CN107311943A - It is a kind of to promote Cyclic dipeptides of angiogenesis and preparation method thereof - Google Patents

It is a kind of to promote Cyclic dipeptides of angiogenesis and preparation method thereof Download PDF

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Publication number
CN107311943A
CN107311943A CN201710544872.8A CN201710544872A CN107311943A CN 107311943 A CN107311943 A CN 107311943A CN 201710544872 A CN201710544872 A CN 201710544872A CN 107311943 A CN107311943 A CN 107311943A
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Prior art keywords
ring
cyclic dipeptides
methanol
water
glycine
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CN201710544872.8A
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Chinese (zh)
Inventor
程永现
刘越飞
王心龙
向斌
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SICHUAN GOOD DOCTOR PANXI PHARMACEUTICAL CO Ltd
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SICHUAN GOOD DOCTOR PANXI PHARMACEUTICAL CO Ltd
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
    • C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
    • C07D241/06—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members
    • C07D241/08—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having one or two double bonds between ring members or between ring members and non-ring members with oxygen atoms directly attached to ring carbon atoms

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Peptides Or Proteins (AREA)

Abstract

The invention provides a kind of Cyclic dipeptides of promotion angiogenesis and preparation method thereof.The Cyclic dipeptides are one kind in ring (alanine-valine), ring (glycine isoleucine), ring (glycine leucine) and ring (tyrosine aspartic acid), and the Cyclic dipeptides are to extract isolated from American cockroach first；Present invention also offers the extraction separation method of the Cyclic dipeptides；Present invention also offers a kind of pharmaceutical composition containing the Cyclic dipeptides；Promoting effect of HUVEC angiogenic activities invention further provides the Cyclic dipeptides, having the advantages that to remarkably promote HUVEC angiogenesis.

Description

It is a kind of to promote Cyclic dipeptides of angiogenesis and preparation method thereof
Technical field
The invention belongs to chemical medicine field, it is related to Cyclic dipeptides and preparation method thereof.
Background technology
Cyclic dipeptides (cyclic dipeptides) are called the derivative of 2,5- diketopiperazines, and such compound is in people, vertebra It is found in animal, invertebrate, plant, bacterium and fungi.Due to Cyclic dipeptides one stable six-membered ring structure of formation, With certain conformation effect of contraction, there are two hydrogen-bond donors and two hydrogen bond receptors, hydrogen bond is drug receptor interaction One of major way, therefore Cyclic dipeptides are an important pharmacophore in pharmaceutical chemistry.Have now been found that many Cyclic dipeptides tools There is strong physiologically active, such as with plant growth regulator effect, antitumor activity, antibacterial and raising immunologic function.
American cockroach (Periplaneta americana) is Insecta Pterigota Blattaria Blattidae Periplaneta elder brother Worm, is commonly called as " cockroach ", is that vitality is most strong, most ancient in the world, one of most successful insect assembly is multiplied so far.American cockroach Traditional Chinese medicine, be used as medicine first recorded in《Sheng Nong's herbal classic》, middle product are listed in, are called " taste is salty, cold ".Conventional numerous research tables Bright, the chemical composition in American cockroach is broadly divided into two major classes.The first kind is that pheromones are that big Lian is waved to having for release in vitro The pheromone component of hair property, mainly includes sex pheromone and aggregation pheromone.Equations of The Second Kind be amino acid and various insect neuropeptides, It is the chemical composition being present in its body.The present inventor is carried out to the chemical composition of the water section of American cockroach ethanol extract Research, separates and identifies a series of Cyclic dipeptides, wherein there is 4 Cyclic dipeptides to be isolated from American cockroach first.Through Research finds that this 4 Cyclic dipeptides have effects that to remarkably promote the generation of HUVEC cellular vasculars.
The content of the invention
Cyclic dipeptides of the present invention are ring (alanine-valine), ring (glycine-isoleucine), ring (glycine-bright Propylhomoserin) and ring (tyrosine-aspartic acid) in any one.
The invention provides the pharmaceutical composition of Cyclic dipeptides, described pharmaceutical composition includes ring (alanine-valine), ring Any one in (glycine-isoleucine), ring (Gly-Leu) or ring (tyrosine-aspartic acid), or its pharmacy Upper acceptable derivates and its esters.
Further, described pharmaceutical composition is pharmaceutical preparation, and it is by ring (alanine-valine), ring (sweet ammonia Acid-isoleucine), ring (Gly-Leu) or ring (tyrosine-aspartic acid) any one, or it can pharmaceutically connect The derivative and its esters received are constituted as active component and pharmaceutically acceptable excipient.
Wherein solid pharmaceutical preparation includes：Tablet, capsule, pill, granule etc.；Semisolid preparation includes ointment, suppository Deng；Liquid preparation includes：Solution, injection, spray etc.；Eye-drops preparations includes：Eye drops, gel for eye etc..
The preparation method that the present invention also disclosed described Cyclic dipeptides is：
Take American cockroach to dry polypide, extracted 2~3 times with hot water after crushing, 1~2h every time, extract solution is recovered under reduced pressure molten Agent obtains total extract.Total extract solution through MCI gel CHP 20P posts gradient elution (methanol/water, 0%~100%) 11 components, Respectively F1~F11.Wherein F4 through RP-18 posts (methanol/water, 0%~100%) 6 components (F4.1~F4.6).F4.2 is passed through Sephadex LH-20(H2O) gel filtration, then row half prepare HPLC (methanol/water, 12%) and obtain compound ring (tyrosine-day Winter propylhomoserin).F6 row MCI gel CHP20P posts (methanol/water, 0%~100%) obtain 10 components (F6.1~F6.10).F6.3 Through Sephadex LH-20 (H2O) gel filtration, then row prepare thin-layer chromatography (chloroform/methanol/ammoniacal liquor, 4:1:1%) made with half Standby HPLC (methanol/water, 8%) obtains compound ring (alanine-valine)；F6.4 is through Sephadex LH-20 (H2O) gel mistake Filter, then row prepare thin-layer chromatography (chloroform/methanol/ammoniacal liquor, 4:1:1%) and partly prepare HPLC (methanol/water, 10%) and obtain compound Ring (glycine-isoleucine) and ring (Gly-Leu).
Further, described Cyclic dipeptides are obtained or obtained by artificial synthesized by being extracted from American cockroach.
Further, application of the described Cyclic dipeptides in terms of angiogenesis is promoted.
Beneficial effects of the present invention：
1. present invention firstly discloses the method that 4 Cyclic dipeptides described in separation are extracted from American cockroach.
2. separation identification is obtained 4 Cyclic dipeptides of the present invention from American cockroach first.
3. 4 Cyclic dipeptides of the present invention have effects that to remarkably promote the generation of HUVEC cellular vasculars.
Brief description of the drawings
Fig. 1 compounds promote HUVECs angiogenic activities result of the test (n=3).*p<0.05, * * p<0.01, * * * p< 0.001。
Embodiment
With reference to embodiment, the invention will be further described.
The extraction separation of the Cyclic dipeptides of the present invention of embodiment 1 and Structural Identification
American cockroach derives from Sichuan Haoyisheng Panxi Pharmaceutical Co., Ltd. American cockroach GAP bases, American cockroach Extract is the main component of Kangfuxin Liquid.
Take American cockroach to dry polypide (25kg), extracted 2~3 times with hot water after crushing, 1~2h every time, extract solution decompression Recycling design obtains total extract.Total extract solution through MCI gel CHP 20P posts gradient elution (methanol/water, 0%~100%) 11 Individual component, respectively F1~F11.Wherein F4 (8g) through RP-18 posts (methanol-water, 0%~100%) 6 components (F4.1~ F4.6).F4.2 (3.2g) is through Sephadex LH-20 (H2O) gel filtration, then row half prepare HPLC (methanol-water, 12%) and obtained Compound ring (tyrosine-aspartic acid) (3.5mg).F6 (20g) row MCI gel CHP 20P posts (methanol/water, 0%~ 100%) 10 components (F6.1~F6.10) are obtained.F6.3 (1.3g) is through Sephadex LH-20 (H2O) gel filtration, then row system Standby thin-layer chromatography (chloroform/methanol/ammoniacal liquor, 4:1:1%) and half prepare HPLC (methanol/water, 8%) compound ring (alanine- Valine) (1.8mg)；F6.4 (1.8g) is through Sephadex LH-20 (H2O) gel filtration, then row prepare thin-layer chromatography (chloroform/ Methanol/ammoniacal liquor, 4:1:1%) and partly prepare HPLC (methanol/water, 10%) and obtain compound ring (glycine-isoleucine) (5.6mg) and ring (Gly-Leu) (28.5mg).
Compound results identification is as follows：
Compound ring (alanine-valine)：White solid,1H NMR(400MHz,methanol-d4)δ:4.04(1H, Q, J=7.0Hz, H-3), 3.84 (1H, m, H-6), 2.27 (1H, m, H-8), 1.45 (3H, d, J=7.0Hz, H-7), 1.05 (3H, d, J=7.1Hz, H-9/H-10), 0.95 (3H, d, J=6.8Hz, H-9/H-10).
Compound ring (glycine-isoleucine)：White powder,1H NMR(400MHz,methanol-d4)δ:4.00 (1H, d, J=17.6Hz, Hb-3), 3.84 (1H, overlap, H-6), 3.84 (1H, overlap, Ha-3), 1.97 (1H, m, H- 7), 1.57 (1H, m, Hb-8), 1.28 (1H, m, Ha-8), 1.03 (3H, d, J=7.0Hz, H-10), 0.97 (3H, t, J= 7.4Hz,H-9)；13C NMR(100MHz,methanol-d4)δ:170.1(C-5),168.8(C-2),61.0(C-6),45.3 (C-3),41.2(C-7),25.5(C-8),12.0(C-9),15.4(C-10)。
Compound ring (Gly-Leu)：Brown solid,1H NMR(400MHz,DMSO-d6)δ:8.27(1H,s,H- 1), 8.02 (1H, s, H-4), 3.82 (1H, d, J=17.3Hz, Hb-3), 3.61 (1H, overlap, H-6), 3.61 (1H, Overlap, Ha-3), 1.74 (1H, m, H-8), 1.51 (2H, t, J=6.8Hz, H-7), 0.86 (6H, overlap, H-9, H- 10)；13C NMR(100MHz,DMSO-d6)δ:168.8(C-5),166.5(C-2),53.0(C-6),44.3(C-3),42.2(C- 7),23.7(C-8),23.0(C-9/C-10),21.9(C-9/C-10)。
Compound ring (tyrosine-aspartic acid)：Faint yellow solid,1H NMR(400MHz,DMSO-d6)δ:8.00(1H, S, 13-OH), 7.62 (1H, s, H-1/H-4), 7.62 (1H, s, H-1/H-4), 6.96 (2H, d, J=8.2Hz, H-11, H-15), 6.66 (2H, d, J=8.2Hz, H-12, H-14), 4.08 (1H, m, H-6), 4.02 (1H, m, H-3), 2.98 (1H, dd, J= 13.7,4.6Hz, Ha-9), 2.81 (1H, dd, J=13.6,4.6Hz, Hb-9), 2.32 (1H, dd, J=15.7,4.0Hz, Ha- 7), 1.52 (1H, dd, J=15.7,8.7Hz, Hb-7)；13C NMR(150MHz,DMSO-d6)δ:171.7(C-8),166.9(C- 2),166.5(C-5),156.2(C-13),131.1(C-11,C-15),126.2(C-10),115.0(C-12,C-14),55.4 (C-6),51.2(C-3),38.2(C-9),37.4(C-7)。
The Cyclic dipeptides of the present invention of embodiment 2 promote HUVEC cellular vasculars generation active testing
The final stage of wound healing is tissue construction, and vascularization has played important function during this, to investigate Whether compound intervenes this process, and angiogenic activity experiment has been carried out to part Cyclic dipeptides.
Human umbilical vein endothelial cells (HUVEC) are presented by Shenzhen University's medical board.M199 culture mediums and hyclone are purchased from Gibco companies.Matrigel glue is purchased from CORNING companies.Matrigel glue is uniformly laid in 96 orifice plates of precooling, per hole 50 μ L, 37 DEG C of incubation 40min.After Matrigel gellings are solid, 1 × 10 is added per hole4HUVEC cells, experimental group adds to be measuredization Compound (20 μM of final concentration) processing, control group not drug containing.Under the conditions of 37 DEG C, 5%CO2, after culture 6h, in sight under microscope Examine and take pictures, picture analyzing is carried out with Image J softwares, each experiment is repeated 3 times.
Angiogenesis rate (%)=drug-treated composition length of tube/control composition length of tube × 100%
As shown in Figure 1, in the Cyclic dipeptides being screened, compound ring (alanine-valine) (numbering 9), ring (glycine- Isoleucine) (numbering 10), ring (Gly-Leu) (numbering 11), ring (tyrosine-aspartic acid) (numbering 21) is at 20 μM When can remarkably promote HUVECs formation tubular structure, show with angiogenesis promote activity.And promote angiogenic activity effect It is really：Ring (Gly-Leu) ＞ ring (alanine-valine) ＞ ring (tyrosine-aspartic acid) ＞ ring (glycine-different Leucine).

Claims (7)

1. a kind of Cyclic dipeptides of promotion angiogenesis, it is characterised in that described Cyclic dipeptides are ring (alanine-valine), ring Any one in (glycine-isoleucine), ring (Gly-Leu) or ring (tyrosine-aspartic acid).
2. the pharmaceutical composition of Cyclic dipeptides as described in claim 1, it is characterised in that described pharmaceutical composition includes ring In (alanine-valine), ring (glycine-isoleucine), ring (Gly-Leu) or ring (tyrosine-aspartic acid) Any one, or its pharmaceutically acceptable derivates and its esters.
3. pharmaceutical composition as claimed in claim 2, it is characterised in that described pharmaceutical composition is pharmaceutical preparation, by ring (third Propylhomoserin-valine), ring (glycine-isoleucine), ring (Gly-Leu) or ring (tyrosine-aspartic acid) it is any One kind, or its pharmaceutically acceptable derivates and its esters are constituted as active component and pharmaceutically acceptable excipient.
4. Cyclic dipeptides according to claim 1, it is characterised in that the preparation method of described Cyclic dipeptides is：
Take American cockroach to dry polypide, extracted 2~3 times with hot water after crushing, 1~2h every time, extract solution is recovered under reduced pressure solvent and obtained Total extract, total extract solution through MCI gel CHP 20P posts gradient elution (methanol/water, 0%~100%) 11 components, respectively For F1~F11, wherein F4 through RP-18 posts (methanol/water, 0%~100%) 6 components (F4.1~F4.6), F4.2 warps Sephadex LH-20(H2O) gel filtration, then row half prepare HPLC (methanol/water, 12%) and obtain compound ring (tyrosine-day Winter propylhomoserin), F6 row MCI gel CHP 20P posts (methanol/water, 0%~100%) obtain 10 components (F6.1~F6.10), F6.3 Through Sephadex LH-20 (H2O) gel filtration, then row prepare thin-layer chromatography (chloroform/methanol/ammoniacal liquor, 4:1:1%) made with half Standby HPLC (methanol/water, 8%) obtains compound ring (alanine-valine), and F6.4 is through Sephadex LH-20 (H2O) gel mistake Filter, then row prepare thin-layer chromatography (chloroform/methanol/ammoniacal liquor, 4:1:1%) and partly prepare HPLC (methanol/water, 10%) and obtain compound Ring (glycine-isoleucine) and ring (Gly-Leu).
5. Cyclic dipeptides according to claim 1, it is characterised in that described Cyclic dipeptides are by being extracted from American cockroach Obtain or obtained by artificial synthesized.
6. application of the Cyclic dipeptides according to claim 1 in terms of angiogenesis is promoted.
7. application of the Cyclic dipeptides in terms of angiogenesis is promoted according to claim 4~5.
CN201710544872.8A 2017-07-05 2017-07-05 It is a kind of to promote Cyclic dipeptides of angiogenesis and preparation method thereof Withdrawn CN107311943A (en)

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Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101979402A (en) * 2010-10-19 2011-02-23 中山大学 Marine cyclic peptide compound and preparation method and use thereof
CN104095851A (en) * 2003-05-15 2014-10-15 安皮奥制药股份有限公司 Treatment of T-cell mediated diseases

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104095851A (en) * 2003-05-15 2014-10-15 安皮奥制药股份有限公司 Treatment of T-cell mediated diseases
CN101979402A (en) * 2010-10-19 2011-02-23 中山大学 Marine cyclic peptide compound and preparation method and use thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
王心龙等: "虫药美洲大蠊水溶性成分及其促血管生成活性", 《天然产物研究与开发》 *

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Application publication date: 20171103