 preparation of Boc-Gly-Phe
4.00G (9.7 mmol) of Boc-Gly-Phe-OBzl are dissolved in 100mL of methanol. To the resulting solution was added 400mg of palladium on carbon. The resulting suspension was stirred at room temperature after air was removed and hydrogenolyzed with hydrogen for 12 hours. TLC showed the disappearance of Boc-Gly-Phe-OBzl, terminating the hydrogenolysis. The suspension was removed with palladium on carbon and the filtrate concentrated under reduced pressure to give 3.10g (99%) of Boc-Gly-Phe as a colourless solid. ESI-MS (m/e): 323[ M+H ] +.
EXAMPLE 5 preparation of Boc-Gly-Phe-Gly-Gly-OBzl
A crude pale yellow Boc-Gly-Phe-Gly-Gly-OBzl was obtained from 3.00g (9.3 mmol) of Boc-Gly-Phe and 2.50g (9.7 mmol) of Tos.Gly-Gly-OBzl by the method of example 1. The crude product was purified by column chromatography (gradient elution with CH 2Cl2 -MeOH; CH 2Cl2/MeOH, 40:1, UV-coloration, rf=0.25) to give 4.39g (90%) of Boc-Gly-Phe-Gly-Gly-OBzl as a colourless solid. ESI-MS (m/e): 562[ M+Cl ] -.
EXAMPLE 6 preparation of HCl.Gly-Phe-Gly-Gly-OBzl
HCl.Gly-Phe-Gly-Gly-OBzl was prepared as pale yellow solid from 4.90g (9.3 mmol) of Boc-Gly-Phe-Gly-Gly-OBzl by the method of example 2. ESI-MS (m/e): 427[ M+H ] +.
EXAMPLE 7 preparation of Boc-Tyr-Gly-Phe-Gly-Gly-OBzl
4.74G (74%) of Boc-Tyr-Gly-Phe-Gly-Gly-OBzl were obtained as colorless solid from 2.60g (9.3 mmol) of Boc-Tyr and 4.30g (9.3 mmol) of HCl-Gly-Phe-Gly-Gly-OBzl by the method of example 1. TLC (CH 2Cl2/MeOH, 20/1; UV-developed; rf=0.20), ESI-MS (m/e): 725[ M+Cl ] -.
EXAMPLE 8 preparation of HCl-Tyr-Gly-Phe-Gly-Gly-OBzl
HCl. Tyr-Gly-Phe-Gly-Gly-OBzl was obtained as pale yellow solid from 4.00g (5.8 mmol) of Boc-Tyr-Gly-Phe-Gly-Gly-Gly-OBzl by the method of example 2, and was used directly in the subsequent reaction. ESI-MS (m/e): 591[ M+H ] +.
EXAMPLE 9 preparation of ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl
0.55G (1.2 mmol) of ursolic acid, 0.46g (1.2 mmol) of 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethylurea hexafluorophosphate and 10mL of anhydrous N, N-dimethylformamide were weighed out and dissolved at room temperature for about 4 hours to obtain a reaction solution A. To the reaction solution A, 0.63g (1.0 mmol) of HCl Tyr-Gly-Phe-Gly-Gly-OBzl was added, and the pH was adjusted to 9 with anhydrous N, N-diisopropylethylamine at 0 ℃. Thereafter, the reaction mixture was stirred at room temperature for 36 hours, the pH was continuously monitored during stirring, and anhydrous N, N-diisopropylethylamine was added to maintain the pH at 8-9. The stirring was terminated. The reaction solution was poured into 100mL of a saturated NaCl solution at 0deg.C, left to stand, and the suspension was washed three times with 30mL of a saturated NaHCO 3 solution, three times with 30mL of a saturated NaCl solution, three times with 30mL of a 5% KHSO 4 solution, and three times with 30mL of a saturated NaCl solution. Thereafter, the mixture was dried over anhydrous Na 2SO4 for 12 hours. Filtering out Na 2SO4, concentrating the filtrate under reduced pressure to obtain a light yellow ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl crude product. The crude product was purified by column chromatography (gradient elution with CH 2Cl2 -MeOH; CH 2Cl2/MeOH, 20:1, UV-developed; rf=0.20) to give 0.38g (35%) of ursolic-Tyr-Gly-Phe-Gly-Gly-OBzl (35%) as a colourless solid. ESI-MS (m/e): 1098[ M+H ] +.
EXAMPLE 10 preparation of ursolic acid-Tyr-Gly-Phe-Gly
Crude ursolic acid-Tyr-Gly-Phe-Gly-Gly-Gly was obtained as colorless solid from 1.00g (1.0 mmol) of ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl by the method of example 4. The solid was purified by column chromatography. The silica gel column was eluted with a gradient of CH 2Cl2/meoh=15/1 plus 2% hco 2 H to give 0.66g (71%) of ursoyl-Tyr-Gly-Phe-Gly as a colorless solid. TLC (ethyl acetate/distilled water/glacial acetic acid, 10/1/1; iodine vapor development; rf=0.30). FT-MS960.55008[ M+Na ] + (960.54624); mp is 185-186 ℃;IR/cm-1＝3303,2925,2868,1649,1514,1453,1377,1226,1105,1028,996,830,744,699,662;1H NMR(300MHz,DMSO-d6)δ/ppm＝12.45(s,1H),9.08(s,1H),8.27(s,1H),8.34(t,J＝5.7Hz,1H),8.10(d,J＝8.1Hz,1H),8.03(t,J＝6.0Hz,1H),7.92(t,J＝5.4Hz,1H),7.27～7.20(m,5H),7.17(d,J＝8.1Hz,1H),7.03(d,J＝8.1Hz,2H),6.60(d,J＝8.1Hz,2H),5.00(s,1H),4.53(m,1H),4.27(s,1H),4.17(m,1H),3.74(m,6H),3.60(dd,J1＝17.1Hz,J2＝5.1Hz,1H),3.16(s,1H),3.06(dd,J1＝13.8Hz,J2＝4.8Hz,1H),2.97(t,J＝7.2Hz,1H),2.85～2.67(m,4H),2.01(d,J＝10.8Hz,1H),1.85(m,1H),1.73～1.57(m,4H),1.49～1.23(m,13H),1.05(d,J＝8.7Hz,1H),0.92～0.58(m,11H),0.78～0.74(m,7H),0.68(s,4H),0.59(d,J＝9.3Hz,1H),0.19(s,3H);13C NMR(125MHz,DMSO-d6)δ/ppm＝176.81,172.46,171.64,171.48,169.42,168.97,156.42,138.62,138.21,130.59,129.59,128.57,128.50,126.71,125.03,115.27,77.38,55.79,55.34,54.87,54.52,52.48,49.07,47.56,46.88,42.33,41.95,41.14,41.11,39.33,39.02,38.84,38.74,38.10,36.93,36.63,36.58,32.77,30.81,28.76,27.67,27.49,23.96,23.71,23.28,21.55,18.37,17.55,16.55,16.44,15.66,0.57.
EXAMPLE 11 evaluation of anti-osteoporosis effect of ursolic acid-Tyr-Gly-Phe-Gly-Gly
Ursolic acid was purchased from Shanghai Ala Biochemical technology Co., ltd. SPF grade ICR strain Male mice (25.+ -.2 g) were purchased from Experimental animal technologies Inc. of Peking Vitre. The experiments adopt a model of osteoporosis caused by bilateral ovariectomy of ICR female mice. The dosage of ursolic acid is 300 mu mol/kg/day and 200 mu mol/kg/day, the dosage of the compound ursolic acid-Tyr-Gly-Phe-Gly-Gly is 100 mu mol/kg/day, 10 mu mol/kg/day and 1 mu mol/kg/day, and the negative control is CMC-Na.
Mice were anesthetized with an anesthesia machine at the time of molding. I.e., the mice were placed on the mouse plate on the bottom, with the mouth and nose aligned to the vent valve, and the mice were continuously anesthetized by isoflurane gas. Then, the lower abdomen of the mouse is disinfected by iodophor and alcohol, the skin of the lower abdomen is cut by surgical scissors, the muscle layer is cut along the white line of the abdomen, the abdominal cavity is exposed, the bladder is found under the fat of the lower abdomen, the Y-shaped uterus is found under the bladder, the oviduct on two sides is found along the uterus, the ovary is found at the blind ends of the oviduct on two sides, the ovary is in a small cauliflower shape and is wrapped by fat, the oviduct is ligated at the position far from the uterus, the ovaries on two sides are removed, a drop of penicillin solution is respectively dropped at the ligation position, after the penicillin solution is put back into the abdominal cavity, a drop of penicillin solution is respectively dropped before and after the muscle layer is sutured, and after the skin layer is sutured, the skin suture position is disinfected by alcohol and iodophor. Penicillin, alcohol, which is an iodophor, acts to prevent infection. After withdrawal of the anaesthetic device, the mice wake up very quickly.
After the sham operation group (sham operation) mice were induced to be anesthetized with an anesthetic machine, the mice were continuously anesthetized by isoflurane gas with their mouths and noses aligned with the vent valves, and placed on the mouse plate. Then, the lower abdomen of the mouse is disinfected by iodophor and alcohol, the skin of the lower abdomen is cut by surgical scissors, the muscle layer is cut along the white line of the abdomen, the abdominal cavity is exposed, the bladder is found under the fat of the lower abdomen, the Y-shaped uterus is found under the bladder, the oviduct on two sides is found along the uterus, the ovary is found at the blind ends of the oviduct on two sides, the ovary is in a floret shape and is wrapped by fat, the oviduct is not ligated, the ovary is not removed, after the oviduct is placed back into the abdominal cavity, a drop of penicillin solution is respectively dropped before and after the muscle layer is sutured, and after the skin layer is sutured, the skin suture is disinfected by alcohol and iodophor. Penicillin, alcohol, which is an iodophor, acts to prevent infection. After withdrawal of the anaesthetic device, the mice wake up very quickly.
Mice were recovered seven days post-surgery and randomized. Wherein ovariectomized mice and sham operated mice were given 5%o CMC-Na solution by intragastric administration daily for 28 consecutive days while recording body weight. Ursolic acid treated mice were given ursolic acid by intragastric administration at doses of 300. Mu. Mol/kg/day and 200. Mu. Mol/kg/day for 28 consecutive days while recording body weight. The ursolic acid-Tyr-Gly-Phe-Gly-Gly treated mice were given 100. Mu. Mol/kg/day, 10. Mu. Mol/kg/day and 1. Mu. Mol/kg/day by intragastric administration of ursolic acid-Tyr-Gly-Phe-Gly-Gly according to body weight, and the administration was continued for 28 days while recording body weight.
Mouse bone trabecular bone density was calculated by scanning the mouse femur with Bruker skyscan 1276 Micro-CT. And selecting a region of interest at the distal end of the femur with the same height, calculating a bone fracture Liang Canshu, and analyzing the bone trabecular structure of the femur. And quantitatively analyzing the bone trabeculae of the ovariectomy mice according to six indexes such as bone trabecula bone density, bone volume fraction, bone trabecula number, bone trabecula thickness, bone trabecula separation degree and bone trabecula mode factor. The results of each index are represented by mean value +/-SD, SD value is subjected to variance analysis through SPSS software, variance uniformity is checked, and t-test is adopted for checking, so that the statistical ratio among groups is performed.
On indexes such as femur trabecular bone density of mice, the osteoporosis of ovariectomized mice can be improved by continuous 28-day gastric lavage administration of ursolic acid at a dosage of 300 mu mol/kg, and the osteoporosis of ovariectomized mice cannot be improved by continuous 28-day gastric lavage administration of ursolic acid at a dosage of 200 mu mol/kg. The osteoporosis of ovariectomized mice can be ameliorated by the administration of ursolic acid-Tyr-Gly-Phe-Gly-Gly by gastric lavage at doses of 100. Mu. Mol/kg and 10. Mu. Mol/kg for 28 consecutive days. Namely, the effective dose of the ursolic acid for treating the osteoporosis is 300 mu mol/kg; the effective dose of the ursolic acid-Tyr-Gly-Phe-Gly-Gly for treating osteoporosis is 10 mu mol/kg, which is 1/30 of that of the ursolic acid. It can be seen that the present invention has unexpected technical effects. The specific results are shown in tables 1 to 6.
TABLE 1 treatment of bone density of the femur trabeculae of mice with ursolic acid and ursolic acid-Tyr-Gly-Phe-Gly-Gly
Gastric lavage administration; sham group n=7, ursolic-Tyr-Gly-Phe-Gly (10 μmol/kg) treatment group n=8, the remaining groups n=10.
A) Ratio P to ovariectomized group <0.01; b) Group ratio P to ursolic acid (300. Mu. Mol/kg) is <0.01; c) The ratio P of the ursolic acid to ovariectomy group is less than 0.01, and the ratio P of the ursolic acid to the ursolic acid (300 mu mol/kg) group is more than 0.05; d) The ratio of P to ovariectomy group is less than 0.01, and the ratio of P to ursoyl-Tyr-Gly-Phe-Gly-Gly (100 mu mol/kg) group is less than 0.05; e) The ratio P to ovariectomy group was >0.01, and the ratio P to ursoyl-Tyr-Gly-Phe-Gly-Gly (10. Mu. Mol/kg) group was <0.01.
TABLE 2 Vaccinic acid and Ursolic acid-Tyr-Gly-Phe-Gly-Gly treating femur bone volume fraction of mice
Gastric lavage administration; sham group n=7, ursolic-Tyr-Gly-Phe-Gly (100 μmol/kg) treatment group n=8, the remaining groups n=10.
A) The ratio of P to ovariectomy group is less than 0.01, and the ratio of P to ursolic acid (200 mu mol/kg) group is less than 0.05; b) Ratio P to ovariectomized group <0.01; c) The ratio of P to ovariectomy group is less than 0.01, and the ratio of P to ursoyl-Tyr-Gly-Phe-Gly-Gly (10 mu mol/kg) group is less than 0.05; d) The ratio of P to ovariectomy group is less than 0.01, and the ratio of P to ursoyl-Tyr-Gly-Phe-Gly-Gly (1 mu mol/kg) group is less than 0.01; e) The ratio P to ovariectomized group was >0.05.
TABLE 3 Ursolic acid and Ursolic acyl-Tyr-Gly-Phe-Gly-Gly treating number of trabeculae of femur in mice
Gastric lavage administration; sham group n=7, ursolic-Tyr-Gly-Phe-Gly (100 μmol/kg) treatment group n=8, the remaining groups n=10.
A) The ratio of P to ovariectomy group is less than 0.05, and the ratio of P to ursolic acid (200 mu mol/kg) group is less than 0.05; b) Ratio P >0.05 to ovariectomized group; c) The ratio P of the ursolic acid to ovariectomy group is less than 0.01, and the ratio P of the ursolic acid to the ursolic acid (300 mu mol/kg) group is more than 0.05; the group ratio P of the ursolic acid-Tyr-Gly-Phe-Gly-Gly (10 mu mol/kg) is less than 0.05; d) The ratio of P to ovariectomy group is less than 0.05, and the ratio of P to ursoyl-Tyr-Gly-Phe-Gly-Gly (1 mu mol/kg) group is less than 0.05; e) The ratio P to ovariectomized group was >0.05.
TABLE 4 Ursolic acid and Ursolic acyl-Tyr-Gly-Phe-Gly-Gly treating thickness of femur trabecula in mice
Gastric lavage administration; sham group n=7, ursolic-Tyr-Gly-Phe-Gly (100 μmol/kg) treatment group n=8, the remaining groups n=10.
A) Ratio P to ovariectomized group <0.05; b) Ratio P to sham surgery group <0.05; c) Group ratio P to ursolic acid (300. Mu. Mol/kg) is <0.05; c) The ratio P to ovariectomized group was <0.05.
TABLE 5 degree of trabecular separation of femur in mice treated with ursolic acid and ursolic acid-Tyr-Gly-Phe-Gly-Gly
Gastric lavage administration; sham group n=7, ursolic-Tyr-Gly-Phe-Gly (100 μmol/kg) n=8, the remaining groups n=10. a) Ratio P to ovariectomized group <0.01; b) Ratio P to ovariectomized group <0.05; c) The ratio P of the artificial operation group to the ursolic acid (200 mu mol/kg) is less than 0.05, and the ratio P of the artificial operation group to the ursolic acid (200 mu mol/kg) is less than 0.05; b) The ratio of P to ovariectomy group is less than 0.05, and the ratio of P to ursoyl-Tyr-Gly-Phe-Gly-Gly (100 mu mol/kg) group is less than 0.01; c) The ratio P to ovariectomy group was >0.05, and the ratio P to ursoyl-Tyr-Gly-Phe-Gly-Gly (10. Mu. Mol/kg) group was <0.01.
TABLE 6 treatment of femoral trabecular model factor in mice with ursolic acid and ursolic acid-Tyr-Gly-Phe-Gly-Gly
Gastric lavage administration; sham group n=7, the remaining groups n=10.
A) The ratio of P to ovariectomy group is less than 0.01, and the ratio of P to ursolic acid (200 mu mol/kg) group is less than 0.05; b) Ratio P >0.05 to ovariectomized group; c) Ratio to ovariectomy group P <0.01, to ursolic acid-Tyr-Gly-Phe-Gly-Gly (10. Mu. Mol/kg) treatment group P <0.05; d) Ratio to ovariectomy group P <0.01, to ursolic acid-Tyr-Gly-Phe-Gly-Gly (1. Mu. Mol/kg) treatment group P <0.05; e) The ratio P to ovariectomized group was >0.05.

Claims (3)

1. Ursolic acid-Tyr-Gly-Phe-Gly-Gly with the structure of the general formula I,
2. The method for preparing the ursolic acid-Tyr-Gly-Phe-Gly of the structure according to claim 1, which is characterized in that the method comprises the following steps:
1) Synthesizing Boc-Tyr-Gly-Phe-Gly-Gly-OBzl;
2) Synthesizing HCl-Tyr-Gly-Phe-Gly-Gly-OBzl;
3) 2- (7-azobenzotriazole) -N, N, N ', N' -tetramethyl urea hexafluorophosphate is used as a condensing agent, and ursolic acid is condensed with HCl.Tyr-Gly-Phe-Gly-Gly-OBzl to prepare ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl;
4) Preparing the ursolic acid-Tyr-Gly-Phe-Gly-Gly-Gly by deprotecting the OBzl of the ursolic acid-Tyr-Gly-Phe-Gly-Gly-OBzl by adopting H 2/Pb.
3. Use of the structure of ursolic acid-Tyr-Gly-Phe-Gly according to claim 1 for the preparation of an anti-osteoporosis medicament.
CN202211069067.1A 2022-05-19 2022-09-01 Ursolic acid-Tyr-Gly-Phe-Gly-Gly, synthesis, activity and application thereof Active CN115403658B (en)

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