NGHIDN CQU - TRRO D6\
DIEU CHE DUtrC CHAT PHONG XA ''C-CHOLINE TAI TRUNG TAM MAY GIA TOC 30 MeV
vu THANH QUANG,
N G U Y I N K H A CTHAT,
N G U Y 6 N VANDINH, PHAM TUAN UNH, BUI THANH RIN, H 6 TRQNG TUNG
Trung t J m M^y gia tSc 3 0 MeV 3$nh v\$r\ Trung uong Quin Ml \0d
P-Hydroxyethyl tri ^'C methyl ammonium (^^C-choline) l i dQ0c chift phdng xg positron dung cho chgp hinh buTc xg positron (Positron Emission Tomography-PET) d l chin doin ung thi/tuylfn tiin not. Quy trinh dilu c h l diTdc chit ^^C-choline g6m cic giai dogn: sin xulft ^X-CO,, dilu ch^ "CH,!
va tdng h^p ^X-choline. Sin phim ^^C-choline dgt tidu chuin thu£»c ti§m.
Production of "C- Choline Radiopharmaceutical at 30 MeV Cyclotron Centre
p-Hydroxyethyl ammonium methyl tri ''C C'C-choline) is the
radiopharmaceutical used for positron emission tomography
(PET) imaging to diagnose prostate cancer. Production procedure of the ^^C-
choline includes:
Production of ^^C- COj, preparation of ''CH3I and ''C-choline
synthesis. The ^^C- choline final product is ready to injection.
I. Gioi thieu
Ung thu tuyi'n tiin Hit l i bOnh thudng gip v i dijfng thijf 2 giy ra XiS vong do ung thu cua dan dng trin 40 tudi d My. Theo Udc tinh, khoing 220.900 din dng My duoc ch^n doin ung thu tuyi'n tiin Hit din nim 2003 va 28.900 ngUdi se chit vi benh niy. Chinh vi viy, cic nha ung thu hpc dang rit cin phit trien cic cdng cu die hiiu d l phat hiin sdm v i dieu tri hiiu qui cin binh niy.
Choline l i thinh phan ri't quan trpng cua phospholipid trong ming te bao.
Choline v i choline kinaza dUdc tim thi'y d mijfc do cao trong cic t i bio ung thi/.
^^C-choline bj bai tie't qua nUdc tiiu la khdng ding k l , vi viy nd phu hdp cho djnh hinh tuyi'n tien Hit d l nghiin cijfu va chin doin ung thu tuyi'n tien Hit (ci u nguyin phat va di can h^ch bgch huyi't [1]). Dg nhgy, dd die hieu, dO chinh xic, gia trj di/
bao dUdng tinh, v i gia tri tiin lUdng im tinh cua ^'C-choiine-PET/CT la 66,81,71, 87, v i 55% tUdng ijfng. Hiin nay, dUdc chit phdng xg niy cd cic ufng dyng trong ky thuit PET d l dd tim ung tha bilu md t l bao gan (hepatocellular carcinoma HCC). Ngoii ra '^C- choline cdn d$c bi$t hiiu qui trong chgp hinh khdi u b trong phii, thi/c quin, tri/c tring, bing quang v i u nao [2,3]
Do thdi gian bin hijy cija hgt nhin carbon-11 chl l i 20 phut, nen dUdc chit nay khdng thi nhip khiu v i khdng thi vin chuyin, phin phit trin mdt khoing each hdn 1 gid XiS ndi sin xui't din ndi cd miy PET. Oe cd dUdc dUOc chat niy phyc vy cho cdng viic nghiin cijfu, chin doin binh thi lya chpn duy nhi't l i ti/ sin xua't.
'^C-choline dUdc dilu chl bdi phin iJfng cCia ''C-Methyl triflate [4] hoac "0- Methyl iodide [5,6] vdi 2-dimethylaminoethanol (DMAE), sau dd tich v i tinh chl bing phUdng phip chiit pha rin (SPE) dgt hi$u suit 60-85% tinh tren cd sd
"C-CO^, ting thdi gian ting hdp l i 15 phut tinh X\l khi ngCfng chii'u xg (EOB), do sgch hoa phdng xg 95-99%. Bii vii't trinh biy cic ki't qui ting h(?p "C-choline tif
"C-Methyl iodide tgi Trung tim Miy gia td'c 30 MeV (Binh viin Trung i/dng Quan ddi 108).
II. Thttc nghiem
1. Hda chit vd v$t ligu
LiAIH^ 0,1 M trong Tetrahydrofuran, HI 57%, 2-dimethylaminoethanol e" 99,5%, Ascarite XiS Aldrich, Accell plus CM XiS Water, Ethanol tuyet ddi, phosphoric acid
HPLC GRADE, naphthalene-2-sulfonic acid HPLC GRADE, choline chloride
84 K H O A H O C ^^*^^"9^ 2012
NGHieNCCrU-TRROOO!
HPLC GRADE, NaOH 99,9%, NaH2PO,.2H20 extra-pure, pharmaceutical grade v i Sicapent -Pp^ dehydrate Xis Merck. Column: Inertsil ODS-2 (6 x 250 mm, APS-Kit8 tif Bioscan).
2. Thiit bj
May gia td'c Cyclotron 30 MeV, bia nhom 50 cm^
hang IBA, Belgium.
- Mel-Plus, hang Bioscan, USA.
ReForm-Plus, hang Bioscan, USA.
Cac thii't bi kilm nghiim: HPLC hang Agilent, German; Gamma counter MCA hang Canberra, pH met...
3. PhUdng phdp thtfc nghidm
Qua trinh ting hdp ''C-choline gom 3 giai dogn chinh:
Giai doan 1: tgo ra '^C-CO^ khi bin bia khi N2 + 0,5%
O2 bdi chum proton da dUdc gia td'c. Sin pham '^C-COg di/dc day ra khdi bia bdi ddng khi quit He va bi bit vao sang phin ti!f.
Giai doan 2: tong hdp ^'C-CHjl tren thii't bi Mel-Plus.
Giai dogn niy gom 3 bUdc cd ban: biy, bit ^^C-COg vio sang phin Xis, tich logi bd khi He, N^ + O2; x i "C-CO^ tCf sang phin tijf va syc vio binh phan Ung vdi LiAIH^ tgo ra mudi Lithium methoxide (LiOCHg) gan Carbon-11 phdng xa; phin ufng giOfa HI v i mudi LiOCH tao ra methyl iodide (^'C-CHgl) phdng xg.
Giai doan 3: tong hdp ^^C-choline trin thii't bj Reform- plus.
"C-CH3 dUdc syc vio binh phin ufng chijfa tien chit DMAE tgo ra ^^C-choline, sau dd dUdc hda tan trong ethanol va bj giO trin cdt Accell plus (DM, cac tgp chit khac di qua cdt vio binh thii. Cac chi't dinh udt tren cot dUdc riia vio binh thii lan thuf 2 bang nUdc. Cudi cung
"C-choline dUdc giii hap phy ra khdi cdt bing djch nUdc muoi sinh ly 0,9% v i dUa vio lo sin pham.
HI Ket qua va thao luan
I.San xuit''C-CO,
Ham kich hogt cija phin ufng ^*U{p,aJY^C[7] ch? ra rang, carbon-11 se dUdc tgo ra vdi hiiu suit khac nhau tuy thuoc nang lUdng chum tia proton trong khoing 4-25 MeV. Trong nghien cufu niy, carbon-11 dUdc tgo ra khi ban bia bdi chum proton cd ning lUdng 16 MeV v i cic dilu kiin sau:
Vat lieu bia: khi N2 99,9999% + 0,05% 0^, - Bia nhdm thi tich: 50 ml,
- Ap suat bia khi ngp: 20 Bar,
Nang li/dng chum proton: 16 MeV, cadng dp
20-30 |LiA,
- Ap suit bia khi bin: 35-40 Bar, Thdi gian bin bia: 5-10 phut
- Hiiu suit dong tich hdp: 150-200 mCi/^iAh.
Khoing 200-300 mCi CCI dUdc tgo ra sau thdi gian bin bia 5-10 phut. Sau do, '^C-CO^ du'dc quit day ra khdi bia bang khi He va bj bit tren sang phan tCf cua module Mel-Plus. Khoing 90% ^^C-CO^ bj bat giO tren sang phin tLf, cac khf khac nhu N^, O2, He v i CQg se dUdc tach ra v i bi bit d biy khi thii.
2. Dieu chi''CMJ
Cic cim bii'n phong xg tgi vj tri ciia sing phin ti!f, binh phin ifng va bay thii se ty ding ghi do v i theo ddi sy thay ddi hogt dp ^hong xg cua "C-COg di chuyin trong he thong. Hinh 1 bieu dien sy thay doi hogt dp phdng xg trong qua trinh tong hop ^^CHgl. Khoing 350 mCi CQg da bj bit giuf tren sang phin tCf. Tii'p theo, nhiet do ciia sang phin tCf dUdc ning len 230°C v i ^^C02 bj day ra khoi sang va sue vao binh phin ufng chufa 150 \i\ LiAIH^. Hogt dd phong xg trong binh phin ufng tang ri't nhanh, tai thdi dilm nay LiO^^CHg dUde tgo thanh. Sau khi bay hdi dung mdi THF, mudi khan LiO^CHj dUde tie dung vdi 350 nHI 57% d nhiet do 115°C tgo thinh ^^CHgl. San pham ''CHJ dUde chung cat va day ra khoi module Mel-Plus d td'c dp 15 ml/phut nhd khi quit N^. Sd lieu ghi nhin tren hinh i eh? ra rang, hieu sua't bit "C-CO cija sang la khoang 86- 90%, hieu sui't tong hdp "CHI la khoang 75-77%.
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Hinh 1: bien thien ho$t dd phong xa trin sing phan td, bM phin dng vi dUdng thii trong qua trinh tdng hdp ^^ChlJ
3. Dieu chi^^C-choline
Khi qua trinh chUng ci't "CH3I bat dau, valve 3 cijfa se chuyin ki't ndi dau ra cua module Mel-Plus vdi dau vao eija module Reform-plus, khoing 80 mCi khi "CH3I dUde din bdi ddng khi H^ vdi lUu lUdng 15 ml/phut syc vao binh phin ufng chufa 10 ^il DMAE trong 200 ^il DMF (hinh 2).
Tai diy, "C-choline dydc tgo thinh theo phin ufng:
HoatdOng S6 thang 4.2012 K H O A H O C 85
NGHI6N CQU - TRRO D6\
''CH3l + HOCH,CH.N(CH3)„^
HOCH2CH2N(CH3V'^CH3 + P
Qua trinh vin chuyin cua nhin phdng xg carbon-11 trong module Reform-plus dUdc ghi nhin trin hinh 3 bdi 2 cim bii'n phdng xg d vj tri cOa binh phin ufng v i CM cartridge. Cic sd li^u cim bii'n ch? ra ring hi$u sui't tdng hdp "C-choline khoing 60-65%).
To W.i-ila
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Hinh 2: kit v6 trung dung 1 lin cua module Reform-plus dung dS diSu chi' ^^C-choline
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Hinh 3: bidn thidn ho$t dd phdng x$ trong binh phin ifng vi trdn CM cartridge trong qui trinh tdng h0p "C-choline
"C-choline v i cic chit cdn Igi trong binh phin ufng duoc hda tan bdi 1 ml ethanol v i dUdc bdm day qua cOt trao dii cation Accell plus CM cartridge. "C-choline bj giuf Igi trin cdt, cic chit khic nhu DMAE, DMF, I- di ra khdi cot vio binh thii Idng. Cdt CM cartridge dUdc rCfa 2 lin nOa bang nUdc cit vd trung d l logi bd cic chi't cdn dinh Udt trong cdt. Cudi cung, "C-choline dUdc ri!fa ra khdi cOt bing 10 ml dich truyln nude mudi 0,9% vio Ip dyng sin pham thdng qua filter 0,22 \im. PhUdng phip tinh chl niy dim bio sin pham cudi cung hoin toin tiit trung v i khdng chufa chit giy sdt.
4. Kiim nghigm ^^C-choline
Do sgch hda phdng xg v i hda hpc ciia "C-choline chloride trong sin pham cudi cung dugic xac dinh trin HPLC Agilent 2000 tich hdp detector do phdng xg Nal v i detector do khuc xg RID. HPLC dung cdt Inertsil ODS-2
(6-250 mm); Dung djch rCfa giii 0,001 M naphthalene- 2-sulfonic acid + 0,05 M H3PO,; td'c dd ddng 1 ml/min.
Authentic choline chloride (Sigma) dung lam chit chuin.
Kit qui phin tich hda phdng xg cua sin phim culi cung ch? cd 1 d?nh ddn d thdi gian lyu 14 phut ch? ra ring, dd sgch hda phdng xg cua "C-choline l i gin nha 100%.
Kit qui ghi nh§n ciia detector khuc xg RID khdng phit hi$n ra DMAE v i ch? cd 1 d?nh d thdi gian luu 13,8 phut cua choline chloride cung chufng td ring sin phim culi cung ch? l i choline. pH cua sin phim dadc xic djnh l i 7-7,5. Kit qui kilm nghiim cung khing djnh, sin phim
"C-choline l i vd triing v i khdng chufa chit giy sdt, dgt tiiu chuin thudc tiim.
IV. Ket luan
Ting hdp "C-choline tgi Trung tim Miy gia tic 30 MeV dUdc thyc hiin trin cic thiit bj dilu khiln ty dOng, chuyin dyng cho nghiin cufu, phit triln cic dugc chit phdng xg. Cic thii't bi niy cho phip ting hdp 2-3 mi/
ngay. Sin phim "C-choline cd dO sgch hda phdng xg dgt
> 99%, khdng phit hien vit tiin chit DMAE trin HPLC.
Dung djch "C-choline cd pH 7-7,5, hoin toin vd trung va khdng chufa chit giy sdt, dgt tiiu chuin thudc tiim theo dUdc diln BP •
Tai lieu tham khao
[1] T. Hara, N. Kosaka and H. Kishi, J. Nud. Med. 39 (1998), p. 990.
[2] T. Hara, N. Kosaka, N. Shinoura and T. Kondo, J. Nud. I^ed. 38 (1997), p. 842.
[3] T. Hara, N. Kosaka, H. KIshi and 0. Kobori, J. Nud. Med. 38 (1997), p. 250P.
[4] B.H. Mock. O.K. Mulholland and M.T. Vavrek, Nud. Med. Biol. 26 (1999), p. 467.
[5] C. Pascal), A. Bogni, R. Iwata, M. Cambie and E. Bombardieri, J.
Labei Compd. Radiophami. 43 (2000), p. 195.
[6] E. Mishani, I. Ben-David and Y. Rozen, Nud. Med. Biol. 29 (2002), p. 359.
[7] IAEA, Cyclotron Produced: Principle and Practice, Technical Reports Series 465. Vienna (2008).