Abstracts Vol. 1 [International Conference on AIDS (11th: 1996: Vancouver, Canada)]

Mo.A.1059 - Mo.A.1064

Monday, July 8, 1996

Mo.A. 1059
THIADIAZOLE DERIVATIVES WERE HIGHLY POTENT INHIBITORS OF HIV-I
Fujiwara Masatoshi', Ijichi K*, Hanasaki Y"*, IdeT**, Katsuura K5,Takayama H**,Aimi
N**', Shigeta *"***, Konno K*, Yokota T*, Baba M i**. "Rational Drug Design
Libor atonres, Tosoh Co. Ltd., "*"Chiba University, '"Fukushima Medical College,
i * g " IKaoshrra Unesty, Japan.
Objective: To e luate whether several thladiazole (TDA) derivatives,e!ctively inhibit the
replication of I--tV Ir ttr
Methods: Ant -HIV activities of TDA denrvatives were examined in vario,, ell Ines.
Inhbitory effects of t he compounds on HIV-I reverse transcriptase (RT) actty t were
determined by a standard assay system. Synergistic effects were evaluated by the median
effect pr inciple and the isoboloi rarn method.We attempted whether TDA der vatives ronpletely suppres sed HIV I rep lication a long term cultureof MT-4 cells.
Results: Amon; the corpounds, RD4-2217 was found to be the most potent inhbitor of
HIV I replication. RD4 221 7 inhibited the replication of HTLV II1B strain in various cell cultures at a iconcentration of narnomolar range. It was also highly inhibitory to clinical isolates
and AZ1 resistant mutants of HIVI. However, nonnucleoside RT inhibitor resistant mutants
and HIV 2 wer lecss sensitive or insensitive to the compound. Studies on their mechanism
of action revealed thatT DA dervatives inhibited the activity of recombinant HIV I RTThe
50% i nhibitory concentration of RD4-2217 was 0.5pM when poly(rC) oligo(dG)12 18 was
used as the terrplate -primer:The combination of RD4 2217 with AZT or the protease
inhibitor A 75925 exerted a synergistic anti -tHIV I activity Studies on the emergence of
drug resist'ant mutants revealed that, although much higher concentrations (I 10 pM) were
requr, edRD 2217 could completely suppress the breakthrough of the virus during its
ong tern treatraent in cll culture.
Conclusion: he TDA der vatyes are worth pursuing as candidate drugs for the
chemotherapy of HIV I infections.
M Fujiwara, 4,M isato, Matsukawai Machl. Fukushima 960 12, Japan TEL: [phone]
FAX: 81 2,5/ 7 555 email: fui(rdL.r p
Mo.A. 1060
ALLOPHENYLNORSTATINE-CONTAINING PEPTIDOMIMETIC HIV PROTEASE
INHIBITORS EXHIBIT BOTH IN VITRO AND IN VIVO ANTIVIRAL ACTIVITIES
Kiso Yoshiak i"imoto *. Kato R*, Mitoguchi T*, Nakata S*, Kimura T*, Ussery M A.***.
*Kyoto Pharmaceutical University Kyoto, Japan; " Japan Energy Co.,Toda, Japan; **U.S.FDA,
Rockvile, MD. U.S.A
Objective: To develop orally potent and small-sized HIV protease inhibitors as anti-HIV drugs.
Methods We desi gned and synthesized a novel class of substrate based peptidomimetic
HIV protease iihibitors containing allophenylnorstatine [Apas; (25,3S)-3 amino-2-hydroxy-4 -phenylbtyricb it cid] wth hydroxyrmethylcaibonyl (HMC) isostere on the basis of transitionstate concept.
Results: An,; them, the tripeptde KNI 272 was a highly selective and superpotent HIV
protease inh bitor (Ki 5.5pM). KNI 272 exhibited potent antiviral activities against both
AZ -senitive nd insensitive clinical HIV I isolates as well as HIV-2 with low cytotoxicity.
After id. administration, bioavlability of KNI 272 was 42.3% in rats. Also, KNI-272 exhibited
in vivo anti -HIV activities in hurman PBMC -SCID mice X ray crystallography and molecular
modeling stud is showed that the HMC group in KNI 272 interacted excellently with the
asp re icci d carboxyl go ups of HIV protease a ctive ste.This result implies that the HMC
isostere is an ideal trinsition state mimic and thus leads to small-sized dipeptide inhibitors
(KNI 413 & -5,19:Fig.).These exhibited antviral actity and improved pharmacokinetic
propertics.
Conclusions: An Apns-conrtaning HIV protease inhib tor has a potential as an orally potent
anti-AIDS drip, and is undergoing clinical trials.
Yoshiaki K so Dept Med. Chem. Kyoro Pharmac. Un v, Yamashina ku, Kyoto 607, Japan
Telephone: [phone] '335 Fax [phone] E mail: PXH0435 niftyserve.op
Mo.A.1061
POTENT ACTIVITY OF THE EXTRACT OF Geum japonicum thunb. FOR THE
PROPHYLAXIS OF CYTOMEGALOVIRUS INFECTION IN AIDS PATIENTS
Kageyami, Sei_ Kurokawa H*, Sate H,Yukawa -T, Ohyama H", Kurimura T#, Namba T*,
Sh aki K ~ayma Medcal & Pharmaceutical University Toyama;'": Showa Shell Sekyu
KK Atig; n5:1 s-aki UaiverhsieyeSoitauJpa
Objettive: A TIdItiOFl hEicibTa C cedTFci G rted Fon euinamrh io R Thank (C) with
tion in immunosuppressed mi ce In the cell culture system, GJ was assayed for its anti-HIV
acteivt. Fthermre, a pried icx ract of Gc was so tested to inquire its anti-HIV activity
b/ the onhbiticon/severs tra n ipsitse PT) tivt/.
Methods: Dried traditional herbal medicine, GJ, was boiled, litered, lyophilized, and resuspended in distiled waster aiit the concentration ofit 20mg/ml. After an addietional boiling, the
extract was orally administered at 5mg/dose three times a day to female ICR mice treated
with 50mg/kg of cyclosporine. Gancic ovir was also administered intraperitoneallyThese
mce were iected intraperntoneall/ with 1,000 PFD oF mcroe CMV (MC V) (Smth
strasin).Vrus ainoectin in the lung mrs assessed on dry 16 after onfection. M,'T-4 cello acutely
c/cected with hI1V- LA mere used For-the evalastior of activity of CJ igainst DIV infection.
PT wctity ma's yed n the presence of the purfed extract of GJ by the conventional
assai/system with pol/(rA),p pdT12 18) as the template.
Results GJ sroil supprecedthe irus ie ung of mouse at a conventina dose
for hunanr. Its art MbCMV activity with the treatment 750mg/kg/day (ore.) of C) corresponds to that with 2mg/kg/day (Lp.) of gancciovir A puriied extract of GJ shows the
strong anti HIV activity (J exhibited partial anti-H V activity against HIV at 2 pg/ml in cell
culture system. RTactivity was reduced,n the presence of thi s, ert. I ie,
of RT were 22, 60, 93% at 2, 20. 200pM, respectively.
Conclusions: Strong anti-CMV activity assessed in mouse rnodel raiy be bero
prophylaxis of retinitis caused by CMV infection in AIDS patents, Jm lv i< e
HIV infection.
Seii Kageyaman 2630 SugitaniToyama, 930 01 Japan Teleaione: 0/,1 34 -8 I8
5020 email:kageyamantoyaman-rit.ac.jp
Mo.A. 1062
ANTI-HIV- I ACTIVITY OF LABIATAE PLANTS, ESPECIALLY AROMATIC PLANTS

Yamasaki K, Nakano M*, Otke 1
'Osaka Prefectural Institute of Pu
Object: We have screened andd
from natural products and define
found some Lcbiarce plants excc
plants(herbs) belonging to La itn
Method: About 40 herbs belong
acetone, ethanol, 70%ethanol a d
replication of HIV I (LAV- I).
Results and Discussion: Acetone
ty but the 70% ethanol extract Sh(
anti-HIV I activity. Especially, Pr u
ni is L. and Perilla (rutescerns Brit v

Kawihata Fr, Mori H Momcto I M e, '

l)c Health, Japan
entified compound ha' ii J If(
i he rechan isms of the int IIV i,ng anti HIV activity we screene
and searched for the ictse
Sto Ln iiutie used in J:an re
vwater: Each extract ys ive it:,rid ethanol extracts of all the 5<<'1
owed light actvity The vate evi
i1 ali Kudl s, owerl m Ir Pl tL. (ot
in ricto KKid( showed vet y potent i(

concentration of 16 mg/ml. By use colaumn clhromatography we are '
ant-HIV I active substances n these herts.
Replication of/the fresh isolated HIV I str iin,anhbtec by l oreIn/ e
weakly inhibited the reverse transcr iptase activity of HIV-. All the
cell formation in co-cultures of HIV I -infected and non- in'cted M t>
these compounds can inhibit virus adsorption or fusion.
Conclusion:Through our study we found many herbs with anti /V
very meaningful that herbs belonging to L Itie plant,i resonale a
show very potent anti-HIV I activity It is very mportant to sear ho
substance in these Lbitae plants

xtr i t, /i> 1
t Ict' Si r i
c]CtlV lt, n', r(,Q,
aC
It 0

Katsuhiro Yamasaki, I-3-69, Nakamiti, Higashinan Ku,
Telephone: 8 I (0)6-972-132 I Fax: 8 I (0)6 972 -239:

Japer

Mo.A. 1063
ANTI-HIV ACTIVITY OF BOROMYCIN
KawahataT*, OtakeT*, Mori H, Moriot M*, Ueba N', KohnoJi '', sh 'M' i n,,,
A**, Komatsubara S**. *Osaka PrefecturIcsttute of P'c Ha tic'iii Id / i CI e4, '
Japan; **Tanabe Seiyaku CO.LTD Lead Generation Rese it I L bory,tTrd,I
Saitama, Japan
We have reported several natural products tithave a p ent rI Iior y i 'vit' 15 IV
replication.The extracts of fermentation broth of miroor anr io1a'ed fronao,.'rie
tested for anti-HIV I activity by the screening assay using r' MT4' r i e produtf
Streptomyces sp. A 3376 was found to have anti-HIV I actity, and den fed i poyether
macrolides antibiotic, boromycin.This compound exhibited potent inhibitor fiet o' the
replication of HIV-I and HIV-2 laboratory strains LAV- ind LAV R-DICS( 7 a
and 0O.06pg/mL respectively) and HIV-I clinical isolate str an KK ca0. ' / / '
did not inhibit virus adsorption, gant cell formation, reverse trnscr pt,se ict sI r, i I te
gration of the HIV genome into the chromoscmal DNA. The nfect ivt of HI i,.,red.o
treated with boromycin, was not reducr'd DHowever, boromsrrv retdi cc It e r-ii t
infectious HIV-I particles from the infected cell s, but the ictvt/ of eers etri r-t J
the culture supernatant did not decrease.
W e concluded that the anti-HIV activity of boromn ycin de i 'ed r ffectrn;a;I tr '.
probably maturity step on replication o HIV molecule
Takuya Kawahata, I3 69, Nakamiti, Higashinar-i-Ku, Osaka, Japan
Telephone: 81 -(0)6 972-1321 Fax: [phone]
Mo.A. 1064
INHIBITION OF CELLULAR ACTIVATION OF LATENTLY HIV-INFECTED
PERIPHERAL BLOOD MONONUCLEAR CELLS BY TRADITIONAL MEDICINE
Haruyo Mori**, InadaY'*, Otake T*, Ueba N*. Osaka Prefecturalin I'tc f P Ir
Health, Osaka, Japan a*St. Luke's-Roosevelt Hospital Center NY USA
The majority of HIV infected CD4+ lymphocytes and macropha es aire It 'itI it i
vivo, particularly the early stage of infection. It is consder-edd t it 'hese tent celle ae
activated by several stimulants such as cytokines and star to produce HI V partcle rut
ng in the progression of disease.
We have investigated the correlation between the response of peripera bloodm rnonu
clear cells (PBMC) f-rom HIV-infected individuals n=44) to stimuato and the m'aure fun
tions. As a result, PBMCs were divided into 4 groups according to the amount of,iI pro
duced after stimulation with cytokines(TNF- u, IL 6) as f llows; ( ) V-,rIot IateCted
(n- 10) (2) Low levels of HIV were produced, but not affected by stir nulatio ( I)!>)
HIV replication was enhanced by stimulation (n=- 10) (4) High Ive otf HIV re 'i aere
observed even though without stimulants (n= 13).The imrurc funtCon t')tetsn
each group progressively decreased from (I) to (4) It s expected, theIe' tr,,t 'attto
of latently infected cells from stimuli can lead to del ay the d -e roresso
We have developed an in vitro drug assay system using quescent PBMs to eval uae the
inhib itory effect on cellular act ation caused by the stim ulation wth cytoknes. PBMCs
obtained from healthy donors were infected with HIV-I clinical isolate and then stimuated
with several cytokines(TNF-, IL-2 and IL-6) in the presence or absence of test m Iater als.
The replication of HIV-I was determined by,T activity in culture supernatant. It was
observed that Sho-saiko-to(TSUMURA & Co., Japan) and Fuscopo i iob qua, the or iental
plant extracts, inhibited the HIV-I replication in latently irfected cells at 50pJ/ml and
40pg/ml, respectively probably due to protection of cells againstcellular actiatlon.
Haruyo Mori, I-3-69, Nakamiti, Higashinari-ku, Osaka 537, Japan Telephone [phone]
Fax: [phone]