Rev. Soc. Bras. Med. Trop. vol.28 número3

R e v is t a d a S o c ie d a d e B r as ile ir a d e M e d ic in a T r o p ic al 2 8 ( 3 ) : 1 7 9 - 1 8 3 , ju l- s e t , 1 9 9 5 .

A N TISCfflSTO SO M A L A CTIV ITY O F A CRID A N O N

E-H YD RA ZO N ES IN

CEBU S

M O N KEYS EXPERIM EN TA LLY

IN FECTED W ITH TH E SJ STRA IN O F

SCHISTO SO M A M A N SO N I

Pau lo M arco s Z ech C o elh o , Leo gen es H o racio Pereira* an d

R om ulo T eixeira d e M ello

In t his st u dy , f o u r c o m p o u n d s w e r e u t iliz e d a t t h e d o s e o f 1 2 .5 m g /kg b o d y w eig ht , p . o . , t o t r e at Cebus m o n k e y s e x p e r im e n t ally in fe c t e d w it h a b o u t 2 0 0 c e r c a r i a e o f Schistosom a m ansoni ( S J s t r ain ) , v ia t r an s c u t an e o u s r o u t e . T he o o g r am s p e r f o r m e d w it h r e c t a l sn ip s, a s w e ll a s s t o o l e x a m in at io n s c a r r i e d o u t p e r io d ic a lly , s h o w e d n o v ia b le e g g s o f t h e p ar as it e , f r o m d a y 2 9 t o 2 2 6 p o s t - t r e at m e n t . T he p e r f u s io n u n d e r t ak e n a f t e r killin g t h e a n im a ls s h o w e d a b s e n c e o f w o r m s in t h e t r e at e d m o n ke y s , w h e r e a s 8 3 w o r m s w e r e r e c o v e r e d f r o m t h e c o n t r o l, t hu s c o r r o b o r at in g t h e resu lt s o b t a i n e d by m e a n s o f o o g r am s a n d c o p r o s c o p y . T hes e re s u lt s c o n fir m t h e e ff i c a c y o f 9 a c r i d a n o n e -h y d r a z o n e s p r e v io u s ly t e s t e d ag a in s t t -h e LE s t r ain o f S. m ansoni. T he lo w c u r at iv e d o s e a n d a p p a r e n t a b s e n c e o f t o x ic it y r e n d e r t h e s e d m g s a n im p o r t an t t h e r ap e u t ic r e s erv e, t ak in g in t o c o n s id e r at io n t h e r e p o r t s o n t h e r e s is t an c e o f S. m ansoni t o t h e m o d e r n d r u g s o x a m n iq u in e a n d p r a z iq u a n t e l.

K e y - w o r d s : Schistosom a m ansoni. Cebus m o n ke y s . A c r id a n o n e - h y d r a z o n e

A lthough much effort has been directed

tow ard the improvement o f the chemotherapy

fo r schistosomiasis mansoni, and despite the

large progress o btained in this field, curative

treatment o f this d isease w ith the already

kno w n standard antischisto somal compounds

presents so me difficulties. The high price o f

drugs, drug sid e effects, cases o f drug

resistance, am o ng o ther p ro blem s, p o int

tow ard the co ntinuous search for new active

substances.

C o elho

and

Pereira4

sho w ed

a

p ro m ising activity o f 9-acridanone-hydrazone

co mpo und s in

monkeys experimentally

infected w ith the LE strain o f

m an s o n i.

However, a questio n arises w henever new

promising co mpo und s against schistosomiasis

appear. Are they effective against all human

D e p artam en to s d e Parasito lo g ia, In stitu to d e C iên cias Bio ló g icas, e d e A nálises C línicas e T o xico ló g icas, Facu ld ad e d e Farm ácia, U niv ersid ade Fed eral d e M inas G erais, Belo H o riz o n te, M G .

T his stud y w as su p p o rted in p art b y C N Pq , FA PEM IG , PRPq /U FM G , Brazil, an d b y W H O , Sw itzerland.

* D eceased 6 Feb ruary, 1 9 9 4

E n d e re ç o p a ra co rres p o ndência - . Pro f. Paulo M arcos Z ech C o elh o , D ep to d e Parasitologia. IC B/U FM G /C aixa Postal 4 8 6 ,3 0 1 6 1 - 9 7 0 Bel o H o riz o n te, M G .

R eceb id o p ara p u b licação em 2 4 / 0 8 / 9 4 .

schistosomiasis, and even if active against a

strain o f a specific sp ecies

fo r

instance), can their effectiv eness be extended

to o ther strains? It is w ell kno w n that

mainly in so me co untries o f Africa,

p resents

d ifferen t

su sc e p tib ilitie s

to

chem otherapy w ith o xamniquine, via oral

route. Thus, in Brazil and West Africa, 15mg/ kg

bo d y w eight is the d o sage co mmonly used,

w hereas in East Africa the effectiv e do se

ranges from 30 to 45mg/ kg, and in Egypt

d o ses o f up to 60mg/ kg given o ver three days

are required 16. On the o ther hand , the

resistance against o xamniquine by

has been d etected in Brazilian patients16. Cioli

et al3 clearly sho w ed that this resistance w as

controlled by a single auto so mal recessive

gene.

In Brazil, three sp ecies o f

mollusks are resp o nsible fo r the natural

transmission o f schistosomiasis mansoni, and

these species are pred ominant as vecto rs in

different areas. Thus, the adaptation betw een

so m e strains o f the p arasite and the

geographical strains o f snails2 81011 has been

demonstrated, as w ell as the variation in the

pathogenicity degree related to strains o f

Coelho PMZ, P ereira LH, M ello RT. A ntischistosomal activity o f acridano ne- hy draz o nes in C eb us monkey s experimentally infect ed with the Sf strain o f S chistosom a m anso ni. Revista da So ciedade Brasileira de M edicina Tro pical 2 8 :1 7 9 - 1 8 3 , jul- set, 1995.

SJ strain o f

S. m ans o ni

(iso lated in the State o f

São Paulo , in an area w here

B io m phalaria

t enago phila

is the snail v ecto r) p resents the

sam e

su scep tib ility

to

the

acrid ano ne-

hy d razo nes utilized against the LE strain o f the

p arasite (iso lated in the State o f Minas Gerais,

in an area w here

B io m p halaria glabrat a

is the

snail v ecto r), using the

C ebus

m o nkey m o d el

to o .

M A TERIA L A N D M ET H O D S

Cap uchin m o nkey s (

C ebus sp),

ad ults o f

b o th sexes, w ere m aintained in ind ivid ual

cag es in the anim al ho use. The m o nkey s w ere

fed o n d aily w ith b red so aked in milk, fruit,

and stand ard p ellet fo o d . They w ere infected

w ith ab o u t 200 cercariae o f

S. m anso úi

(SJ

strain), transcutaneo usly . The life cy cle o f the

trem ato d e w as m aintained at the labo rato ry

thro ug h ham ster -

B io m phalaria t enago phila

-ham ster

p assag es.

A crid ano ne-hy d raz o ne

drugs w ere g iv en after the m ature infectio n

had b e e n stablished . The m o nkey s that w ere

fo und to b e p o sitiv e (p resenting larg e num bers

o f schisto so m e eg g s in the sto o ls and in rectal

snip s) w ere selected fo r the exp erim ents. The

co m p o und s Ro -15-5458/ 000, Ro-15.8843/ 000,

Ro-15.9268/ 000 and Ro -16.2308/ 000 (Fig ure 1)

w ere ad m inistered at the d o sag e o f 12.5mg/ kg

b o d y w eig ht, by o ral ro ute, sing le d o se. The

infected m o nkey E-2 w as kep t as co ntro l. The

quantitativ e o o g ram o f rectal snip s9, sto o l

exam inatio n4, and p erfusio n o f p o rtal system

fo r w o rm reco v ery l2 w ere used as criteria fo r

d eterm inatio n o f p arasito lo g ical cure.

RESU LTS

It

m ust

b e

em p hasiz ed

that

the

c o p ro sco p ies

p erfo rm ed

co nfirm ed

the

o o g ram in rectal snip s in all cases.

The drug effects ag ainst the SJ strain o f

S.

m anso ni

are sum m arized in Tab le 1. The sto o l

exam inatio n and p erfusio n o f p o rtal system o f

the m o nkey s co nfirm ed the find ings o btained

w ith the rectal snip s. So , w hen all the treated

m o nkey s w ere exam ined (26 exam inatio ns p er

m o nkey ) no eggs co uld b e d etected in sto o l

exam inatio ns. O n the o ther hand , the p resence

o f v iable eg g s w as d etected in all the sto o l

exam inatio ns o f the co ntro l m o nkey . No

w o rm s co uld b e d etected thro ug h p erfusio n

into the treated m o nkey s, w hereas 83 w o rm s

(24 fem ales and 59 m ales) w ere reco v ered

fro m the co ntro l m o nkey (E-2).

D ISCU SSIO N

So m e 9-acrid ano ne-hy d razo nes hav e b een

p ro v ed v ery effectiv e in the p reclinical

treatm ent fo r schisto so m iasis, w hen no nhum an

Co elho PM Z, P ereira LH, M ello RT. A ntischisto so m ál activ ity o f acridano ne- hy draz o nes in C eb u s m o nkey s

experim ent ally infect ed w ith t he Sf st rain o f S chisto so m a m an so n i. Rev ista da So ciedade B rasileira d e

M edicina Tro pical 2 8 :1 7 9 - 1 8 3 , jul- set, 1 9 9 5 .

p rim ate m o d els, as b ab o o n s14 and

C ebus

m o nkey s4, w ere used . Very g o o d results have

b een rep o rted b y C o elho and Pereira4 using

C ebus

m o nkey s infected w ith the LE strain o f

S. m anso ni,

and treated w ith so m e acrid ano ne-

hy d razo nes that sho w ed curative activity at

d o ses as lo w as 25m g, so m etim es 12.5m g, p er

o s, sing le ad m inistratio n. These autho rs

sho w ed that the results o btained thro ugh

p erio d ical

rectal

b io p sies

and

sto o l

C o e lh o PM Z , P e r e i r a LH , M e llo RT. A n t is c h is t o s o m al ac t iv it y o f a c r id a n o n e - h y d r a z o n e s in

Cebus

Schistoso ma manso ni.

o ccasio n, it w as verified that the curative

d o ses, besid es being quite low , and singly

orally administered, did no t sho w visible side

effects o n the primates.

In the same w ork, it has been o bserv ed a

marked activity o f all the co mpo unds studied

against the SJ strain o f S. m an s o n i, at a lo w er

d o se (12.5mg/ kg), thus confirming the results

o btained by Co elho and Pereira4 w ith the LE

strain. These results lead the authors to think

on the possibility o f the same compounds

being successful in a lo w er curative do se (less

than 12.5mg/ kg bo d y w eig ht in

m o nkey s). It is w ell kno w n that many

schisto so micid es in the monkey require near

five times more do sing than that used fo r man.

Thus it seems that d o ses lo w er than 3mg/ kg

bo d y w eight co uld po ssibly be used for man,

although it must be proved w hether the

to xicity o f these co mpo unds do es no t prevent

their use in humans.

The repo rt o n drug resistance related to the

use o f o xam niquine, the most utilized drug in

the treatment for schistosomiasis in Brazil17,

w id en the po ssibility o f co nsid ering the drugs

here studied as an important reserve supply to

be used, perhaps, in the near future.

RESUMO

N o p r e s e n t e t r ab alh o , q u a t r o c o m p o s t o s f o r a m u t iliz ad o s n a d o s e d e 1 2 ,5 m g ^ kg d e p e s o , p o r v ia o r al, e m m a c a c o s in fe c t ad o s t r an s c u t a n e a m e n t e c o m c e r c a d e 2 0 0 c e r c á r i a s d e Schistosom a m ansoni. O s o o g r a m as r e a liz a d o s c o m fr a g m e n t o s d e m u c o s a r e t a l e o s e x a m e s d e f e z e s r e aliz ad o s , p e r i o d ic a m e n t e , d e m o n s t r a r am a a u s ê n c i a d e o v o s v iáv e is d o p a r a s i t o a p a r t i r d o 2 9 - a t é o 2 2 6 a d i a p ó s - t r at a m e n t o . A p e r f u s ã o , a p ô s s a c r ifí c io d o s a n im a is ti' atados , n ã o d e t e c t o u v e rm es, e n q u a n t o q u e d o m a c a c o c o t z t r o le 8 3 v e r m e s f o r a m r e c u p e r ad o s , c o n f i r m a n d o as s im o s r e s u lt ad o s d o s o o g r a m a s e d a c o p r o s c o p i a . E s t e s r e s u lt a d o s c o n f i r m a m a e f i c á c i a d a s 9 a c r i d a n o n a s -h y d r a z o n a s j á o b s e r v ad a an t e r io r m e n t e c o n t r a a c e p a LE d e S. m ansoni. A b a i x a d o s ag e m c u r at iv a e a p a r e n t e a u s ê n c i a d e t o x ic id a d e c o l o c a m e s t as d r o g as c o m o u m a r e s e r v a t e r ap ê u t ic a im p o r t an t e , t e n d o e m v is t a o r e lat o d e r e s is t ê n c ia d o S. m ansoni à s d r o g a s m o d e r n a s o x a m n iq u ín a e p r a z iq u a n t e l.

P alav r as - c h av e s : Schistosom a m ansoni. M ac ac o s

Cebus.

A CKNOW LEDGEMENTS

To Dr. H.R. Stohler (F. Hoffmann-La Ro che

& Co. Ltd., Basle, Sw itzerland) fo r providing

the co mpo unds used in this w o rk, as w ell as

for financial support and advice; Dr. Faiçal

Simon and Dr. Adayr Mafuz Saliba (Fund ação

Parque Z o o ló gico de São Paulo) for the supply

o f

for translating the manuscript; Mr. A delino

Ferreira (in memoriam), Mr. Maurício V. Costa

(in memoriam), Mr. A lberto G. Santos and Miss

Zenir de Souza for technical assistance.

REFERENCES

1. A raújo N, Katz N, D ias EF; Souza E Susceptibility to ch em o therap eu tic agents o f strains o f

S c h is t o s o m a m a n s o n i isolated from treated and untreated p atients.T he A m erican Journal o f Tropical M edicine and H ygiene 2 9 :8 9 0 - 8 9 4 ,1 9 8 0 .

2. B arb o sa FS, B arre to A C . D if f e re n ce s in

susceptibility o f Brazilian strains o f A u s t r alo r b is g la b r at u s to S c h is t o s o m a m an s o n i. Experim ental Parasitology 9 :1 3 7 - 1 4 0 , I960.

3. Cioli D, Pica-M attoccia L, M oroni R. S c h is t o s o m a m an s o n i: hyeanthone/oxam niquine resistance is controlled b y a single autosom al recessive gene. Experim ental Parasitological 7 5 :4 2 5 - 4 3 2 ,1 9 9 2 .

4. Coelho PM Z, Pereira LH. S c h is t o s o m a m a n s o n i.: Preclinical studies w ith 9-acridanone-hydrazones

in C e b u s m onkeys exp erim entally infected . Revista do Instituto de M edicina Tropical de São Paulo 3 3 :5 0 - 5 7 ,1 9 9 1 .

5. C oelho PM Z, Raso P, M ello RT, Toppa N H. D imensões do granuloma hepático produzido

p o r ovos de duas linhagens geográficas do S c h is t o s o m a m a n s o n i, n o cam und ongo. M emórias do Instituto O swaldo Cruz 8 4 :2 1 3 - 2 1 7 ,

1989.

6. D ias LCS, G onçalves ER. S c h is t o s o m a m a n s o n i diz não às drogas. Ciência H oje 1 4 :2 2 - 2 5 ,1 9 9 2 .

7. D ias LCS, Pedro RJ, D eberaldini ER. U se of praziquantel in patients w ith schistosom iasis

mansoni previously treated w ith oxam niquine and/or hycanthone: resistance o f S c h is t o s o m a m a n s o n i t o schistosomicidal agents. Transactions o f the Royal Society o f Tropical M edicine and

H ygiene 7 6 :6 5 2 - 6 5 9 ,1 9 8 2 .

8. Files VS, Cram EB. A study on the com parative susceptib ility o f snails v e cto r to strains of

S c h is t o s o m a m an s o n i. Journal o f Parasitology 3 5 :5 5 5 - 5 6 0 ,1 9 4 9 .

9. K atz N, Pellegrino J, Pom péu-M em ória JM . Q uantitative oogram m ethod in C e b u s monkeys exp erim entally infected w ith S c h is t o s o m a

C o e lh o PM Z , P e r e ir a LH , M e llo RT. A n t is c h is t o s o m al ac t iv it y o f a c r id a n o n e - h y d r a z o n e s in

Cebus

Schistosoma mansoni.

m an s o n i. Journal o f Parasitology 52:917- 919, 1966.

10. N ew ton W L. The inheritance o f susceptibility to inf ection w ith S c h is t o s o m a m a n s o n i in A u s t r alo r b is g lab r at u s . Experim ental Parasitology 2; 2 4 2 - 2 5 7 ,1 9 5 3 .

11. Paraense W L, Correa R. V ariation in susceptibility o f populations o f A u s t r alo r b is g la b r at u s to a strain o f S c h is t o s o m a m a n s o n i. Revista do Instituto de M edicina Tropical de São Paulo

5:15-2 5:15-2 ,1 9 6 3 .

12. Pellegrino J, Siqueira AP. T écnica de perfusão para

colheita de S c h is t o s o m a m a n s o n i em cobaias experim entalm ente infestadas. Revista Brasileira de M alariologia 8: 5 8 9 - 5 9 7 ,1 9 5 6 .

13. Saoud M FA .The infectivity and pathogenicity of geograp hic strains o f S c h is t o s o m a m an s o n i. Transactions o f the Royal Society o f Tropical M edicine and H ygiene 6 0 :5 8 5 - 6 0 0 ,1 9 6 6 .

14. Sturrock RF, Bain J,W eb be G, D oenhoff MJ, Stohler H. Parasitological evaluation o f curative and

subcurative doses o f 9-A cridanone-hydrazone drugs against S c h is t o s o m a m a n s o n i in baboons, and observations in serum levels o f anti-egg antibodies d etected by ELISA. Transactions o f the Royal Society o f Tropical M edicine and H ygiene

8 1 :1 8 8 - 1 9 2 ,1 9 8 7 .

15. W arren KS. C om p arison o f Pu erto Rican, Brazilian, Egyptian and Tanzanian strains of

S c h is t o s o m a m a n s o n i in m ice: penetration o f cercariae, m aturation on schisto so m es and production o f liver disease. Transactions of the Royal Society o f Tropical M edicine and H ygiene 6 1 :7 9 5 - 8 0 2 ,1 9 6 7 .

16. W orld H ealth O rganization. T he ro le o f ch em o therap y in schistosom iasis co n tro l. W H O / S ch isto / 83.70, Revue. 1. W orld H ealth