CN111423411B

CN111423411B - Novel renin inhibitor

Info

Publication number: CN111423411B
Application number: CN202010252949.6A
Authority: CN
Inventors: 张聿
Original assignee: Nanjing Life Source Medical Technology Co ltd
Current assignee: Nanjing life source Medical Technology Co.,Ltd.
Priority date: 2020-04-02
Filing date: 2020-04-02
Publication date: 2021-04-16
Anticipated expiration: 2040-04-02
Also published as: GB2589531A; CN111423411A; GB202104473D0

Abstract

A novel renin inhibitor containing pyrrolidine-3-carboxylic acid-3-piperidine ester structure can be used for treating diseases related to blocking RAS system, including hypertension, heart disease, etc. In particular to a compound shown as a formula (I) or pharmaceutically acceptable salt thereof.

Wherein R represents a lower alkane or cycloalkane, aryl or substituted aryl group, R₁、R₂Respectively represent C₁‑C₄Alkyl, aryl or substituted aryl, fused aryl or substituted fused aryl of (A), R₃Represents lower alkyl or cycloalkyl, substituted benzyl.

Description

Novel renin inhibitor

Technical Field

The present invention relates to a novel renin inhibitor. The invention particularly relates to pyrrolidine-3-formic acid-3-piperidine ester compounds, a preparation method and an effect of the compounds as renin inhibitors.

Background

Chronic sustained activation of the renin-angiotensin-aldosterone system (RAAS) is an important factor in the development and progression of cardiovascular disease, renal disease, and diabetes. Thus, RAAS has become a major pathophysiological target for intervention in the above-mentioned diseases. Blocking the pathophysiological effects of RAAS can occur from three sites: angiotensin Converting Enzyme Inhibitors (ACEIs) reduce the conversion of angioangiotensinogen to angiotensin i (angi); angiotensin ii receptor Antagonists (ARBs) block the action of angiotensin ii (Ang 11); the renin inhibitor can block RAAS from the source to effectively reduce the generation of AngI and AngII. Previous studies have shown that ACEI or ARB blockade of RAAS leads to a compensatory increase in plasma renin activity and accumulation of AngI, which in turn activates RAAS leading to the "escape of angiotensin II". Inhibition of renin, in turn, can block and reduce overall RAAS activity and can be a rational treatment for potential benefit.

Renin is a caspase that is synthesized mainly by the cells beside the kidney sphere, and has species specificity. The kidney pericentral cells first synthesize biologically inactive prorenin (prorenin), then undergo intracellular processing to remove a number of amino acids and convert them into single-chain active renin, which is stored in secretory granules and released into blood or tissues when being stimulated from the outside. The chronic activation state of RAAS promotes accelerated prorenin switch to renin. Renin release is affected by the following factors: hypotension, hyponatremia, or decreased body volume can stimulate renin release by cells beside the kidney sphere; renal sympathetic nerve excitation (mediated via the renal β 1 receptor) increases renin release; angiotensin ii negatively fed back through AT receptors inhibits renin release. Renin has previously been thought to function only to catalyse the hydrolysis of angiotensinogen to generate AngI. However, with the discovery of the renin receptor, it was recognized that renin, in addition to acting enzymatically in free form, promotes angiotensin production, and triggers a new signaling pathway distinct from that of angiotensin ii receptor. When renin is bound to the renin receptor, the ability to catalyze the hydrolysis of angiotensinogen to produce AngI is significantly enhanced. It has been recently found that inactive prorenin also binds to renin receptor, undergoes conformational change, and has the same enzymatic activity as renin. Thus, the renin receptor has been renamed as the prorenin/renin receptor. Prorenin/renin receptors are widely distributed in tissues such as kidney, heart, liver, pancreas, placenta, retina, vascular smooth muscle cells, etc. Prorenin and prorenin/renin receptor binding activates mitogen-activated protein (MAP) kinases ERK1(p44) and ERK2(p42) phosphorylation, upregulates expression of profibrotic molecules such as transforming growth factor beta (TGF- β), plasminogen activation inhibitor-1 (PAI-1), type I collagen, and fibronectin, agonizes the p38MAP kinase/heat shock protein 27 pathway, leading to altered actin dynamics. Activation of these renin receptors independently of Ang ii mechanisms leads to tissue fibrosis and cellular hypertrophy, which can lead to cardiovascular and renal pathologies. Recently, ACEI and ARB, which effectively block RAAS, have been widely used in diseases such as hypertension, coronary heart disease, myocardial infarction, diabetic nephropathy, and systolic insufficiency heart failure. However, after the ACEI or ARB treatment, the renin activity is increased sharply due to the negative feedback effect, and the escape phenomenon of angiotensin II and aldosterone appears. Treatment with the combination of ACEI and ARB was attempted to completely potentiate blockade of RAAS, but no significant clinical effect was found. Renin is a protease at the initiation and rate-limiting of RAAS, and renin activity is a key link in regulating the status of the entire RAAS, playing a role in regulating blood pressure and cardiovascular function. The activity of directly inhibiting renin can counteract the compensatory increase of renin activity caused by ACEI or ARB, and has little interference with other physiological pathways, thus achieving the purpose of comprehensively strengthening and blocking RAAS.

Although the use of 3, 4-substituted piperidine compounds in the manufacture of renin inhibitors is disclosed in International publication No. 06/069788. However, no details of pyrrolidine-3-carboxylic acid-3-piperidine esters are mentioned in this publication.

Disclosure of Invention

The present invention provides a novel compound having an excellent renin inhibitory action.

The invention relates to a novel renin inhibitor, in particular to a substituted pyrrolidine-3-formic acid-3-piperidine ester derivative with a general formula (I) or a pharmaceutically acceptable salt thereof.

In the formula:

r can be represented as: 1) a hydrogen atom; 2) c₁-C₆Alkyl groups of (a); 3) c₁-C₆Alkenyl of (a); 4) c₃-C₆Cycloalkyl groups of (a); 5) phenyl or substituted phenyl; 6) benzyl or substituted benzyl; 7) pyridyl or substituted pyridyl; 8) quinolinyl or substituted quinolinyl; 9) indolyl or substituted indolyl.

R₁Can be represented by 1) C₁-C₄Linear alkyl of (2), phenyl or substituted phenyl, 3) pyridyl or substituted pyridyl, 4) pyrazolyl or substituted pyrazolyl, 5) indolyl or substituted indolyl, 6) benzofuranyl or substituted benzofuranyl, 7) quinolinyl or substituted quinolinyl, 8) chromanyl or substituted chromanyl, 9) dihydrobenzofuranPyranyl or substituted dihydrobenzofuranyl, 10) indazolyl or substituted indazolyl, 11) pyrrolopyridyl or substituted pyrrolopyridyl, 12) benzisoxazolyl or substituted benzisoxazolyl, 13) indolinyl or substituted indolinyl, 14) quinazolinyl or substituted quinazolinyl, 15) dihydroquinazolinyl or substituted dihydroquinazolinyl, 16) furopyridyl or substituted furopyridyl, 17) isoquinolinyl or substituted isoquinolinyl, 18) pyrrolopyrimidyl or substituted pyrrolopyrimidyl, 19) tetrahydroquinolinyl or substituted tetrahydroquinolinyl, 20) tetrahydroindazolyl or substituted tetrahydroindazolyl, 21) tetrahydrocyclopentapyrazolyl or substituted tetrahydrocyclopentapyrazolyl, 22) pyrrolyl which may be substituted, 23) imidazolyl or substituted imidazolyl, 24) pyrazolyl or substituted pyrazolyl, pyrazolyl, 25) Pyrrolyl or substituted thienyl, 26) thiazolyl or substituted thiazolyl, 27) triazolyl, 28) pyrimidinyl, 29) pyrazinyl or substituted pyrazinyl, 30) imidazopyridinyl or substituted imidazopyridinyl, 31) pyrrolopyrazinyl, 32) a hydrogen atom.

The substituted radicals can be selected from one or two of the following groups 1) to 12), which are identical or different, i.e.:

1) a methyl group; 2) an ethyl group; 3) propyl; 4) an alkoxy group; 5) an acyl group; 6) a cyano group; 7) an ester group; 8) a cycloalkyl group; 9) an amino group; 10) a sulfonyl group; 11) a nitro group; 12) halogen element

Wherein halogen is fluorine, chlorine, bromine and iodine.

R₂Can be represented by 1) C₁-C₄Linear alkyl of (a), 2) phenyl or substituted phenyl, 3) pyridyl or substituted pyridyl, 4) pyrazolyl or substituted pyrazolyl, 5) indolyl or substituted indolyl, 6) benzofuranyl or substituted benzofuranyl, 7) quinolinyl or substituted quinolinyl, 8) chromanyl or substituted chromanyl, 9) dihydrobenzofuranyl or substituted dihydrobenzofuranyl, 10) indazolyl or substituted indazolyl, 11) pyrrolopyridyl or substituted pyrrolopyridyl, 12) benzisoxazolyl or substituted benzisoxazolyl, 13) indolinyl or substituted indolinyl, 14) quinazolinyl or substituted quinazolinyl, 15) dihydroquinazolylOxazolinyl or substituted dihydroquinazolinyl, 16) furopyridinyl or substituted furopyridinyl, 17) isoquinolinyl or substituted isoquinolinyl, 18) pyrrolopyrimidinyl or substituted pyrrolopyrimidinyl, 19) tetrahydroquinolinyl or substituted tetrahydroquinolinyl, 20) tetrahydroindazolyl or substituted tetrahydroindazolyl, 21) tetrahydrocyclopentapyrazolyl or substituted tetrahydrocyclopentapyrazolyl, 22) pyrrolyl which may be substituted, 23) imidazolyl or substituted imidazolyl, 24) pyrazolyl or substituted pyrazolyl, 25) pyrrolyl or substituted thienyl, 26) thiazolyl or substituted thiazolyl, 27) triazolyl, 28) pyrimidinyl, 29) pyrazinyl or substituted pyrazinyl, 30) imidazopyridinyl or substituted imidazopyridinyl, 31) pyrrolopyrazinyl, 32) hydrogen atom.

Wherein halogen is fluorine, chlorine, bromine and iodine.

R₃Can be expressed as: 1) a hydrogen atom; 2) c₁-C₆Alkyl groups of (a); 4) c₃-C₆Cycloalkyl groups of (a); 5) a substituted benzyl group.

The general preparation method of the invention comprises the following steps:

the method comprises the following steps:

adding Tetrahydrofuran (THF) into a reactor as a solvent at a low temperature, then adding reactant amine, finally slowly dropping malonyl chloride, stirring at the low temperature for 1-5 hours, and reacting for 0.5-5 hours after heating to the room temperature. The compound II is obtained by reduced pressure distillation.

Step two:

under the condition of low temperature, THF is added into a reactor as a solvent, a reactant pyrrolidine-3-formic acid is added, then a certain amount of triethylamine is added, finally the compound shown in the structural formula II is slowly added, the mixture is stirred for 1-5 hours at low temperature, and the reaction is continued for 0.5-5 hours after the temperature is raised to room temperature. The reaction was monitored by Gas Chromatography (GC) and when the reaction was complete, the compound of formula III was obtained by distillation under reduced pressure.

Step three:

under the protection of inert gas, adding toluene as a solvent into a reactor, then sequentially adding 3, 5-dihydroxypiperidine and sodium hydride, stirring for 1 hour at room temperature, slowly adding halogenated hydrocarbon into the reactor, and reacting for 1-5 hours under the heating condition. After the reaction is completed, the catalyst is quenched. And finally, separating by using column chromatography to obtain the compound with the structural formula VI.

Step four:

and at a low temperature, sequentially adding a compound with a structural formula III, toluene, a compound with a structural formula IV and triethylamine into a reactor, stirring for 0.5-1 hour, slowly heating to 40-120 ℃, and monitoring the reaction process by TLC. After the reaction is finished, the target product is obtained by a recrystallization method.

Pharmaceutically acceptable salts include those salts of formula (I) which are non-toxic to living organisms formed with inorganic or organic acids, for example hydrochloric, sulphuric, nitric, phosphoric, formic, acetic, propionic, citric, lactic, tartaric, oxalic, malic, citric, ascorbic, benzoic, salicylic, caffeic, malonic, succinic, methanesulphonic, p-toluenesulphonic and the like.

Preferred compounds are compounds of the general formula (IA) and salts thereof:

wherein R is₁、R₂、R₃Are groups as defined above for compounds of formula (I).

In preferred compounds, the first step is further:

adding THF (tetrahydrofuran) as a solvent into a reactor at a low temperature, then adding reactant amine, finally slowly dropping malonyl chloride, stirring at the low temperature for 1-2 hours, and reacting for 0.5-2 hours after heating to the room temperature. Vacuum distillation is utilized to obtain the compound of the structural formula (IIA).

In a preferred compound, said second step is further:

adding THF (tetrahydrofuran) serving as a solvent into a reactor at a low temperature, adding a reactant pyrrolidine-3-formic acid, slowly adding a compound shown in a structural formula II, stirring at the low temperature for 1-2 hours, and heating to room temperature to continue reacting for 0.5-2 hours. The reaction was monitored by Gas Chromatography (GC) and when the reaction was complete the compound of formula (IIIA) was isolated by column chromatography.

In a preferred compound, said step three is further:

under the protection of inert gas, adding toluene as a solvent into a reactor, then sequentially adding 3, 5-dihydroxypiperidine and a catalyst, stirring for 1 hour at room temperature, slowly adding halogenated hydrocarbon into the reactor, and reacting for 1-2 hours under a heating condition. After the reaction is complete, the catalyst is quenched. And finally, separating by using column chromatography to obtain the compound with the structural formula (VA).

In preferred compounds, said step four is further:

and at a low temperature, sequentially adding a compound with a structural formula V, toluene, a compound with a structural formula IV and triethylamine into the reactor, stirring for 0.5-1 hour, slowly heating to 40-60 ℃, and monitoring the reaction process by TLC. After the reaction is finished, the target product is obtained by a recrystallization method.

Among the above conditions:

the low temperature is-25 ℃ to 5 DEG C

Room temperature is 15-30 DEG C

TLC is thin layer chromatography

The inert gas is argon or nitrogen

Use of structural formula IA or a pharmaceutically acceptable salt thereof for the treatment of renin.

The invention has the beneficial effects

The invention relates to a novel renin inhibitor containing a pyrrolidine-3-formic acid-3-piperidine ester structure, which has simple synthetic process and is suitable for industrial production.

Has obvious inhibition effect on the renin, long action effect duration and small toxic action on human bodies.

Examples

Example 1

Chemical formula 1:

the synthesis process is as follows:

(1) step 1A three-necked round-bottomed flask was charged with 25mL of Tetrahydrofuran (THF) at-20 ℃ and then with 25.3mL of a 2M solution of methylamine in tetrahydrofuran as a reactant, and finally 7.1g of malonyl chloride was slowly added dropwise thereto, followed by stirring at low temperature for 1 hour and warming to room temperature for 0.5 hour. Methylamine and malonyl chloride are distilled off by reduced pressure distillation.

(2) Step2, under the condition of-20 ℃, adding 20mL of THF into a distilled three-neck round-bottom flask, adding 10 mol% of triethylamine, finally slowly adding 5.75g of pyrrolidine-3-formic acid, stirring at low temperature for 1-2 hours, heating to room temperature, and continuing to react for 1 hour. The progress of the reaction was monitored by GC and when the reaction was over 9.6g of intermediate compound III were obtained by distillation with a yield of 90%.

(3) Step3 adding 30mL of toluene as a solvent to a reactor under an inert gas atmosphere, then adding 5.3g of 3, 5-dihydroxypiperidine, slowly adding 1.1g of sodium hydride in three portions at a low temperature, stirring at room temperature for 1 hour, then slowly adding 2.5mL of a monochloromethane ether solution to the reactor, and reacting for 1.5 hours. After the reaction is finished, slowly dropwise adding an ammonium chloride aqueous solution for quenching. Finally, 5.8g of intermediate compound VI are isolated in 98% yield.

(4) Step 4A three-necked round-bottomed flask was charged with 35mL of toluene, 5.8g of intermediate compound VI and 9.5g of intermediate compound III in this order at-10 ℃ and 2 mol% of tetrapropyl titanate was added thereto, and the mixture was stirred for 1 hour, slowly warmed to 100 ℃ and heated to distill off water. The progress of the reaction was monitored by GC. After the reaction was complete, the reaction was quenched with n-hexane: recrystallization from dichloromethane ═ 5:1 gave a white solid. After drying, 12.7g of a white solid were obtained in 87.5% yield. m.p.420-421 ℃,¹H NMR(CDCl₃,400M):δ5.87(s,1H),4.92(s,1H),4.02(s,1H),3.94(s,1H),3.77(s,1H),3.57(d,J＝12.3Hz,2H),3.50(s,1H),3.34–3.30(m,3H),3.11(s,1H),3.06(t,J＝7.5Hz,3H),2.86(s,1H),2.82–2.78(m,3H),2.75(s,1H),2.44(s,1H),2.35(s,1H),2.19(s,1H),1.57(s,1H),1.21(s,1H).¹³C(CDCl₃,100M):δ172.1,168.8,161.5,75.0,69.1,56.4,50.8,49.9,48.0,46.8,44.7,37.5,36.0,28.9,26.6.APCI-MS m/z：328.38[M+H]+

example 2

Chemical formula 2:

the synthesis process is as follows:

(1) step1 Tetrahydrofuran (THF) 25mL was added to a three-necked round-bottomed flask at-20 ℃ followed by addition of the reactant (4.7 g) aniline and dropwise addition of 7.1g malonyl chloride slowly, followed by stirring at low temperature for 1 hour and warming to room temperature for 0.5 hour. After the reaction was complete, 9.4g of the compound of formula II was obtained by recrystallization.

(2) Step2 adding 20mL of THF into a three-neck flask at-20 ℃, adding 9.0g of compound II and 10 mol% of triethylamine, finally slowly adding 5.47g of pyrrolidine-3-carboxylic acid, stirring at low temperature for 1-2 hours, heating to room temperature, and continuing to react for 1 hour. The progress of the reaction was monitored by GC and when the reaction was over 11.8g of intermediate compound III were obtained by column chromatography in 90% yield.

(4) Step 4A three-necked round-bottomed flask was charged with 35mL of toluene, 5.6g of intermediate compound VI and 11.8g of intermediate compound III in this order at-10 ℃ and 2 mol% of tetrapropyl titanate was added thereto, and the mixture was stirred for 1 hour, slowly warmed to 100 ℃ and heated to distill off water. The progress of the reaction was monitored by GC. After the reaction is finished, separating by using column chromatography to obtain 14.8g of white solid with the yield of 89 percent and the temperature of m.p.503-504 ℃,¹H NMR(CDCl₃,500M):δ7.56–7.43(m,33H),7.39–7.32(m,49H),7.10(s,16H),4.90(s,14H),4.20–4.16(m,32H),3.96(d,J＝61.7Hz,26H),3.89(s,5H),3.71(d,J＝58.4Hz,30H),3.64(d,J＝5.7Hz,4H),3.55(d,J＝4.6Hz,30H),3.35–3.31(m,48H),3.10(d,J＝9.1Hz,31H),2.86(s,13H),2.75(s,12H),2.44(s,12H),2.35(s,12H),2.19(s,12H),1.84(s,12H),1.25(s,16H).¹³C(CDCl₃,125M):δ172.1,165.4,161.8,137.8,129.4,129.2,123.7,121.6,121.4,74.8,69.0,56.4,49.8,49.0,47.9,45.2,45.0,42.6,36.7,28.6.APCI-MS m/z：390.45[M+H]⁺

example 3

Chemical formula 3:

the synthesis process is as follows:

(1) step1 Tetrahydrofuran (THF) 25mL was added to a three-necked round-bottomed flask at-20 ℃ followed by the addition of the reactant (3.0 g n-propylamine) and finally the dropwise addition of 7.1g malonyl chloride slowly, followed by stirring at low temperature for 1 hour and warming to room temperature for 0.5 hour. After the reaction, malonyl chloride and n-propylamine were distilled off.

(2) Step2, under the condition of-20 ℃, adding 20mL of THF into a distilled three-neck round-bottom flask, adding 10 mol% of triethylamine, finally slowly adding 5.8g of pyrrolidine-3-formic acid, stirring at low temperature for 1-2 hours, heating to room temperature, and continuing to react for 1 hour. The progress of the reaction was monitored by GC and when the reaction was over 10.0g of intermediate compound III was obtained by distillation in 88% yield.

(4) Step 4A three-necked round-bottomed flask was charged with 35mL of toluene, 5.8g of intermediate compound VI and 10.0g of intermediate compound III in this order at-10 ℃ and 2 mol% of tetrapropyl titanate was added thereto, and the mixture was stirred for 1 hour, slowly warmed to 100 ℃ and heated to distill off water. The progress of the reaction was monitored by GC. After the reaction is finished, separating by using column chromatography to obtain 12.7g of white solid with the yield of 85 percent and the m.p.432-433 ℃,¹H NMR(CDCl₃,500M):δ4.81(s,1H),4.66(s,1H),4.02(s,1H),3.77(s,1H),3.55(dd,J＝14.6,9.2Hz,4H),3.34–3.30(m,3H),3.27(s,1H),3.09(dd,J＝15.6,4.4Hz,4H),2.88(d,J＝19.3Hz,2H),2.75(s,1H),2.45(s,1H),2.20(d,J＝14.4Hz,2H),1.38(s,1H),1.26–1.22(m,3H),1.17(s,1H).¹³C(CDCl₃,125M):δ172.1,166.8,161.6,75.0,69.1,56.43,55.8,49.9,48.0,46.8,44.7,41.4,37.2,36.0,28.9,14.23.APCI-MS m/z：342.41[M+H]⁺

example 4

Chemical formula 4:

the synthesis process is as follows:

(1) step1 Tetrahydrofuran (THF) 25mL was added to a three-necked round-bottomed flask at-20 ℃ followed by the addition of the reactant (5.4 g) of benzylamine and finally the dropwise addition of malonyl chloride (7.05 g), followed by stirring at low temperature for 1 hour and warming to room temperature for 0.5 hour. After the reaction was complete, 10.1g of formula II were obtained by recrystallization.

(2) Step2 adding 20mL of THF into a three-neck flask at-20 ℃, adding 10.1g of compound II and 10 mol% of triethylamine, finally slowly adding 5.47g of pyrrolidine-3-carboxylic acid, stirring at low temperature for 1-2 hours, heating to room temperature, and continuing to react for 1 hour. The progress of the reaction was monitored by GC and when the reaction was over 12.7g of intermediate compound III was obtained by column chromatography in 92% yield.

(4) Step4 into a three-necked round-bottomed flask at-10 ℃ were added 35mL of toluene, 5.7g of intermediate Compound VI, 12.7g of intermediate Compound III, and 2 mol% tetrapropyl titanateAfter stirring for 1 hour, the temperature is slowly raised to 100 ℃, and water is evaporated by heating. The progress of the reaction was monitored by GC. After the reaction is finished, separating by using column chromatography to obtain 13.8g of white solid with the yield of 78 percent and the m.p.514-515 ℃,¹H NMR(CDCl₃,500M):δ7.34–7.25(m,5H),7.20(s,1H),5.72(s,1H),4.93(s,1H),4.60(s,1H),4.51(s,1H),4.02(s,2H),3.96(s,1H),3.77(s,1H),3.34–3.30(m,4H),3.11(s,1H),3.06(t,J＝7.2Hz,4H),2.86(s,1H),2.75(s,1H),2.44(s,1H),2.36(s,1H),2.19(s,1H),1.58(s,1H),1.22(s,1H).¹³C(CDCl₃,125M):δ172.1,167.0,161.8,139.9,128.5,128.3,127.7,127.3,126.9,74.8,69.0,56.4,49.8,49.0,47.9,45.2,45.0,44.0,42.4,36.7,28.6.APCI-MS m/z：404.48[M+H]⁺

example 5

Chemical formula 5:

the synthesis process is as follows:

the preparation process refers to example 1.

13.5g of white solid, the yield is 85 percent, m.p.496-467 ℃,¹HNMR(CDCl₃,500M):δ9.76(s,1H),4.99(s,1H),4.02(s,3H),3.83–3.57(m,4H),3.48(s,1H),3.13(d,J＝4.3Hz,1H),3.12(s,2H),3.12–2.88(m,4H),2.45(s,1H),2.14(dd,J＝54.1,3.0Hz,2H),2.07–2.05(m,2H),1.92(s,1H),1.73–1.69(m,3H),1.61(s,2H),1.43–1.34(m,3H),1.30–1.26(m,4H).¹³C(CDCl₃,125M):172.1,166.8,161.6,75.5,72.4,69.1,50.4,49.9,48.0,46.80,44.7,41.4,37.2,34.8,31.4,31.4,28.9,25.9,24.6,24.6,14.3.APCI-MS m/z：410.26[M+H]⁺

example 6

Chemical formula 6:

the synthesis process is as follows:

the preparation process refers to example 1.

14.2g of white solid, 88 percent of yield, m.p.526-527 ℃,¹H NMR(CDCl₃,500M):δ7.34–7.13(m,5H),4.97(s,1H),4.88(s,1H),4.66–4.59(m,2H),4.02(s,1H),3.74(t,J＝17.2Hz,3H),3.56(d,J＝35.8Hz,2H),3.31(s,1H),3.15–2.94(m,5H),2.86(d,J＝0.8Hz,2H),2.44(s,1H),2.23(d,J＝38.2Hz,2H),1.99(s,1H),1.31–1.13(m,4H).¹³C(CDCl₃,125M):172.1,166.8,161.8,138.0,128.3,128.2,128.1,127.8,73.4,71.0,69.0,49.6,49.0,47.9,45.2,45.0,42.4,37.2,36.0,28.6,14.3.APCI-MS m/z：418.23[M+H]⁺

example 7

Chemical formula 7:

the synthesis process is as follows:

the preparation process refers to example 1.

15.3g of white solid, the yield is 93 percent, m.p.432-433 ℃,¹HNMR(CDCl₃,500M):δ5.80(s,1H),4.91(s,1H),4.02(s,1H),3.83(s,1H),3.78(d,J＝6.5Hz,2H),3.56(s,1H),3.55–3.43(m,3H),3.39(s,1H),3.06(t,J＝7.4Hz,3H),2.86(s,1H),2.82–2.78(m,3H),2.75(s,1H),2.60(s,1H),2.45(s,1H),2.20(s,1H),1.96(s,1H),1.29(s,1H),1.23–1.19(m,4H).¹³C(CDCl₃,125M):δ172.1,165.36,161.78,135.18,134.6,130.1,128.8,127.6,127.3,126.6,124.7,121.9,117.4,74.2,69.0,65.0,49.6,49.0,47.9,45.2,45.0,42.6,36.0,28.6,15.5.APCI-MS m/z：342.20[M+H]+

example 8

Chemical formula 8:

the synthesis process is as follows:

the preparation process refers to example 1.

12.5g of white solid, the yield is 70 percent, m.p.726-727 ℃,¹H NMR(CDCl₃,500M):δ7.89–7.71(m,4H),7.54(d,J＝6.4Hz,2H),7.37(d,J＝16.1Hz,2H),4.93(s,1H),4.19–4.15(m,2H),4.02(s,1H),3.76(d,J＝7.6Hz,2H),3.71(s,1H),3.55(d,J＝11.1Hz,2H),3.50–3.42(m,2H),3.11(d,J＝1.8Hz,2H),2.86(s,1H),2.75(s,1H),2.44(s,1H),2.38(s,1H),2.18(s,1H),1.86(s,1H),1.26(s,1H),1.22–1.18(m,3H)。¹³C(CDCl₃,125M):172.1,168.2,161.8,134.0,129.3,121.6,120.7,120.1,111.4,95.4,74.2,69.0,65.0,49.6,49.0,47.9,45.2,45.0,42.7,36.0,26.6,15.5.APCI-MS m/z：454.54[M+H]⁺

example 9

Chemical formula 9:

the synthesis process is as follows:

the preparation process refers to example 1.

White solid, 10.8g, yield 68%, m.p.608-609 ℃,¹H NMR(CDCl₃,500M):δ7.34–7.26(m,2H),7.26–7.17(m,7H),7.08(s,1H),6.59(s,1H),4.67(s,1H),4.65–4.61(m,2H),4.58(s,1H),4.48(s,1H),4.02(s,1H),3.77(d,J＝3.6Hz,2H),3.70(s,1H),3.55(s,1H),3.12(d,J＝13.5Hz,2H),3.09–3.05(m,2H),3.00(s,1H),2.86(s,1H),2.75(s,2H),2.43(s,2H),2.27(s,1H),2.18(s,2H),1.74(s,1H).13C(CDCl₃,125M):172.1,167.0,161.8,139.9,138.0,128.4,128.4,128.3,128.3,128.2,128.2,127.8,127.6,127.6,126.9,73.4,71.0,69.0,49.6,49.0,47.9,45.2,45.0,44.0,45.2,45.0,44.0,42.4,35.4,23.6.APCI-MS m/z：480.24[M+H]⁺

example 10

Chemical formula 10:

the synthesis process is as follows:

the preparation process refers to example 1.

White solid, 11.6g, 76% yield, m.p.693 to 694 ℃,1H NMR (cdcl3,500m): δ 7.31(t, J ═ 20.9Hz,3H), 7.27-7.19 (m,4H),7.16(s,1H),6.90(s,1H),6.78(s,1H),6.62(s,1H),4.97(s,1H), 4.65-4.61 (m,2H), 4.19-4.15 (m,2H), 4.10-4.06 (m,2H),4.02(s,1H),3.94(s,1H),3.76(d, J ═ 8.5Hz,2H),3.59(s,1H),3.31(s,1H),3.10 (cld, J ═ 8.0, 2H), 3.45H, 1H, 19(s,1H), 3.49H, 1H, 19(s,1H), 3.45H, 1H), 3.49H, 1H, 3.0, 3.45H, 1H, 3.47, 1H, 49H, 0, 1H, 3.45H, 3, 42.6,36.0,28.6.APCI-MS m/z: 481.24[ M + H]⁺

Example 11

Chemical formula 11:

the synthesis process is as follows:

the preparation process refers to example 1.

White solid, 13.8g, 85% yield, m.p.348 to 349 ℃,1H NMR (cdcl3,500m) δ 4.92(s,1H),4.07(s,1H),4.02(s,1H),3.77(s,1H),3.59(t, J ═ 14.2Hz,3H), 3.34-3.30 (m,3H),3.11(s,1H), 3.09-3.04 (m,3H), 2.88-2.83 (m,7H),2.75(s,1H),2.45(s,1H),2.35(s,1H),2.20(s,1H),1.59(s,1H),1.21(s,1H), 13C (cdcl3,125m):172.1,170.2,161.0,75.0,69.1,56.4,50.8, 9.48, 8.46, 7.36, 7.8, 36.46, 7.36, 7.8, 36.8, 7.8, 36, 36.8, 7.8, 7,36, 7 z: 342.2[ M + H]⁺

Example 12

Chemical formula 12:

the synthesis process is as follows:

the preparation process refers to example 1.

White solid, 12.6g, yield 80%, m.p.370 to 371 ℃, 1HNMR (cdcl3,500m): δ 4.91(s,1H),4.02(s,1H),3.82 to 3.69(m,4H),3.57(d, J ═ 10.9Hz,2H),3.44(s,1H),3.34 to 3.30(m,3H),3.26 to 3.13(m,2H),3.11(s,1H),3.06(t, J ═ 8.9, 3H),2.86(s,1H),2.75(s,1H),2.44(s,1H),2.34(s,1H),2.19(s,1H),1.57(s,1H),1.24 to 1.18(m,7H), 13C (s, 13 m) 3569, 0.38, 25.19, 19(s,1H),1.57(s,1H),1.24 to 1.18(m,7H), 13C (m,3, 13 m, 25.19, 19, 13, 19, 13 m, 19, 13, 19, 13 m, 19, 13, 19, 8: 370.23[ M + H]⁺

Test example 1

Assay for angiotensin II receptor antagonistic Activity

The assay was carried out according to the method disclosed by Wangxiaowei et al (determination of the activity of angiotensin II receptor antagonist, journal of Beijing medical university, Vol.30, No. 4, p.370, 1998).

Adding about 100 μ g liver membrane protein and fixed amount into 0.35ml rat liver cell membrane receptor reaction solution¹²⁵Mixing I-angiotensin II receptor (about 5,000 count rate/min) with non-labeled angiotensin II receptor (0.045 ng-30 ng), adding 1 μ g angiotensin II receptor in non-specific tube, reacting at 25 deg.C for 70min, stopping reaction in water bath, collecting the bound angiotensin II receptor with a multi-head collector¹²⁵The I-angiotensin II receptor is collected on glass fiber filter paper (pre-saturated with 1mg/L angiotensin II receptor), and each tube is washed with 5ml of washing liquid for 3-4 times. Measuring radioactivity with gamma counter, calculating IC₅₀The value is obtained.

The results of the experiments show that the IC's of the compounds 1 to 12 of the above examples of the present invention₅₀≤2μmol。

Test example 2

Human renin inhibition

The synthesized compound was mixed with the peptide compound Nma-KHPFH LVIHK (Dnp) -NH2, and the fluorescence intensity before the start of the reaction was measured (excitation wavelength 365nm, measurement wavelength 435 nm). Subsequently, human recombinant renin was added, cultured at 37 ℃ for 3.5 hours, and the fluorescence intensity after the reaction was measured with a fluorescence photometer. Inhibitory Activity of Compounds expressing IC₅₀Equal to the fluorescence intensity before reaction minus the fluorescence intensity after reaction. Examples Compounds IC described in this specification₅₀As shown in the table below.

TABLE 1

Note: the compound of the control example was aliskiren.

The experiment shows that the synthesized compound containing pyrrolidine-3-formic acid-3-piperidine ester has obvious inhibition effect on renin.

Test example 3

Depressurization test

The ability of the compounds of the invention to lower blood pressure was evaluated in conscious, Spontaneously Hypertensive Rats (SHRs). Groups of SHRs (250-300g) were given equimolar doses of either the control compound or the compound of the invention for 3 consecutive days. After administration, Systolic Blood Pressure (SBP) and heart rate were monitored at different time points by teleassays. The results are shown in table 2 below:

TABLE 2 Effect of the Compounds of the invention on systolic blood pressure (mmHg)

Compound (I)	Basal blood pressure	30min	24h	48h	72
						Example 1	150	128	123	123	130
Example 2	150	127	124	124	129
						Example 3	150	128	123	123	132
Example 4	150	123	123	123	131
						Example 5	150	126	124	124	128
Example 6	150	125	123	123	129
						Example 7	150	128	125	125	130
Example 8	150	124	123	123	128
						Example 9	150	126	123	123	131
Example 10	150	127	122	123	129
						Example 11	150	128	123	123	133
Example 12	150	129	124	124	131
						Control Compound 1	150	135	128	135	147
Control Compound 2	150	132	126	135	148

Note: control Compound 1 is captopril and control Compound 2 is losartan Potassium

Experiments have shown that compounds 1 to 12 according to the invention all exhibit similar hypotensive properties as described above. Compared with the control, the compounds 1 to 12 of the present invention all showed the ability to continuously and strongly lower blood pressure to normal level, with long duration of drug effect, and did not cause large fluctuation of blood pressure, avoiding the occurrence of severe hypotension, compared with the control compounds 1 to 2, during the whole treatment period.

Claims

1. A compound (I) shown below or a pharmaceutically acceptable salt thereof:

wherein

R is: 1) a hydrogen atom;

R₁is 1) C₁-C₄Alkyl of 2) phenyl or substituted phenyl, 4) a hydrogen atom; r₁The substituent of the substituted phenyl group is 1) methyl; 2) an ethyl group; 3) propyl; 4) an amino group; 5) a nitro group; 6) halogen, wherein halogen is fluorine, chlorine, bromine and iodine;

R₂is 1) C₁-C₄Alkyl of 2) phenyl or substituted phenyl, 4) a hydrogen atom; r₂Wherein the substituent of the substituted phenyl group is 1)A methyl group; 2) an ethyl group; 3) propyl; 4) an amino group; 5) a nitro group; 6) halogen, wherein halogen is fluorine, chlorine, bromine and iodine;

R₃comprises the following steps: 1) hydrogen atom, 2) C₁-C₆Alkyl of 3) C₃-C₆Cycloalkyl of (4), benzyl.

2. The compound of claim 1, wherein R₁Is 1) C₁-C₄Alkyl group of (2), phenyl group, 3) hydrogen atom.

3. The compound of claim 1, wherein R₂Is 1) C₁-C₄Alkyl group of (2), phenyl group, 3) hydrogen atom.

4. The compound of claim 1, wherein R₃1) methyl, 2) ethyl, 3) benzyl.

5. The compound according to claim 1, selected from: