CN110551109B - Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof - Google Patents

Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof Download PDF

Info

Publication number
CN110551109B
CN110551109B CN201810564285.XA CN201810564285A CN110551109B CN 110551109 B CN110551109 B CN 110551109B CN 201810564285 A CN201810564285 A CN 201810564285A CN 110551109 B CN110551109 B CN 110551109B
Authority
CN
China
Prior art keywords
diketopiperazine
butyl
methyl
indole
boc
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.)
Expired - Fee Related
Application number
CN201810564285.XA
Other languages
Chinese (zh)
Other versions
CN110551109A (en
Inventor
赵明
王玉记
张筱宜
彭师奇
吴靖
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Capital Medical University
Original Assignee
Capital Medical University
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Capital Medical University filed Critical Capital Medical University
Priority to CN201810564285.XA priority Critical patent/CN110551109B/en
Publication of CN110551109A publication Critical patent/CN110551109A/en
Application granted granted Critical
Publication of CN110551109B publication Critical patent/CN110551109B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/04—Antineoplastic agents specific for metastasis
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/06—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing only aliphatic carbon atoms

Landscapes

  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Oncology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The invention discloses a 3R- (indole-3-methyl) -6R- [4- (Lys-aminomethyl cycloamido n-butyl) with the following structure]-2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl)]-2, 5-diketopiperazine. Discloses a preparation method of the compounds and an anti-tumor and anti-metastasis effect of the compounds, and clarifies the application of the compounds in preparing anti-tumor and anti-metastasis medicaments.
Figure DDA0001684127830000011

Description

Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof
Technical Field
The present invention relates to 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine. Relates to a preparation method thereof, relates to the anti-tumor and anti-metastasis effects thereof, and relates to the application thereof in preparing anti-tumor and anti-metastasis medicaments. The invention belongs to the field of biological medicine.
Technical Field
The lung cancer is the most common malignant tumor at present, the high morbidity and the high mortality of the lung cancer in China are positioned at the first position of the malignant tumor, 90 percent of lung cancer patients die from tumor metastasis rather than primary tumor, and the inventor discloses that diketopiperazines with S, S-, R, R-, R, S-and S, R-four configurations can inhibit migration and invasion of HCCLM3 (highly metastatic human liver cancer cells) at the concentration of 0.5 mu M. Later, the inventors further disclosed that R, R-configured diketopiperazines at a minimum effective dose of 5. mu. mol/kg/day inhibited tumor metastasis to the lung in C57BL/6 mice and tumor proliferation in ICR mice. To reduce the minimum effective dose, the inventors have developed various modifications to the butylamino group of the diketopiperazine in the R, R-configuration. After 3 years of exploration, it was found that acylation of the 4-amino n-butyl group of the diketopiperazine of R, R-configuration with the amino acids L-Lys and L-Thr reduced the minimum effective antitumor dose to 0.5. mu. mol/kg/day. The 10-fold reduction of the lowest effective dose indicates that the structural modification has outstanding technical effects. Based on these findings, the inventors have proposed the present invention.
Disclosure of Invention
The first aspect of the present invention provides 3R- (indol-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indol-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine of the following structures.
Figure BDA0001684127810000011
A second aspect of the present invention provides a process for the preparation of 3R- (indol-3-methyl) -6R- [4- (Lys-carbamoylamino-n-butyl) ] -2, 5-diketopiperazine and 3R- (indol-3-methyl) -6R- [4- (Thr-carbamoylamino-n-butyl) ] -2, 5-diketopiperazine, which process comprises:
(1) reacting D-Trp-OBzl with Boc-D-Lys (Cbz) to obtain Boc-D-Lys (Cbz) -D-Trp-OBzl;
(2) removing Boc from hydrogen chloride in ethyl acetate solution (4M) with ice bath to obtain D-Lys (Cbz) -D-Trp-OBzl;
(3) Boc-D-Lys (Cbz) -D-Trp-OBzl is automatically cyclized under a weak alkaline environment to generate 3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino n-butyl) -2, 5-diketopiperazine (1);
(4)3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino n-butyl) -2, 5-diketopiperazine (1) is subjected to catalytic hydrogenolysis to remove benzyloxycarbonyl to obtain 3R- (indole-3-methyl) -6R- (4-amino n-butyl) -2, 5-diketopiperazine (2);
(5) reacting 3R- (indole-3-methyl) -6R- (4-amino-n-butyl) -2, 5-diketopiperazine (2) with Boc-tranexamic acid to produce 3R- (indole-3-methyl) -6R- [4- (Boc-tranylcarboxamido-n-butyl) ] -2, 5-diketopiperazine (3);
(6) removing Boc from the 3R- (indole-3-methyl) -6R- [4- (Boc-carbamimidoyl n-butyl) ] -2, 5-diketopiperazine (3) in ethyl acetate solution of hydrogen chloride (4M) under ice bath to obtain 3R- (indole-3-methyl) -6R- [4- (carbamimidoyl n-butyl) ] -2, 5-diketopiperazine (4);
(7) condensing 3R- (indole-3-methyl) -6R- [4- (carbamic acid amino n-butyl) ] -2, 5-diketopiperazine (4) with Boc-L-Lys or Boc-L-Thr to obtain a compound 3R- (indole-3-methyl) -6R- [4- (Boc-Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5a) or 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5 b);
(8) removing Boc in ethyl acetate solution of hydrogen chloride under ice bath (4M)3R- (indole-3-methyl) -6R- [4- (Boc-Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5a) or 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5b) to obtain 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (6a) or 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (6 b).
The third aspect of the present invention is to evaluate the inhibitory effect of 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine on the tumor growth of S180 mice and the anti-lung cancer metastasis activity of C57BL/6 mice.
Drawings
FIG. 1.3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl)]-2, 5-diketopiperazine (6a) and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl)]-a synthetic route for 2, 5-diketopiperazines (6 b); i) n, N-Dicyclohexylcarbodiimide (DCC), N-hydroxybenzotriazole (HOBt), N-methylmorpholine (DMM), Tetrahydrofuran (THF); ii)4N hydrogen chloride-ethyl acetate reagent; iii) ethyl acetate, 5% sodium bicarbonate; iv) methanol, N, N-Dimethylformamide (DMF), Pd/C, H2(ii) a v) N, N-Dicyclohexylcarbodiimide (DCC), N-hydroxybenzotriazole (HOBt), N-methylmorpholine (DMM) anhydrous N, N-Dimethylformamide (DMF).
Detailed Description
To further illustrate the invention, a series of examples are given below. These examples are purely illustrative and are intended to be a detailed description of the invention only and should not be taken as limiting the invention.
EXAMPLE 1 preparation of Boc-D-Lys (Cbz) -D-Trp-OBzl
1.930g (5mmol) Boc-D-Lys (Cbz) was suspended in 100mL dry Tetrahydrofuran (THF) in an ice bathThen, 0.747g (5.5mmol) of 1-hydroxybenzotriazole (HOBt) and 1.243g (6.0mmol) of N, N-Dicyclohexylcarbodiimide (DCC) were added to the suspension in this order, followed by stirring for 30 minutes. Then, 1.620g (5.2mmol) of D-Trp-OBzl HCl was added. The reaction mixture was adjusted to pH 8-9 by dropwise addition of N-methylmorpholine (NMM). The reaction mixture was stirred first for 1h under ice bath and then at room temperature for 12 h. The reaction mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was dissolved in 150mL of ethyl acetate. The mixture was washed with a saturated 5% aqueous solution of potassium hydrogensulfate and a saturated aqueous solution of sodium chloride, respectively, and extracted three times. The ethyl acetate layer was dried over anhydrous sodium sulfate for 12 hours. Filtering, and concentrating under reduced pressure to dryness. The resulting yellow syrup was separated and purified by silica gel column chromatography (ethyl acetate: petroleum ether ═ 1:3 to 1:1) to obtain 2.423 (73.2%) g of the title compound as a pink solid powder. ESI-MS (M/e):657[ M + H]+。
EXAMPLE 2 preparation of D-Lys (Cbz) -D-Trp-OBzl
1.9g (2.5mmol) Boc-D-Lys (Cbz) -D-Trp-OBzl was slowly mixed with 40mL of ethyl acetate (4M) of hydrogen chloride with stirring in an ice bath. The resulting solution was stirred in an ice bath for 3 hours. After that, the reaction mixture was dried under reduced pressure, and the residue was sufficiently stirred with dry ethyl acetate and then dried by suction, and repeated three times. The residue was triturated with dry ether three times to give 1.7g (93%) of the title compound as a dark red solid powder. ESI-MS (M/e):557[ M + H ]]+。
EXAMPLE 3 preparation of 3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino-n-butyl) -2, 5-diketopiperazine (1)
1.7g (3.1mmol) of D-Lys (Cbz) -D-Trp-Obzl were dissolved in 100mL of ethyl acetate. The resulting solution was washed three times with 5% aqueous sodium bicarbonate solution, the ethyl acetate layer was concentrated under reduced pressure and stirred at room temperature for 12 hours, after white solid was sufficiently precipitated, it was filtered to obtain 0.76g (46.5%) of the title compound. ESI-MS (M/e):449[ M + H ]]+。
EXAMPLE 4 preparation of 3R- (indole-3-methyl) -6R- (4-amino-n-butyl) -2, 5-diketopiperazine (2)
To 0.900g (2mmol) of 3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino-N-butyl) -2, 5-diketopiperazine (1) was added 10mL of methanol and 10mL of N, N-Dimethylformamide (DMF) to dissolve, and then90mg of Pd/C was added to the above solution, the air in the flask was evacuated, and hydrogen gas was introduced to the flask to stir the reaction at room temperature for 12 hours. Pd/C was filtered off, and the filtrate was concentrated to dryness under reduced pressure to give the title compound as a white solid powder 0.600 (95%). ESI-MS (M/e) 315[ M + H ]]+。
EXAMPLE 5 preparation of 3R- (indole-3-methyl) -6R- [4- (Boc-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (3)
From 0.668g (2.6mmol) of Boc-tranexamic acid and 0.813g (2.6mmol) of 3R- (indole-3-methyl) -6R- (4-amino-n-butyl) -2, 5-diketopiperazine (2) by the method of example 1, 0.77g (53.8%) of the title compound are obtained as a pale yellow powder. ESI-MS (M/e):554[ M + H]+；1HNMR(300MHz,DMSO-d6)δ/ppm＝10.88(s,1H),8.05(s,1H),7.94(s,1H),7.58(d,J＝7.8Hz,1H),7.50(t,J＝4.8Hz,1H),7.30(d,J＝7.8Hz,1H),7.03(s,1H),7.02(m,1H),6.93(t,J＝7.4Hz,1H),6.81(t,J＝5.6Hz,1H),4.11(m,1H),3.50(m,1H),3.25(dd,J1＝4.0Hz,J2＝14.2Hz,1H),3.00(dd,J1＝4.0Hz,J2＝14.4Hz,1H),2.76(m,4H),1.97(m,1H),1.68(m,4H),1.37(s,9H),1.27(m,3H),0.96(m,3H),0.83(m,2H),0.58(m,3H)。
EXAMPLE 6 preparation of 3R- (indole-3-methyl) -6R- [4- (amino n-butyl tranexamate) ] -2, 5-diketopiperazine (4)
Following the procedure of example 2, from 0.75g (1.35mmol) of 3R- (indole-3-methyl) -6R- [4- (Boc-carbamoylamino n-butyl)]-2, 5-diketopiperazine (3) gave 0.65g (98.3%) of the title compound as a pale yellow solid. ESI-MS (m/e): 454;1HNMR(300MHz,DMSO-d6)δ/ppm＝10.94(s,1H),8.04(s,1H),8.02(br,3H),7.94(s,1H),7.58(m,2H),7.31(d,J＝8.0Hz,1H),7.04(s,1H),7.03(m,1H),6.93(t,J＝7.2Hz,1H),4.11(m,1H),3.50(m,1H),3.24(dd,J1＝3.4Hz,J2＝14.2Hz,1H),3.02(dd,J1＝3.4Hz,J2＝14.4Hz,1H),2.76(m,2H),2.63(m,2H),2.02(m,1H),1.76(m,4H),1.53(m,1H),1.32(m,2H),0.97(m,5H),0.60(m,3H)。
EXAMPLE 7 preparation of 3R- (indole-3-methyl) -6R- [4- (Boc-Lys (Boc) -carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5a)
In accordance with the method of example 1,from 0.76g (2.2mmol) Boc-Lys (Boc) and 0.99g (2.0mmol)3R- (indole-3-methyl) -6R- [4- (tranexamic acid amino-n-butyl)]-2, 5-diketopiperazine (4) gave 1.03g (65.2%) of the title compound as a pale yellow solid powder. ESI-MS (M/e):783[ M + H]+；1HNMR(300MHz,DMSO-d6)δ/ppm＝11.00(s,1H),8.07(s,1H),7.95(s,1H),7.79(m,1H),7.58(m,2H),7.30(d,J＝7.8Hz,1H),7.03(s,1H),7.01(m,1H),6.91(m,1H),6.77(m,1H),4.10(m,1H),3.81(m,1H),3.48(m,1H),3.24(dd,J1＝2.3Hz,J2＝14.4Hz,1H),3.02(dd,J1＝3.2Hz,J2＝14.4Hz,1H),2.86(m,4H),2.74(m,2H),1.98(m,1H),1.68(m,4H),1.49(m,2H),1.36(s,18H),1.30(m,7H),0.95(m,3H),0.83(m,2H),0.56(m,3H)。
EXAMPLE 8 preparation of 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (5b)
Following the procedure of example 1, starting from 0.49g (2.2mmol) Boc-Thr and 1.02g (2.1mmol)3R- (indole-3-methyl) -6R- [4- (carbamic acid amino n-butyl)]-2, 5-diketopiperazine (4) gave 0.42g (24.6%) of the title compound as a pale yellow solid powder. ESI-MS (M/e):655[ M + H]+；1HNMR(300MHz,DMSO-d6)δ/ppm＝10.97(s,1H),8.05(s,1H),7.94(s,1H),7.76(m,1H),7.57(m,2H),7.30(d,J＝7.8Hz,1H),7.04(s,1H),7.01(m,1H),6.92(m,1H),6.32(m,1H),4.78(m,1H),·4.10(m,1H),3.88(m,1H),3.80(m,1H),3.49(m,1H),3.24(dd,J1＝3.8Hz,J2＝14.6Hz,1H),3.02(dd,J1＝4.2Hz,J2＝14.2Hz,1H),2.91(m,2H),2.74(m,2H),1.99(m,1H),1.70(m,4H),1.38(s,9H),1.27(m,3H),1.03(d,3H,J＝5.8Hz),0.98(m,3H),0.82(m,2H),0.59(m,3H)。
EXAMPLE 9 preparation of 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl)]-2, 5-Diketopiperazine (6a) following the procedure of example 2, from 0.204g (0.26mmol) of 3R- (indole-3-methyl) -6R- [4- (Boc-Lys (Boc) -carbamoylamino n-butyl)]-2, 5-diketopiperazine (5a) gave 0.163g (95.4%) of the title compound as a pale yellow powder. ESI-MS (M/e):583[ M + H]+；
Figure BDA0001684127810000052
＝+34.68,(C 0.13,CH3OH)；IR(cm-1):3491,3450,3218,3050,2926,2859,1656,1628,1614,1556,1455,1329,1264,1095,1010,736；1HNMR(300MHz,DMSO-d6)δ/ppm＝11.02(s,1H),8.76(m,1H),8.38(s,3H),8.23(s,3H),8.08(s,1H),7.96(s,1H),7.66(m,1H),7.57(d,J＝8.0Hz,1H),7.30(d,J＝8.0Hz,1H),7.03(s,1H),7.02(m,1H),6.91(m,1H),4.10(m,1H),3.80(m,1H),3.49(m,1H),3.23(dd,J1＝3.7Hz,J2＝14.7Hz,1H),2.99(m,3H),2.73(m,4H),2.02(m,1H),1.72(m,6H),1.61(m,2H),1.33(m,5H),0.95(m,5H),0.58(m,3H)。
EXAMPLE 10 preparation of 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine (6b)
Following the procedure of example 2, from 0.120g (0.18mmol) of 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamoylamino n-butyl)]-2, 5-diketopiperazine (5b) gave 0.098g (90.8%) of the title compound as a white powder. ESI-MS (M/e) 555[ M + H ]]+；
Figure BDA0001684127810000051
＝+9.17,(C 0.16,CH3OH)；IR(cm-1):3235,3084,2928,2851,1654,1556,1454,1331,1247,1095,1011,924,745；1HNMR(300MHz,DMSO-d6)δ/ppm＝11.00(s,1H),8.69(m,1H),8.18(m,3H),8.07(s,1H),7.95(s,1H),7.62(m,1H),7.57(d,J＝7.8Hz,1H),7.30(d,J＝7.8Hz,1H),7.03(s,1H),7.02(m,1H),6.91(m,1H),5.60(m,1H),4.10(m,1H),3.89(m,1H),3.61(m,1H),3.48(m,1H),3.24(dd,J1＝3.9Hz,J2＝14.3Hz,1H),3.01(dd,J1＝4.5Hz,J2＝14.3Hz,1H),2.91(m,2H),2.74(m,2H),2.01(m,1H),1.72(m,4H),1.27(m,3H),1.13(d,3H,J＝6.3Hz),0.95(m,5H),0.58(m,3H)。
Experimental example 1 evaluation of antitumor proliferative Activity of Compounds 6a, b
1) The compounds 6a, b were dissolved in physiological saline, doxorubicin was dissolved in physiological saline as a positive control, and physiological saline as a negative control;
2) the oral dosage of the compound 6a and the compound b is 0.5 mu mol/kg, the oral dosage of physiological saline is 0.2mL/20g, and the intraperitoneal injection dosage of the adriamycin is 2 mu mol/kg. The administration was started 5 days after tumor inoculation and continued for 10 days for a total of 10 administrations.
3) The experimental animals were ICR male mice (clean grade) weighing 20 + -2 g, 12 mice per group.
4) The tumor source is mouse S180 sarcoma purchased from animal experiment center of department of medicine of Beijing university and maintained by self passage.
5) Inoculating S180 ascites tumor liquid which grows vigorously under aseptic condition, diluting the liquid into liquid (1:2) by using normal saline, fully mixing, staining tumor cell suspension by using freshly prepared 0.2% trypan blue, uniformly mixing, counting according to a white cell counting method, wherein the blue stained cells are dead cells, and the non-stained cells are live cells. The cell concentration is 4-large-grid viable cell number/4 × 104The cell concentration was calculated by x dilution factor ═ cell number/mL. The cell survival rate was calculated as the number of live cells/(number of live cells + number of dead cells) × 100%.
Preparing tumor solution with survival rate of more than 90% into 2.0 × 10 by homogenizing method7One cell/mL of the cell suspension was inoculated subcutaneously to the mouse axilla, and 0.2 mL/mouse was prepared as S180 tumor-bearing mice. Tumor inoculation was administered 5 days later. The mice in the treatment group were dosed with compound 6a, b orally at 0.5 μmol/kg daily. The daily oral physiological saline dose of the blank group of mice is 0.2mL/20 g. The dose of the positive control group of mice injected with adriamycin into the abdominal cavity every day is 2 mu mol/kg. The administration was continued for ten days, and on the eleventh day, the mice were weighed, anesthetized with ether, sacrificed by removing the cervical vertebrae, then the right axillary tumor growth sites of the mice were fixed with forceps, the skin was cut open, the tumors were exposed, blunt-stripped, and weighed. Experimental data were expressed as (mean ± SD g) tumor weights using t-test and analysis of variance. The results are shown in Table 1. As can be seen from Table 1, compound 6a, b effectively inhibited tumor regrowth in mice at an oral dose of 0.5. mu. mol/kg. The invention has obvious technical effect.
TABLE 1 antitumor Activity of Compounds 6a, b
Figure BDA0001684127810000061
a) P <0.05 in the saline group; n is 12.
Experimental example 2 evaluation of antitumor metastasis Activity of Compounds 6a, b
The assay model was inoculated with Lewis mouse lung carcinoma cells (LLC, purchased from ATCC) in DMEM medium (containing 10% inactivated fetal bovine serum, 1X 10)5U/L penicillin and 100mg/L streptomycin), and the cells are enriched by passage every two days according to an adherent cell culture method. Digesting the cells when the cells are in good growth state and in logarithmic growth phase, and adjusting the cell density to 1 × 10 with physiological saline7one/mL. Staining with placental blue to count viable cells>95 percent. Inbred C57BL/6 male mice (SPF grade, body weight 20. + -.2 g) were taken and left-handed mice fixed. The right anterior limb axillary skin of the mouse was disinfected with 75% ethanol. The LLC tumor cell suspension is injected subcutaneously into the axilla of a mouse with a 1mL sterile syringe held in the right hand, and 0.2mL is injected into each mouse. After the mice are inoculated for 10 days, tumors with the diameter of about 4-5mm grow out, namely the tumor source. The Lewis lung cancer tumor-bearing mice are inoculated for 10 days and anesthetized by ether, and then the cervical vertebrae are removed for killing. Soaking in 75% ethanol for 10min, sterilizing, and removing tumor on clean bench. Well-grown tumor tissue was selected, minced in a sterile plate, and placed in a tissue homogenizer made of glass. Adding physiological saline with the temperature of 4 ℃ according to the ratio of the tumor mass to the volume of the physiological saline of 1 to 3(g to mL), and lightly grinding to prepare the cell suspension. The cell suspension is screened by 200-mesh cells to prepare single cell suspension. Adjusting the cell density of the single cell suspension to 1.5X 10 with physiological saline7one/mL. Staining with placental blue to count viable cells>95 percent. Left-handed inbred C57BL/6 male mice were fixed and their right anterior limb axillary skin was disinfected with 75% ethanol. The tumor cell suspension was injected subcutaneously into the mouse axilla with a 1mL sterile syringe in the right hand, 0.2mL each. 10 days after inoculation, the mice developed tumors of 4-5mm in diameter, and the inoculated mice were randomly grouped by the measured tumor volume. Each group had 12 mice. On day 11 of tumor inoculation, mice were injected intraperitoneally with a normal saline solution of the putative antitumor metastatic peptide RGDS (at a dose of 20. mu. mol/kg/day) or with a normal saline solution of compounds 6a, b orally (at a dose of 0.5. mu. mol/kg/day) or with a normal saline orally (at a dose of 10 mL/kg/day), given 1 dose per day for 12 consecutive days, and tumor volumes were measured and recorded every two days. Tumor volume was measured the next day of last administration, ether anesthesiaThe cervical vertebrae are removed and killed, the tumor of the mouse is weighed, the lung of the mouse is taken, and the tumor node number of the tumor lung metastasis is calculated. Statistical analysis of the data was performed using the t-test. The results are shown in Table 2. Compound 6a, b was not only effective in inhibiting tumor lung metastasis at a dose of 0.5 μmol/kg/day (i.e., 10-fold reduction of the lowest effective dose), but the activity was not significantly different from RGDS at the 20 μmol/kg/day dose. These data indicate that the present invention has significant technical effects.
TABLE 2 antitumor metastasis Activity of Compounds 6a, b
Figure BDA0001684127810000071
a) P <0.01 to saline, p >0.05 to RGDS; b) p is less than 0.01 to normal saline, p is less than 0.05 to RGDS; n is 12.

Claims (3)

1.3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine of the following structures,
Figure FDA0002775596240000011
2. a process for the preparation of 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine of claim 1, which comprises the steps of:
(1) reacting D-Trp-OBzl with Boc-D-Lys (Cbz) to obtain Boc-D-Lys (Cbz) -D-Trp-OBzl;
(2) removing Boc from Boc-D-Lys (Cbz) -D-Trp-OBzl in ethyl acetate solution of hydrogen chloride with the concentration of 4M in ice bath to obtain D-Lys (Cbz) -D-Trp-OBzl;
(3) an ethyl acetate solution of D-Lys (Cbz) -D-Trp-OBzl is washed by a 5% sodium bicarbonate water solution and cyclized to generate 3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino n-butyl) -2, 5-diketopiperazine;
(4)3R- (indole-3-methyl) -6R- (4-benzyloxycarbonylamino n-butyl) -2, 5-diketopiperazine is subjected to catalytic hydrogenolysis to remove benzyloxycarbonyl to obtain 3R- (indole-3-methyl) -6R- (4-amino n-butyl) -2, 5-diketopiperazine;
(5) reacting 3R- (indole-3-methyl) -6R- (4-amino-n-butyl) -2, 5-diketopiperazine with Boc-tranexamic acid to produce 3R- (indole-3-methyl) -6R- [4- (Boc-carbamoylamino-n-butyl) ] -2, 5-diketopiperazine;
(6) removing Boc from 3R- (indole-3-methyl) -6R- [4- (Boc-carbamimidoyl n-butyl) ] -2, 5-diketopiperazine in 4M ethyl acetate solution of hydrogen chloride under ice bath to obtain 3R- (indole-3-methyl) -6R- [4- (carbamimidoyl n-butyl) ] -2, 5-diketopiperazine;
(7) condensing 3R- (indole-3-methyl) -6R- [4- (carbamic acid amino n-butyl) ] -2, 5-diketopiperazine with Boc-L-Lys or Boc-L-Thr to obtain a compound 3R- (indole-3-methyl) -6R- [4- (Boc-Lys-carbamic acid cycloamido n-butyl) ] -2, 5-diketopiperazine or 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamic acid cycloamido n-butyl) ] -2, 5-diketopiperazine;
(8) removing Boc from 3R- (indole-3-methyl) -6R- [4- (Boc-Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine or 3R- (indole-3-methyl) -6R- [4- (Boc-Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine in 4M ethyl acetate solution of hydrogen chloride under ice bath to obtain 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine or 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazines.
3. The use of 3R- (indole-3-methyl) -6R- [4- (Lys-carbamoylamino n-butyl) ] -2, 5-diketopiperazine and 3R- (indole-3-methyl) -6R- [4- (Thr-carbamoylamino n-butyl) ] -2, 5-diketopiperazine of claim 1 for the preparation of anti-tumor and anti-metastatic drugs.
CN201810564285.XA 2018-06-04 2018-06-04 Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof Expired - Fee Related CN110551109B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201810564285.XA CN110551109B (en) 2018-06-04 2018-06-04 Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201810564285.XA CN110551109B (en) 2018-06-04 2018-06-04 Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof

Publications (2)

Publication Number Publication Date
CN110551109A CN110551109A (en) 2019-12-10
CN110551109B true CN110551109B (en) 2021-02-12

Family

ID=68735925

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201810564285.XA Expired - Fee Related CN110551109B (en) 2018-06-04 2018-06-04 Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof

Country Status (1)

Country Link
CN (1) CN110551109B (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105503833A (en) * 2014-10-20 2016-04-20 首都医科大学 Indole piperazine dione compound, and preparation and treatment effect thereof
CN106349148A (en) * 2015-07-13 2017-01-25 首都医科大学 Novel indoles compound having anti-tumor metastasis activity and anti-inflammatory activity, as well as synthesis and application of novel indoles compound

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105503833A (en) * 2014-10-20 2016-04-20 首都医科大学 Indole piperazine dione compound, and preparation and treatment effect thereof
CN106349148A (en) * 2015-07-13 2017-01-25 首都医科大学 Novel indoles compound having anti-tumor metastasis activity and anti-inflammatory activity, as well as synthesis and application of novel indoles compound

Also Published As

Publication number Publication date
CN110551109A (en) 2019-12-10

Similar Documents

Publication Publication Date Title
CN110551109B (en) Amino acid and tranexamic acid modified diketopiperazine, preparation and application thereof
CN108929320B (en) 3R-indolylmethyl-6R-oxazolidinone modified piperazine-2, 5-dione, synthesis, activity and application thereof
CN110551175B (en) Amino acid and tranexamic acid modified diketopiperazine, preparation, activity and application thereof
CN108948137B (en) 3S-indolylethyl-6S-polar amino acid modified piperazine-2, 5-diketone and synthesis, activity and application thereof
CN108976201B (en) 3R-indolylmethyl-6S-aromatic amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN110551174B (en) Amino acid and tranexamic acid modified diketopiperazines, their preparation, activity and use
CN110551107B (en) Amino acid and tranexamic acid modified diketopiperazines, their preparation, activity and use
CN110577516B (en) Amino acid and tranexamic acid modified diketopiperazines, their preparation and use
CN110577518B (en) Methyl indole and amide side chain amino acid modified diketopiperazine, synthesis and application thereof
CN110577568B (en) Methyl indole and amide side chain amino acid modified diketopiperazine, synthesis, activity and application thereof
CN108947979B (en) 3R-indolylmethyl-6R-polar amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108947975B (en) 3S-indolylethyl-6S-fatty amino acid modified piperazine-2, 5-diketone and synthesis, activity and application thereof
CN110577517B (en) Methyl indole and aromatic amino acid modified 2, 5-diketopiperazine, synthesis, activity and application thereof
CN108947976B (en) 3S-indolylmethyl-6S-aromatic amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN110551108B (en) 3R-indolylmethyl-6R-amino acid modified 2, 5-diketopiperazines, synthesis, activity and use thereof
CN108976204B (en) 3S-indolylmethyl-6R-aspartic acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108929313B (en) 3R-indolylmethyl-6R-aliphatic amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108929369B (en) 3R-indolylmethyl-6R-acidic amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108947978B (en) 3R-indolylmethyl-6R-Tyr modified piperazine-2, 5-diketone and synthesis, activity and application thereof
CN108976277B (en) 3R-indolylmethyl-6S-polar amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108948141B (en) 3R-indolylmethyl-6S-Pro modified piperazine-2, 5-dione, synthesis, activity and application thereof
CN109456311B (en) 3R-indolylmethyl-6R-aromatic amino acid modified piperazine-2, 5-diketone and synthesis, activity and application thereof
CN108976211B (en) 3S-indolylmethyl-6R-aliphatic amino acid modified piperazine-2, 5-dione, and synthesis, activity and application thereof
CN108947977B (en) Piperazine-2, 5-diketone modified by 3R-indolylmethyl-6R-amide side chain amino acid, and synthesis, activity and application thereof
CN108929315B (en) 3R-indolylmethyl-6S-methionine modified piperazine-2, 5-dione, and synthesis, activity 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
CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20210212