Nghien CLFU - Ky thuat
thy tuyin vii ngydi MCF-7 trong thy nghiem invitro (Kristikarlos&CS,1989)''''-
K§t luan
- TO' can chiit ethylacetat cua djch len men ehung nim Trichoderma konilangbra bing phyong phap sic ki lap lai nhieu lin tren cdt silicagel vdi he dung mdi thieh hgp thu dygc cae hgp chit Bing cac phyong phap nhy pho hdng ngoai, pho NMR, HMQC va HMBC xae dinh eau true hda hpe eua hgp ehit KS14-5 la ergosterol peroxid.
- Kit qua hoat tinh khang VSVKD cho thiy chit nay cd hoat tinh khang 7/8 chung kiem djnh vdi gia trj MIC tyong ifng la lOOjig /ml.
- Hgp ehit nay bieu hien hoat tinh dde t i bao manh vdi ca 3 ddng te bao thy la ung thy gan, ung thy mang ty eung va ung thy mang tim vdi gia trj IC50 tyong irng la 6,25;7,5 va 6,8.
Summary
By combination of various chromatographic techniques some compounds were isolated from etylaxetate extracts of the endophytic fungus Trichoderma konilangbra. Of these, one was identified as ergosterol peroxide, and showed the strong activities against bacteria, yeasts and moulds. In addition, it also exhibited strong
cytotoxicity on three human cancer cell lines: Fl - Fibril sarcoma of Uteus ; RD - Rhabdosareoma;
and Hep- G2 - Hepatocellular carcinoma.
Tai lieu tham khao
1. Pham Thi Hong Minh, (2002), Lu$n an tien sT, Vien Hoa hoc, Vien Khoa hoc va Cong nghe Viet Nam, Ha Noi.
2.. Bamathan, G. et al.( 1998), Phytochemistry 47, pp.761-765.
3. Goad L.J., Akihisa T.(1997)," Analysis of sterol", Blackie academy and Professional, Chapman and Hall.
4 Kahlos Kirsti, Kangas L., Hiltunen R. (1989),
"Ergosterol Peroxide, an active compound from Inonotus radiatus" Planta Medlca 55, pp.389-390.
5. Krzyczkowski W., Malinowska E., Suchocki P., Kleps J., Herold P., Olejnik M, and Turlo J. (2007),
"Isolation of ergosterol peroxide from the mycelium of Hericlum Erinaceum (bull. Ex. Fr.) Pers.", Herbal Po/in/ca 53(2), pp.181.
6. Likhitayawuid K., Angerhofer CK. (1993), 'Cytotoxic and antimalarial bisbenzylisoquinoline alkaloids from Sephania evecta", Jounal of natural products 56 0). PP- 30-38.
70hta, S.etal. (1981), Chem.Pharni.Bull29, pp. 2762 8. Vanden Bergher D. A., Vlietlink A. J. (1991), "
Screening methods for antibacterial and antiviral agents from hight plants". Methods in Plant Biochemistry 6, pp. 47-68
Nghien ciru che tao va danh gia mot so dac tinh cua he nano piroxicam
Nguyen Thi Mai Anh, Nguyen Van Long, Nguyen Trutmg Son Truang dgi hoc Duac Hd Noi
Dat van de
Cdng nghe nano ra dd'i v^o cuoi nhifng nam 60 cua the ky 20 va da tirng byde irng dung trong ITnh vyc dygc pham. Cdng nghe nay gdp phin tao ra mpt sd dang bao chi hien dai vdi nhifng yu diem vygt trpi ve kha nang hip thu.
Oac biet nd edn cd y nghTa trong viee md ra hydng phat then mdi cho eac thude tac dung tai dich '^' ^\ Tuy nhien, hieu qua dieu trj eua eae dang thudc bao chi tif nguyen lieu nano phu
thudc ho^n toSn v^o kich thud'c vi cau true eua c^e tieu phan trong he. Piroxicam thudc nhdm giam dau chdng viem khdng sterpid, tuy cd nhiiu yu diem trong diiu trj nhirng do dae tinh it tan trong nyde va thim kem nen hieu qua dieu tri cua cae thudc dung ngoai cdn han che. De lam tang hap thu piroxicam qua da, niem mac vi gdp phin dya cdng nghe nano v^o irng dung san xuit thudc trong nyd'C ehiing tdi thyc hien de ii\:
Nghien cifu ehe tao vi danh gia mdt so dac tfnh cua he nano piroxicam.
50 TAP CHi DU'QC HQC - 8/2009 (SO 400 NAM 49)
Nghien CLFU - Ky thuat
Nguyen lieu, thiet bj va phu'O'ng phap
Nguyen lieu:
piroxicam (Trung Quoc), tieu chuan USP 24, Eudragid (S 100, RL 100, RS 100) va aleol polyvinie (Diyc), tieu chuan nha san xuit. Mdt so dung mdi, hoa chit tinh khiet hda hpe va diing cho HPLC.
Thiit bj:
May khuiy Braul 4185 (See), may ddng nhit hda Homogenizer FBF 101 (Italia), may khuiy tif IKA-WERKE (Diyc), may ly tam Sigma 3-18 K Satorius (Difc), kinh hien vi dien tCf quet FESEM Hitachi S-4800 (Nhat), kfnh hien vi dien ty truyin qua JEM 1010 (JOEL), may xae dinh phan bo kich thyde hat HORIBA LA-950, may sieu am Ultrasonic LC 60 H, may do quang pho UV-VIS U1800 HITACHI (Nhat), may HPLC Spectra System Thermo, may do t h i Zeta Phoremeter IV- CAD, may phan tich nhiet vi sai SETARAM DSC 131 (Australia), may do quang pho hong ngoai RKIN ELME. Mdt SO thiet bi b^o che, kiem nghiem khac.
Phu'O'ng phap nghien cu'u
Che tao he nano piroxicam bang phwang phap nhu hoa va boc hai dung moi ^^K
- Hda tan piroxicam va Eudragit trong dicloromethan, phoi hgp vdi dung djch alcol polyvinie (PVA) ndng dp tif 0,5 den 5,0% d nhiet dp 5-10°C. Khuiy trpn tde dp eao de tao nhu tyong, dong nhit hda nhtJ tyong b l n g may dong nhit hda.
- Loai dicloromethan b i n g each khuiy trdn lien tuc d nhiet dp phdng trong 24 gid'.
- Loai nyde bing each ly tam d 14.000 vdng/phiiL Rya tua 3 l i n vdi nyde c i t , sau dd, de khd d nhiet dp phdng trong 48 gid'.
Danh gia mgt so dac tinh cua he nano Hinh dang, ciu triic, kich thw&c vi bi mat tiiu phan (KTTP):
Sy dung kinh hien vi dien t y truyen qua (TEM) vdi eac thdng so: dp phdng dai M = 50x- 600000X, dp phan giai 5=3A°, dien ap gia tdc U=
40-100 kV vi kinh hien vi dien t y quet (SEM) vdi dp phdng dai M=20x-800000x; dp phan giai 5=1,Onm; dien ap gia tde U = 0,5-30 kV.
Kich thw&c vi phin bd kich thw&c:
Kic djnh bing phyong phap tan xa laser (LS).
(may Horiba LA-95). D i i u kien do: he sd tan xa anh sang \i 1,390.
The zeta
Sif dung phyong phap dien di (EP) trong dieu kien nhiet dp: 25 ± 2°C, dp d i n dien: 0,017 mS/cm, pH: 7.
Dinh gii thay ddi tinh chit vat ly
Phygng phap phan tieh nhiet vi sai (DSO).
Nhiet dp quet tif 20°O - 220°C, vdi toe dp 20°C/phut
Djnh Iwg'ng dwac chit
Dung phyong phap HPLC vdi cdt 018, 25cmx4,6em, hat nhoi kich thyde 5pm, pha ddng la hdn hgp methanol va dem (7,72 gam acid citric khan va 5,35 gam dinatri hydro phosphat khan trong 1000 ml nyde) vdi ty le 55:
45. Toe dp ddng 1,2 ml/phut, detector UV byde sdng 254 nm, nhiet dp cot 25°0 ± 1, the tich tiem 25pl.
K§t qua va ban luan
Khao sat anh hu'o'ng cua ta du'O'c d i n qua trinh c h i tao he nano
Cae m i u dygc e h i tao theo phyong phap trinh bay d mue 2.4.1, sy dung eae ehit mang khac nhau la Eudragit S, RL100, RS 100 (vd'i cac ty le dygc ehit : c h i t mang la 1:2,5, 1: 5, 1: 10), dung djch PVA d y g c diing vd'i eac nong dp 0,5- 1-3-5%. Sau khi ehe tao, eae m i u d y g c danh gia hinh dang, c i u triic, kich thyd'c hat b i n g phyong phap trinh bay d muc 2.4.2. Phan tieh hinh anh tieu phan eho thay khi tang ndng dp dung dieh PVA trong qua trinh tao nhu tygng, trong san pham cuoi cung cac hat hlnh thanh ed hinh c i u deu dan hon, b i mat nhin mjn hon va polyme bao deu hat d y g c chit, dae biet la kieh thyd'c hat nhd hon (bieu do hinh 1). Dieu nay cd the giai thich la do nong dp ehit dien hoat tang, kha nang nhu hda tang len, cac gipt c h i t long trong nhu tyong nhd di, he on djnh hon trong qua trinh bay hoi pha d i u de tao hat
Hinh 1. Biiu dd so sinh kich thw&c tiiu phin trong cic mau v&i ndng dd PVA khic nhau Ddi vdi ehit mang polyme, trong so cae Eudragit da khao sat, eae m i u sau khi ehe tao d i u cd dang hinh c i u , be mat nhin, polyme bao ngoai
TAP CHI DU'QC HQC - 8/2009 (SO 400 NAM 49) 51
Nghien CLFU - Ky thuat
dygc ehit nhyng ty le d y g c ehit (DC) va c h i t mang (CM) anh hyd'ng nhieu d i n c i u triic va kich thyde hat trong he (bang 1).
Bang 1 : Anh hw&ng ciia ty le polyme din hlnh dang, ciu triic va kich thw&c hat nano
Miu
Piroxieam:Eud RS100 = 1:2,5
(PERS2,5) Piroxicam:Eud
RS 1:5 (PERS5) Piroxieam:Eud
RS1:10 (PERS10)
Hinh thai, c i u true, kich thu'd'e trung binh hat (KTTB) Hinh e i u , b i mat nhin, it tinh the. KTTB 1000,75 nm, 49,8%
hat dyd'i lOOOnm
Hinh e i u d i u , b i mat n h i n , khdng cd tinh the, it ed dam hat k i t tu vdi nhau, KTTB 725,6 nm, 78,6% hat dyd'i 10OOnm Hinh c i u , be mat nhin, ed dam polyme k i t dinh, KTTB 573,25 nm, 79,2% hat dyd'i lOOOnm, nhiiu hat polymer t y do
Khi lygng polyme thap (DC:CM=1:2,5), kich thyde hat khdng dong d i u , polyme khdng bao d i u ngoai dygc chit. Khi ty le polyme qua eao (DC:CM=1:10), trong m i u ton tai mdt ty le Idn eae hat polyme t y do vdi kich thyd'c thay doi lam anh hydng d i n kich thyd'c trung binh tieu phan va phan bd kieh thyd'c cua ea he. Hon nira cac hat polyme ty do nay edn anh hydng den phan bo dygc chit trong m i u va khd dam bao ham lygng dygc ehit khi dya vao dang bao c h i . Ty le DC va CM la 1:5 tao ra d y g c san pham ed nhieu yu diem hon ea ve hinh dang, c i u true va kich thyde hat.
Trong 3 polyme da sy dung de e h i tao he nano thi Eudragit RS 100 tao hat tot hon so vd'i hai polyme cdn lai va qua trinh c h i tao d i dang hon. Vi vay, he gdm Piroxicam va Eudragit RS 100 ty le 1:
5 dygc sy dung cho cae nghien eiru tiip theo.
Danh gia mot s6 dac tinh cua he nano Hinh dang, kich thwac tieu phan
Chon cic mau: Piroxicam nguyen lieu, m i u tring (Eudragit RS 100); M i u chirng (Px-DC); he nano Piroxicam:Eudragit RS 100 1:5 de so sanh va danh gia.
; ^ r , .
" * l ! m ' l ' " H I
(c) (d) Hinh 2: Anh SEM cic tiiu phin trong cic miu dinh gii
(a) Piroxicam: Eudragit RS 1:5; (b) Mau tring (Eudragit RS 100); (c) Miu chirng (Px-DC);
(d) Piroxicam nguyen lieu (Px-NL)
Phan tieh hinh anh SEM (hinh 2) nhan thay m i u piroxicam: Eudragit RS 1:5 va m i u tring Eudragit RSI00 hat cd hinh c i u , d i u , be mat nhin va da so hat cd kich thyde nhd hon 1000 nm. M i u Piroxicam nguyen lieu (Px-NL) hat hinh khoi chQ' nhat, kich thyde Id'n (khoang 10 pm).
M i u ehirng (Px-DC) ed hat hinh oval, khdng cd gdc canh, be mat khdng n h i n , kich thyde khoang 1000 nm. Mat khae, qua hinh anh TEM eua m i u piroxicam:Eudragit RS 1:5 (Hlnh 3) ed the thiy rd da sd cac tieu phan ed kieh thyde nho hon 1000 nm, hinh trdn deu dan c i u triic gdm hai phin: vd ngoai sang mau va p h i n nhan s i m mau. Nhy vay sau qua trinh ehe tao, piroxicam k i t hgp vd'i polyme tao ra eae tieu phan ed eiu triic polyme bao quanh d y g c ehit.
•Jti
%
Hinh 3: Anh TEM ciia tliu phin trong miu Piroxicam: Eudragit RS 1:5
K i t qua nay tygng t y vd'i nghien ciru eua Khosro Adibkia v i he nano piroxicam vdi Eudragit RSI00'"'.
52 TAP CHi DU'QC HQC - 8/2009 (SO 400 NAM 49)
Nghien
CLFU- Ky thuat
The zeta cua tieu phan nano
Cic m i u sau khi ehe tao d y g c phan tan trong dung dich ding tryong pH 7,4 cd chira 0,5%
HPMC E606, do the zeta theo phyong phap trinh bay d mue Danh gii mot sd die tinh ciia he nano, dygc k i t qua ghi trong bang 2.
Bang 2: Thi zeta ciia tiiu phan nano
Mlu UmV)
EuRS-DC -42,31
Px-DC -0,88
PERS5 -39,12
D i dang nhan thiy ring n i u chi ed mdt minh piroxicam thi sau qua trinh d i i u che the zeta cua cac tieu phan r i t nhd edn khi che tao vdi Eudragit RS 100 tao ra nhO'ng hat cd c i u true polyme bao ngoai DC, t h i zeta tang len nhiiu (-39,12mV).
Dac tinh nay se khiin he nano on djnh hon vi dien the be mat ngan can sy k i t tu eua nhtfng tieu phSn. K i t qua nSy tyong t y nhy k i t qua nghien Khosro Adibkia v i he nano piroxicam vdi Eudragit (the zeta cua he nano trong khoang 35- 42 mV) '^'. va phu hgp vdi quan diem cua mpt so tae gia khae eho ring tieu phan nano vdi mdt the zeta tren (+/-) 30 mV coi nhy on djnh trong hdn djeh '"1.
Mgt so dac diem khac
M i u piroxicam: Eudragit = 1:5 d y g c dem phan tich nhiet vi sai, pho DSC eua m i u so vdi nguyen lieu khdng thiy x u i t hien pie thu nhiet (Hinh 4).
11
\'i
Hinh 4: Phd phan tich nhiet vi sai
Nhy v$y, sau khi ehe tao giira piorxieam va Eudragit ed tyong tae lam thay doi tinh ehit vat ly eua he. Tuy nhiSn giu'a ehiing khdng ed tygng tae hda hpc, diiu n^y the hien tren pho hdng ngoai eua m i u phan tich, pho nay ed djnh dang gidng het pho hong ngoai cua nguyen lieu, khdng thiy x u i t hien c^e pie la eua nhdm chirc md'i. Hon nifa, s i c ky do cua m i u khi dinh lygng bing HPLC
cung khdng thay x u i t hien eae pie la so vd'i s i e ky do eua nguyen lieu. Nhan djnh nay tyong t y vd'i k i t qua eua Ram N. Prajapati va cpng sy khi nghien ciru ve dendrimer piroxicam '*'.
K§t luan
Tif eac k i t qua thye nhiem d tren, nhdm nghien eiru da lya chpn Eudragit RS 100 la ta d y g c cd nhiiu y u diem hon ea de e h i tao he nano piroxicam vdi ty le DC:CM la 1:5. He nano sau khi d i i u e h i cd kich thyd'c tieu phan giam h i n so vdi nguyen lieu ban d i u , khoang 70 % hat nhd hon 1000 nm. Cae hat ed c i u triie polyme bao ngoai d y g c chit, b i mat n h i n min va the zeta khoang -39 mV. GiO'a DC va CM cd tyong tae vat ly nhyng khdng t h i y ed tyong tae hda hpe.
Summary
The nanoparticles of piroxicam was prepared by solvent evaporation/extraction technique (the also called single emulsion technique) using ratio 1:5 of piroxicam: Eudragit RS 100. The nano particles were characterized by SEM, TEM, DSC, EP and LS... The examinations resulted in spherical shape with smooth surface for the drug and polymer nanoparticles, about 70% particles are less than 1000 nm. Zeta potential is about -39 m V.
Tai lieu tham khao
1. Haririshna Devalapally (2007), "Role of nanotechnology in pharmaceutical product development", J. Pharm. Sci.. 96, p.2547-2565.
2. James Swarbrick (2006), "Nanoparticle technology for drug delivery". Taylor & Francis Group 270 IVIadison Avenue New York. NY 10016, p.1-197.
3. Khosro Adibkia et al (2007), "Piroxicam nanoparticles for ocular delivery; Physieoehemieal charaeterization and implementation in endotoxin- induced uvitis", J. Drug Target., 15, 6, p.407-416,
4. Mohanraj VJ and Y Chen (2006), "Nanoparticles - A review", Trop., J. Phann. Res.5,1, p 561 - 573.
5. Ram N. Prajapati et al (2009), "Dendimer- mediated solubilization, formulation development and in vitro, in vivo assessment of piroxicam", Mol.
Pharm..,3, p. 9 4 0 - 9 5 0 .
TAP CHI DUQC HQC - 8/2009 (SO 400 NAM 49) 53
