CN114671920A - Amide penta-acetyl geniposide derivative and preparation method and application thereof - Google Patents
Amide penta-acetyl geniposide derivative and preparation method and application thereof Download PDFInfo
- Publication number
- CN114671920A CN114671920A CN202210384149.9A CN202210384149A CN114671920A CN 114671920 A CN114671920 A CN 114671920A CN 202210384149 A CN202210384149 A CN 202210384149A CN 114671920 A CN114671920 A CN 114671920A
- Authority
- CN
- China
- Prior art keywords
- geniposide
- derivative
- nmr
- mmol
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H17/00—Compounds containing heterocyclic radicals directly attached to hetero atoms of saccharide radicals
- C07H17/04—Heterocyclic radicals containing only oxygen as ring hetero atoms
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/06—Antigout agents, e.g. antihyperuricemic or uricosuric agents
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
The invention belongs to the technical field of new compound preparation, and particularly relates to an amide penta-acetyl geniposide derivative, and a preparation method and application thereof. The amide penta-acetyl geniposide derivative provided by the invention is prepared by hydrolyzing lipid group at C-4 position of iridoid in geniposide to obtain geniposide acid, acetylating 5 hydroxyls at 2 ', 3', 4 ', 6' and 11 positions on sugar to obtain penta-acetyl geniposide acid (2 a), and finally reacting with RNH2Reacting to obtain the penta-acetyl geniposide derivative. The geniposide derivative prepared by the invention has obvious XOD (X-ray diffraction) inhibition activity and kidney damage improvement activity, and can effectively reduce the accumulation of uric acid in the kidney. Reduce inflammationSymptomatic reaction, thereby reducing the level of inflammatory factors; renal fibrosis in hyperuricemia mice can also be reduced by inhibiting the expression of the inflammatory factor TGF-beta.
Description
Technical Field
The invention belongs to the technical field of new compound preparation, and particularly relates to an amide pentaacetyl geniposide derivative, and a preparation method and application thereof.
Background
Hyperuricemia is a metabolic disease caused by metabolic disorder of purine substances in vivo. In recent years, the number of patients with hyperuricemia has increased, and this has become the second largest metabolic disease. Long-term hyperuricemia can cause a plurality of complications, particularly kidney inflammation, and can cause kidney fibrosis in severe cases, thereby seriously damaging the kidney function. Xanthine Oxidase (XOD) plays an important role in uric acid metabolism in vivo, and is a key enzyme for regulating uric acid production. Therefore, most of the current uric acid lowering drugs target XOD, such as: allopurinol, febuxostat, etc. However, most of these XOD inhibitors have significant toxic side effects, which can cause kidney damage. Therefore, in addition to having a higher XOD inhibitory activity, novel XOD inhibitors should also be considered to reduce kidney damage.
In recent years, natural products have been increasingly used in the medical field, and some natural products have been found to have uric acid lowering activity. The fructus Gardeniae is Gardenia jasminoides Ellis of RubiaceaeGardenia jasminoides Elli) belongs to a medicine and food dual-purpose resource, wherein geniposide is a main active ingredient in gardenia. Geniposide has activities of resisting inflammation, reducing uric acid, reducing blood sugar, reducing blood pressure and the like, but cannot be directly applied to clinic due to low activity of geniposide. How to further improve the activity of geniposide is an important subject in current research.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides an amide pentaacetyl geniposide derivative with high activity.
The invention also provides a preparation method of the amide penta-acetyl geniposide derivative.
The invention also aims to provide the application of the amide pentaacetyl geniposide derivative in preparing the medicament for reducing uric acid.
The technical scheme adopted by the invention for realizing the purpose is as follows:
the invention provides an amide penta-acetyl geniposide derivative, the molecular structural formula of which is as follows:
r is
、
、
、
、
、
、
、
、
、
、
、
The invention also provides a preparation method of the amide penta-acetyl geniposide derivative, which comprises the following steps:
(1) adding the geniposide derivative 2a into DMF, stirring the solution, adding EDCI, adding HOBT and DIPEA, and stirring for 2 h at room temperature;
(2) after the stirring is finished, RNH is added2(30.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) and stirring at room temperature for 8-10 h;
(3) after the reaction is finished, adding ice water, and extracting with dichloromethane; the combined organic solvents were washed with 1M aqueous diluted hydrochloric acid and saturated brine; organic phase in anhydrous Na2SO4Dried overnight, distilled under reduced pressure, and purified by column chromatography (eluent v/v: petroleum ether/ethyl acetate = 10/1-1/1) to give the product as a pale yellow powder.
Further, in the step (1), the concentration of the geniposide derivative 2a in DMF is 0.11-0.12 mmol/mL; the molar ratio of the geniposide derivative 2a to EDCI is 0.34: 0.41; the molar ratio of HOBT to EDCI is 1: 1; the molar ratio of DIPEA to HOBT is 1.02: 0.41.
further, in the step (2), the RNH2And geniposide derivative 2a at a molar ratio of 0.37: 0.34; the molar ratio of DMAP to amino compound is 0.51: 0.37.
the above RNH2Specifically 1-aminopyrrole, 1-aminoindazole, 1-aminooxazole, 1-aminobiphenyl, 1-aminoindole, 1-aminopropionitrile, 1-amino-4-bromonaphthalene, p-aminobenzoic acid, 3-fluoroaniline, 4-aminobenzonitrile, 4-aminomethylpyridine, 5-aminotetrazole, 4-aminoisoquinoline and 4-aminomauric acid.
Further, the geniposide derivative 2a has a structural formula as follows:
the preparation method of the geniposide derivative 2a comprises the following steps:
(1) sequentially adding 0.257 mmol of geniposide and 10 mL of 4% NaOH solution into a round-bottom flask, refluxing and stirring at 65 ℃ for reaction, tracking by TLC (thin layer chromatography) until a raw material point disappears, neutralizing by using 1M hydrochloric acid until the pH is =7 after the reaction is finished, refining, concentrating the obtained reaction solution under reduced pressure to dryness, and passing through a column to obtain white powder geniposidic acid 1 a;
(2) sequentially adding geniposide derivative 1a0.026 mmol and triethylamine 5 mL into round-bottom flask, cooling with ice water bath, slowly dropwise adding acetic anhydride 5 mL while stirring, removing ice water bath after dropwise adding, reacting at room temperature, tracking by TLC until the raw material point disappears, and reacting with saturated NaHCO3Solution 20 mL of neutralized solution to pH =7.0, washed with deionized water, combined organic phases, anhydrous Na2SO4Drying overnight, suction filtering to remove Na2SO4Then, the mixture is distilled under reduced pressure and is subjected to column chromatography to obtain light yellow powder pentaacetyl geniposidic acid 2 a.
The structural formula of the raw material geniposide 1a used by the invention is as follows:
further, in the step (3), the eluent used for the column chromatography purification is composed of petroleum ether/ethyl acetate = 10/1-1/1.
The invention also provides application of the amide pentaacetyl geniposide derivative prepared by the preparation method in preparation of a medicament for reducing uric acid, and the amide pentaacetyl geniposide derivative is used as a novel XOD inhibitor.
The amide penta-acetyl geniposide derivative is prepared by hydrolyzing C-4 lipid group of iridoid in geniposide to obtain geniposide acid, acetylating 5 hydroxyls at 2 ', 3', 4 ', 6' and 11 positions on sugar to obtain penta-acetyl geniposide acid (2 a), and mixing with RNH2Reacting to obtain the penta-acetyl geniposide derivative, wherein the specific synthetic route is as follows:
(i) 4% NaOH, 60℃; (ii) acetic anhydride, Et3N, 0℃ to r.t.
(iii) BOP reagent, Et3N, DMF-CH2Cl2, HOBT, DMAP, DIPEA, EDCI, 0℃ to r.t.
the invention has the beneficial effects that:
(1) the geniposide derivative prepared by the invention has obvious XOD (X-ray diffraction) inhibition activity and kidney damage improvement activity, and can effectively reduce the accumulation of uric acid in the kidney.
(2) The geniposide derivative prepared by the invention reduces inflammatory reaction by regulating the expression of TLR 4/IkB alpha/NF-kB signal channel, thereby reducing the level of inflammatory factors; renal fibrosis in hyperuricemia mice can also be reduced by inhibiting the expression of the inflammatory factor TGF-beta.
Drawings
FIG. 1 is a pathological section of kidney tissue of each group of mice.
FIG. 2 is a graph showing the staining of the kidney tissue masson in each group of mice.
FIG. 3 shows CCK-8 cell viability assay.
Figure 4 is a graph of the effect of geniposide derivative 6c on levels of inflammatory factors.
FIG. 5 is a graph comparing the deposition of urate in mouse kidney under polarized light.
FIG. 6 is a compound1a 1H NMR and13c NMR spectrum.
FIG. 7 shows Compound 2a1H NMR and13c NMR spectrum.
FIG. 8 is Compound 6a1H NMR and13c NMR spectrum.
FIG. 9 shows Compound 6b1H NMR and13c NMR spectrum.
FIG. 10 is Compound 6c1H NMR and13c NMR spectrum.
FIG. 11 shows Compound 6d1H NMR and13c NMR spectrum.
FIG. 12 is Compound 6e1H NMR and13c NMR spectrum.
FIG. 13 is Compound 6f1H NMR and13c NMR spectrum.
FIG. 14 shows Compound 6g1H NMR and13c NMR spectrum.
FIG. 15 shows Compound 6h1H NMR and13c NMR spectrum.
FIG. 16 is Compound 6i1H NMR and13c NMR spectrum.
FIG. 17 is Compound 6j1H NMR and13c NMR spectrum.
FIG. 18 is compound 6k1H NMR and13c NMR spectrum.
FIG. 19 shows Compound 6l1H NMR and13c NMR spectrum.
FIG. 20 shows Compound 6m1H NMR and13c NMR spectrum.
FIG. 21 is Compound 6n1H NMR and13c NMR spectrum.
Detailed Description
The technical solution of the present invention is further explained and illustrated by the following specific examples.
Example 1 Synthesis of geniposide lead Compound, geniposide
(1) Synthesis of geniposide (1 a)
Geniposide (100.0 mg, 0.257 mmol) and 10 mL of 4% NaOH solution were sequentially added to a round-bottomed flask, the reaction was stirred at 65 ℃ under reflux, and TLC followed until the starting material spot disappeared (developer v/v: dichloromethane/methanol = 5/1). After completion of the reaction, the reaction mixture was neutralized with 1M hydrochloric acid to pH =7, purified, and the resulting reaction mixture was concentrated to dryness under reduced pressure and subjected to column chromatography (eluent v/v: dichloromethane/methanol = 8/1-1/1) to obtain 61.5 mg of a white powder, mp.250.9-251.6 ℃ in a yield of 91.3%, which was determined to be geniposide (1 a) by analysis such as NMR and HR-MS. The spectrum is shown in FIG. 6.
1H NMR (400 MHz, CD3OD) δ 7.19 (d, J = 11.5 Hz, 1H), 5.01 (dd, J = 9.1, 5.4 Hz, 1H), 4.64 (dd, J = 12.4, 8.4 Hz, 1H), 4.31 – 4.08 (m, 1H), 3.83 – 3.78 (m, 1H), 3.71 – 3.65 (m, 1H), 3.58 (d, J = 3.7 Hz, 1H), 3.33 (t, J = 6.4 Hz, 2H), 3.26 (dt, J = 3.0, 1.5 Hz, 1H), 2.78 (dd, J = 16.2, 7.8 Hz, 1H), 2.70 – 2.64 (m, 1H), 1.99 (dd, J = 17.1, 8.8 Hz, 5H). 13C NMR (100 MHz, CD3OD) δ 174.98, 147.47, 143.98, 127.14, 117.50, 99.75, 96.70, 78.10, 77.48, 74.20, 71.54, 61.94, 60.11, 46.03, 38.75, 36.29. HR-MS (ESI): calcd for C16H22O10 m/z: 374.1213, found: 397.1045 [M+Na]+.
Example 2 Synthesis of geniposide derivative Pentaacetyl geniposide (2 a)
(1) Synthesis of Pentaacetylgeniposidic acid (2 a)
Sequentially adding geniposide derivative 1a (100.0 mg, 0.026 mmol) and triethylamine 5 mL into round-bottom flask, cooling with ice-water bath, slowly dropwise adding acetic anhydride 5 mL while stirring, removing ice-water bath after dropwise adding, reacting at room temperature, tracking by TLC to the point of raw materialDisappearance (developing solvent v/v: petroleum ether/ethyl acetate =1/1, 1 drop of formic acid). After the reaction is finished, the reaction solution is saturated NaHCO3The solution (20 mL) was neutralized to pH =7.0, washed with deionized water (20 mL × 3), and the organic phases were combined, anhydrous Na2SO4Drying overnight, suction filtering to remove Na2SO4Then, the mixture was subjected to distillation under reduced pressure and subjected to column chromatography (eluent v/v: petroleum ether/ethyl acetate = 8/1-1/1) to obtain 88.2 mg of a pale yellow powder, mp.125.7-126.5 ℃ and a yield of 71.2%, and the product was analyzed by NMR, HR-MS and the like to determine that the product was pentaacetylgeniposide (2 a). The spectrum is shown in FIG. 7.
1H NMR (400 MHz, CDCl3) δ 7.32 (s, 1H), 5.85 (s, 2H), 5.25 (t, J = 9.5 Hz, 2H), 5.11 (d, J = 2.3 Hz, 1H), 5.01 (d, J = 1.4 Hz, 1H), 4.91 (s, 1H), 4.70 (dd, J = 13.9 Hz, 1H), 4.32 (s, 1H), 4.18 (dd, J = 2.4 Hz, 1H), 2.88 (dd, 1H), 2.69 (d, J = 6.2 Hz, 2H),2.54 (d, 1H), 2.09 (d, J = 14.0 Hz 1H), 2.02 (m, 15H).
13C NMR (100 MHz, CDCl3)δ 170.90, 170.79, 170.29, 170.15, 169.50, 169.33, 167.80, 163.36, 136.84, 132.20, 131.26, 130.98, 128.80, 96.82, 96.13, 72.45, 71.97, 70.67, 68.20, 65.60, 61.94, 61.59, 46.29, 39.97, 38.43, 36.91, 33.68, 31.77, 30.49, 29.64, 20.84, 20.64, 20.55, 20.42, 19.13. HR-MS (ESI): calcd for C26H32O15 m/z: 584.1741, found: 607.1642 [M+Na]+.
EXAMPLE 3 Synthesis of Pentaacetylgeniposide-1 '' -Pyrrolinamide (6 a)
Pentaacetylgeniposide-1 '' -pyrrole amide (6 a) structural formula:
the synthesis method comprises the following steps:
(1) the geniposide derivative 2a (200.0 mg, 0.34 mmol) DMF (3 mL), EDCI (78.0 mg, 0.41 mmol) in solution was stirred, HOBT (55.0 mg, 0.41 mmol) and DIPEA (0.2 mL, 1.02 mmol) were added and stirred at room temperature for 2 h;
(2) after stirring, 1-aminopyrrole (30.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added, and stirring was continued at room temperature overnight;
(3) after the reaction was completed, the solution was added to ice water (30 mL) and extracted with dichloromethane (20 mL. times.3). The combined organic solvents were washed with 1M aqueous diluted hydrochloric acid (20 mL. times.3) and saturated brine (20 mL. times.3). Organic phase in anhydrous Na2SO4Dried overnight, distilled under reduced pressure, and purified by column chromatography (eluent v/v: petroleum ether/ethyl acetate = 10/1-1/1) to give 59.9 mg of a pale yellow powder, mp.127.6-128.3 ℃, yield 26.8%, determined by analysis by NMR, HR-MS, etc., as pentaacetylgeniposide-1 "-pyrrole amide (6 a). The spectrum is shown in FIG. 8.
1H NMR (400 MHz, CDCl3) δ 7.64 (dt, J = 7.1, 3.6 Hz, 1H), 7.48 – 7.44 (m, 1H), 6.55 (d, J = 2.2 Hz, 2H), 6.09 – 6.07 (m, 2H), 5.77 (s, 1H), 5.20 – 5.14 (m, 1H), 5.07 (d, J = 5.7 Hz, 1H), 5.04 (d, J = 9.6 Hz, 1H), 4.94 (dd, J= 9.6, 8.1 Hz, 1H), 4.83 – 4.80 (m, 1H), 4.63 (d, J = 6.3 Hz, 2H), 4.23 (t, J= 6.7 Hz, 2H), 2.95 (d, J = 7.3 Hz, 2H), 2.12 (d, J = 7.7 Hz, 1H), 2.03 – 1.95 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15, 169.50, 168.42, 166.77, 147.27, 120.74, 120.61, 120.44, 111.74, 106.64, 95.77, 94.55, 76.25, 76.04, 75.83, 64.58, 60.69, 44.34, 29.50, 28.67, 20.60, 20.53, 20.50, 19.85. HR-MS (ESI): calcd for C30H36N2O14S m/z: 648.2167, found: 671.2042 [M+Na]+.
Example 4 Synthesis of Pentaacetylgeniposide-1 "-indazolamide (6 b)
Pentaacetylgeniposide-1 "-indazolamide (6 b) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 1-aminoindazole (49.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight.
50.27 mg of light yellow powder is obtained in the step (3), mp.129.6-131.3 ℃, the yield is 21.0 percent, and the product is determined to be pentaacetyl geniposide-1 '' -indazole amide (6 b) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 9.
1H NMR (400 MHz, CDCl3) δ 9.70 (s, 1H), 8.00 (dd, J = 32.5, 9.5 Hz, 1H), 7.65 (s, 1H), 7.46 (d, J = 3.3 Hz, 2H), 7.19 (s, 2H), 6.37 (d, J = 88.1 Hz, 1H), 5.28 (d, J = 3.7 Hz, 2H), 5.25 (d, J = 7.2 Hz, 2H), 5.21 (s, 1H), 4.89 (dd, J = 38.4, 14.0 Hz, 1H), 4.64 (d, J = 29.3 Hz, 1H), 4.25 (s, 1H), 4.05 (dd, J = 14.3, 7.1 Hz, 1H), 2.73 – 2.68 (m, 1H), 2.28 (t, J = 7.4 Hz, 2H), 2.17 – 2.14 (m, 1H), 1.98 (s, 1H), 1.71 – 1.61 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15, 169.50, 167.75, 132.25, 130.94, 129.91, 128.83, 96.91, 77.23, 76.80, 72.37, 68.10, 61.45, 35.92, 34.17, 22.69, 20.89, 20.60, 20.53, 20.50. HR-MS (ESI): calcd for C33H37N3O14S m/z: 699.2276, found: 722.5331 [M+Na]+.
EXAMPLE 5 Synthesis of Pentaacetylgeniposide-1 '' -oxazolamide (6 c)
Pentaacetylgeniposide-1 '' -oxazoleamide (6 c) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 1-aminooxazole (31.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added, and stirring was continued at room temperature overnight.
(3) Thus, 46.7 mg of a pale yellow powder was obtained, mp.136.3-137.3 ℃ and the yield was 21.3%, and the powder was analyzed by NMR, HR-MS and the like to determine that the powder was pentaacetylgeniposide-1' -oxazolylamide (6 c). The spectrum is shown in FIG. 10.
1H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.73 – 7.72 (m, 1H), 7.54 (d, J = 2.4 Hz, 1H), 7.27 (s, 1H), 5.88 (s, 1H), 5.84 (s, 1H), 5.46 – 5.42 (m, 1H), 5.35 (t, J = 4.8 Hz, 1H), 5.03 – 5.01 (m, 1H), 4.86 (d, J = 8.1 Hz, 1H), 4.70 – 4.68 (m, 1H), 4.32 (s, 1H), 4.09 (s, 1H), 2.93 – 2.88 (m, 1H), 2.81 (d, J = 7.8 Hz, 1H), 2.36 – 2.32 (m, 1H), 2.14 (d, J = 4.5 Hz, 1H), 2.10 – 2.03 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15,170.67, 167.75, 132.25, 130.94, 129.91, 128.83, 96.91, 77.23, 76.80, 72.37, 68.10, 61.45, 35.92, 34.17, 22.69, 20.89, 20.60, 20.53, 20.50. HR-MS (ESI): calcd for C29H34N2O14 m/z: 650.1959, found: 673.3450 [M+Na]+.
Example 6 Synthesis of Pentaacetylgeniposide-1 '' -benzidine (6 d)
Pentaacetylguanipin-1 '' -benzidine (6 d) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 1-aminobiphenyl (62.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight.
(3) 70.5 mg of light yellow powder is obtained, mp.147.6-148.3 ℃, the yield is 28.2 percent, and the powder is determined to be pentaacetyl geniposide-1 '' -benzidine (6 d) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 11.
1H NMR (400 MHz, CDCl3) δ 8.34 (d, J = 8.2 Hz, 1H), 8.03 (d, J = 7.1 Hz, 1H), 7.43 (dd, J = 18.6, 11.0 Hz, 3H), 7.36 (d, J = 7.4 Hz, 1H), 7.33 – 7.30 (m, 2H), 7.23 (s, 1H), 7.18 – 7.15 (m, 1H), 7.11 (dd, J = 10.6, 4.2 Hz, 1H), 5.59 (s, 1H), 5.26 (d, J = 18.5 Hz, 1H), 5.15 (t, J = 9.5 Hz, 1H), 5.00 (d, J = 5.2 Hz, 1H), 4.92 (dd, J = 9.6, 8.1 Hz, 1H), 4.77 (d, J = 8.1 Hz, 1H), 4.62 (d, J = 3.2 Hz, 1H), 4.18 (dd, J = 12.4, 4.4 Hz, 1H), 4.08 (dd, J = 12.3, 2.5 Hz, 1H), 2.75 (dd, J = 13.5, 8.1 Hz, 1H), 2.36 – 2.30 (m, 1H), 2.27 (d, J = 7.6 Hz, 1H), 2.16 (dd, J = 13.7, 6.3 Hz, 1H), 2.03 – 1.93 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 169.61, 169.56, 169.15, 168.38, 168.20, 163.23, 147.05, 137.08, 135.97, 133.92, 131.11, 129.09, 128.80, 128.33, 128.28, 127.57, 127.21, 122.95, 119.78, 113.71, 95.65, 94.06, 71.39, 71.06, 69.58, 67.19, 60.55, 45.73, 36.70, 32.28, 21.67, 19.81, 19.67, 19.57, 19.48. HR-MS (ESI): calcd for C33H41NO14 m/z: 735.2527, found: 758.3514 [M+Na]+.
Example 7 Synthesis of Pentaacetylgeniposide-1' -indoleamide (6 e)
The structural formula of pentaacetyl geniposide-1' -indoleamide (6 e) is as follows:
the synthesis method is basically the same as example 3, except that:
step (2): 1-aminoindole (49.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 68.2 mg of light yellow powder is obtained, mp.157.6-158.3 ℃, the yield is 28.1 percent, and the powder is determined to be pentaacetyl geniposide-1 '' -indole amide (6 e) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 12.
1H NMR (400 MHz, CDCl3) δ 8.31 (s, 1H), 7.68 (dd, J = 6.9, 1.4 Hz, 1H), 7.39 (s, 1H), 7.30 (s, 1H), 7.19 (s, 1H), 7.15 – 7.09 (m, 2H), 6.44 (s, 1H), 5.79 (s, 1H), 5.29 – 5.26 (m, 1H), 5.23 (s, 1H), 5.19 (d, J = 4.3 Hz, 1H), 5.05 (t, J = 9.7 Hz, 1H), 4.96 (dd, J = 9.6, 8.1 Hz, 1H), 4.66 (d, J = 5.6 Hz, 2H), 4.24 – 4.19 (m, 1H), 4.11 (dd, J = 12.4, 2.3 Hz, 1H), 3.00 (d, J= 15.4 Hz, 1H), 2.85 (dd, J = 16.3, 7.6 Hz, 1H), 2.32 (d, J = 16.3 Hz, 1H), 2.16 (d, J = 7.5 Hz, 1H), 2.08 – 1.92 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15,170.67,167.80, 132.30, 130.95, 130.19, 130.13, 130.04, 129.74, 128.85, 115.08, 110.22, 107.28, 107.10, 77.23, 65.62, 33.70, 31.94, 30.57, 29.71. HR-MS (ESI): calcd for C34H38N2O14 m/z: 698.2323, found: 711.3537 [M+Na]+.
EXAMPLE 8 Synthesis of Pentaacetylgeniposide-1 '' -propionitrile amide (6 f)
The structural formula of pentaacetyl geniposide-1 '' -propionitrile amide (6 f) is as follows:
the synthesis method is basically the same as example 3, except that:
step (2): 1-aminopropionitrile (25.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring was continued at room temperature overnight;
(3) 32.6 mg of pale yellow powder is obtained, mp.121.6-122.3 ℃, the yield is 15.0 percent, and the powder is determined to be pentaacetyl geniposide-1 '' -propionitrile amide (6 f) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 13.
1H NMR (400 MHz, CDCl3) δ 8.06 – 8.02 (m, 1H), 7.45 (s, 1H), 5.28 (dd, J = 9.8, 5.0 Hz, 1H), 4.92 (dd, J = 25.6, 9.4 Hz, 1H), 4.81 (dd, J = 8.0, 6.1 Hz, 1H), 4.62 (s, 1H), 4.38 – 4.34 (m, 1H), 4.02 (d, J = 6.7 Hz, 1H), 3.59 (dd, J = 8.5, 3.8 Hz, 1H), 3.10 (s, 1H), 2.64 (dd, J = 31.5, 5.9 Hz, 1H), 2.59 – 2.56 (m, 1H), 2.47 – 2.44 (m, 1H), 2.14 (s, 1H), 2.07 (s, 1H), 2.05 – 1.92 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15,170.67,166.73, 131.23, 129.92, 128.99, 128.71, 127.81, 76.20, 75.99, 75.78, 64.56, 37.64, 34.87, 30.90, 30.88, 18.15. HR-MS (ESI): calcd for C29H36N2O14 m/ z: 636.2167, found: 659.2057 [M+Na]+.
EXAMPLE 9 Synthesis of Pentaacetylgeniposide-4 '' -bromonaphthylamide (6 g)
Pentaacetylgeniposide-4 '' -bromonaphthylamide (6 g) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 1-amino-4-bromonaphthalene (80.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 51.3 mg of light yellow powder is obtained, mp.145.6-146.3 ℃, the yield is 18.8 percent, and the powder is determined to be pentaacetyl geniposide-4 '' -bromonaphthylamide (6 g) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 14.
1H NMR (600 MHz, CDCl3) δ 8.11 (d, J = 13.2 Hz, 1H), 8.01 (s, 1H), 7.72 (dd, J = 5.7, 3.3 Hz, 1H), 7.53 (dd, J = 5.7, 3.3 Hz, 2H), 7.43 (s, 1H), 7.29 (d, J = 1.9 Hz, 1H), 6.66 (d, J = 8.0 Hz, 1H), 5.36 – 5.34 (m, 4H), 4.69 (s, 1H), 4.31 (t, J = 6.7 Hz, 3H), 4.21 (d, J = 8.2 Hz, 1H), 3.39 – 3.36 (m, 1H), 3.31 – 3.27 (m, 1H), 2.22 (d, J = 3.4 Hz, 1H), 2.04 (s, 1H), 1.55 (s, 15H). 13C NMR (151 MHz, CDCl3) δ 170.90, 170.79, 170.29, 170.15,170.67,166.73,130.93, 128.85, 110.00, 65.58, 31.94, 30.58, 29.71, 29.37, 22.70, 19.20. HR-MS (ESI): calcd for C36H38BrNO14 m/z: 788.1476, found: 811.1402 [M+Na]+.
EXAMPLE 10 Synthesis of Pentaacetylgeniposide-4 '' -benzoic acid amide (6 h)
Pentaacetylguanipin-4 '' -benzoic acid amide (6 h) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): p-aminobenzoic acid (51.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) obtain light yellow powder 60.1 mg, mp.157.6-158.3 deg.C, yield 25.0%, and analysis by NMR, HR-MS, etc. determined to be pentaacetyl geniposide-4 '' -benzoic acid amide (6 h). The spectrum is shown in FIG. 15.
1H NMR (400 MHz, CDCl3) δ 8.05 (d, J = 8.7 Hz, 1H), 7.73 – 7.70 (m, 1H), 7.33 (s, 1H), 5.84 (s, 1H), 5.26 (d, J = 4.8 Hz, 1H), 5.24 – 5.22 (m, 1H), 5.01 (d, J = 10.2 Hz, 1H), 4.71 (d, J = 7.6 Hz, 1H), 4.65 (d, J = 13.7 Hz, 1H), 4.37 (s, 1H), 3.75 (d, J = 2.5 Hz, 1H), 2.97 (s, 1H), 2.86 (d, J = 6.9 Hz, 1H), 2.76 (t, J = 7.3 Hz, 1H), 2.13 (s, 1H), 2.09 (dd, J = 7.2, 4.3 Hz, 15 H). 13C NMR (100 MHz, CDCl3) δ 172.03, 171.65, 171.36, 170.85, 170.80, 170.72, 170.68, 170.63, 170.21, 169.63, 169.53, 169.44, 167.77, 165.16, 153.06, 152.90, 147.66, 142.75, 142.62, 132.29, 131.49, 131.18, 130.94, 130.56, 128.85, 124.83, 119.19, 111.21, 96.60, 94.94, 70.85, 70.80, 68.65, 62.40, 46.98, 36.65, 34.40, 20.87, 20.59. HR-MS (ESI): calcd for C33H41NO14 m/z: 703.2112, found: 726.2005 [M+Na]+.
Example 11 Synthesis of Pentaacetylgeniposide-3 '' -F-benzamide (6 i)
The structural formula of pentaacetyl geniposide-3 '' -F-benzamide (6 i) is as follows:
the synthesis method is basically the same as example 3, except that:
step (2): 3-fluoroaniline (41.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring was continued at room temperature overnight;
(3) 43.1 mg of pale yellow powder was obtained, mp.177.6-178.3 ℃, yield 18.0%, and the powder was analyzed by NMR, HR-MS, and the like to determine pentaacetyl geniposide-3 '' -F-benzamide (6 i). The spectrum is shown in FIG. 16.
1H NMR (400 MHz, CDCl3) δ 8.21 (s, 1H), 7.53 (d, J = 2.4 Hz, 1H), 7.37 (s, 2H), 7.22 (s, 1H), 6.78 (s, 1H), 5.46 – 5.43 (m, 2H), 5.35 (s, 2H), 4.88 (d, J = 7.9 Hz, 1H), 4.69 (s, 2H), 4.32 (s, 1H), 4.31 (s, 1H), 4.29 (s, 1H), 3.09 (d, J = 7.3 Hz, 1H), 2.64 (s, 1H), 2.38 (s, 1H), 2.09 (d, J = 3.1 Hz, 1H), 2.06 – 1.97 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 169.64, 169.45, 169.13, 168.47, 168.41, 163.64, 146.17, 135.99, 135.51, 129.52, 128.72, 122.93, 121.74, 119.10, 114.52, 110.77, 106.86, 97.45, 95.56, 93.78, 86.32, 71.37, 71.10, 69.71, 67.25, 60.62, 60.56, 46.09, 37.12. HR-MS (ESI): calcd for C32H36FNO14 m/z: 677.2120, found: 700.2042 [M+Na]+.
EXAMPLE 12 Synthesis of Pentaacetylgeniposide-4 '' -carbonitrile amide (6 j)
The structural formula of pentaacetyl geniposide-4 '' -carbonitrile amide (6 j) is as follows:
the synthesis method is basically the same as example 3, except that:
step (2): 4-aminobenzonitrile (41.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 32.8 mg of light yellow powder is obtained, mp.157.6-158.3 ℃, the yield is 14.0 percent, and the product is determined to be pentaacetyl geniposide-4 '' -formonitrile amide (6 j) through analysis of NMR, HR-MS and the like. The spectrum is shown in FIG. 17.
1H NMR (400 MHz, CDCl3) δ 8.04 (s, 1H), 7.67 (d, J = 8.1 Hz, 2H), 7.44 (d, J = 7.8 Hz, 2H), 7.34 (d, J = 8.5 Hz, 1H), 5.77 (s, 1H), 5.74 (s, 1H), 5.28 (t, J = 4.6 Hz, 1H), 5.19 (d, J = 2.5 Hz, 1H), 4.97 (d, J = 2.2 Hz, 1H), 4.80 (d, J = 2.2 Hz, 1H), 4.64 (s, 2H), 4.20 (d, J = 4.2 Hz, 1H), 4.11 (d, J= 2.9 Hz, 1H), 3.12 (d, J = 6.7 Hz, 1H), 2.52 (s, 1H), 2.32 (s, 1H), 2.16 (s, 1H), 2.04 – 2.02 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 169.59, 169.59, 169.22, 169.22, 169.22, 161.61, 150.82, 138.68, 138.68, 138.68, 131.25, 131.25, 128.86, 128.86, 128.86, 98.96, 98.96, 95.76, 95.64, 71.08, 71.08, 69.66, 69.66, 50.28, 30.89, 28.68, 19.93, 19.85, 19.75, 19.66, 19.58. HR-MS (ESI): calcd for C33H36N2O14 m/z: 684.2167, found: 708.1923 [M+Na]+.
Example 13 Synthesis of Pentaacetylgeniposide-4 '' -methylpyridine amide (6 k)
Pentaacetylguanipin-4 '' -picolinamide (6 k) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 4-Aminomethylpyridine (66.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 46.3 mg of pale yellow powder was obtained, mp.157.6-159.3 ℃ and the yield was 13.8%, and the powder was analyzed by NMR, HR-MS and the like to determine pentaacetylgeniposide-4 '' -picolinamide (6 k). The spectrum is shown in FIG. 18.
1H NMR (400 MHz, CDCl3) δ 8.47 (s, 2H), 8.02 (d, J = 7.1 Hz, 1H), 7.60 (d, J = 7.8 Hz, 1H), 7.22 (d, J = 10.2 Hz, 1H), 5.95 (d, J = 2.4 Hz, 1H), 5.73 (s, 1H), 5.16 (t, J = 9.5 Hz, 1H), 5.08 – 5.05 (m, 1H), 5.02 (d, J = 9.6 Hz, 1H), 4.92 (dd, J = 9.6, 8.1 Hz, 1H), 4.79 (d, J = 8.1 Hz, 1H), 4.67 – 4.57 (m, 2H), 4.41 (dd, J = 15.1, 5.8 Hz, 1H), 4.09 (dd, J = 12.3, 2.3 Hz, 1H), 3.13 (dd, J = 13.3, 7.6 Hz, 1H), 2.89 (t, J = 5.9 Hz, 1H), 2.69 (dd, J = 16.5, 8.0 Hz, 1H), 2.12 (d, J = 16.5 Hz, 1H), 2.03 – 1.84 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 166.77, 166.67, 166.38, 166.22, 166.06, 165.98, 151.95, 150.57, 149.95, 140.64, 135.22, 133.78, 132.40, 128.56, 117.43, 110.42, 95.52, 92.61, 63.37, 62.08, 50.52, 50.32, 45.85, 43.91, 40.81, 37.82, 35.43, 33.84, 22.98, 22.12, 21.84, 21.67, 21.56. HR-MS (ESI): calcd for C32H38NO14 m/z: 674.2323, found: 675.2461 [M+H]+.
EXAMPLE 14 Synthesis of Pentaacetylglucosamine 5 '' -tetrazolamide (6 l)
Pentaacetylguanipin-5 '' -tetrazole amide (6 l) has the following structural formula:
the synthesis method is basically the same as example 3, except that:
step (2): 5-Aminotetrazole (38.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 47.3 mg of light yellow powder is obtained, mp.167.6-168.3 ℃, the yield is 17.2 percent, and the powder is determined to be pentaacetyl geniposide-5 '' -tetrazole amide (6 l) through analysis such as NMR, HR-MS and the like. The spectrum is shown in FIG. 19.
1H NMR (400 MHz, CDCl3) δ 8.07 – 7.99 (m, 1H), 7.20 (s, 2H), 5.66 (s, 2H), 5.35 – 5.12 (m, 6H), 4.98 (d, J = 55.7 Hz, 13H), 4.62 (s, 9H), 4.25 – 4.00 (m, 8H), 3.73 (s, 5H), 2.70 (s, 6H), 2.53 (s, 6H), 2.40 – 2.17 (m, 4H), 2.15 – 1.83 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 169.61, 169.56, 169.15, 168.38, 168.20, 163.23, 147.05, 137.08, 135.97, 133.92, 131.11, 129.09, 128.80, 128.33, 128.28, 127.57, 127.21, 122.95, 119.78, 113.71, 95.65, 94.06, 71.39, 71.06, 69.58, 67.19, 60.55, 45.73, 36.70, 32.28, 21.67, 19.81, 19.67, 19.57, 19.48. HR-MS (ESI): calcd for C27H33N5O14 m/z: 651.2524, found: 674.1897 [M+Na]+.
EXAMPLE 15 Synthesis of Pentaacetylgeniposide-4 '' -isoquinolinamide (6 m)
The structural formula of pentaacetyl geniposide-4 '' -isoquinoline amide (6 m) is as follows:
the synthesis method is basically the same as example 3, except that:
step (2): 4-aminoisoquinoline (60.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) 47.3 mg of pale yellow powder was obtained, mp.187.6-188.3 ℃, yield 14.2%, and it was confirmed by NMR, HR-MS, and other analyses to be pentaacetyl geniposide-4 "-isoquinoline amide (6 m). The spectrum is shown in FIG. 20.
1H NMR (400 MHz, CDCl3) δ 8.81 (s, 1H), 7.96 (d, J = 8.1 Hz, 1H), 7.78 (d, J = 8.3 Hz, 1H), 7.71 (t, J = 7.6 Hz, 1H), 7.59 (t, J = 7.5 Hz, 1H), 7.49 (s, 1H), 7.38 (s, 1H), 5.83 (s, 1H), 5.46 – 5.31 (m, 1H), 5.07 (dd, J = 16.0, 6.3 Hz, 1H), 4.98 – 4.93 (m, 1H), 4.85 (d, J = 8.0 Hz, 1H), 4.67 (d, J = 7.5 Hz, 1H), 4.22 (dd, J = 12.3, 4.4 Hz, 1H), 4.12 (dd, J = 12.2, 2.3 Hz, 1H), 3.04 (s, 1H), 2.89 (dd, J = 16.0, 8.1 Hz, 1H), 2.37 (d, J = 16.2 Hz, 1H), 2.29 – 2.17 (m, 1H), 2.03 – 1.91 (m, 15H). 13C NMR (100 MHz, CDCl3) δ 169.65, 169.63, 169.13, 168.43, 168.41, 164.64, 149.10, 147.25, 137.42, 136.13, 129.86, 129.63, 129.43, 127.78, 127.29, 126.91, 126.55, 119.57, 113.70, 95.78, 94.09, 71.35, 71.11, 69.76, 67.21, 60.61, 60.54, 46.07, 37.31, 32.56, 28.68, 19.85, 19.70, 19.58, 19.45. HR-MS (ESI): calcd for C35H38N2O14 m/z: 710.2323, found: 733.2360 [M+Na]+.
EXAMPLE 16 Synthesis of Pentaacetylguanidine-4 '' -hippuramide (6 n)
The synthesis method is basically the same as example 3, except that:
step (2): 4-Aminohippuric acid (60.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) were added and stirring continued at room temperature overnight;
(3) obtain light yellow powder 62.4 mg, mp.154.6-155.3 deg.C, yield 24%, and is determined to be pentaacetyl geniposide-4 '' -hippuric acid amide (6 n) by NMR, HR-MS and other analyses. The spectrum is shown in FIG. 21.
1H NMR (400 MHz, CDCl3) δ 8.04 (s, 2H), 7.76 – 7.70 (m, 2H), 7.57 – 7.52 (m, 2H), 7.45 (s, 1H), 5.78 – 5.77 (m, 1H), 5.59 – 5.55 (m, 1H), 5.45 (s, 1H), 5.32 – 5.31 (m, 1H), 5.01 (s, 1H), 4.84 – 4.80 (m, 1H), 4.62 (s, 1H), 4.35 – 4.32 (m, 1H), 4.13 – 4.11 (m, 1H), 3.00 – 2.97 (m, 1H), 2.58 – 2.54 (m, 1H), 2.44 – 2.41 (m, 1H), 2.17 (s, 1H), 2.13 (s, 1H), 2.04 – 1.93 (m, 15H). 13C NMR (150 MHz, CDCl3) δ 175.33, 169.87, 169.59, 168.73, 168.51, 167.93, 165.49, 164.13, 150.39, 143.87, 132.94, 129.90, 128.56, 127.69, 127.49, 118.42, 109.16, 95.71, 94.35, 71.65, 70.84, 69.49, 67.24, 64.52, 61.50, 48.15, 35.78, 33.06, 21.85, 21.06. HR-MS (ESI): calcd for C35H40NO17 m/z: 760.2327, found: 783.2289 [M+Na]+.
Effects of the embodiment
Screening of geniposide derivative in vitro enzyme inhibition activity
XOD and xanthine were first dissolved in PBS to prepare XOD solution (100U/L) and xanthine solution (0.5 mM). Geniposide derivatives 6a-6n were accurately weighed and the mother liquor concentration was made to 1mM, and the geniposide derivatives to be tested were further diluted to different concentrations (200, 100, 50, 25, 12.5, 6.25, 3.125 μ M) with PBS. Meanwhile, PBS was used instead of the test compound to set a blank control, and allopurinol was used to set a positive control. Reactions were performed in 96-well plates with 3 replicates per concentration setting. Data were analyzed using Microsoft Excel 2021, and the OD value changes of the blank group and the test compound group were calculated from the inhibition (%) and IC was calculated using GraphPad Prism 8.050Values to evaluate the XOD inhibitory activity of the test compounds, the results of the experiments are given in table 1.
TABLE 1
In vivo uric acid reduction and kidney protection activity screening of (II) geniposide derivative
Experimental Kunming mice (60 males, 20 + -5 g in weight, certificate number: SCXK [ Yu ] 2020-. Mice were first acclimated to the laboratory environment for 1 week prior to the start of the experiment, during which time the mice were fed normally and monitored for body weight. Subsequently, all mice were randomly divided into two groups, 10 in the blank group and 50 in the model group. The model group establishes a hyperuricemia mouse model by means of combined administration of adenine and potassium oxonate, receives drug feeding once a day, randomly divides the mice in the model group into 8 groups (6 mice in each group) after one week of drug feeding, and randomly selects 5 mice from the blank group as blank control. Each group received one time of Potassium Oxonate + adenine feeding and corresponding geniposide derivative treatment every day, and the dosage was strictly calculated according to the human and mouse dosage conversion formula for two weeks:
blank group: 1% CMC-Na (10 mL/kg)
Model group: 1% CMC-Na (10 mL/kg) + adenine (200 mg/kg) + Potassium Oxonate (500 mg/kg)
Allopurinol group (10 mg/kg + 0.5% CMC-Na, 10 mL/kg)
Geniposide group (200 mg/kg + 0.5% CMC-Na, 10 mL/kg)
About 0.4 mL of blood is collected from the retroorbital venous plexus after two weeks (a small amount of heparin sodium is added into a blood collection tube), the blood is kept still for 30 min at room temperature to allow the blood to coagulate naturally, the blood is centrifuged at 3000 rpm for 5 min, serum is separated, and the contents of SUA, Cr and BUN are detected (the specific operation is carried out according to the kit specification). After blood sampling was completed, the experimental mice were sacrificed by cervical dislocation, and then abdominal dissection was performed: carefully cutting off liver leaves, putting the liver leaves into a 2 mL freezing tube, and storing the liver leaves in a refrigerator at the temperature of-20 ℃; carefully separating the left kidney and the right kidney with surgical forceps, carefully removing the envelope on the kidney, taking off the left kidney, putting the left kidney into a 2 mL freezing tube, and storing the left kidney in a refrigerator at the temperature of-20 ℃. After the liver tissue and the left kidney tissue are collected, the liver tissue and the left kidney tissue are intensively transferred to a refrigerator at minus 80 ℃ for freezing storage, the right kidney is fixed in a paraformaldehyde solution with the volume being about 10 times of that of the kidney tissue and stored in the refrigerator at 4 ℃, and the organ tissue is used for subsequent experiments.
Measuring SUA by adopting an enzyme dynamic colorimetry; detecting Cr by adopting a sarcosine oxidase method; the BUN was measured using the urease method and the results are shown in table 2.
TABLE 2
Note: in comparison with the normal control group, # # p <0.01, # # p < 0.001; p <0.05, p <0.01, p <0.001, compared to the hyperuricemia group
The 4 geniposide derivatives showed higher uric acid reducing activity compared with mice in hyperuricemic group (SUA = 318.16 μ M, Cr = 87.20 μ M, BUN = 9.87 mM) and mice in positive control allopurinol group (SUA = 233.62 μ M, Cr = 63.35 μ M, BUN = 6.30 mM), wherein the geniposide derivatives 6a, 6c, 6j all showed similar uric acid reducing activity to allopurinol. Except the geniposide derivative 6j, the compound shows certain kidney protection effect. Geniposide derivative 6c inhibited XOD activity and repaired kidney injury activity best (SUA = 236.03 μ M, Cr = 40.85 μ M, BUN = 4.14 mM) compared to 4 geniposide derivatives, and activity was comparable to allopurinol overall.
(III) performing histological detection on the mice in the step (II), wherein specific results are shown in fig. 1, and it can be seen from fig. 1 that HE staining results show that a large number of renal tubules of the mice in the hyperuricemia group are expanded, renal tubular epithelial cells are vacuolated, the renal tubular epithelial cells of the mice are unclear, connective tissue hyperplasia is accompanied, and neutrophil punctate infiltration is caused, so that necrosis of a small number of renal tubular epithelial cells, nuclear fragmentation and brush border shedding of the renal tubular epithelial cells are observed, and severe renal inflammatory reaction is caused in the mice in the hyperuricemia group, so that the kidneys are seriously damaged; in addition, the kidney of the allopurinol group mouse has obvious pathological changes such as vacuolation and the like. As shown in figure 2, Masson staining results show that the kidney of the mouse with high uric acid can obviously dilate the renal tubules, a plurality of local inflammatory cells infiltrate into the kidney, and the interstitial part can obviously fibrosis the kidney; the kidney interstitial fibrosis of the allopurinol mice is obvious, a large number of tissue vacuoles are generated, and the kidney is excessive to generate inflammation. The treated mice have the characteristics of uniform staining of kidney tissues, normal glomerular morphological structure, clear renal tubular structure, close arrangement of renal tubular epithelial cells and more lymphocyte focal infiltration around local blood vessels, which shows that the geniposide derivatives 6a, 6c, 6g and 6j have certain protection effect on kidney injury caused by hyperuricemia, and the protection effect takes the geniposide derivative 6c as the strongest.
Toxicity test of (tri) geniposide derivatives on HK-2 cells
In order to verify that high-concentration uric acid has harmful stimulation on renal tubular epithelium HK-2 cells and judge the toxic effect of geniposide derivatives on HK-2 cells, the activity of the cells of each experimental group is researched by a CCK-8 method. As shown in FIG. 3, the cell viability of HK-2 cells gradually decreased with increasing uric acid concentration, and was less than 80% at 800. mu.M. The influence of the geniposide derivative 6c on the cell viability of HK-2 cells is very little, and the geniposide derivative 6c still does not obviously show the cytotoxicity to the HK-2 cells until the concentration of the geniposide derivative 6c reaches 100 mu M.
Results and discussion of anti-inflammatory Activity of (tetra) geniposide derivative 6c
And (3) selecting 600 mu M uric acid for carrying out inflammatory induction on HK-2 cells, and carrying out drug intervention by adopting 12.5, 25, 50 and 100 mu M geniposide derivative 6 c.
The geniposide derivative 6c acts on HK-2 cells induced by 600 mu M uric acid, and the content of IL-1 beta, TNF-alpha and TGF-beta in the cells is measured. As can be seen from FIG. 4, after the normal cell group is induced by uric acid, the contents of IL-1 beta, TNF-alpha and TGF-beta are obviously increased, the contents of the IL-1 beta, TNF-alpha and TGF-beta induced by uric acid can be obviously reduced after the action of geniposide derivatives with different concentrations, and the higher the compound concentration is, the better the inhibition effect is. And under the same concentration, the geniposide derivative 6c has stronger inhibiting effect on IL-1 beta, TNF-alpha and TGF-beta generated by uric acid induction than the parent nucleus compound and the positive control. This suggests that geniposide derivative 6c can improve the inflammatory response induced by hyperuricemia by inhibiting the expression of inflammatory factors IL-1 β, TNF- α, and TGF- β.
(penta) geniposide derivative 6c reduces kidney urate deposition
Uric acid deposition was detected using polarized light microscopy in hyperuricemia renal rats. As shown in fig. 5, the kidneys of hyperuricemia mice had significant uric acid accumulation compared to the blank group. Allopurinol 10 mg/kg can effectively reduce the accumulation of uric acid in the kidney of a mouse. In addition, compared with the hyperuricemia group, different doses (5, 10 and 20 mg/kg) effectively reduce the uric acid accumulation in the kidney, and show that the uric acid accumulation level is obviously reduced after the geniposide derivative 6c is treated, and the dose-dependent effect is shown.
Claims (10)
1. An amide penta-acetyl geniposide derivative is characterized in that the molecular structural formula is as follows:
r is
、
、
、
、
、
、
、
、
、
、
、
2. The preparation method of the amide pentaacetyl geniposide derivative according to claim 1, characterized by comprising the following steps:
(1) adding the geniposide derivative 2a into DMF, stirring the solution, adding EDCI, adding HOBT and DIPEA, and stirring for 2 h at room temperature;
(2) after the stirring is finished, RNH is added2(30.0 mg,0.37 mmol) and DMAP (63.0 mg, 0.51 mmol) and stirring at room temperature for 8-10 h;
(3) after the reaction is finished, adding ice water, and extracting with dichloromethane; the combined organic solvents were washed with 1M aqueous diluted hydrochloric acid and saturated brine; organic phase in anhydrous Na2SO4Dried overnight, distilled under reduced pressure, and purified by column chromatography to give the product as a pale yellow powder.
3. The preparation method according to claim 2, wherein in step (1), the concentration of the geniposide derivative 2a in DMF is 0.11-0.12 mmol/mL; the molar ratio of the geniposide derivative 2a to EDCI is 0.34: 0.41; the molar ratio of HOBT to EDCI is 1: 1; the molar ratio of DIPEA to HOBT is 1.02: 0.41.
4. the method according to claim 2 or 3, wherein in the step (2), the RNH is2And geniposide derivative 2a at a molar ratio of 0.37: 0.34; the DMAP and RNH2In a molar ratio of 0.51: 0.37.
5. the method of claim 4, wherein the RNH is2Specifically 1-aminopyrrole, 1-aminoindazole, 1-aminooxazole, 1-aminobiphenyl, 1-aminoindole, 1-aminopropionitrile, 1-amino-4-bromonaphthalene, p-aminobenzoic acid, 3-fluoroaniline, 4-aminobenzonitrile, 4-aminomethylpyridine, 5-aminotetrazole, 4-aminoisoquinoline and 4-aminomauric acid.
6. The preparation method according to any one of claims 2 to 5, wherein the geniposide derivative 2a has a structural formula:
7. the preparation method according to claim 6, wherein the geniposide derivative 2a is prepared by a specific method comprising:
(1) sequentially adding 0.257 mmol of geniposide and 10 mL of 4% NaOH solution into a round-bottom flask, refluxing and stirring at 65 ℃ for reaction, tracking by TLC (thin layer chromatography) until a raw material point disappears, neutralizing by using 1M hydrochloric acid until the pH is =7 after the reaction is finished, refining, concentrating the obtained reaction solution under reduced pressure to dryness, and passing through a column to obtain white powder geniposidic acid 1 a;
(2) sequentially adding geniposide derivative 1a0.026 mmol and triethylamine 5 mL into round-bottom flask, cooling with ice water bath, slowly dropwise adding acetic anhydride 5 mL while stirring, removing ice water bath after dropwise adding, reacting at room temperature, tracking by TLC until the raw material point disappears, and reacting with saturated NaHCO3Solution 20 mL of neutralized solution to pH =7.0, washed with deionized water, combined organic phases, anhydrous Na2SO4DryingOver night, filtering by suction to remove Na2SO4Then, the mixture is distilled under reduced pressure and is subjected to column chromatography to obtain light yellow powder pentaacetyl geniposidic acid 2 a.
8. The preparation method according to claim 7, wherein the formula of the geniposide 1a is as follows:
9. the preparation method according to claim 1, wherein in the step (3), the eluent used for the column chromatography purification is composed of petroleum ether/ethyl acetate = 10/1-1/1.
10. An application of the amide penta-acetyl geniposide derivative prepared by the preparation method according to any one of claims 2-9 in preparing a medicament for reducing uric acid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210384149.9A CN114671920B (en) | 2022-04-13 | 2022-04-13 | Amide pentaacetyl geniposide derivative as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210384149.9A CN114671920B (en) | 2022-04-13 | 2022-04-13 | Amide pentaacetyl geniposide derivative as well as preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114671920A true CN114671920A (en) | 2022-06-28 |
| CN114671920B CN114671920B (en) | 2023-06-20 |
Family
ID=82077897
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210384149.9A Active CN114671920B (en) | 2022-04-13 | 2022-04-13 | Amide pentaacetyl geniposide derivative as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114671920B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115785044A (en) * | 2023-02-08 | 2023-03-14 | 山东省华泰营养健康产业技术研究院有限公司 | Iminogenipin with uric acid reducing activity and preparation method and application thereof |
| CN116173054A (en) * | 2023-04-27 | 2023-05-30 | 中国中医科学院中药研究所 | Application of geniposide acid in preparation of uric acid reducing and/or kidney injury treating medicines |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875617A (en) * | 2012-09-12 | 2013-01-16 | 安徽医科大学 | Geniposide derivative, preparation method thereof and application of geniposide derivative to inflammation resistance |
| CN110437291A (en) * | 2019-08-16 | 2019-11-12 | 山东省农业科学院农产品研究所 | One kind having active five acetyl Gardenoside Cyclohexamide of anti-trioxypurine and preparation method thereof, application |
| CN113768945A (en) * | 2021-08-12 | 2021-12-10 | 宁夏医科大学 | Application of geniposide in preparation of medicine for treating oligospermia |
| CN114315929A (en) * | 2022-01-21 | 2022-04-12 | 山东省农业科学院 | Geniposide derivative with xanthine oxidase inhibitory activity, and preparation method and application thereof |
-
2022
- 2022-04-13 CN CN202210384149.9A patent/CN114671920B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875617A (en) * | 2012-09-12 | 2013-01-16 | 安徽医科大学 | Geniposide derivative, preparation method thereof and application of geniposide derivative to inflammation resistance |
| CN110437291A (en) * | 2019-08-16 | 2019-11-12 | 山东省农业科学院农产品研究所 | One kind having active five acetyl Gardenoside Cyclohexamide of anti-trioxypurine and preparation method thereof, application |
| CN113768945A (en) * | 2021-08-12 | 2021-12-10 | 宁夏医科大学 | Application of geniposide in preparation of medicine for treating oligospermia |
| CN114315929A (en) * | 2022-01-21 | 2022-04-12 | 山东省农业科学院 | Geniposide derivative with xanthine oxidase inhibitory activity, and preparation method and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115785044A (en) * | 2023-02-08 | 2023-03-14 | 山东省华泰营养健康产业技术研究院有限公司 | Iminogenipin with uric acid reducing activity and preparation method and application thereof |
| CN116173054A (en) * | 2023-04-27 | 2023-05-30 | 中国中医科学院中药研究所 | Application of geniposide acid in preparation of uric acid reducing and/or kidney injury treating medicines |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114671920B (en) | 2023-06-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN114671920B (en) | Amide pentaacetyl geniposide derivative as well as preparation method and application thereof | |
| EP2427416B1 (en) | Substituted aromatic compounds and pharmaceutical uses thereof | |
| EA023804B1 (en) | Oxabicycloheptanes, their preparation and use | |
| HUE035073T2 (en) | A NOVEL CHOLESTEROL METABOLITE, 5-CHOLESTEN, 3beta-25-DIOL, DISULFATE (25HCDS) FOR THERAPY OF METABOLIC DISORDERS, HYPERLIPIDEMIA, DIABETES, FATTY LIVER DISEASES AND ATHEROSCLEROSIS | |
| WO2014154131A1 (en) | Caffeoyl substituted pentacyclic triterpene derivative and use thereof | |
| EP3313816B1 (en) | Metabolically robust analogs of cyp-eicosanoids for the treatment of cardiac disease | |
| CN109651199B (en) | Histone deacetylase and microtubule double-target inhibitor and preparation method thereof | |
| CN108349911A (en) | Releasable nitric oxide production prodrugs | |
| CN110627774A (en) | Anti-hepatic fibrosis compound, preparation method and application | |
| CN114736253A (en) | Ester penta-acetyl geniposide derivative obtained through esterification reaction, and preparation method and application thereof | |
| WO2015188609A1 (en) | New use of a benzopyran derivative in preparation of medicament for treating hyperuricemia | |
| KR20100097162A (en) | Dioxoanthracene sulphonate derivatives | |
| Xu et al. | Design, synthesis and in vitro biological evaluation of marine phidianidine derivatives as potential anti-inflammatory agents | |
| US10947179B2 (en) | 2-(3,4-dihydroxyphenyl)ethyl 3-hydroxybutanoate, composition, and method for improving function of aortic endothelial cell | |
| KR102433731B1 (en) | Analogs of CYP-eicosanoids for use in the treatment or prevention of disorders associated with angiogenesis and/or inflammation | |
| WO2015196323A1 (en) | New derivatives of 2-(3-cyano-4-isobutyloxyphenyl)-4-methylthiazole-5-formate, preparation method therefor and application thereof | |
| US20060111327A1 (en) | Derivatives of triptolide having a high immunosuppressive effect and high water solubility, and uses thereof | |
| CN114292226B (en) | Compound with hydroxamic acid structure and preparation method and application thereof | |
| CN111417632B (en) | Thiazole-5-formic acid derivative and preparation method and application thereof | |
| CN100551364C (en) | The purposes of 3-(3-nitro-4-hydroxyl) phenyl-ethyl acrylate in the medicine of preparation inhibition xanthine oxidase | |
| CN110078668B (en) | Phenyl imidazole XOR inhibitor, preparation and application | |
| CN1966484B (en) | New phenoxy eicosanoic acid derivative and its medical use | |
| CN102018688A (en) | PTP1B inhibitor and synthesis and application thereof in preparation of medicine used for treating type II diabetes | |
| CN114989233B (en) | Ester pentaacetylgeniposide derivative obtained through acyl chlorination reaction, and preparation method and application thereof | |
| CN107880094B (en) | Ursolic acid neutral cholesterol ester hydrolase inhibitor and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |