Başlık: Triazene Derivatives. I. Sulfaguanidine as a Carrier for Substituted Triazeno Group : Triazen Türevleri I. Sülfaguanidin Molekülünde Substitüe Triazen Yapılarının OluşturulmasıYazar(lar):NOYANALPAN, Ningur;ÖZDEN, Seçkin;ÖZDEN, Tuncel Cilt: 7 Sayı:

7 . 1 0 4 ( 1 9 7 7 ) 7 . 1 0 4 ( 1 9 7 7 )

Triazene Derivatives. I. Sulfaguanidine as a Carrier

for Substituted Triazeno Group*

Triazen Türevleri I. Sülfaguanidin Molekülünde Substitüe

Triazen Yapılarının Oluşturulması

N i n g u r N O Y A N A L P A N * * S e ç k i n Ö Z D E N * * T u n c e l Ö Z D E N * *

Most compounds which contain a di-2-chloroethyl amino group

possessing a certain level of chemical reactivity and a number of

substituted triazeno derivatives are cytotoxic to proliferating tissues

'

However a number of such compounds have limited use as

che-motherapeutic agents because of the lack of specific action on

malig-nant growths, also many normal proliferating tissues are equally

affected by most of the derivatives.

It was shown by D A N I E L L I (Nature 170, 863, 1952) that the

cells may be highly selective in the types of molecule which they

con-cantrate within their plasma membranes. Following this many

aut-hors tried to incorporate the di-2-choloroethyl amino group into

molecules of different physical and chemical properties varying in

their anionic, cationic, liphophilic and hydrophilic character as

well as trying to have different structures permitting the chlorine atom

to hydrolize in different rates

- Manv others have tried to

incorpo-rate triazeno moiety into suitable carrier molecules and some others

have tried to combine triazeno structure with di-2-chloro ethyl

moietv to increase the activity

'

Redaksiyona verildiği tarih: 22 Nisan 1977

* This work is the first of a series of research with triazene derivatives. ** Farmasötik K i m y a Kürsüsü, Eczacılık Fakültesi, Ankara Üniversitesi

Besides the compounds such as Nitrogen Mustard (NSC- 762),

Cyclophosphamide (NSC-26271), Melphalan (NSC-8806),

Chlo-rambucil (NSC- 3088) Imidazole Mustard (NSC- 82196), B C N U

(NSC-409962), C C N U (NSC-79037), many others have been prepared

and tried for antitumor activity among which only the ones carrying

either sulfamide structure or triazenc structure concern the present

research.

Triazeno derivatives of various ring systems such as substituted

imidazoles

and pyrazoles

both carrying either carboxamide or

carboxilic acid groups have been synthesized and a number of these

have displayed antineoplastic activity among which Imidazole

Mus-tard (NSC - 82196) has been clinically accepted. E L K S and H E Y

have prepared a number of l-aryl-3,3-dimethyl triazenes from

aniline, m - a n d p-nitro aniline, anthranilic acid, methyl anthranilate

2-naphtylamine,4-amino phtalimide, methyl-4-amino phtalate

and 5amino quinoline. L E C O C Q , has synthesized some c o m

pounds of the type sulfanilic acid and thioanisidine, carrying d i 2

choloroethyl amino function at the para position and also p d i

-N, N-2-chloroethyl or hydroxy ethyl sulfamoil aniline. The

anti-bacterial activity of these compounds were found inferior than that of

sulfanilamide. The analogues of the latter compound have also been

been prepared by Ross and Wilson

replacing the p-amino function

with various groups. Again by replacing p- amino function by several

others Boucherle et al

and Di M o d i c a and Angeletti

have prepared

several sulfonic and sulfanilic acid derivatives. Benn et al.

have

pre-pared derivatives of p-di-2-chloroethyl amino substituted sulfanilic

acid esters and sulfonamides. Pearison, Holland and M i d g e t

using

the method of Elks and Hey, have synthesized some triazeno

deriva-tives of sulphanilamide carrying 3,3-di(2— hydroxyethyl), diethyl,

cyclopentyl, piperidine, ethoxyethyl, morpholino groups and also

triazeno derivatives of sulfadiazine carrying diethyl and cyclopentyl

groups. Then assayed the antibacterial activities of these compounds

coming to the conclusion that the antibacterial affect is slight in c o m

-parison with sulfanilamide.

Considering that the antineoplastic activity- is born by the

subs-tituted triazeno groups but heterocyclic moiety provides the molecules

C o m p o u n d R R2 m . p . C°

I 2 - h y d r o x y ethyl 2—hydroxy ethyl 152

I I ethyl ethyl 2 3 6 III — H 2 - h y d r o x y ethyl 193 I V — H morpholino 198 V — H N - p i p e r i d i n o 183 V I — H cyclohexyl 2 2 2 V I I — H benzyl 2 1 8 V I I I — H propyl 2 2 9

I X cyclohexyl cyclohexyl chars at 3 0 0

X — H phenyl 2 1 0

X I - H 2 - b u t y l 195

X I I — H t - b u t y l 2 2 0 X I I I n - b u t y l n - b u t y l 2 1 5

X I V methyl methyl 2 0 2

with the fact that either amide or ester moiety increases the

antileu-kemic activity when tested in standard mouse L-1210 leukemia assay,

it has been though to incorporate triazeno group into some suitable

molecule which has already been proved to involve in metabolic

processes. Since sulfonamides posses the amide moiety and have been

proved to involve in metabolic processes with little toxic side effects

they meet the requirements mentioned above. Additionally Ross et

al.

have shown that sulfonamides deposit selectively in the neoplastic

tissues because the pH of such tissues is lower then most of the normal

tissues due to the high rate production of lactic acid from glucose under

under aerobic conditions

After all the considerations mentioned above, one of the

sulfo-namides, sulfaguanidine was selected as a carrier for substituted

tria-zeno group.

The compounds synthesized are tabulated in Table. I along with

their melting points.

T a b l e I. — T h e triazeno derivatives of Sulfaguanidine

R

R

Generally the triazeno derivatives of sulfaguanidine are highly

insoluble products both in polar and nonpolar solvents but sparingly

soluble in ethanol. They are insoluble in aqueous sodium carbonate

but but soluble in aqueous sodium hydroxide developing a brown color.

They are also soluble in acid medium giving a dark color, but nmr

spectrum taken in trifluoroacetic acid has shown decomposition of

the structure and loss of alkyl groups situated at the 3 position of

triazeno moiety. In the ir spectra of triazeno sulfaguanidines there

are two sets of bands. A set of them are steady in all spectra but

characteristic of sulfamide structure and the other set of bands are

characteristic of triazene structure. These bands are listed in Table.II.

T a b l e I I . T h e ir bands of the derivatives of triazeno sulfaguanidine

Steady bands in all derivatives: (in c m- 1)

3 2 0 0 — 3 5 0 0 Generally a trident 1 6 0 0 — 1 6 4 0 a fork 1550 s 1250 s 1 1 4 0 — 1150 s 1080 — 1100 ms 8 2 0 — 8 5 0 m 5 5 0 m

Characteristic bands of triazenes: 1 4 5 0 — 1 4 7 0 a spike 1 4 0 0 — 1 4 1 0 a spike 1 0 5 0 — a small spike or a dent

7 3 0 — 7 4 0 a small spike or a dent 6 5 0 — a spike, a fork or a triple band

The UV spectra of the compounds synthesized are very similar.

Generally the bands shift very slightly. The values of the bands are:

208-210 nm the bands shift very slightly. The values of the bands

are: 2 0 8 - 210 max., 226-228 nm min., 236-250 nm max., 268-278

nm min., 350-366 nm max. The tlc of all the derivatives on Silicagel

plates with n-Bu O H : A c O H : Wa ( 5 : 2: 3) allways gave one spot

having an Rf value of about 0.50 - 0.66. M a n y of these compounds

are being biologically tested. The results are to be published later.

E X P E R I M E N T A L

All the solvents used were technical grade except those used

for recystallization and spectrometry for which only analytical grade

and spectral grade solvents were used. Melting points have been taken

with a Mettler FP - 5 instrument. All melting points are uncorrected.

UV spectra were taken with a Pye-Unicam SP - 1700 instrument

and ır spectra were taken with a Pye - Unicam SP-1100 instrument,

nmr spectra were taken with a Perkin Elmer R 32, 90 M H z instrument.

For the synthesis of all the derivatives basicly the same ractions have

been applied.

General Method:

Reaction A- In a 1 It flask 40.00 g (0.187 Mol) Sulfaguanidine

was dissolved in 170 ml (conc.) HC1 , to this solution 400 ml water

was added. The flask was kept in an ice-bath. When the inner

tem-perature lowered to 0-5°C sodium nitrite solution (13 g / 50 ml in

water and cooled to 0°C) was added dropwise. This addition was

car-ried on till it gave reaction with the KI-starch paper.

Reaction B- When the diazotization reaction was over, 0.3 M o l

of the amine derivative which was dissolved in the mininum quantity

of water and cooled to 0°C was added to this mixture for the

coup-ling reaction. The product which precipitated was filtered and washed

with cold water. During the entire reaction the temperature was

kept below 5°C. When possible, the product was recrystallized from

ethanol/water and dried in vacuum over sulphuric acid.

D I S C U S S I O N

One of the major problems in cancer chemotherapy is the

diffi-culty of finding a compound that acts differentially on the cancer

tissues but not on normal proliferating tissues. In order to realize such

a discrimination it was thought to benefit from the differences

bet-ween normal and neoplastic tissues considering their biochemical

properties, particularly the p H . The fundamental metabolic property

of all cancer cells is the production of lactic acid from glucose under

aerobic conditions. Consequently the pH of neoplastic tissues has a

lower value than most of normal tissues. Therefore one of the ways

in which this lower pH of tumour tissue can be exploited is to utilize

this property to accumulate an effective carcinostatic chemical gruop

via a carrier of suitable PKa value.

In this research,expecting a favorable carcinostatic activity,

sulfaguanidine molecule has been chosen as carrier and substituted

triazeno group has been chosen as carcinostatic moiety and they were

incorporated.

S U M M A R Y

One of the major biochemical property of neoplastic tissues is

their low pH value. This property has been taken into consideration

to incorporate a well known carcinostatic group, substituted triazeno

moiety onto a suitable carrier, sulfaguanidine.

By substitution of triazeno moiety fourteen different molecules

have been synthetized. All of the compounds have been characterized

by their melting points, Rf value, uv, ir and nmr spectra.

These compounds are being tested for antineoplastic activity,

the data are to be published later.

Ö Z E T

Kanserli hücrelerin temel biyokimyasal özelliklerinden birisi de

düşük pH değerleridir. Bu özellik dikkate alınarak iyi bilinen bir

karsinostatik grup, sübstitüe trizeno grubu uygun bir taşıyıcıya,

sül-faquanidine takılmıştır.

Triazen yapısının sübstitüsyonu ile 14 değişik molekül sentez

edilmiştir. T ü m bileşikler E.n. ları, Rf değerleri, U V , İR ve N M R

spektrumları ile tanımlanmışlardır.

Bu bileşikler henüz antineoplastik etki yönünden

deneylenmek-tedir. Elde edilen bilgiler daha sonra yayınlanacaktır.

R E F E R E N C E S

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1 9 . S k i p p e r , H . E . , S c h a b e l , F . M . J r . , W i l c o x , W . S., Cancer Chemotherapy Rept., 3 5 , 1 (1964)

2 0 . S k i p p e r , H . E . , S c h a b e l , F . M . J r . , W i l c o x , W . S.,Cancer Chemotherapy, Rept., 4 5 , 5 ( 1 9 6 5 ) .

I. T e z l e r : 1 . V e t . H e k . S e v i n ç T ü r k e r . " E t v e B a l ı k K u r u m u A n k a r a K o m b i n a s ı n d a k e s i l e r e k p i y a s a y a v e r i l e n P a r ç a - P a k e t e t l e r i n H i j y e n i k K a l i t e l e r i n i n M i k r o b i y o l o j i k y ö n d e n a n a l i z i " . D o k t o r a T e z i ( 1 9 7 7 ) . 2 . Ecz. B i l g e Ş e n e r " O r t h o r u s h e t e r o c a r p u s ( B o i s s ) Juz. b i t k i s i n i n k ö k l e r i ü z e r i n d e F a r m a k o g n o z i k araş-t ı r m a l a r " D o k araş-t o r a T e z i ( 1 9 7 7 ) . 3 . Ecz. G ü l d e n S e z i k " T ü r k i y e ' d e y e t i ş e n H e l i c h r y s u m t ü r l e r i ü z e r i n d e F a r m a s ö t i k B o t a n i k y ö n ü n d e n a r a ş ­ t ı r m a l a r " D o k t o r a T e z i ( 1 9 7 7 ) . 4 . E c z . M u a l l a Y e n e n " B o r e a v a o r i e n t a l i s J a u b . e t S p a c h . b i t k i s i n i n m e y v a l a r ı ü z e r i n d e F a r m a k o g n o z i k a r a ş -t ı r m a l a r " D o k -t o r a T e z i ( 1 9 7 7 ) . 5. E c z . N i l ü f e r Kafalı " G l i b e n k l a m i d t a b l e t l e r i n i n ç ö z ü n m e h ı z ı n d a y a r d ı m c ı m a d d e l e r ile z a m a n , s ı c a k l ı k v e n e m i n e t k i l e r i n i n i n c e l e n m e s i " D o k t o r a T e z i ( 1 9 7 7 ) . 6 . E c z . Z e y n e p M u t a c e d d e d " M i k r o s p e k t r o f o t o m e t r i k y ö n t e m l e C O H b t a y i n i n s t a n d a r d i z a s y o n u v e b u y ö n t e m ­ l e m e s l e k s e l o l a r a k C O ' e m a r u z k a l a n l a r d a C O i n h a l a s y o n u n u n s a p t a n m a s ı " D o k t o r a T e z i ( 1 9 7 7 ) . 7 . E c z . N a z i r e Ö z k a l " G l y c r r h i z a g l a b r a L . b i t k i s i n i n T ü r k i y e ' d e y e t i ş m e k t e olan v a r y e t e l e r i n i n F a r m a k o g -n o z i k k a r ş ı l a ş t ı r ı l m a s ı " D o k t o r a T e z i ( 1 9 7 7 ) . 8 . D r . M e v l ü t E r t a n " B a z ı 1,2,4-triazoller v e N - g l i k o z i t l e r i ü z e r i n d e s e n t e z ç a l ı ş m a l a r ı " D o ç e n t l i k T e z i ( 1 9 7 7 ) . I I . S e m i n e r v e K o n f e r a n s l a r : 1 . P r o f . R . B O U R D O N , 2 3 . M a y ı s - 3 . H a z i r a n . 1 9 7 7 t a r i h l e r i a r a s ı n d a f a k ü l t e m i z d e bir s e r i k o n f e r a n s v e s e m i n e r v e r m i ş t i r .

2 . P r o f . D r . E . N E U Z İ L , 9 1 7 . K a s ı m . 1 9 7 7 tarihleri a r a s ı n d a f a k ü l t e m i z d e bir seri k o n -f e r a n s v e s e m i n e r v e r m i ş t i r .

I I I . D e r s K i t a p l a r ı :

1 . " V i t a m i n l e r v e E n z i m l e r " P r o f . D r . G a z a n f e r B İ N G Ö L , A . Ü . Ecz. Fak. yayınları N o :

ve 3 5 8 sayılı K a r a r ı ile Fakülte M e c m u a s ı n d a yayınlanacak yazılar için tebit edilen esaslar

1 ) D e r g i d e , b a ş k a bir m e c m u a d a aynı isimle v e aynı t a r z d a n e ş r e d i l m e m i ş orijinal ça­ lışmalar y a y ı n l a n ı r .

2 ) Yazılar K o m i s y o n a v e r i l d i ğ i t a r i h sırasıyla y a y ı n l a n ı r .

3 ) M e t i n 1 5 d a k t i l o sayfasını g e ç m e m e k ü z e r e T ü r k ç e v e y a y a b a n c ı dilde yazılabilir. M e t i n başlığı v e ö z e t i T ü r k ç e v e y a b a n c ı dilde yazılacaktır.

Y a b a n c ı dilde y a z ı l m ı ş başlık, m e t i n v e ö z e t l e r i n dil k u r a l l a r ı n a u y g u n o l m a s ı n ı n t e m i ­ ni, y a z a r a aittir.

4) Yazılar, k â ğ ı d ı n bir y ü z ü n e , d a k t i l o ile ve n o r m a l aralıkla y a z ı l m a l ı , italik yazılacak k e l i m e l e r i n altı çizilmeli, klişesi yapılacak g r a f i k , ş e m a , f o r m ü l g i b i ş e k i l l e r , çini m ü r e k k e p ile, a y d i n g e r k â ğ ı d ı n a ç i z i l m e l i ; f o t o ğ r a f l a r p a r l a k k â ğ ı d a v e k o n t r a s l ı o l a r a k ç e k i l m e l i d i r . Ş e k i l l e r i n h e r biri a y r ı k â ğ ı t l a r d a o l m a l ı v e k â ğ ı d ı n ü z e r i n d e y a z a r ı n a d ı , kaçıncı şekil o l d u ğ u , r e s i m altı yazılması i s t e n e n i b a r e k a y d e d i l m e l i d i r .

5 ) Yazı plânı a ş a ğ ı d a k i ş e k i l d e o l m a l ı d ı r : K o n u n u n t a k d i m i , b u l g u l a r , denel k ı s ı m , m ü n a k a ş a , T ü r k ç e ö z e t , y a b a n c ı dilde ö z e t , literatür.

K o n u n u n t a k d i m i 2 d a k t i l o sahifesini g e ç m e m e l i ; m a t e r y a l , m e t o t v e yapılan a m e l i y e ­ ler " d e n e l k ı s ı m " d a y e r almalı, " m ü n a k a ş a " k ı s m ı , g e r e k l i ise k o n m a l ı d ı r .

L i t e r a t ü r , m e t i n d e p a r e n t e z i ç i n d e k i n u m a r a l a r l a b e l i r t i l m e s i v e m e t i n s o n u n d a b u n u m a r a l a r a u y g u n o l a r a k s ı r a l a n m a l ı d ı r . Sırasıyla y a z a r ı n s o a y d ı , a d ı n ı n ilk harfi, m e c m u a ­ n ı n milletlerarası kullanılan kısaltılmış i s m i , cilt n u m a r a s ı (italik), sayfa ve p a r e n t e z içinde t a r i h y a z ı l m a l ı d ı r .

6) T a s h i h l e r y a z a r tarafından yapılacaktır. 7 ) Y a z a r a 5 0 a y r ı b a s k ı verilir.