| Epimedium sagittatum(Sieb.et Zucc.)Maxim is a plant of the genus Epimedium in the Berberidaceae family.It is recorded in the Chinese Pharmacopoeia as one of the varieties of Epimedium.The main functions of E.sagittatum include tonifying kidney Yang,strengthening muscles and bones,and eliminating rheumatism.It can be used for the treatment of kidney Yang deficiency,impotence and spermatorrhea,weakness of muscles and bones,rheumatism,arthralgia,numbness and contracture.Isoprenyl substituted flavonoids are the main chemical components of E.sagittatum.These compounds exhibit a wide range of biological activities,including promoting cartilage repair,anti-aging,anti-fatigue,anti-tussive and anti-asthmatic,hypoglycemic,anti-tumor,etc.Most flavonoid glycosides from E.sagittatum shared a common aglycone,icaritin,which has been approved as a new drug for the treatment of liver cancer.In order to obtain isoprenyl flavonoids with good biological activity and novel structure,the chemical constituents of E.sagittatum were systematically studied.The ethanol extract of E.sagittatum was separated by silica gel column chromatography,Sephadex LH-20 column chromatography,C18 reverse-phased silica gel column chromatography,MCI column chromatography and semi preparative HPLC.20 compounds were obtained and the structures were identified by NMR and MS,Here includes a new compound,a flavonoid glycoside compound,named as sagittasine D(1).The known compounds include 12 flavonoid glycosides:3"-O-demethylsapinorhamnoside(2),sagittasine A(3),kaempferide 3-O-neohesperidoside(4),icarisid N(5),epimedin C(6),icariin(7),baohuaoside I(8),quercetin 3-O-glucoside(9),ikarisoside A(10),hexandraside E(11),icariside(12),and epimedoside C(13);five flavonoids:yingyanghuo A(14),apigenin(15),quercetin(16),β-anhydroicaritin(17)and icaritin(18);one simple aromatic compounds:anisic acid(19);and a steroid compound:β-sitosterol(20).Flavonoids are the main pharmacological components of Epimedium,among which native glycosides such as icariin and epimedin C account for about 52%of the total flavonoids in Epimedium.These native glycosides are poorly absorbed in the small intestine.However,their hydrolysates such as baohuoside I and sagittatoside A,as well as icaritin(aglycone),have better lipid solubility,can be better absorbed by the human intestinal tract,and exhibit better biological activity.For example,baohuoside I and icaritin have better effects than icariin in inhibiting tumor proliferation,but their content is lower.In this paper,the preparation of icariin and baohuoside I by enzymatic hydrolysis of icariin was studied.Icariin was prepared by hydrolysis of icariin with snail enzyme and baohuoside I was prepared by hydrolysis of icariin with β-glucosidase.Using the conversion rate as an index,the effects of pH,temperature,enzyme dosage,reaction time,and other factors on the conversion rate were investigated through single factor analysis to optimize the preparation conditions.The results showed that the optimized conditions for snail enzyme conversion of icariin were as follows:snail enzyme dosage of 15 mg(per 10 mg of icariin),enzymatic hydrolysis time of 60 h,buffer pH of 6.0,temperature of 37℃.The optimized conditions for the conversion of icariin by glucosidase are as follows:the dosage of β-glucosidase is 60 μL(6000 U/mL),enzymatic hydrolysis time is 6 hours,buffer pH is 6.0,and temperature is 55℃.Morus alba L.is a plant of the Moraceae family.The functional indications recorded in the Chinese Pharmacopoeia are purging the lungs,relieving asthma,and eliminating water and swelling.For lung heat,asthma,cough,edema,fullness,urine loss,and puffiness of the face and skin.Isoprenyl substituted flavonoids and Diels-Alder type adducts are the main components of M.alba L.In order to obtain isopentenyl substituted compounds with diverse structures,we isolated the components of the root bark of M.alba L.We have systematically isolated and identified the chemical components of the root of M.alba L.,and a total of 20 compounds have been isolated.Among them,there is a new compound:leachianone J(S1).In addition,19 known compounds were isolated from the root bark of Morus alba L.The known compounds include four Diels-Alder adducts:cathaynon B(S2),cathaynin B(S3),sanggenon C(S4)and sanggenon D(S5);6 flavonoids:sanggenol G(S6),sanggenon A(S7),cyclocommunol(S8),morachalcone A(S9),sanggenon H(S10)and 2-(3,5-dihydroxyphenyl)-7-hydroxy-4H-1-benzopran-4-one(S11);six 2-arylbenzofurans:moracin P(S12),isomoracin D(S13),moracin O(S14),moracin C(S15),moracin M(S16)and moracin D(S17);as well as three triterpenoids:lupeol(S18),betulinic acid(S19)and ursolic acid(S20).Four isoprenyl substituted compounds obtained with large quantities from E.sagittatum were selected to evaluate the proliferation inhibitory effect on pancreatic cancer cells by MTT method.Among them,compound 6 and 7 had extremely weak anti-proliferation effect.Compound 18 also showed extremely weak inhibitory effect due to poor solubility.Compound 8 showed moderate inhibitory effect on pancreatic cancer cell proliferation.Further evaluation showed that compound 8 also had proliferation inhibitory activity on lung cancer cells and could inhibit the migration of lung cancer cells.Four Diels-Alder adducts obtained from M alba were evaluated for their inhibitory effects on lung cancer cell proliferation.Taken together,40 compounds were obtained through systematic chemical study of the two traditional Chinese medicines,29 isoprenyl substituted compounds were included.Eight of them were screened for tumor cell proliferation inhibition effect,and it was found that compound 8 had a moderate proliferation inhibition activity on pancreatic cancer and lung cancer cells.This study provides a compound reserve and research basis for further research on the anti-tumor effects of isoprenyl substituted compounds. |
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