CN104971086A - Application of uraria for promoting osteogenesis or providing neuro-protection - Google Patents
Application of uraria for promoting osteogenesis or providing neuro-protection Download PDFInfo
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Abstract
The invention provides a method of promoting osteogenesis, increasing bone quality or increasing strong bone growth rate. The method includes a step of feeding uraria, or a part of the uraria, or an extract of the uraria to an individual. The invention also provides a method of treating and/or preventing osteoporosis and a method of providing neuro-protection, wherein the methods includes feeding the uraria, or a part of the uraria, or the extract of the uraria to the individual.
Description
Technical field
The present invention uses plant product to promote osteogenesis about a kind of or provide the method for neuroprotective.Particularly, the present invention uses rabbit tail grass genus (Uraria) promote osteogenesis or provide neuroprotective.
Background technology
Bone is the structural material of health skeleton, and for keeping required bone amount and structure.Bone contains calcium ion (Ca2+) and calcium amount is played an important role in maintenance blood.For this reason, the growth of bone is the metabolic balance between the osteoblast of bone remoulding circulation and the activity of osteoclast.Balance between bone resorption and bone formation is destroyed, and the osseous tissue amount that osteoblast is replaced mismatches the osseous tissue amount that osteoclast absorbs, and thus causes osteoporosis, a kind of common disease causing bone density or bone amount to lose.Osteoblast is bone formation cell, and it is derived from bone marrow interstital stem cell (MSCs).Osteoblast plays the part of very important role in creation and maintenance skeletal architecture.Bone formation relates to the event of series of complex, comprises osteoblastic propagation and differentiation, finally causes the mineralising of extracellular matrix.In each stage that osteoblast is grown, specific gene is sequentially showed or is showed, and such as, H4 (histone4) is a proliferation marker, alkali phosphatase (alkaline phosphate; ALP) for differentiation marker and Bone Gla protein (osteocalcin) are mineralising labelling.In human health, bone formation at the skeleton normal growth of immunoenzyme technics, in the loss of climacteric women prevention bone amount, and the healing of fracture patient and to reinvent be an important clinical concern.In order to prevent bone lesion with and increase bone formation, need nutrition and pharmaceutical preparation.Therefore, the external model that osteoblast is just cultivated for cell (osteoblast primary cell) has been widely used in the novel drugs found inducing osteoblast differentiation, Matrix Mineralization and form new bone.There are many parameters affecting the performance of osteoblasts in vitro in cell culture.
Osteopathia, as osteoporosis, is common in middle age or elderly woman.Therapeutic agent example for osteoporosis comprises, but be not limited to, diphosphonate product (Aaron's phosphonate (alendronate), etidronate (etidronate)), hormone product (raloxifene (raloxifene)), vitamin D product, calcitonin (calcitonin) product and calcium product.But diphosphonate product has low absorptivity and the dispensing problem of making us perplexing, and may bring out esophagitis.Hormone product needed is offerd medicine throughout one's life and is had potential side effect, as breast carcinoma, and uterus carcinoma, cholelithiasis and thrombosis.Vitamin D product is expensive and be not very effective.Calcitonin product also has the high problem with being difficult to offer medicine of cost.Calcium product has lower side effect, but is only limitted to prevention instead for the treatment of.Therefore, the osteoporosis therapies that needs are new is still had.
Plant amedica therapy has been considered to the feasible alternative medicine of various disease.The U.S.'s No. 5478549 patent relates to a kind of method entering Mammalian bone tissue for oral induction and promotion calcium absorption, and its flavone glycoside aglucon glucosides (flavonol aglycone glucoside) comprising dispensing effective dose is in conjunction with nutrition calcium.The U.S.'s No. 6340703 patent provides a kind of method for the treatment of and prevention of osteoporosis disease, comprise the isoflavone formononetin (isoflavone formononetin) of the experimenter's effective dose needing this treatment, optionally give one or more pharmaceutically acceptable adjuvant, supporting agent and/or excipient.The U.S.'s No. 7122214 patent uses Rhizoma Drynariae extract (RDE) as osteoporosis therapy agent.The U.S.'s No. 7350914 patent provides a kind of medicine or pharmaceutical preparation, comprise Semen sojae atricolor (Glycine max), hair larynx sheath stamen (Coleus forskohlii), Camellia sinensis (Camellia sinensis), Bulbus Allii (Allium sativum), Withania somnifera (Withania somnifera), Herba Hedyotidis Diffusae (Boerhavia diffusa), with the plant mixture of Rhizoma Curcumae Longae (Curcumalonga), or extraction is from the active ingredient mixture of they plant.The U.S.'s No. 8153167 patent provides a kind of compositions derived from 6 plant material: (i) Herba Epimedii (Herba epimedii), (ii) Fructus Psoraleae (Fructus Psoraleae), (iii) Radix Rehmanniae (Radix Rehmanniae preparatae) (Radix Rehmanniae Preparata), (iv) Cortex Eucommiae (Cortex Eucommiae), (v) Fructus Cnidii (Fructus Cnidii), (vi) compositions of the Radix Astragali (Radix Astragali), said composition is used for the treatment of the patient's condition relevant with osteoblast and/or osteoclast, as osteoporosis with about the patient's condition of bone amount or menopause, obesity, the not resistance to disease of glucose and diabetes.Radix Puerariae (Puerariae radix) (Wang is comprised to the previous research of other Chinese traditional herbs of bone formation tool possibility effect, X., Wu, J., Chiba, H., Yamada, K., & Ishimi, Y. (2005) .Puerariae radix prevents bone loss in castratedmale mice.Metabolism, 54, 1536-1541.), Rhizoma Drynariae (Drynariae rhizome) (Jeong, J.C., Lee, J.W., Yoon, C.H., Lee, Y.C., Chung, K.H., Kim, M.G., & Kim, C.H. (2005) .Stimulative effects of Drynariae Rhizoma extracts on the proliferation anddifferentiation of osteoblastic MC3T3-E1 cells.Journal of Ethnopharmacology, 96, 489-495.) and Herba Epimedii (Epimedium pubescens) (Hsieh, T.P., Sheu, S.Y., Sun, J.S., Chen, M.H., & Liu, M.H. (2010) .Icariin isolated from Epimedium pubescensregulates osteoblasts anabolism through BMP-2, SMAD4, and Cbfa1 expression.Phytomedicine, 17, 414-423).
Rabbit tail grass (Uraria crinita (L.) Desv.Ex DC. (Fabaceae)) is a kind of traditional edible plants medicine, is extensively distributed in whole India, Thailand, Indonesia, SOUTHERN CHINA and Taiwan.Existing report is pointed out effectively to suppress pressure ulcer.The genus nitric oxide production removing of display of rabbit tail grass and in vitro antioxidation (Luo, C., Liu, A.M., Xing, W.Q., Shi, G., Cao, Y., Pang, J.X., & Qiu, Y.C. (2011) .Antioxidanteffect of flavonoids from Uraria crinita.Zhongguo Shiyan Fangjixue Zazhi, 17,198-201.).The purposes of its treatment fear of cold, swelling, stomachache and ulcer may be due to its anti-inflammatory activity.In Taiwan, people also drink its taste, fragrance, sweet taste with decocting in water as Radix Ginseng tea, and quench the thirst; Therefore it is also called as " Taiwan Radix Ginseng " always.In addition, and the bone distortion that motion relevant bad for diet supplement therapy children's skeleton development in Chinese medicine of its root, sprain and pull (Machida, K., Sakamoto, S., & Kikuchi, M. (2009) .Twonew neolignan glycosides from leaves of Osmanthus heterophyllus.Journal ofNatural Medicines, 63,227-231).
But, do not report display rabbit tail grass relevant with skeletal diseases.
Summary of the invention
The invention provides a kind of osteogenetic method of promotion, the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provides a kind of rabbit tail grass genus or its part or its extract for the manufacture of the purposes promoting osteogenetic medicament.
The present invention also provides a kind of method treating and/or preventing osteopathia, and the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provide a kind of rabbit tail grass to belong to or its part or its extract for the manufacture of the purposes of medicament treating and/or preventing osteopathia.
The present invention further provides a kind of method providing neuroprotective, the method comprises the rabbit tail grass casting effective dose and belongs to its part or its extract extremely individuality.The present invention further provides a kind of rabbit tail grass belong to or its part or its extract for the manufacture of the purposes of medicament providing neuroprotective.
Accompanying drawing explanation
Fig. 1 shows rabbit tail grass (Uraria crinite; UC) ethanolic extract and distribution portion (partitionfraction) ALP in HOb cell is active.HOb cell is inoculated in 96 porose discs.After 24 hours, discard old culture medium, and with tested extract process cell (100 mcg/ml) 72 hours in osteoblast differentiation culture medium.P-nitrophenyl disodium hydrogen phosphate substrate (p-nitrophenyl phosphate disodium substrate) is used to assess ALP activity.Data show with meansigma methods ± standard deviation (n=3).Compared to matched group group (n=3), * P < 0.05 and * * P < 0.01.
Fig. 2 shows rabbit tail grass (UC) ethanolic extract and the mineralising (mineralization) of distribution portion (partition fraction) in HOb cell is active.HOb cell is laid in 24 porose discs, discard old culture medium after three days, and in Mineralized Culture base, process cell (containing the osteoblast differentiation culture medium of ascorbic acid (50 mcg/ml) with β-phosphoglycerol (10 milli mole)) 12 days with tested extract (100 mcg/ml).Experiment at the end of, by culture with 75% ethanol fix, and Mineral nodules (mineralized nodule) is formationed alizarin red-S (Alizarin Red-S) dye assess.Carry the product dyed thereby of combination with the punching of 10% cetylpyridinium chloride (cetylpyridinium chloride) solution, and use microtiter plate reader (microtiter plate reader) quantitative.Data show with meansigma methods ± standard deviation (n=3).Compared to matched group group (n=3), * P < 0.05 and * * P < 0.01.
The schematic diagram of the chemical constitution of the composition that Fig. 3 display is separated from rabbit tail grass.
Fig. 4 demonstrates apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside (the HMBC dependency (H → C) of apigenin6-C-(β-D-apiofuranosyl (1 → 2)-α-D-xylopyranoside) (3).
The cell survival rate of compound in HOb cell that Fig. 5 display separation belongs to from rabbit tail grass.HOb cell is inoculated in 96 porose discs.After 24 hours, discard old culture medium, and in osteoblast differentiation culture medium, process cell 72 hours with tested compounds (100 μMs).MTT is used to analyze (MTT assay) assess and determine cytoactive.Data show with meansigma methods ± standard deviation (n=3).Compared to matched group group (n=3), * P < 0.05 and * * P < 0.01.
Fig. 6 display separation is active from the ALP of compound in HOb cell of rabbit tail grass.HOb cell is inoculated in 96 porose discs.After 24 hours, discard old culture medium, and in osteoblast differentiation culture medium, process cell 72 hours with tested compounds (100 μMs).P-nitrophenyl disodium hydrogen phosphate substrate (p-nitrophenyl phosphate disodium substrate) is used to assess ALP activity.Data show with meansigma methods ± standard deviation (n=3).Compared to matched group group (n=3), * P < 0.05 and * * P < 0.01.
Fig. 7 display separation is active from the mineralising of compound in HOb cell (mineralization) of rabbit tail grass.HOb cell is laid in 24 porose discs, discard old culture medium after three days, and in Mineralized Culture base, process cell (containing the osteoblast differentiation culture medium of ascorbic acid (50 mcg/ml) with β-phosphoglycerol (10 milli mole)) 12 days with tested extract (100 mcg/ml).Experiment at the end of, by culture with 75% ethanol fix, and Mineral nodules (mineralizednodule) is formationed alizarin red-S (Alizarin Red-S) dye assess.Carry the product dyed thereby of combination with the punching of 10% cetylpyridinium chloride (cetylpyridinium chloride) solution, and use microtiter plate reader (microtiter plate reader) quantitative.Data show with meansigma methods ± standard deviation (n=3).Compared to matched group group (n=3), * P < 0.05 and * * P < 0.01.
Detailed description of the invention
The present invention surprisingly finds that rabbit tail grass belongs to or its extract increases osteogenesis and can stimulate bone growth and reparation, and provides the effect of neuroprotective.
Unless otherwise defined, all technology used herein have the implication identical with the usual understanding of those skilled in the art with scientific terminology.Although any method and similar or be equal to material described herein can be used for implement or test the present invention, preferably method and material are described.For object of the present invention, following term is defined as follows.
Person as used herein, term " " and " one " refer to the grammar object of this article one or more than one (that is, at least one).
Person as used herein, apply for a patent model be in term "or" mean " with (and)/or ", to be only unless expressly stated or, unless or should or mutual exclusion.
Person as used herein, term " promotion ", " enhancement " or " improvement " refer to the enhancement of activity, reaction, the patient's condition, disease or other biological parameter.
Person as used herein, term " individuality " comprises biology alive, such as people, monkey, cattle, sheep, horse, pig, cattle, goat, Canis familiaris L., cat, mice, rat, cultured cells and genetically modified organism.In a preferred embodiment, individuality is people.
Person as used herein, term " dispensing " (or cast, use) comprises route of administration, allows rabbit tail of the present invention grass to belong to and performs its expectation function.
Person as used herein, term " treatment " refers to and palliates a disease or the method for impact of disease.Treatment also can refer to the method for the reason reducing disease or disease itself.Treatment also can be any reduction from natural horizontal, and can be but be not limited to disease, disease, or the symptom of disease or disease is eliminated completely.
Person as used herein, term " prevention " refers to and suppresses or alleviate the symptom relevant to osteoporosis.
Person as used herein, term " effective dose " refers to the amount effectively treating and/or preventing osteoporosis or provide the rabbit tail grass of neuroprotective to belong to.
Person as used herein, term " osteoblast " or " skeletonization (osteogenesis) " refer to from undifferentiated stem cell and osteoblast system propagation be osteoblast and osseous tissue (such as, the synthesis of new bone matrix and accumulation).Skeletonization also refers to that CFU-GM (progenitor) or precursor (precursor cell) differentiation or transdifferentiation are osteocyte (that is, osteoblast).CFU-GM or precursor can be pluripotent stem cells, comprise, such as, and interstital stem cell.CFU-GM or precursor can be the cell of preboarding osteoblast system (such as, pre-osteoblast (pre-osteoblast cells)) or the not predetermined cell (such as, PECTORAL LIMB SKELETON or sarcoplast (myoblast)) forming osteoblast system.
Person as used herein, term " osteopathia " refers to and loses relevant disease with osseous tissue, as osteoporosis.
Promote osteogenesis and treat and/or prevent method or the purposes of osteoporosis
On the one hand, the invention provides a kind of osteogenetic method of promotion, the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provides a kind of rabbit tail grass genus or its part or its extract for the manufacture of the purposes promoting osteogenetic medicament.
On the other hand, the invention provides a kind of method increasing bone amount, the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provides a kind of rabbit tail grass genus or its part or its extract for the manufacture of the purposes of the medicament of increase bone amount.
On the other hand, the invention provides a kind of method of raising or enhance bone growth speed, the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provide a kind of rabbit tail grass to belong to or its part or its extract for the manufacture of improving or the purposes of medicament of enhance bone growth speed.
In another, the invention provides a kind of method treating and/or preventing osteopathia, the method comprises the rabbit tail grass genus or its part or its extract to one individuality that cast effective dose.The present invention also provide a kind of rabbit tail grass to belong to or its part or its extract for the manufacture of the purposes of medicament treating and/or preventing osteopathia.
In vitro or in vivo, any method as known in the art can be used to detect osteogenetic induction; Such as, by the performance being detected as osteocyte specific protein, detect the performance of bone idiosyncratic transcription factor, and detect the change of bone density.Osteoblast Specific protein comprises, such as, and alkali phosphatase (alkaline phosphatase; ALP), I-type collagen, Bone Gla protein (osteocalcin), and make bone protein (osteoponin) (Olsen et al., Annu.Rev.Cell.Dev.Biol.16:191 (2000)).At some specific embodiments, the performance of alkali phosphatase is detected as osteogenetic index.Bone idiosyncratic transcription factor comprises, such as, and Cbfa1/Runx2, GSC, D1x1, D1x5, Msx1, Cart1, Hoxa1, Hoxa2, Hoxa3, Hoxb1, rae28, Twist, AP-2, MF1, Pax1, Pax3, Pax9, TBX3, TBX4, TBX5 and Brachyury (Olsen et al, 2000, supra).
Method of the present invention can be used for treating osteopathia.According to the present invention, treatment osteopathia can be carried out by increase bone mass or osteogenesis, includes but not limited to, osteoporosis, arthritis, osteoarthritis, periodontal disease, alveolus bone-loss, osteotomy bone-loss, Childhood idiopathic bone-loss, the loss of curvature spinal column and height.Can treat destructive osteopathia according to the present invention, include but not limited to, osteoporosis, osteoarthritis and osteolytic lesion, such as those are by tumor disease, radiotherapy, or the pathological changes that chemotherapy causes.
Osteopathia can by being characterized as low bone mass, Cranial defect, or caused by the situation of cartilage defect.Term " bone amount (bone mass) " refers to the bone amount of per unit volume.Feature to be the patient's condition of Low BMD be wherein bone amount level lower than the normal level of given age, as " Assessment of Fracture Risk and Its Application to Screening forPostmenopausal Osteoporosis; " Report of a World Health Organization Study Group, World Health Organization Techhical Series 843 (1994) " in defined.Cranial defect is the proportional imbalance of bone formation to bone resorption, and so, as unmodified, individual cognition presents the skeleton lower than expecting, or the skeleton of individuality can be more imperfect than what expect.Cranial defect also can by fracturing, getting involved or cause from tooth or periodontal from operation.Knitting includes, but not limited to repairing bone defect, as betided, such as, closing, opening and not healedmyocardial fracture.Cartilage defect is the cartilage of damage, than the few cartilage of expection, or comparatively expects incomplete cartilage.
Comprising the patient's condition being characterized as low bone mass is, but is not limited to, constitutional and secondary osteoporosis, periodontal disease, alveolus bone-loss, and osteotomy bone runs off, childhood period idiopathic bone-loss.The patient's condition being characterized as low bone mass also includes, but not limited to the long-term complications of osteoporosis as rachiocamposis, the loss of height and prosthetic surgery.
Osteoporosis or porous bone are a kind of diseases, and feature is that the bone mass net flow of per unit volume is lost.This kind of bone mass runs off and the consequence of fracture that causes is the destruction of skeleton, and with the support structure providing health enough, the structure degradation of Low BMD and osseous tissue causes bone fragility and increases the fracture of hip joint, vertebra and wrist.There is bone-loss and there is no symptom.Osteoporosis comprises " secondary osteoporosis ", as the osteoporosis (glucocorticoid-induced osteoporosis) of glucocorticoid inducible, the osteoporosis that hyperthyroidism causes, the osteoporosis that immobilization causes, the osteoporosis that heparin brings out or the osteoporosis that immunosuppressant is brought out.Suffering from the sufferer of osteoporosis, bone can become fragility and make unexpected anxiety can cause fracture or cause vertebrae collapse.Current most osteoporosis treatment stops the bone-loss continuing, but cannot strengthen bone formation and therefore the quality of bone is still very poor, but can not become even worse.
At a specific embodiment, any said method of the present invention comprises the additional step casting osteogenesis promoter.Medicament in any such use of the present invention separately comprises osteogenesis promoter.Preferably, described additional active agent system is selected from diphosphonate (bisphosphonates).Preferably diphosphonate includes but not limited to, tiludronic acid (tiludronic acid), alendronic Acid (alendronic acid), zoledronic acid (zoledronic acid), ibandronic acid (ibandronicacid), risedronic acid (risedronic acid), etidronic acid (etidronic acid), Crow phosphonic acids (clodronicacid), and pamidronic acid (pamidronic acid) and their pharmaceutically acceptable salt.Those skilled in the art will know, these compounds represent with its ionic species usually, such as, Tiludronate (tiludronate), fosamax (alendronate), zoledronate (zoledronate), ibandronate (ibandronate), Risedronate (risedronate), etidronic acid (etidronate), Crow phosphonate (clodronate) and pamldronate (pamidronate).Special preferably diphosphonate comprises fosamax and Risedronate.
Method or the purposes of neuroprotective are provided
On the other hand, the invention provides the method for providing neuroprotective, the method comprises the rabbit tail grass casting effective dose and belongs to its part or its extract extremely individuality.The present invention also provide a kind of rabbit tail grass to belong to or its part or its extract for the manufacture of the purposes of medicament providing neuroprotective.
Neuroprotective refers to for acute illness (as apoplexy or brain or nervous system injury/wound; anoxia; spinal cord injury or peripheral nerve injury) after prevent nervous system midbrain/neuronal damage or degeneration or prevent chronic neurodegenerative disease (as parkinson; A Zihai Mo's disease, multiple sclerosis) Mode and policy of result.The object of neuroprotective reduces neuron dysfunction/death after central nervous system injury, and attempt the integrity maintaining the interactional maximum possible of brain cell, thus produce undisturbed function of nervous system.By providing neuroprotective, rabbit tail grass belongs to or its part or its extract can be used for treatment nerve injury (as apoplexy, especially cerebral infarction, brain injury, anoxia, spinal cord injury or peripheral nerve injury), and treatment or prevention neurodegenerative disease.
Rabbit tail grass for the inventive method or purposes belongs to or its part or its extract
It is pulse family that rabbit tail grass belongs to (Uraria), a genus of flowering plant in Papilionaceae.It belongs to Papillionoideae (Faboideae).At a specific embodiment, it is be selected from following group that rabbit tail grass belongs to: rabbit tail grass (Uraria crinita (L.) Desv.ex DC.), great Ye rabbit tail grass (Uraria lagopodioides (L.) Desv.ex DC.), leatherleaf rabbit tail grass (Uraria picta (Jacq.) Desv.ex DC.), roundleaf rabbit tail grass (Uraria neglecta Prain), Urariaacaulis Schindl, Uraria acuminata Kurz, Uraria balansae Schindl., Uraria barbataLace, Uraria campanulata (Benth.) Gagnep., Uraria candida Backer, Uraria clarkeiGagnep., Uraria cochinchinensis Schindl., Uraria cordifolia Wall., Uraria crinita (L.) DC., Uraria cylindracea Benth., Uraria fujianensis Y.C.Yang & P.H.Huang, Uraria gossweileri Baker f., Uraria kurzii Schindl., Uraria lacei Craib, Urarialagopodoides (L.) DC., Uraria lagopus DC., Uraria longibracteata Y.C.Yang & P.H.Huang, Uraria pierrei Schindl., Uraria poilanei Phon, Uraria prunellifoliaBaker, Uraria rotundata Craib, Uraria rufescens (DC.) Schindl. and Uraria sinensis (Hemsl.) Franch.Preferably, rabbit tail grass belongs to for rabbit tail grass, great Ye rabbit tail grass, leatherleaf rabbit tail grass or roundleaf rabbit tail grass.More preferably, rabbit tail grass belongs to for rabbit tail grass.Preferably, it is rabbit tail grass that rabbit tail grass belongs to extract, great Ye rabbit tail grass, the extract of leatherleaf rabbit tail grass or roundleaf rabbit tail grass.
All parts of whole rabbit tail grass platymiscium or rabbit tail grass platymiscium can use in the present invention.At a specific embodiment, the root of rabbit tail grass platymiscium is used in the present invention.
At a specific embodiment, the extract that rabbit tail grass belongs to can be used for the present invention.Preferably, rabbit tail grass, great Ye rabbit tail grass, leatherleaf rabbit tail grass or roundleaf rabbit tail grass are used for being extracted thing.Preferably, this rabbit tail grass belongs to for rabbit tail grass.More preferably, rabbit tail grass is for obtaining this extract.Preferably, described extract is the organic solvent extraction thing of whole rabbit tail grass platymiscium, and preferably extract is the organic solvent extraction thing that rabbit tail grass belongs to root.Preferably, described extract is methanol extraction thing, ethanolic extract, propanol extract, butanol extract or hexane extraction thing.More preferably, described extract is ethanolic extract.According to a specific embodiment of the present invention, organic solvent extraction thing can use hexane further, and the distribution (partition) of ethyl acetate and butanols, obtains hexane extraction thing, acetic acid ethyl ester extract and butanol extract.Therefore, the invention provides a kind of rabbit tail grass and belong to extract, it is produced by following steps: belong to alcohol extraction rabbit tail grass and obtain ethanolic extract, and obtain acetic acid ethyl ester extract or butanol extract with ethyl acetate or butanol, before immunoassay ethanolic extract.Acetic acid ethyl ester extract can Further Division, obtains 9 parts.3rd part of these parts uses Sephadex LH-20 tubing string chromatographic analysis (chromatograph) abstraction and purification further, and obtains 17 parts with methanol/water extraction.Gained part is separated with chromatographic analysis further.Therefore, a kind of new compound can be obtained, apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside.Accordingly, the invention provides a kind of rabbit tail grass and belong to extract, it belongs to by extraction rabbit tail grass obtaining ethanolic extract, and with ethyl acetate or butanol, before immunoassay ethanolic extract to obtain acetic acid ethyl ester extract or butanol extract.
Any abstraction technique known in the art can be used for preparing extract of the present invention.The extract obtained can further with chromatographic analysis Further Division.Preferably chromatographic analysis adopts solvent to rush the liquid phase chromatographic analysis proposed.Preferably, described liquid phase chromatographic analysis is high-efficient liquid phase color layer analysis (HPLC) or anti-phase HPLC.
Be separated the noval chemical compound from rabbit tail grass genus or its part or its extract
The invention provides a kind of following noval chemical compound be separated from rabbit tail grass genus or its part or its extract:
Apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside (apigenin6-C-(β-D-apiofuranosyl (1 → 2)-α-D-xylopyranoside)
Therefore, the invention provides a kind of formula (I) compound,
Wherein R
1it is furan celery glycosyl-α-L-arabopyranose base (apiofuranosyl-α-L-arabinopyranosyl), furan celery glycosyl-β-D-Glucose pyranose (apiofuranosyl-β-D-glucopyranosyl), furan celery glycosyl-β-D-galactopyranosyl glycosyl (apiofuranosyl-β-D-galactopyranosyl), furan celery glycosyl-β-D-xylopyranosyl (apiofuranosyl-β-D-xylopyranosyl), furan celery glycosyl-β-D-arabopyranose base (apiofuranosyl-β-D-arabinopyranosyl), furan celery glycosyl-β-D-ribopyranose base (apiofuranosyl-β-D-ribopyranosyl), furan celery glycosyl-β-D-pyrans lysol glycosyl (apiofuranosyl-β-D-lyxopyranosyl), furan celery glycosyl-β-D-pyrans ribulose base (apiofuranosyl-β-D-ribulopyranosyl), furan celery glycosyl-β-D-xylopyranose ketose (apiofuranosyl-β-D-xylulopyranosyl), furan celery glycosyl-β-D-A Luo pyrans glycosyl (apiofuranosyl-β-D-allopyranosyl), furan celery glycosyl-β-D-pyrans altrose base (apiofuranosyl-β-D-altropyranosyl), furan celery glycosyl-β-D-mannopyranose base (apiofuranosyl-β-D-mannopyranosyl), furan celery glycosyl-β-D-pyrans gulose base (apiofuranosyl-β-D-gulopyranosyl), furan celery glycosyl-β-D-pyrans idose base (apiofuranosyl-β-D-idopyranosyl), furan celery glycosyl-β-D-pyrans talose base (apiofuranosyl-β-D-talopyranosyl), furan celery glycosyl-α-L-glucopyranosyl (apiofuranosyl-α-L-glucopyranosyl), furan celery glycosyl-α-L-galactopyranosyl glycosyl (apiofuranosyl-α-L-galactopyranosyl), furan celery glycosyl-α-L-xylopyranosyl (apiofuranosyl-α-L-xylopyranosyl), furan celery glycosyl-α-L-arabopyranose base (apiofuranosyl-α-L-arabinopyranosyl), furan celery glycosyl-α-L-ribopyranose base (apiofuranosyl-α-L-ribopyranosyl), furan celery glycosyl-α-L-pyrans lysol glycosyl (apiofuranosyl-α-L-lyxopyranosyl), furan celery glycosyl-α-L-pyrans ribulose base (apiofuranosyl-α-L-ribulopyranosyl), furan celery glycosyl-α-L-xylopyranose ketose base (apiofuranosyl-α-L-xylulopyranosyl), furan celery glycosyl-α-L-A Luo pyrans glycosyl (apiofuranosyl-α-L-allopyranosyl), furan celery glycosyl-α-L-pyrans altrose base (apiofuranosyl-α-L-altropyranosyl), furan celery glycosyl-α-L-mannopyranose base (apiofuranosyl-α-L-mannopyranosyl), furan celery glycosyl-α-L-pyrans gulose base (apiofuranosyl-α-L-gulopyranosyl), furan celery glycosyl-α-L-Chinese mugwort Du's pyrans glycosyl (apiofuranosyl-α-L-idopyranosyl) or furan celery glycosyl-α-L-pyrans talose base (apiofuranosyl-α-L-talopyranosyl).
Preferably, R1 is furan celery glycosyl-α-L-arabopyranose base, furan celery glycosyl-β-D-arabopyranose base, furan celery glycosyl-β-D-glucopyranosyl, furan celery glycosyl-β-D-galactopyranosyl glycosyl, furan celery glycosyl-β-D-xylopyranosyl or furan celery glycosyl-β-D-arabopyranose base.More preferably, R1 is furan celery glycosyl-α-L-arabopyranose base.
The compound of the different sugar group of R1 can (Org.Biomol.Chem. according to procedures known in the art in tool general formula (I), 2010,8,4451-4462), modify from apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside and obtain.The various glycosyls connecting R1 are as follows to the synthetic schemes of apigenin (apigenin):
Composite and medication administration method
Rabbit tail grass belongs to, its part or its extract can with pharmaceutically acceptable supporting agent, excipient, diluent and/or salt are allocated as medical composition for administration or medicine together.
Pharmaceutically acceptable supporting agent, diluent, excipient and/or salt refer to supporting agent, diluent, excipient and/or salt must be compatible with other compositions in preparation, can not adversely affect the treatment benefit that rabbit tail grass belongs to its part or its extract, and its receiver can not be made to be injured.
Belong to for implementing rabbit tail of the present invention grass, the casting of its part or its extract or its medical composition can be any method that general and/or local (such as, in fracture, the position of osteotomy or orthomorphia) send compound.These methods comprise oral route, parenteral approach, intraduodenal route etc.
In topical application, rabbit tail grass belongs to, its part or its extract or its medical composition are applied to fracture, the position of osteotomy or graft, such as, by injection suitable solvent (such as, oil-based solvent is as Oleum Arachidis hypogaeae semen) in rabbit tail grass belong to, its part or its extract are in fracture site or knitting position, or when open surgery, by topical application, such as Suitable carriers (such as bone wax, demineralized bone end, the bone cement of polymerization, bone sealant, polylactic acid, polyglycolic acid, polylactic acid-polyglycolic acid etc.) in such as bone wax, decalcification bone powder, the bone cement of polymerization, bone sealant, polylactic acid, polyglycolic acid, polylactic acid, its part such as polyglycolic acid or its extract.Or, topical application can mat be applied in solution supporting agent rabbit tail grass belong to, the solution of its part or its extract or dispersion liquid are in surface or the solid or the semisolid implant that are incorporated to conventional use in Orthopeadic Surgery, as terylene mesh sheet (Dacron-mesh), gel foam and Kiel bone (kiel bone), or prosthese (proctheses).
For local application, rabbit tail grass belongs to, and its part or its extract or adjustable the making of its medical composition to suspend containing active component or to be dissolved in the suitable ointment of one or more supporting agents.Belong to for rabbit tail grass of the present invention, the supporting agent of the topical of its part or its extract includes, but not limited to mineral oil, Albolene, white vaseline, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifing wax, and sugar, as lactose and water.Or described medical composition can be mixed with containing suspension or the rabbit tail grass genus being dissolved in one or more pharmaceutically acceptable supporting agents, the Emulsion of its part or its extract or emulsifiable paste.Suitable supporting agent includes, but not limited to mineral oil, sorbitol monostearate, polysorbate60, spermaceti ester type waxes, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
According to concrete plan disposition, disease or disease, additional therapeutic agent can belong to rabbit tail grass, its part or the administration together of its extract.These extra therapeutic agents belong to containing rabbit tail grass, and the compositions of its part or its extract, can the administration successively of any order, as the part (continuously or intermittently administration) of the multiple dose course for the treatment of.Or those medicaments can be a part for single dosage form, belong to rabbit tail grass, its part or its extract are woven into single compositions (administration simultaneously) together.
For oral administration, can be solution for medical composition of the present invention, suspension, lozenge, pill, capsule, powder, granule, semi-solid, slow releasing preparation, elixir, the patterns such as aerosol.Lozenge contains various excipient as sodium citrate, and calcium carbonate and calcium phosphate and various disintegrating agent, as starch, be preferably Rhizoma Solani tuber osi or tapioca and some composition silicate, and bonding agent are as polyvinylpyrrolidones, sucrose, gelatin and arabic gum.In addition, lubricant is as magnesium stearate, and sodium lauryl sulphate and Talcum are usually very useful for briquetting object.Similar solid composite is also used as the filler of soft hard-filled gelatin capsule, and preferably material also comprises lactose or nougat and high molecular weight polyethylene glycol in this respect.When waterborne suspension and/or elixir are used for oral administration, rabbit tail grass of the present invention belongs to, and its part or its extract can in conjunction with various sweeting agents, flavoring agent, coloring agent, emulsifying agent and/or suspending agent, and diluent, such as, as water, ethanol, propylene glycol, glycerol and various types of seemingly combination thereof.
Many factors is depended in the selection of preparation, the bioavailability of such as drug administration pattern (such as, in oral administration, preparation is lozenge, and the form of pill or capsule is preferably) and drug substance.
Term used herein " parenteral " refers to administering mode, comprises intravenous, intramuscular, intraperitoneal, in breastbone, subcutaneous, with intra-articular injection and infusion in marrow.Medical composition for parenteral injection can comprise pharmaceutically acceptable sterilized water or non-aqueous solution, dispersion liquid, suspension or emulsion, and is again mixed with the sterilized powder of aseptic injectable solution or dispersion liquid before using.Aqueous solution is specially adapted to intravenous, intramuscular, the object of subcutaneous and peritoneal injection.In this respect, aseptic aqueous medium used is all easily obtained by the standard technique of those skilled in the art.Suitable aqueous and non-aqueous supporting agent, diluent, the example of solvent or excipient comprises water, ethanol, and polyhydric alcohol (as glycerol, propylene glycol, Polyethylene Glycol etc.), carboxymethyl cellulose and suitable mixture thereof, vegetable oil (as olive oil), and injectable organic ester is as ethyl oleate.Suitable mobility can be kept, such as, by use coating material as lecithin, by maintaining required particle diameter when dispersion liquid, and by use surfactant.
Also can adjuvant be comprised for medical composition of the present invention, such as, but not limited to, antiseptic, wetting agent, emulsifying agent and dispersant.Prevent microbial action can such as, by comprising various antibacterial and antifungal is guaranteed, p-Hydroxybenzoate, methaform, phenol, sorbic acid etc.It also can comprise isotonic agent, such as sugar, sodium chloride etc.The prolongation of injectable drug form absorbs and can realize by comprising the material such as aluminum monostearate and the gelatin that postpone to absorb.
Slow administered by infusion in sheath or epidural route particularly useful.Pu is helped to be known in the art for sending the many implantable of compound or can installing in the body of the speed of adjustment.See, such as, the U.S.'s No. 4619652 patent.
Suspension, in addition to the active compound, can comprise suspending agent, such as, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and Isosorbide Dinitrate, microcrystalline Cellulose, inclined aluminium hydroxide, bentonite, agar, and Tragacanth (tragacanth), and their mixture.
In order to percutaneous (as local) administration, preparation dilution aseptic aqueous solution or part the aqueous solution concentration of about 0.1% to 5% (usually), be similar to above-mentioned parenteral solution.
Also can by nose aerosol or inhalation for medical composition of the present invention.This kind of compositions can according to the known technology preparation in field of pharmaceutical preparations, and can be prepared as saline solution, adopt benzyl alcohol or other suitable antiseptic, absorption enhancer is to improve bioavailability, fluorocarbon, and/or other conventional solubilizing agent or dispersant.
Compositions for rectum or vagina administration is preferably suppository, can belong to by mixing rabbit tail grass of the present invention, its part or its extract and suitable nonirritant excipient or supporting agent, as cocoa butter, Polyethylene Glycol or suppository wax (are at room temperature solid and are liquid under body temperature, therefore dissolve in rectum or vaginal canal and discharge medicine), prepare.
Other pharmaceutically acceptable supporting agent comprises, but be not limited to, avirulent solid, semisolid or liquid filling agent, diluent, the formulation auxiliary agents of cover material or any type, include but not limited to, ion-exchanger, aluminium oxide, aluminium stearate, lecithin, serum albumin, as human serum albumin, buffer substance is as phosphate, glycine, sorbic acid, potassium sorbate, saturated vegetable fatty acid, water, salt or electrolytical partial glyceride mixture, as protamine sulfate, sodium hydrogen phosphate, hydrogen hydrogen orthophosphate, sodium chloride, zinc salt, silica sol, magnesium trisilicate, polyvinylpyrrolidone, based on cellulosic material, Polyethylene Glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene-polyoxypropylene block polymer, Polyethylene Glycol and lanoline.
Solid pharmaceutical excipients includes, but not limited to starch, cellulose, Talcum, glucose, lactose, sucrose, gelatin, Fructus Hordei Germinatus, rice, flour, Chalk, silica gel, magnesium stearate, sodium stearate, glyceryl monostearate, sodium chloride, defatted milk powder etc.Liquid and semisolid excipient can be selected from glycerol, propylene glycol, water, and ethanol and various oil are selected, and comprise those oil, animal, plant or synthesis source, such as, and Oleum Arachidis hypogaeae semen, soybean oil, mineral oil, Semen Sesami wet goods.Preferably liquid carrier, especially for Injectable solution, comprises water, saline, D/W and ethylene glycol.
Preparation is known containing the method for the various medical compositions of a certain amount of active component, or according to this exposure, will be apparent to those this areas skill personage.Other suitable drug excipient and preparation system thereof are described in Remington ' sPharmaceutical Sciences, edited by E.W.Martin, Mack Publishing Company, 19thed. (1995).
The rabbit tail grass containing 1%-100% (by weight) for medical composition of the present invention belongs to, and its part or its extract, be preferably 10%-100%, 10%-95% or 20%-95% (by weight).Under any circumstance, compositions to be administered or preparation are by the treatment of effective dose according to the present invention, and the situation of the experimenter be treated, disease or disease, determine that contained rabbit tail grass belongs to, the amount of its part or its extract.
Those of ordinary skill in the art will be understood that, pharmaceutically the rabbit tail grass of effective dose belongs to, and its part or its extract can be determined by rule of thumb, and can in a pure form (when such form exists), with pharmaceutically acceptable salt, the form of ester or prodrug uses.This medicament can be applied to patient as the medical composition with one or more pharmaceutically acceptable excipient composition.Will be understood that when being applied to, such as, human patients, the usage total every day of medicament of the present invention or compositions will be determined by attending doctor within the scope of reliable medical judgment.Concrete treatment effective dose, for any particular patient, will depend on many factors: the cell effect type that reach and degree, the concrete medicament of use or the activity of compositions, the concrete reagent used or compositions, age, body weight, general health situation, the sex of patient and diet, administration time, the discharge rate of route of administration and medicament, the persistent period for the treatment of, the medicine of the particular agent combinationally used or simultaneously use, and similar factor known in medical domain.
Throw agent and also can arrange specific mode in patients, to provide the predetermined concentration of medicine in blood, as determined by the technology accepted and conventional technology.
Or rabbit tail grass belongs to, and its part or its extract can use food stage supporting agent, excipient, and diluent and/or salt are as health food or dietary supplement.The excipient of above-mentioned supporting agent, diluent and/or salt can use in health food of the present invention or dietary supplement.Health food or dietary supplement can exist in a variety of manners, include but not limited to lozenge, capsule, capsule shape lozenge, powder, and beverage, comprises milk shake, and solid food comprises snack bar etc.
For those of ordinary skill in the related art, other suitable amendment and adjustment will be apparent, can in the spirit and scope not departing from the present invention or its any embodiment with methods and applications as herein described.There is provided the following example to illustrate but not limit the present invention.
Embodiment
Materials and methods
vegetable material
Peasant association of the root Shi Congmingjian township meeting of rabbit tail grass (Uraria crinite), Nantou County, Taiwan Province is collected, and by Mr. Guo Zhaolin, Chinese pharmacy and natural resources of Chinese medicinal materials system, China Medical University, in platform, Taiwan is identified.One specimen (M-343) is stored in pharmacognostical study institute of Taipei Medical University, Taiwan.
reagent and chemicals
DMSO (dimethyl sulfoxine), tetrazolium bromide tetrazole bromide (MTT), triton x-100 (Triton X-100), p-nitrophenyl disodium hydrogen phosphate substrate, alizarin red-S (Alizarin Red-S), cetylpyridinium chloride monohydrate (cetylpyridinium chloride monohydrate), ascorbic acid, β-phosphoglycerol disodium salt hydrate is purchased from Sigma-Aldrich company (Sigma, St.Louis, Missouri, USA).Phosphate buffered saline (PBS) (PBS) and trypsin are purchased from Gibco (Gibco Canada Inc., Burlington, Ont., Canada).Osteoblasticly revolve brightness and measure with JASCO P-1020 numerical digit polariscope (JASCO P-1020 digital polarimeter).Ultraviolet spectrogram Hitachi U-2800U-2800 ultraviolet/vis spectroscopy luminance meter (Hitachi U-2800UV/visspectrometer) measures.1D-and 2D-nuclear magnetic resonance, NMR (NMR) spectrum uses dimethyl sulfoxine-d6 with Bruker AM-500 spectrogrph, MOH-d4, and D
2o measured in solution.HRESI-MS and ESI-MS analyzes and measures with the VG Platform Electrospray ESI/MS purchased from Cell Applications (Cell Applications, Inc., San Diego, CA, USA).All chemicals for this research and reagent are high-grade commercial product.Other materials comprises BCA (dihomocinchonine acid; Bicinchoninic acid) Protein Assay Kit (Thermo Scientific, Rockford, IL, and pNPP hexahydrate (4-nitrophenyl phosphate disodium salthexahydrate) (Alfa Aesar USA), Ward Hill, Massachusetts, USA).
general experimental technique
Revolve brightness to measure with JASCO P-1020 numerical digit polariscope.Ultraviolet spectrogram Hitachi U-2800U-2800 ultraviolet/vis spectroscopy luminance meter measures.1D-and 2D-nuclear magnetic resonance, NMR (NMR) spectrum uses dimethyl sulfoxine-d6 with Bruker AM-500 spectrogrph, MOH-d4, and D
2o measured in solution.HRESI-MS and ESI-MS analyzes with VG PlatformElectrospray ESI/MS.
the preparation and purification of thick extract
Fresh rabbit tail grass roots (14.7 kilograms) aquiferous ethanol (95%v/v, totally 120 liters of litres) extracts 3 times.After filtration, filtrate evaporation is also dry under vacuo.Dried extract (601.4 grams) is suspended in H
2o, and successively with normal hexane, ethyl acetate (EtOAc) and n-butyl alcohol (n-BuOH) divide, and obtain 78.4,32.0 respectively, and the extract of 133.9 grams.At EtOAc and n-butyl alcohol extract, there is maximum Matrix Mineralization activity (matrix mineralizationactivities) (Fig. 2).EtOAc extract is gathered through Portugal candy gel Sephadex LH-20 tubing string with 95% ethanol punching carry, obtain 9 parts (UCE-1 to UCE-9).UCE-3 part is separated further with Sephadex LH-20 col-umn chromatography, and with MeOH/H
2o (
) rush and carry, obtain 17 parts (UCE-3-1 to UCE-3-17).UCE-3-7 part is with MCI CHP-20 col-umn chromatography H
2the punching of O-MeOH gradient is carried, and obtains 14 second part (UCE-3-7-1 to UCE-3-7-14).Compound 2 (98.1 milligrams) UCE-3-7-4 part is through MeOH/H
2obtain after O recrystallize.UCE-4 part Sephadex LH-20 col-umn chromatography is separated further, and with MeOH/H
2o (50 to 100%) punching is carried, and obtains 17 parts (UCE-4-1 to UCE-4-17).UCE-4-3 and UCE-4-10 part through MCI CHP-20 tubing string, and uses MeOH/H
2o (30 to 100%) punching carry, obtain respectively 8 (
) and 11 second part (UCE-4-10-1 to UCE-4-10-11).Compound 11 (12.0 milligrams) prepares C-18 reverse tubing string through HPLC in half from UCE-4-3-2 part, obtains after rushing extraction solvent system purification using 10%MeOH.Compound 4 (6.9 milligrams) prepares C-18 reverse tubing string through HPLC in half from UCE-4-10-2 part, obtains after rushing extraction solvent system purification using 25%MeCN.UCE-5 part MCI CHP-20 col-umn chromatography, with H
2the punching of O-MeOH gradient is carried, and obtains 24 second part (UCE-5-1 to UCE-5-24).UCE-5-15 and UCE-5-16 part gathers candy gel Sephadex LH-20 tubing string through Portugal, with MeOH/H
2o (60%) punching is carried, and obtains 6 (UCE-5-15-1 to UCE-5-15-6) and 10 second part (UCE-5-16-1 to UCE-5-16-10) respectively.Compound 3 (12.2 milligrams) and compound 1 (184.5 milligrams) prepare C-18 reverse tubing string through HPLC in half from UCE-5-15-1 and UCE-5-16-9 part, obtain after rushing extraction solvent system purification using 55%MeOH.N-butanol extract through Diaion HP-20 tubing string, with H
2the punching of O-MeOH gradient is carried, and obtains 6 components (UCB-1 to UCB-6).UCB-3, UCB-4 and UCB-5 part is by respective Sephadex LH-20 tubing string, further separation is proposed with 95%EtOH punching, obtain respectively 6 (UCB-3-1 to UCB-3-6), 9 (UCB-4-1 to UCB-4-9) and 6 (UCB-5-1 to UCB-5-6) parts.UCB-3-3 and UCB-5-2 part through MCI CHP-20 col-umn chromatography, with H
2o-MeOH gradient punching propose separation, obtain respectively 6 (
) and 8 (UCB-5-2-1 to UCB-5-2-8) second part.Compound 7 (38.3 milligrams) and compound 8 (15.3 milligrams) obtain from UCB-3-3-2 part.Compound 5 (58.6 milligrams) and compound 6 (7.6 milligrams) prepare C-18 reverse tubing string with respective HPLC in half from UCB-3-3-3 and UCB-5-2-1 part, obtain after rushing extraction solvent system purification using 20%MeOH.UCB-4-3 and UCB-4-4 part through MCI CHP-20 tubing string, and with MeOH/H
2o (is respectively
with
) separation and purification, obtain 4 (UCB-4-3-1 to UCB-4-3-4) and 9 (UCB-4-4-1 to UCB-4-4-9) second part respectively.Compound 9 (38.5 milligrams) prepares C-18 reverse tubing string through HPLC in half from UCB-4-3-3 part, obtains after rushing extraction solvent system purification using 20%MeOH.Compound 10 (13.5 milligrams) prepares C-18 reverse tubing string through HPLC in half from UCB-4-4-3 part, obtains after rushing extraction solvent system purification using 40%MeOH.
the acid hydrolysis of compound 3
The compound 3 of 1 milligram is hydrolyzed 6-8 hour with 6N HCl with heat block at 80 DEG C.Mixture is cooled, and evaporation is to remove disacidify, and then before analyzing, settling flux in milli-Q water, and passes through Millipore-GX nylon membrane.The monosaccharide of compound 3 and polysaccharide hydrolysate analyses (HPAEC) system (Dionex, Sunnyvale, CA) with high performance anion exchange and anion exchange column (Carbopac PA-10,4.6x250mm) is separated.The degree 18mM sodium hydroxide solutions such as the analysis use of monosaccharide carry out at ambient temperature.
Apigenin 6-C-(β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside (3): yellow solid; [α] 22D+21 (c0.5, MeOH); UV (MeOH) λ max (log ε): 272 (4.36), 329 (4.35) nm; 1H-NMR (DMSO-d6,500MHz) and13C-NMR (DMSO-d6,125MHz), ginseng table 1; HRESI-MS m/z535.1459 [M+H]+(C25H27O13,535.1452 value of calculation).
cell culture
Human osteoblast (HOB) is the normal femur of the Caucasian female deriving from 63 years old and derives from the primary cell of CellApplications (San Diego, CA, USA).HOb Growth of Cells in Oesteoblast growth culture medium (Osteoblast Growth Medium), and is maintained at 37 DEG C, 5%CO
2moist environment.Culture medium is every other day changed.For maintaining exponential growth, cell every 7 days Secondary Culture.Hob cell is used for cell survival, and differentiation and mineralising measure.
cell survival detects
The cell survival of rabbit tail grass extract and part thereof uses MTT to detect to measure.HOb cell is with every porose disc 4 × 10
3the density of individual cell is inoculated in 96 porose discs.Inoculate after 24 hours, discard old culture medium, and with tested extract or compound treated cells.After the cultivation of 72 hours, discard old culture medium and to add in MTT reagent to each hole and to cultivate 4 hours.Then discard culture medium, and the DMSO adding 100 μ l porose to institute in, and fully mixing to dissolve Dark blue crystals.The suction brightness at 600nm place is recorded in ELISA reader (ELISA reader).
alkali phosphatase (ALP) detects
Alkali phosphatase (ALP) activity of rabbit tail grass extract and part thereof measures using p-nitrophenyl phosphate (p-nitrophenylphosphate) as substrate.HOb cell is with every porose disc 4 × 10
3the density of individual cell is inoculated in 96 porose discs.Inoculate after 24 hours, discard old culture medium, and with tested extract or compound treatment the cell in osteoblast differentiation culture medium (Osteoblast Differentiation Medium).Through the cultivation of 72 hours, discard culture medium, the cold PBS of cell is rinsed twice, and with lysis buffer (PBS, the Triton X-100 containing 0.1%) cell lysis.Cellular protein concentration in supernatant, measures after 1 hour with BCA (dihomocinchonine acid) protein reagent process at 37 DEG C, adds 1M NaOH stopped reaction and measures the suction brightness of 560nm.In addition, the alkali phosphatase in supernatant at 37 DEG C, in NaHCO3-Na2CO3 buffer (pH10, the MgSO containing 0.1%Triton X-100,2mM of 0.1M
4and 6mM p-nitrophenyl phosphate) in measure after 1 hour active.Add 1M NaOH stopped reaction and be determined at the suction brightness at 405nm place.Each value is normalized to protein concentration.
mineralized dentin matrix is formed analyzes (Mineralized Matrix Formation Assay)
The mineralization activity of rabbit tail grass extract and part thereof is dyeed by the alizarin red-S of calcium to measure.HOb cell is with every porose disc 7 × 10
4the density of individual cell is inoculated in 96 porose discs.Inoculate after 3 days, discard old culture medium, and with tested extract or compound treatment the cell in the Mineralized Culture base (Osteoblast Differentiation Medium) containing ascorbic acid (50 mcg/ml) and β-phosphoglycerol (10mM) 12 days.At the end of experiment, culture is fixed on the ethanol of 75%, and doped calcium at room temperature uses alizarin red-S solution (40mM, pH value 4.2) to dye 15 minutes.Propose the dyeing of combination with 10% (w/v) cetylpyridinium chloride solution punching in 10mM sodium phosphate (pH7.0), and carry out quantitatively at 550nm place with ELISA reader.
statistical analysis
Student t check (Student ' s t test) for measure each experimental group value between statistically-significant difference.Data represent with meansigma methods ± standard deviation and P value < 0.05 is statistically significant difference.
Example 1
rabbit tail grass promotes osteoblast differentiation and Matrix Mineralization (matrix mineralization) in Hob cell
95% ethanol (EtOH) extract of rabbit tail grass (UC) root uses the method for biological level to carry out extracting and chromatography.The thick extract of ethanol of 95% is suspended in H
2o with normal hexane, ethyl acetate and n-butyl alcohol sequentially divide, obtains 4 parts.Thick extract adopts mtt assay to assess at the cell proliferation of Hob cell.Result display without any thick extract be found to cause significant cytotoxicity or HOb cell in the propagation (data are not shown) of 100 mcg/ml or following concentration.ALP activity is the labelling of main osteoblast differentiation.By measuring the ALP Activity determination extract of HOb cell to the effect of osteoblast differentiation.In the process of Hob cell after 72 hours, result shows the ALP activity (Fig. 1) that the thick extract of ethanol of 95% of 100 mcg/ml and aqueous fractions present increase by 106.51 ± 0.44 and 119.36 ± 5.64% respectively.In addition, test extract is formed on the impact of osteoblast mineralization by the bone tuberosity being determined at HOb cell.At the 12nd day, histochemistry Alizarin red staining is carried out to Mineral nodules.Result shows, and at the thick extract of ethanol of 95% of 100 mcg/ml, ethyl acetate portion and n-butanol fraction present increase by 130.45 ± 1.51,123.11 ± 3.28 respectively, and the mineralization activity of 146.73 ± 2.85% (Fig. 2).
Example 2 guides separation andpreconcentration reactive compound from rabbit tail grass is biological
This active acetate ethyl ester and n-butanol fraction are imported into various col-umn chromatography and high performance liquid chroma-tography purification, and produce new compound (3), and 10 known compounds, 2 ', 4 ', 5, 7-tetrahydroxy isoflavanone (dalbergioidin) (1) (Durango, Quinones, Torres, Rosero, Gil & Echeverri, 2002), salicylic acid (2) (Scott, 1972), vitexin (vitexin) (4) (Krafczyk & Glomb, 2008), tryptophan (tryptophan) (5) ((Lee & Phillips, 1992)), maltol-3-O-β-D-pyranglucoside (maltol-3-O-β-D-glucopyranoside) (6) (Ono, Masuoka, Tanaka, Ito & Nohara, 2001), gland nucleoside (adenosine) (7) (Moyroud & Strazewski, 1999), spatholosinesideA (8) (Yin, Liu, Wang, Tu, Liang & Zhao, 2008), byzantionoside B (9) (Matsunami, Otsuka & Takeda, 2010), (7R, 8R)-threo-guaiacylglycerol-8-O-4 '-sinapylether7-O-β-D-pyranglucoside ((7R, 8R)-threo-guaiacylglycerol-8-O-4 '-sinapylether7-O-β-D-glucopyranoside) (10) (Machida, Sakamoto & Kikuchi, 2009) and thymus pyrimidine (thymine) (11) (Pouchert, 1993) (Fig. 3).These known compounds by the data corresponding with document or compare its spectroscopic data with authentic sample and identify (UV,
1h-NMR,
13c-NMR, ESI-MS).
New compound 3 is yellow powder.The molecular formula of 3 is by its front HRESI-MS M/Z:535.1459 [M+H]
+(be calculated as C
25h
27o
13 +, 535.1452) and be confirmed as C
25h
26o
13, nuclear magnetic resonance, NMR (NMR) data consistent (table 1) of this and it.Ultraviolet spectra is presented at the absorption band of λ max272 and 329nm, and NMR spectrum display δ
h6.75 (1H, s, H-3) and δ
cthe signal of 102.8 (C-3), 163.5 (C-2) and 181.8 (C-4), this represents the existence of flavone group.At A
2x
2in Coupling System, at δ
hthe signal of 6.91 (2H, d, J=8.8Hz, H-3 ', H-5 ') and 7.92 (2H, d, J=8.8Hz, H-2 ', H-6 '), and δ in NMR spectrum
c116.0 (C-3 ', and C-5 ' ") and 128.5 (C-2 ', there is mono-substituted aromatic portion (ring B) in the existence suggestion 3 of C-6 ' ") signal.At relevant (the Heteronuclear MultipleBond Correlation of heteronuclear multiple-bond; HMBC) spectrum, due to respectively with C-6,
, long-chain even summation aromatic signals of C-9, δ
h6.51 (1H, s, H-8) are specified in H-8.These data display 3 has the skeleton (5,7,4 '-trihydroxyflavone (apigenin) skeleton) (Fig. 4) of 5,7,4-trihydroxyflavone (apigenin).NMR spectrum also shows two Sugar signal δ
h2.52 to 5.25 and δ
c64.5 to 109.1.After the acid hydrolysis of 3, water layer is separated by HPLC, and compares with standard substance, only obtain a glucosides: apiose (apiose).Mat NMR under the existence of other sugar, HSQC, COSY and HMBC spectrum is determined.Above-mentioned evidence is pointed out, 3 is the apigenins (apigenin) replaced through xylose and apiose (apiose) group.The 6-C-glycosylation of apigenin and xylose is at field domain (the field) (δ of comparatively luteolin (leutolin)
c100.1) low about 8ppm and being identified, and C-1 " signal in NMR analyzes near δ
cthere are (Dubois, Zoll, Bouillant & Chopin, 1982) in 72.1 places.α-the configuration of D-xyloside and the beta configuration of D-apioside (D-apioside) can be derived by the coupling constant of anomeric proton (anomeric proton) H-1 " with H-1 ' ".Position (C-1 ' " → C-2 ") the mat HMBC that β-D-furan apioside (β-D-apiofuranoside) is connected with α-D-xylopyranoside (α-D-xylopyranoside) relevant (HMBC correlation) is being positioned at δ
cthe C-1 ' of 198.1 " and be positioned at δ
hthe H-2 of 4.27 (1H, brs) " and be identified (Fig. 4).Based on above-mentioned data, the structure of 3 is confirmed as apigenin 6-C-(β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside.
Table 1 compound 3
1h and
13nMR data (the DMSO-d of C
6, 500 and 125Hz).
Example 3 is separated from the compound of rabbit tail grass the survival of HOb cell
HOb cell cultivates 72 hours to be separated from the compound of rabbit tail grass, and adopts mtt assay to detect cell survival.Result display only have compound (5) 100 μMs with 80.43 ± 1.11% value slightly T suppression cell survival.Its result as shown in Figure 5.
Example 4 is separated from the compound of rabbit tail grass active to the ALP of HOb cell
In order to study the compound of separation from rabbit tail grass to the impact of osteoblast differentiation, the alkali phosphatase (ALP) measuring HOb cell is active.Plant component, daidzein (daidzein), as positive controls.In these compounds, 4 compounds, dalbergioidin (1), apigenin 6-C-(β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside (3), gland nucleoside (7) and byzantionoside B (9), be found in HOb cell, and in 100 μMs time, ALP activity significantly increases by 114.10 ± 4.41 respectively, 109.68 ± 2.20,108.70 ± 4.14 and 114.81 ± 0.18% (Fig. 6).
Example 5 is separated from the compound of rabbit tail grass the mineralising of HOb cell
Alizarin red staining, be widely used in and detect the existence of calcium in cell, for studying the compound of separation from rabbit tail grass in the effect (in tested compounds, the existence of ascorbic acid and β-phosphoglycerol under carry out) of the cultivation HOb cell of 12 days to bone mineralising.Result display noval chemical compound 3, dalbergioidin (1), gland nucleoside (7) and byzantionoside B (9) significantly increase the formation of mineralising bone tuberosity in HOb cell at 100 μMs, data are respectively 150.10 ± 0.80%, 107.09 ± 2.80%, 125.21 ± 3.75% and 129.21 ± 6.13% (Fig. 7).These results display noval chemical compound 3 presents the most effective mineralization activity of human osteoblast.
The neuroprotective of example 6 rabbit tail grass extract
NGF (nerve growth factor) PC 12 cells Induced by differentiation detects and is used to nerve injury test model, and 6-hydroxy dopamine (6-hydroxydopamine) is used to inducing neural damage.As previously mentioned and preparation rabbit tail grass roots extract be used to detect.Just find that acetic acid ethyl ester extract and n-butyl alcohol extract have excellent neuroprotective.Cell survival rate through the PC12 cell of 6-hydroxy dopamine damage is increased to 62.88+/-1.10% (cell with acetic acid ethyl ester extract process) and 61.54+/-1.48% (cell of n-butyl alcohol extract process) from 0.55%.
Claims (13)
1. rabbit tail grass belong to or its part or its extract for the manufacture of promoting osteogenesis or treating and/or preventing osteopathia or provide the purposes of medicament of neuroprotective.
2. purposes according to claim 1, wherein said rabbit tail grass belongs to the group that system is selected from following composition: rabbit tail grass (Urariacrinita (L.) Desv.ex DC.), great Ye rabbit tail grass (Uraria lagopodioides (L.) Desv.ex DC.), leatherleaf rabbit tail grass (Uraria picta (Jacq.) Desv.ex DC.), roundleaf rabbit tail grass (Uraria neglecta Prain), Uraria acaulis Schindl, Uraria acuminata Kurz, Uraria balansae Schindl., Urariabarbata Lace, Uraria campanulata (Benth.) Gagnep., Uraria candida Backer, Urariaclarkei Gagnep., Uraria cochinchinensis Schindl., Uraria cordifolia Wall., Uraria crinita (L.) DC., Uraria cylindracea Benth., Uraria fujianensis Y.C.Yang & P.H.Huang, Uraria gossweileri Baker f., Uraria kurzii Schindl., Uraria laceiCraib, Uraria lagopodoides (L.) DC., Uraria lagopus DC., Uraria longibracteataY.C.Yang & P.H.Huang, Uraria pierrei Schindl., Uraria poilanei Phon, Urariaprunellifolia Baker, Uraria rotundata Craib, Uraria rufescens (DC.) Schindl. and Uraria sinensis (Hemsl.) Franch.
3. purposes according to claim 1, wherein said rabbit tail grass belongs to for rabbit tail grass (Uraria crinita (L.) Desv.ex DC.), great Ye rabbit tail grass (Uraria lagopodioides (L.) Desv.ex DC.), leatherleaf rabbit tail grass (Urariapicta (Jacq.) Desv.ex DC.) or roundleaf rabbit tail grass (Uraria neglecta Prain).
4. purposes according to claim 1, the part that wherein said rabbit tail grass belongs to is the root that rabbit tail grass belongs to.
5. purposes according to claim 1, it is ethanolic extract that wherein said rabbit tail grass belongs to extract.
6. purposes according to claim 1, wherein said rabbit tail grass belongs to extraction system and is obtained by the following step: belong to alcohol extraction rabbit tail grass and obtain ethanolic extract, and obtain acetic acid ethyl ester extract or butanol extract with ethyl acetate or this ethanolic extract of butanol, before immunoassay.
7. purposes according to claim 1, it is the acetic acid ethyl ester extract or the butanol extract that derive from rabbit tail grass genus ethanolic extract that wherein said rabbit tail grass belongs to extract.
8., according to the purposes of claim 1, wherein said rabbit tail grass belongs to the acetic acid ethyl ester extract or butanol extract that extract is the ethanolic extract deriving from the root that rabbit tail grass belongs to.
9. purposes according to claim 8, wherein said acetic acid ethyl ester extract contains apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside.
10. purposes according to claim 1, wherein said osteopathia is osteoporosis, arthritis, osteoarthritis, periodontal disease, alveolus bone-loss, osteotomy bone-loss, Childhood idiopathic bone-loss, curvature spinal column, loss, osteoarthritis or the osteolytic lesion of height.
11. purposes according to claim 1, wherein said neuroprotective is apoplexy or brain or nervous system injury/wound, anoxia, spinal cord injury, peripheral nerve injury, parkinson, A Zihai Mo's disease or multiple sclerosis.
12. 1 kinds of formula (I) compounds,
Wherein R
1it is furan celery glycosyl-α-L-arabopyranose base (apiofuranosyl-α-L-arabinopyranosyl), furan celery glycosyl-β-D-Glucose pyranose (apiofuranosyl-β-D-glucopyranosyl), furan celery glycosyl-β-D-galactopyranosyl glycosyl (apiofuranosyl-β-D-galactopyranosyl), furan celery glycosyl-β-D-xylopyranosyl (apiofuranosyl-β-D-xylopyranosyl), furan celery glycosyl-β-D-arabopyranose base (apiofuranosyl-β-D-arabinopyranosyl), furan celery glycosyl-β-D-ribopyranose base (apiofuranosyl-β-D-ribopyranosyl), furan celery glycosyl-β-D-pyrans lysol glycosyl (apiofuranosyl-β-D-lyxopyranosyl), furan celery glycosyl-β-D-pyrans ribulose base (apiofuranosyl-β-D-ribulopyranosyl), furan celery glycosyl-β-D-xylopyranose ketose (apiofuranosyl-β-D-xylulopyranosyl), furan celery glycosyl-β-D-A Luo pyrans glycosyl (apiofuranosyl-β-D-allopyranosyl), furan celery glycosyl-β-D-pyrans altrose base (apiofuranosyl-β-D-altropyranosyl), furan celery glycosyl-β-D-mannopyranose base (apiofuranosyl-β-D-mannopyranosyl), furan celery glycosyl-β-D-pyrans gulose base (apiofuranosyl-β-D-gulopyranosyl), furan celery glycosyl-β-D-pyrans idose base (apiofuranosyl-β-D-idopyranosyl), furan celery glycosyl-β-D-pyrans talose base (apiofuranosyl-β-D-talopyranosyl), furan celery glycosyl-α-L-glucopyranosyl (apiofuranosyl-α-L-glucopyranosyl), furan celery glycosyl-α-L-galactopyranosyl glycosyl (apiofuranosyl-α-L-galactopyranosyl), furan celery glycosyl-α-L-xylopyranosyl (apiofuranosyl-α-L-xylopyranosyl), furan celery glycosyl-α-L-arabopyranose base (apiofuranosyl-α-L-arabinopyranosyl), furan celery glycosyl-α-L-ribopyranose base (apiofuranosyl-α-L-ribopyranosyl), furan celery glycosyl-α-L-pyrans lysol glycosyl (apiofuranosyl-α-L-lyxopyranosyl), furan celery glycosyl-α-L-pyrans ribulose base (apiofuranosyl-α-L-ribulopyranosyl), furan celery glycosyl-α-L-xylopyranose ketose base (apiofuranosyl-α-L-xylulopyranosyl), furan celery glycosyl-α-L-A Luo pyrans glycosyl (apiofuranosyl-α-L-allopyranosyl), furan celery glycosyl-α-L-pyrans altrose base (apiofuranosyl-α-L-altropyranosyl), furan celery glycosyl-α-L-mannopyranose base (apiofuranosyl-α-L-mannopyranosyl), furan celery glycosyl-α-L-pyrans gulose base (apiofuranosyl-α-L-gulopyranosyl), furan celery glycosyl-α-L-Chinese mugwort Du's pyrans glycosyl (apiofuranosyl-α-L-idopyranosyl) or furan celery glycosyl-α-L-pyrans talose base (apiofuranosyl-α-L-talopyranosyl).
13. compounds according to claim 12, it is apigenin 6-C-β-D-furan celery glycosyl (1 → 2)-α-D-xylopyranoside of tool following formula:
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107456476A (en) * | 2016-06-03 | 2017-12-12 | 崇仁医护管理专科学校 | Lift active component content, medicinal material chemical composition composition uniformity and/or method and the application for the treatment of benefit of rabbit tail grass platymiscium |
| TWI608841B (en) * | 2016-06-03 | 2017-12-21 | Chung Jen Junior College Of Nursing Health Sciences And Management | Method for improving the content of active ingredients, consistency of chemical compositions, and/or therapeutic benefits of medicinal plants of the genus Martinia and its products and applications |
| CN108299367A (en) * | 2018-01-26 | 2018-07-20 | 南阳师范学院 | A kind of celery aglycon carbamate compound, preparation method and application |
| CN111372586A (en) * | 2017-11-20 | 2020-07-03 | 诺麦塔制药有限公司 | Composition for preventing, ameliorating or treating bone loss disease comprising CHP |
| CN116064767A (en) * | 2022-08-23 | 2023-05-05 | 南京医科大学 | LncRNA marker related to osteoarthritis and application thereof |
-
2014
- 2014-04-09 CN CN201410140673.7A patent/CN104971086B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| 赵秀贞: "《青草药彩色图谱》", 30 April 1997, 福建科学技术出版社 * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107456476A (en) * | 2016-06-03 | 2017-12-12 | 崇仁医护管理专科学校 | Lift active component content, medicinal material chemical composition composition uniformity and/or method and the application for the treatment of benefit of rabbit tail grass platymiscium |
| TWI608841B (en) * | 2016-06-03 | 2017-12-21 | Chung Jen Junior College Of Nursing Health Sciences And Management | Method for improving the content of active ingredients, consistency of chemical compositions, and/or therapeutic benefits of medicinal plants of the genus Martinia and its products and applications |
| CN111372586A (en) * | 2017-11-20 | 2020-07-03 | 诺麦塔制药有限公司 | Composition for preventing, ameliorating or treating bone loss disease comprising CHP |
| CN111372586B (en) * | 2017-11-20 | 2023-07-25 | 诺麦塔制药有限公司 | Composition for preventing, improving or treating bone loss diseases comprising CHP |
| CN108299367A (en) * | 2018-01-26 | 2018-07-20 | 南阳师范学院 | A kind of celery aglycon carbamate compound, preparation method and application |
| CN108299367B (en) * | 2018-01-26 | 2022-04-12 | 南阳师范学院 | Celery aglycone carbamate compound, preparation method and application thereof |
| CN116064767A (en) * | 2022-08-23 | 2023-05-05 | 南京医科大学 | LncRNA marker related to osteoarthritis and application thereof |
| CN116064767B (en) * | 2022-08-23 | 2024-06-04 | 南京医科大学 | LncRNA marker related to osteoarthritis and application thereof |
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