CN109810173B - Dihydroisoquinoline-3-formyl-TARGD(aa)aa, its preparation, anti-venous thrombotic activity and application - Google Patents

Abstract

The invention discloses 1, 2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp (aa) -aa in the formula, wherein aa is Phe, Ser or Val residue, and discloses a preparation method and anti-venous thrombosis activity thereof, so that the invention discloses application thereof in preparing anti-venous thrombosis medicaments.

Description

Dihydroisoquinoline-3-formyl-TARGD(aa)aa, its preparation, anti-venous thrombotic activity and application

technical field

The present invention relates to 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa)-aa, to their preparation method, to their anti-venous thrombosis activity, and thus the present invention The invention relates to their application in the preparation of anti-venous thrombosis drugs. The invention belongs to the field of biomedicine.

Background technique

Thrombosis has become a disease with high morbidity and mortality. The number of patients with venous thrombosis, including deep vein thrombosis and pulmonary embolism, exceeds the total number of myocardial infarction and stroke, and is higher than the total number of deaths caused by breast cancer and AIDS. The incidence of thrombosis increases exponentially with age, which is a particularly serious threat to people's health in an aging country like my country. Taking into account the population base, venous thrombosis has a particularly serious negative impact on the national economy and people's livelihood in my country. The prevention and treatment of venous thrombosis has always been one of the focuses of the medical field. Although warfarin has been used clinically since 1941, its safety window is narrow. Low doses can cause pulmonary embolism, and high doses can cause fatal bleeding. Although a lot of effort has been devoted to the invention of safe anti-venous thrombosis drugs for more than 50 years, the results are very small. In the study of antithrombotic drugs, the inventors once disclosed that N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Arg- Gly-Asp(Ser)-Ser,N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Arg-Gly-Asp(Val)-Val and N -[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Arg-Gly-Asp(Phe)-Phe can effectively inhibit arterial thrombosis in rats, but has no effect on venous thrombosis. Therapeutic effect. The inventors found 1,2-dihydroisoquinoline-3-carboxyl-Thr-Arg-Gly-Asp(Ser)-Ser,1,2-dihydroisoquinoline- 3-Formyl-Thr-Arg-Gly-Asp(Val)-Val and 1,2-dihydroisoquinoline-3-formyl-Thr-Arg-Gly-Asp(Phe)-Phe (see transformation below Mode). In follow-up studies, the inventors further found that 1,2-dihydroisoquinoline-3-carboxyl-Thr-Arg-Gly-Asp(Ser)-Ser,1,2-dihydroisoquinoline-3-carboxyl Acyl-Thr-Arg-Gly-Asp(Val)-Val and 1,2-dihydroisoquinoline-3-carboxyl-Thr-Arg-Gly-Asp(Phe)-Phe have excellent anti-venous thrombotic activity. Therefore, the inventors propose the present invention.

SUMMARY OF THE INVENTION

The first aspect of the present invention is to prepare N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp( aa)-aa (where aa is a Ser, Val or Phe residue).

The second content of the present invention is to convert N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(aa)-aa (where aa is a Ser, Val or Phe residue) quantitatively dehydrogenated into 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa) in mouse serum -aa (where aa is a Ser, Val or Phe residue).

The third aspect of the present invention is to evaluate 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa)-aa (where aa is a Ser, Val or Phe residue) ) anti-venous thrombotic activity and observed hemorrhagic side effects.

Description of drawings

Figure 1. The synthetic route of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa)-aa (where aa is a Ser, Val or Phe residue).

Implementation

In order to further illustrate the present invention, a series of examples are given below. It must be pointed out that these examples are purely illustrative. The purpose of giving these embodiments is to fully clarify the meaning and content of the present invention, and will never limit the present invention in any form.

Example 1 Preparation of N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(aa)-aa

N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(aa)-aa (wherein aa is a Ser, Val or Phe residue).

Example 2 Preparation of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp-(Ser)-Ser(10a)

Put 50 mg (0.04 mmol) of N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Ser)-Ser in Dissolved with 1 mL mouse serum at 37°C, the resulting solution was shaken at 37°C for 4 hours, and the disappearance of the raw materials was monitored by TLC. 2 mL of methanol was added to the serum solution, and the mixture was shaken at 37°C for 10 minutes, and the resulting mixed solution was centrifuged at 3000 rpm for 10 minutes. The residue obtained by centrifugation was fully extracted with ultrapure water, and the extract was separated and centrifuged. The supernatant obtained by centrifugation was concentrated under reduced pressure at 37°C, the residue was fully extracted with ultrapure water, and the extract was separated. The combined extracts were lyophilized to give 33 mg (96%) of the title compound. ESI(-)-FT-MS: 848.37150 [MH] ^- .Mp 131-132°C. [α] ^D ₂₅ = -13.1 (c=1.3, CH ₃ OH). IR (cm ^-1 ) 3671, 3442, 3235 , 2924, 2362, 1645, 1542, 1453, 1390, 1178, 1122, 1034, 643, 512, 426. ¹ H NMR(300MHz, DMSO)δ/ppm=11.10(s,2H),8.55(s,1H),8.54(s ,1H),8.05(m,7H),7.31(m,1H),7.22(m,2H),7.16(m,1H),6.67(s,1H),5.02(br,1H),4.65(m, 1H), 4.33(m, 1H), 4.26(m, 3H), 4.12(m, 1H), 4.05(t, J=5.6Hz, 1H), 3.60(m, 2H), 3.07(m, 1H), 2.65(dd,J＝6.4Hz,J＝6.4Hz,1H),1.52(m,1H),1.50(m,2H),1.25(q,J＝5.6Hz,3H),1.08(d,J＝5.6 Hz, 3H).

Example 3 Preparation of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(Val)-Val(10b)

According to the method of Example 1, 50 mg (0.04 mmol) of N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Val )-Val yielded 33 mg (96%) of the title compound. ESI(+)-FT-MS: 874.44535 [M+H] ⁺ .Mp 160-161°C. [α] ^D ₂₅ = -12.5 (c=1.6, CH ₃ OH). IR (cm ^-1 ) 3444, 3253 , 2964, 2384, 1652, 1543, 1456, 1394, 1255, 1173, 1028, 763, 642, 518, 438. ¹ H NMR(300MHz, DMSO)δ/ppm=11.04(s,2H),8.55(s,1H),8.54(s ,1H),8.04(m,7H),7.26(m,1H),7.21(m,2H),7.19(m,1H),6.66(s,1H),4.62(m,1H),4.31(m, 4H), 4.17(m, 3H), 4.14(m, 2H), 3.97(m, 1H), 2.58(m, 1H), 2.02(m, 2H), 1.71(m, 1H), 1.54(m, 1H) ), 1.50(m, 3H), 1.26(t, J=7.2Hz, 3H), 1.09(d, J=5.6Hz 3H). 0.85(m, 12H).

Example 4 Preparation of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(Phe)-Phe(10c)

According to the method of Example 1, 50 mg (0.04 mmol) of N-[(3S)-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Phe )-Phe yielded 38 mg (97%) of the title compound. ESI(+)-FT-MS: 970.06400 [M+H] ⁺ .Mp 122-123°C. [α] ^D ₂₅ =-14.2 (c=1.1CH ₃ OH).IR(cm ^-1 )3449,2935, 2366, 1742, 1643, 1518, 1459, 1392, 1274, 1029, 674, 510, 442. ¹ H NMR (300MHz, DMSO) δ/ppm=10.99(s, 2H), 8.56(s, 1H), 8.55(s, 1H) ,8.04(m,7H),7.27(m,1H),7.20(m,1H),7.25(m,2H),7.34-7.42(m,10H),6.66(s,1H),4.81(m,2H) ), 4.62(m, 1H), 4.51(m, 1H), 4.45(q, J=8.0Hz, 1H), 4.28(m, 4H), 4.12(m, 1H), 3.15(m, 3H), 3.04 (m,1H),2.64(dd,J=6.3Hz,J=6.9Hz,1H),2.55(dd,J=6.3Hz,J=6.9Hz,1H),2.01(m,3H),1.97(m , 1H), 1.77 (m, 2H), 1.55 (m, 2H), 1.49 (d, J=7.8Hz, 3H), 1.22 (d, J=8.2Hz, 3H).

Example 5 Evaluation of the anti-venous thrombotic activity of 10a-c

Male SD rats (250±20g) were acclimated to the environment and fasted for one day. They were anesthetized by intraperitoneal administration of 20% urethane solution 2 minutes before the operation and fixed on the plate. 2mL of blood was taken from the carotid artery for the determination of blood-related indexes. The rat abdominal skin was prepared, sterilized, and the abdominal cavity was opened along the linea alba (down to the coagulation gland, up to the exposed corner of the liver). Intra-abdominal organs such as the small intestine were removed and wrapped with gauze soaked with normal saline. The perivascular connective tissue was bluntly dissected to expose the inferior vena cava and its branches. The abdominal aorta and inferior vena cava were dissected below the renal vein, and then the inferior vena cava was ligated at the junction of the inferior vena cava and the left renal vein with saline-soaked suture. Move the intestines and other organs back to the abdominal cavity according to the anatomical position, and suture the abdominal cavity layer by layer with sutures. 10a-c saline solution was injected from the tail vein at a dose of 1 nmol/kg, the dose of warfarin in the positive control was 4.87 μmol/kg, and the negative control was saline. After circulating at 25-28℃ for 4 hours, the abdominal cavity of the rat was opened, its branches were ligated one by one, and the 2cm inferior vena cava was taken out from the ligation at the junction of the inferior vena cava and the left renal vein, and the thrombus was taken out. The thrombus was weighed and the results were counted by t test. The operation was performed alternately with two in each group, 12 in each group. Thrombus weighing is shown in Table 1. In addition, the effective dose of 10a-c is 1/4870 of that of warfarin without the bleeding side effects of warfarin. The present invention has unexpected technical effects.

Table 1 Anti-venous thrombotic activity of 10a-c

compound dose Thrombus wet weight (mean ± SD mg) normal saline 3ml/kg 25.35±1.41 Warfarin 4.87μmol/kg 13.13±3.71 10a 1nmol/kg 13.00±2.87^a 10b 1nmol/kg 12.76±2.53^a 10c 1nmol/kg 13.11±2.68^a

a) Compared with normal saline, p<0.01, and compared with warfarin, p>0.05; n=12.

Claims (3)

1. 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa)-aa of the following formula, where aa is a Ser, Val or Phe residue,

2. the preparation method of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp (aa)-aa of claim 1, the method steps are as follows: 1) Prepare N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Ser)- Ser; 2) Prepare N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Val)- Val; 3) Prepare N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Pro-Ala-Arg-Gly-Asp(Phe)- Phe; 4) N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Ser)- Ser is converted to 1,2-dihydroisoquinoline-3-carboxyl-Thr-Ala-Arg-Gly-Asp(Ser)-Ser; 5) N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Val)- Val is converted to 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(Val)-Val; 6) N-[(3S)-N-1,2,3,4-tetrahydroisoquinoline-3-formyl]-Thr-Ala-Arg-Gly-Asp(Phe)- Phe was converted to 1,2-dihydroisoquinoline-3-carboxyl-Thr-Ala-Arg-Gly-Asp(Phe)-Phe. 3. The application of 1,2-dihydroisoquinoline-3-formyl-Thr-Ala-Arg-Gly-Asp(aa)-aa of claim 1 in the preparation of anti-venous thrombosis drugs.

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