CN105343041A

CN105343041A - Composition and application thereof to drugs for treating or preventing renal fibrosis

Info

Publication number: CN105343041A
Application number: CN201510881848.4A
Authority: CN
Inventors: 王卓婷
Original assignee: Nanjing Fuhai Aosai Medicine Science & Technology Co Ltd
Current assignee: Nanjing Fuhai Aosai Medicine Science & Technology Co Ltd
Priority date: 2015-12-03
Filing date: 2015-12-03
Publication date: 2016-02-24

Abstract

The invention relates to the field of organic synthesis and medicinal chemistry, particularly relates to a composition, a preparation method and application thereof to preparation of drugs for preventing or treating renal fibrosis; the invention discloses the composition and the preparation method thereof. The pharmacology experiment indicates that the composition provided by the invention has the effect of preventing or treating renal fibrosis, and further has a value in development of drugs for preventing or treating renal fibrosis.

Description

A kind of compositions and the application in treatment or preventing renal fibrosis medicine thereof

Technical field

The present invention relates to organic synthesis and medicinal chemistry art, be specifically related to compositions, preparation method and its usage.

Background technology

Renal fibrosis (comprising kidney region fibrosis and glomerular sclerosis) is the main pathological basis of the kidney damage final stage that a variety of causes causes, renal fibrosis mechanism is comparatively complicated, relevant with many factors, the wherein main and celliferous propagation of extracellular matrix-cell and activation, the conversion of vaso-active substance, cytokine and extracellular matrix is unbalance relevant, and kidney region fibrosis is almost so former or secondary kidney disease proceed to the common pathway of end stage renal failure.Be badly in need of the anti-renal fibrosis medicine of research and development high-efficiency low-toxicity.

There is the problem that toxicity is large, safety is low in the current existing medicine for the treatment of of renal fibrosis, finds compound or lead compound and carry out structural modification to obtain its derivant from natural product, thus the potential drug obtaining high-efficiency low-toxicity has important value.

The Compound I that the present invention relates to is one and delivers (AyumiOhsakietal. in 2011,2011.SalviskinoneA, aditerpenewithanewskeletonfromSalviaprzewalskii.Tetrahed ronLetters52 (2011) 1375 – 1377) compound, we have carried out structural modification to Compound I, obtain two new derivants and compound III and compound IV, and prepared compositions by compound III and compound IV and the anti-renal fibrosis activity of said composition is evaluated, it is active that it has anti-renal fibrosis.

Summary of the invention

The invention discloses a new compositions, said composition is made up of compound III and compound IV, and in said composition, the mass percent of compound III and compound IV is respectively 80% and 20%.

Compositions disclosed by the invention can make pharmaceutically acceptable salt or pharmaceutically acceptable carrier.

Pharmacodynamic experiment shows, compositions of the present invention has good anti-renal fibrosis effect.Pharmaceutically acceptable salt of the present invention has same drug effect.

The present invention is further detailed explanation by the following examples, but protection scope of the present invention is not by any restriction of specific embodiment, but be limited by claim.

Detailed description of the invention

The preparation of embodiment 1 compound S alviskinoneA

Document (the AyumiOhsakietal. that the people such as the preparation method reference AyumiOhsaki of compound S alviskinoneA (I) deliver, 2011.SalviskinoneA, aditerpenewithanewskeletonfromSalviaprzewalskii.Tetrahed ronLetters52 (2011) 1375 – 1377) method.

The synthesis of the O-bromoethyl derivant (II) of embodiment 2SalviskinoneA

By Compound I (312mg, 1.00mmol) be dissolved in 15mL benzene, add in solution tetrabutyl ammonium bromide (TBAB) (0.08g), 1,50% sodium hydroxide solution of 2-Bromofume (3.760g, 20.00mmol) and 6mL.Mixture stirs 12h at 35 degrees Celsius.After 12h, reactant liquor is poured in frozen water, use dichloromethane extraction twice immediately, merge organic phase solution.Then use water and saturated common salt water washing 4 times successively to organic phase solution, then use anhydrous sodium sulfate drying, last concentrating under reduced pressure is removed solvent and is obtained product crude product.Product crude product purification by silica gel column chromatography (mobile phase is: petroleum ether/acetone=100:1.5, v/v), collects brown concentrated elution band and flings to the brown ceramic powder (327mg, 78%) that namely solvent obtains Compound II per.

¹HNMR(500MHz,DMSO-d ₆)δ6.63(s,1H),6.37(s,1H),5.81(s,1H),4.51(s,2H),3.84(s,1H),3.79(s,2H),2.15(s,1H),2.04(s,1H),1.91(s,1H),1.65(s,1H),1.39(s,3H),1.08(s,6H),0.99(s,6H).

¹³CNMR(125MHz,DMSO-d6)δ188.07(s),183.70(s),154.38(s),147.57(s),140.21(s),136.40(s),134.71(s),131.25(s),128.40(s),118.83(s),72.12(s),45.49(s),37.54(s),33.58(s),31.78(s),26.17(s),25.12(s),24.66(s),23.51(s),23.17(s),22.65(s).

HRMS(ESI)m/z[M+H] ⁺calcdforC ₂₂H ₂₈BrO ₃:419.1222；found419.1220.

The synthesis of the O-(two hydroxyethylamines) ethyl derivative (III) of embodiment 3SalviskinoneA

Compound II per (209mg, 0.5mmol) is dissolved in the middle of 30mL acetonitrile, adds Anhydrous potassium carbonate (690mg wherein, 5.0mmol), potassium iodide (252mg, 1.5mmol) and diethanolamine (1051mg, 10mmol), mixture reflux 9h.After reaction terminates, reactant liquor is poured in 25mL frozen water, with equivalent dichloromethane extraction 2 times, merge organic facies.Organic facies after merging with water and saturated common salt water washing successively, then use anhydrous sodium sulfate drying, concentrating under reduced pressure is removed solvent and is obtained product crude product.Product crude product purification by silica gel column chromatography (mobile phase is: petroleum ether/acetone=100:0.5, v/v), collects yellow concentrated elution band and flings to the faint yellow solid (159.5mg, 72%) that namely solvent obtains compound III.

¹HNMR(500MHz,DMSO-d6)δ6.52(s,1H),6.40(s,1H),5.71(s,1H),4.15(s,2H),3.70(s,1H),3.37(s,4H),3.00(s,2H),2.50(s,4H),2.07(s,1H),1.96(s,1H),1.83(s,1H),1.57(d,J＝1.4Hz,3H),1.30(s,3H),0.99(s,6H),0.90(s,6H).

¹³CNMR(125MHz,DMSO-d6)δ188.01(s),183.52(s),154.08(s),147.15(s),140.11(s),136.23(s),134.42(s),130.84(s),128.31(s),118.62(s),68.93(s),58.73(s),56.61(s),53.94(s),45.20(s),37.13(s),33.49(s),25.96(s),24.84(s),24.26(s),23.43(s),22.97(s),22.33(s).

HRMS(ESI):m/z[M+H] ⁺calcdforC ₂₆H ₃₈NO ₅:444.2750；found:444.2746。

The synthesis of the O-(bischloroethylamines base) ethyl derivative (IV) of embodiment 4SalviskinoneA

Compound III (0.222g, 0.5mmol) is dissolved in 6mL chloroform, dropwise adds thionyl chloride (0.238g, 2mmol), reactant reflux 1h.Reactant is cooled to room temperature, drips the thionyl chloride that Methanol Decomposition is excessive, concentrating under reduced pressure is except desolventizing.Product, through purification by silica gel column chromatography (petroleum ether/acetone 100:0.3, v/v), obtains the faint yellow solid (170.0mg, 71%) of compound IV.

¹HNMR(500MHz,Chloroform-d1)δ6.51(s,1H),6.22(s,1H),5.72(s,1H),4.16(s,1H),3.66(s,1H),3.44(s,4H),3.01(s,2H),2.80(s,2H),2.64(s,2H),2.04(s,1H),1.98(s,1H),1.85(s,1H),1.59(s,1H),1.28(s,3H),0.97(s,6H),0.93(s,6H).

¹³CNMR(125MHz,DMSO-d6)δ187.90(s),183.41(s),153.97(s),147.04(s),140.00(s),136.12(s),134.31(s),130.73(s),128.20(s),118.51(s),68.82(s),55.57(s),53.94(s),45.20(s),39.10(s),37.02(s),33.39(s),25.87(s),24.76(s),24.19(s),23.33(s),22.88(s),22.25(s).

HRMS(ESI):m/z[M+H] ⁺calcdforC ₂₆H ₃₆C _l2NO ₃:480.2072；found:480.2070。

Embodiment 5 compositions is on the impact of Rats Undergoing Unilateral Urethral Ligation kidney region fibrosis

1.1 material

Benazepril, Novartis Pharma AG produces; Hydroxyproline (HYP) test kit, biotech firm is built up in Nanjing; Fibronectin (FN) test kit is purchased from Shanghai institute of Biological Products.

The preparation of compositions: loaded by the powder of 20mg compound IV crossing 200 order nets after crossing the powder of the 80mg compound III of 200 order nets and grinding after grinding to mix in tubule with cover and with turbine stirring instrument and namely obtain 100mg compositions, obtains the solution of compositions by the compositions of this 100mg of water dissolution during use.

Laboratory animal: regular grade Wistar rat, male, body weight 150-200g, SD rat.

1.2 test methods and result

Rat 60, random point 6 groups, i.e. sham operated rats, model group, benazepril gavage 10mg/kg group, compositions gavage 10mg/kg group, compound III gavage 10mg/kg group, compound IV gavage 10mg/kg group, animal feeding 1 week, each rat is with after 10% chloral hydrate 3.0mL/kg intraperitoneal injection of anesthesia, by fixing for Rat Right lateral position with on operating-table, with iodine tincture after cropping, 75% alcohol disinfecting field of operation, row left side abdomen otch, successively cut skin, muscle and each layer of stomach wall, expose and be separated left side ureter, sham operated rats only cuts abdominal cavity and free left side ureter, but not ligation and cutting off, other are group rat 4-0 silk thread ligation twice respectively, upper one ligation point is positioned at left inferior pole of kidney level, then ureter is cut off at twice ligation point, layer-by-layer suture, each treated animal is put to death after postoperative 10 days 10% chloral hydrate anesthesia, get blood, measure by Fibronectin and illustrate that mensuration illustrates fibre asphalt mixture (FN).Normal saline leaves and takes left kidney after lavation repeatedly, and nephridial tissue is fixed through the paraformaldehyde buffer of 4%.Cut appropriate nephridial tissue, illustrate by hydroxyproline kit measurement and measure hydroxyproline.

Routine pathology examines 1. perusal: sham operated rats kidney color is scarlet, shows smooth, and peplos gloss, without adhesion.Model control group Kidney Volume increases, and color is pale, shows in graininess, similar human body branny kidney, the adhesion of a few regions kidney peplos.2. light microscopy checking: sham operated rats nephron result is clear, glomerular capsule is without expansion or cell infiltration.The large stretch of renal tubular necrosis of model control group, renal interstitial fibroblast proliferation, tubular ectasia, inside has the epithelial cell that a large amount of brown color shading material or necrosis come off, glomerule decreased number, part glomerule fibrosis and with the adhesion of bowman's capsule wall, blister cavities disappears.Compositions administration group pathological changes and model control group similar, but all have morphology in various degree to improve, compare with model control group and have notable difference.And compound III administration group and compound IV administration group and model control group there was no significant difference.

Table 1 compositions is on the impact of Rats Undergoing Unilateral Urethral Ligation kidney region fibrosis

* p<0.05, compared with model group

T inspection is carried out to each group of FN, HYP.The results are shown in Table 1, compositions gavage 10mg/kg significantly reduces FN, HYP level (comparing with model control group, P<0.05); And compound III and compound IV gavage 10mg/kg fail significantly to reduce FN, HYP level.

Conclusion: compositions significantly can reduce the rising of kidney region fibrosis FN, HYP level, suppresses kidney region fibrosis, can be used for preparing anti-renal fibrosis medicine.Compound III and compound IV significantly can not reduce the rising of kidney region fibrosis FN, HYP level, can not suppress kidney region fibrosis, are not available to prepare anti-renal fibrosis medicine.

The preparation of embodiment 6 composition tablet involved in the present invention

Get 2 grams of compositionss, add the customary adjuvant 18 grams preparing tablet, mixing, conventional tablet presses makes 100.

The preparation of embodiment 7 composition capsule involved in the present invention

Get 2 grams of compositionss, add prepare capsule customary adjuvant as starch 18 grams, mixing, encapsulatedly makes 100.

Claims

1. a compositions, it is characterized by said composition and be made up of compound III and compound IV, in said composition, the mass percent of compound III and compound IV is respectively 80% and 20%,

2. the preparation method of compositions as claimed in claim 1, is characterized by: the powder of compound III and the powder of compound IV are respectively 80% and 20% according to mass percent and fully mix.

3. the application of compositions as claimed in claim 1 in treatment renal fibrosis medicine.

4. the application of compositions as claimed in claim 3 in treatment renal fibrosis medicine, is characterized by: described renal fibrosis is kidney region fibrosis.

5. the application of compositions as claimed in claim 4 in treatment renal fibrosis medicine, is characterized by: described kidney region fibrosis is the kidney region fibrosis that Unilateral Ureteric Obstruction is formed.

6. the application of compositions as claimed in claim 3 in treatment renal fibrosis medicine, is characterized by: described compositions reduces the rising of the Fibronectin FN that renal fibrosis causes.

7. the application of compositions as claimed in claim 3 in treatment renal fibrosis medicine, is characterized by: described compositions reduces the rising of the hydroxyproline that renal fibrosis causes.