DIPEPTIDE DERIVATIVES AND THEIR PRODUCTION

DERIVES DIPEPTIDIQUES ET PROCEDE DE PRODUCTION

English Abstract

A B S T R A C T
Dipeptides represented by the formula:
<IMG>
wherein R represents hydrogen, C1-C6 alkyl group, C2-C7
cyanoalkyl group, or C3-C10 dialkylaminoalkyl group; R1
represents hydrogen, methyl, isopropyl, isobutyl, benzyl or
phenyl; R2 represents C1-C6 alkyl group, C7-C14 aralkyl group,
glycyl group or glycylglycyl group; R3 represents hydrogen or
C1-C6 alkyl group, group R2-N-R3 optionally represents phthalimide,
piperidino group 4-hydroxy-4-(p-halogenophenyl)-piperidino,
morpholino or piperazino group substituted by alkyl or phenyl
group; A ring represents benzene ring or pyridine ring optionally
substituted by halogen and B ring represents halo-benzene ring
and its acid accition salt, compounds are useful as anxiolytics,
sedatives, anticonvulsants, hypnotics or muscle relaxants.

Claims

THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE PROPERTY
OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A process for preparing a compound represented by the
formula:
<IMG>
wherein R represents hydrogen, C1-C6 alkyl group, C2-C7
cyanoalkyl group, or C3-C10 dialkylaminoalkyl group, R1 repre-
sents hydrogen, methyl, isopropyl, isobutyl, benzyl or phenyl;
R2 represents hydrogen, C1-C6 alkyl group, C7-C14 aralkyl group,
glycyl group, or glycylglycyl group; R3 represents hydrogen,
C1-C6 alkyl group or amino-protecting group, group R2-N-R3
optionally represents phthalimido, piperidino group, 4-hydroxy-4
-(p-halogenophenyl)-piperidino, morpholino or piperazino group
substituted by C1-C6 alkyl group or phenyl group; A ring repre-
sents benzene ring or pyridine ring optionally substituted by
halogen, and B ring represents halo-benzene ring, said process
comprising one of the following steps:
1) when R2 is other than hydrogen and R is other
than amino-protecting group, subjecting the amine
represented by the formula:
<IMG>
29

wherein R, A ring and E ring are as defined above
and the glycine derivative represented by the formula:
<IMG>
wherein R1, R2 and R3 are each as deinfed above to
the amido bonding formation in a conventional manner
for the peptide condensation;
2) when R2 is other than hydrogen and R3 is other
than amino-protecting group, subjecting the glycylamide
represented by the formula:
<IMG>
wherein R, A ring and B ring are as defined above and
the amino acid represented by the formula:
<IMG>
wherein R1, R2 and R3 are each as defined above to the
amido bonding formation;
3) reacting the glycylamide represented by the formula:
<IMG>

wherein R, A ring and B ring are as defined above
with the halogenoacetyl halide represented by the
formula:
<IMG>
wherein X and X1 represent each halogen and R1 is
as defined above to give the halogenoacetyl-glycylamide
represented by the formula:
<IMG>
wherein R, R1, X, A ring and B ring are each as defined
above and then reacting the latter with ammonia, alkali
phthalimide or amine represented by the formula:
R2NHR3
wherein R2 and R3 are each as defined above;
4) subjecting the methylol compound represented by the
formula:
<IMG>
31

wherein R, A ring and B ring are as defined above
and the glycine derivative represented by the formula:
<IMG>
wherein R1, R2 and R3 are each as defined above to
the amido bonding formation to give the peptide repre-
sented by the formula:
<IMG>
wherein R, R1, R2, R3, A ring and B ring are each as
defined above and then oxidizing the latter;
5) subjecting the 2-aminomethylindole represented
by the formula:
<IMG>
wherein R, A ring and B ring are as defined above but
B ring is benzene ring and the amino acid represented
by the formula:
<IMG>
32

wherein R1, R2 and R3 are each as defined above to
the amido bonding formation to give the amide repre-
sented by the formula:
<IMG>
wherein R, R1, R2, R3, A ring and B ring are each
as defined above and then oxidizing the latter; or
6) subjecting a compound represented by the formula:
<IMG>
wherein R represents hydrogen, C1-C3 alkyl group,
C2-C7 cyanoalkyl group, C3-C10 dialkylaminoalkyl group
or halogen, R2-?-R3 represents phthalimido group; R4
represents halogen and R5 represents hydrogen or
halogen to hydrazinolysis to give a compound represented
by the formula:
<IMG>
wherein R, R4 and R5 are as defined above.
33

2. A process of claim 1 for preparing compounds represented
by the formula:
<IMG>
wherein any one of said steps 1 through 5, R represents hydrogen,
C1-C3 alkyl group, C2-C5 cyanoalkyl or C3-C8 dialkylaminoalkyl
group; R1 represents hydrogen or methyl, isopropyl, isobutyl,
benzyl or phenyl; R2 represents C1-C3 alkyl group, glycyl group
or glycylglycyl group; R3 represents hydrogen or C1-C3 group,
R2-?-R3 optionally represents phthaloyl group; R4 represents
halogen and R5 represents hydrogen or halogen and in steps 3,
4 or 5, R2 is additionally hydrogen and R3 is additionally amino-
protecting group.
3. A process of claim 2, wherein any one of said steps 1
through 5, R is hydrogen or methyl; R1 is benzyl; R2 and R3
are each C1-C3 alkyl group, and R4 and R5 are each halogen, and
in steps 3, 4 or 5, R2 and R3 are additionally each hydrogen.
4. A process of claim 1 for preparing 2-benzoyl-4-chloro-
N-methyl-N.alpha.-glycyl-glycinanilide, wherein any one of said steps
3 through 5, R is methyl; R1, R2 and R3 are each hydrogen; said
A ring is a benzene ring and said B ring is substituted at the
4-position with chloro.
34

5. A process of claim 1 for preparing 2-o-chloro-benzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein any one of
said steps 3 through 5, R is methyl; R1, R2 and R3 are each
hydrogen and said A and B rings are benzene rings substituted
respectively at the 2-position with chloro and at the 4-position
with chloro.
6. A process of claim 1 for preparing 2-o-fluorobenzoyl-
4-chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein any one of
said steps 3 through 5, R is methyl; R1, R2 and R3 are each
hydrogen and said A and B rings are benzene rings substituted
respectively at the 2-position with fluoro and at the 4-position
with chloro.
7. A process of step 3 of claim 1 for preparing 2-o-chloro-
benzoyl-4-chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein
2-o-chlorobenzoyl-4-chloro-N-methyl-N.alpha.-haloacetylglycinanilide
is reacted with ammonia.
8. A process of step 3 of claim 1, for preparing 2-o-
chlorobenzoyl-4-chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein
2-o-chlorobenzoyl-4-chloro-N-methyl-N.alpha.-iodoacetylglycinanilide
is reacted with ammonia.
9. A process of step 6 of claim 1, wherein R is hydrogen,
cyanomethyl or diethylaminoethyl; R4 is chlorine and R5 is
hydrogen, chlorine or fluorine.
10. A process of step 6 of claim 1 for preparing 2-o-fluoro-
benzoyl-4-chloro-N-diethylaminoethyl-N.alpha.-glycyl-glycinanilide,
wherein R is diethylaminoethyl; R4 is chloro and R5 is fluoro.

11. A process of step 6 of claim 1 for preparing 2-o-chloro-
benzoyl-4-chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein R
is methyl, R4 and R5 are chloro.
12. A process of step 6 of claim 1, for preparing 2-o-fluoro-
benzoyl-4-chloro-N-methyl-N.alpha.-glycyl-glycinanilide, wherein R
is methyl, R4 is chloro and R5 is fluoro.
13. A process of step 6 of claim 1, for preparing 2-benzoyl-
4-chloro-N-cyanomethyl-N.alpha.-glycyl-glycinanilide, wherein R is
cyanomethyl, R4 is hydrogen and R5 is chloro.
14. A process of step 6 of claim 1, for preparing 2-o-chloro-
benzoyl-4-chloro-N-cyanomethyl-N.alpha.-glycyl-glycinanilide, wherein
R is cyanomethyl and R4 and R5 are chloro.
15. A process of steps 3, 4 or 5 of claim 1 for preparing a
compound represented by the formula:
<IMG>
wherein R is hydrogen, C1-C3 alkyl group, C12-C15 cyanoalkyl
group, or C3-C8 dialkylaminoalkyl group; R1 represents hydrogen,
methyl, isopropyl, isobutyl, benzyl or phenyl; R4 represents
halogen and R5 represents hydrogen or halogen which comprises
cleaving the amino protecting group from the compound of the
formula prepared by a selected one of said steps 3, 4 or 5:
36

<IMG>
wherein R3 represents said amino protecting group and R, R1, R4
and R5 are as defined above.
16. A compound represented by the formula:
<IMG>
wherein R represents hydrogen, C1-C6 alkyl group, C2-C7
cyanoalkyl group, or C3-C10 dialkylaminoalkyl group, R1 repre-
sents hydrogen, methyl, isopropyl, isobutyl, benzyl or phenyl;
R2 represents hydrogen, C1-C6 alkyl group, C7-C14 aralkyl group,
glycyl group, or glycylglycyl group; R3 represents hydrogen,
C1-C6 alkyl group or amino-protecting group, group R2-?-R3
optionally represents phthalimido, piperidino group, 4-hydroxy-4-
(p-halogenophenyl)-piperidino, morpholino or piperazino group
substituted by C1-C6 alkyl group or phenyl group; A ring repre-
sents benzene ring or pyridine ring optionally substituted by
halogen, and B ring represents halo-benzene ring, when prepared
by steps 3, 4, 5 and 6 of claim 1 and when R2 is other than
hydrogen and R3 is other than amino protecting group, by steps
1 and 2 of claim 1.
37

17. A compound represented by the formula:
<IMG>
wherein R represents hydrogen, C1-C3 alkyl group, C2-C5
cyanoalkyl or C3-C8 dialkylaminoalkyl group; R1 represents
hydrogen, methyl, isopropyl, isobutyl, benzyl or phenyl;
R2 represents C1-C3 alkyl group, glycyl group or glycylglycyl
group; R3 represents hydrogen or C1-C3 alkyl group; R2-?-R3
optionally represents phthalimido; R4 represents halogen and
R5 represents hydrogen or halogen, when prepared by the process
of claim 2.
18. A compound according to claim 17, wherein R is hydrogen
or methyl; R1 is benzyl, R2 and R3 are each C1-C3 alkyl group
and R4 and R5 are each hydrogen, when prepared by the process
of claim 3.
19. A compound of claim 16, namely 2-benzoyl-4-chloro-N-
methyl-N.alpha.-glycyl-glycynanilide, when prepared by the process
of claim 4.
20. A compound of claim 16, namely 2-o-chloro-benzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 5.
21. A compound of claim 16, namely 2-o-fluorobenzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 6.
38

22. A compound of claim 16, namely 2-o-chloro-benzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 7.
23. A compound of claim 16, namely 2-o-chloro-benzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 8.
24. A compound of the formula:
<IMG>
wherein R represents hydrogen, C1-C3 alkyl group, C2-C7 cyano-
alkyl group or C3-C10 dialkylaminoalkyl group; R1 represents
hydrogen; R4 represents halogen and R5 represents hydrogen or
halogen, when prepared by the process of claim 1, step 6.
25. A compound of claim 24, whrein R is hydrogen, cyano-
methyl or diethylaminoethyl; R1 is hydrogen, R4 is chlorine and
R5 is hydrogen, chlorine or fluorine, when prepared by the
process of claim 9.
26. A compound of claim 24, namely 2-o-fluorobenzoyl-4-
chloro-N-diethylaminoethyl-N.alpha.-glycyl-glycinanilide, when
prepared by the process of claim 10.
27. A compound of claim 24, namely 2-o-chlorobenzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 11.
39

28. A compound of claim 24, namely 2-o-fluorobenzoyl-4-
chloro-N-methyl-N.alpha.-glycyl-glycinanilide, when prepared by the
process of claim 12.
29. A compound of claim 24, namely 2-benzoyl-4-chloro-N-
cyanomethyl-N.alpha.-glycyl-glycinanilide, when prepared by the process
of claim 13.
30. A compound of claim 24, namely 2-o-chlorobenzoyl-4-
chloro-N-cyanomethyl-N.alpha.-glycyl-glycinanilide, when prepared by
the process of claim 14.
31. A compound of claim 17 of the formula:
<IMG>
wherein R is hydrogen, C1-C3 alkyl group; C2-C5 cyanoalkyl
group or C3-C8 dialkylaminoalkyl group; R1 represents hydrogen,
methyl, isopropyl, isobutyl, benzyl or phenyl; R4 represents
halogen and R5 represents hydrogen or halogen, when prepared by
the process of claim 15.

Description

.. 1/~9~6SZ
The present invention relates to dipeptide derivatives ana
their production. More particularly, this invention relates to
dipeptide derivatives represented by the formula:
R R R2
NCOCH2NHCOCHN \ 3 (I)
C ~ ~ R
wherein R represents hydrogen, cl-C6alkyl group, c2-c7alken~1
group, C2-C7cyanoalkyl group, C2-C7carbamoylalkyl group, C3-C10
dialkylaminoalkyl group or cyclopropylmethyl group, R represents
hydrogen, cl-c6alkyl group, C7-C14aralkyl group, c7-cl4hydroxyar-
alkyl group, C6-C12aryl group, C2-C7carbamoylalkyl group, C2-C7
carboxyalkyl gr~up, cl-c6aminoalkyl group, C4-ClOguanidylalkyl
group, Cl-C6mercaptoalkyl group, c2-c7alkylthioalkyl group, Cg-
C15indolylalkyl group or C4-Cgimidazolylalkyl group, R represents
hydrogen, Cl-C6alkyl group, c7-C14aralkyl group, C6-C12 aryl
group, glycyl group or glycyl-glycyl group, R3 represents hydro-
gen, Cl-C6alkyl group or amino-protecting group, R and R
optionally combine to form c2-C4alkylene group, group R -N-R
optionally represents phthalimido group, piperidino group, 4-
hydroxy-~-(p-halogenophenyl)piperidino,morpholino, or piperazino
group substituted by Cl-C6alkyl group or phenyl group, A ring
represents benzene ring or pyridine ring optionally substituted
by halogen and B ring represents benzene ring or thiophene ring
optionally substituted by halogen, trifluoromethyl group, methyl-
sulfonyl group, nitro group or Cl-C6alkyl group and their acid
addition salts, being useful as anxiolytics, sedatives, antico-
vulsants, hypnotics, muscle relaxants or their synthetic inter-
mediates.
Illustrative explanation is given to the above definition
as follows: alkyl group (e.g. methyl, ethyl, isopropyl, butyl,
., 1 ~

1~91652
pentyl), alkenyl group (e.g. allyl, butenyl, pentadienyl),
cyanoalkyl group (e.g. cyanomethyl, cyanoethyl, cyanopropyl,
cyanobutyl), carbamoylalkyl group (e.g. carbamoylmethyl,
carbamoylethyl, carbamoylpropyl), carboxyalkyl group (e.g.
carboxymethyl, carboxyethyl, carboxypropyl), aminoalkyl group
(e.g. aminomethyl, aminoethyl, aminopropyl, aminobuty]),
hydroxyalkyl group (e.g. hydroxymethyl, hydroxyethyl, hydroxy-
propyl, hydroxybutyl), guanidylalkyl group (e.g. guanidylmethyl,
guanidylethyl, guanidylpropyl), mercaptoalkyl group (e.g~
mercaptomethyl, mercaptoethyl, mercaptopropyl, mercaptobutyl),
alkylthioalkyl group (e.g. methylthiomethyl, ethylthiopropyl,
methylthiobutyl), indolylalkyl group (e.g. indolylmethyl,
indolylethyl, indolylpropyl), aralkyl group (e.g. benzyl, phen-
ethyl, phenylpropyl), hydroxyaralkyl group (e.g. hydroxybenzyl,
hydroxyphenethyl), aryl group (e.g. phenyl, naphthyl), halogèn
(e.g. chlorine, bromine, fluorine, iodine), alkylene group (e.g.
dimethylene, trimethylene, tetramethylene), amino-protecting
group (e.g. carbobenzoxy, methoxycarbonyl, t-butoxycarbonyl,
p-methoxybenzyloxycarbonyl, o-nitrophenylsulfenyl, chlorobenzyl-
oxycarbonyl, trityl), and dialkylaminoalkyl group (e.g. dimethyl-
aminoethyl, diethylaminoethyl, diethylaminopropyl).
The dipeptide derivatives (I) can be prepared as shown in
the ~ollowing scheme~
_

!L~165Z
R2 R
3~ NCHCONHCH2COOH (III)
NHR
-- CO ~ Route A .
(II)
IR
NCOCH2NH2
CO ~ (salt)
( IV) \ NCHCOOH
\ R ~
Route B\ (V)
Rl
XCHC3X
Route C NCOCH2NHCOCH ~ NCOCH2NHCOCHN
CO ~ R NHR ~ ~ CO ~
(VIII) ~__J ~ ~ (I)
Oxidation
/ 1 2 Oxidation
~HCOCH2NHCOCN ~ 3 NCH NHCOC N-
(XII)
NCHCONHCH2COOH Route D ~ -~`NCHCOOH (V)
3~ Route E R3//
R (III) OH NHR
CH ~ 2 2
(IX)
3 _ (XI)

91652 1 2
wherein x and X represents each halogen and R, R , R , R , A
ring ànd B ring each is as defined above but B ring is XI and XII
is benzene ring.
Route A
-
This route is effected by su~jecting the starting amine (II)
and the glycine derivative (III) to the amido bonding formation
in a conventional manner for the peptide condensation. The
amido bonding formation substantially involves the condensation
of amino group on the amine (II) with carboxy group on the gly-
cine derivative (III) to forM the peptide bonding and also other
accessory treatments for attaining this object, inclusive of
treatment ~or converting the carboxy group of the glycine deri-
vative (III)into its reactive derivatives in advance of the
amido bonding formation, treatment for previous protection of
active group (e.g. amino group, carboxy group) which should not
participate in the reaction and treatment for removin~ such pro-
tecting groups after the amido bonding formation. The conversion
of the glycine derivative (III) into the reactive derivative in-
volves halogenation, anhydride formation, azide formation, active
ester formation, etc. Introduction and removal of such protecting
groups can be effected in a conventional manner. Illustrating
an example about amino group, the arnino group of the peptide can
be protected by treating with carbobenzoxy chloride in the pre-
sence of an alkali, and this amino-protecting group o~ the final
product (I~ can be removed by treating with such an acid as
hydrobromic acid, hydrofluoric acid or trifluoroacetic acid or
by hydrogenating or raducing with liquid ammonia/metallic
sodium. Trityl group can be introduced by treating with trityl
chloride in the presence of a base and can be removed by treating
with dilute acetic acid, and phthalyl group can be removed by
i `
...... .. . .. . .. . . ... ... .

;)9~65Z
treating with hydrazine hydrate. This step is cJenela]ly carried
out in an insert solvent (e.g. methylene chloride, dilnethylforma-
mide, dimethylsulfoxid~, hexamethylphosphoric triamide, chl~ro-
form, dioxane, benzene, tetrahydrofuran, a mixture ~hereof) at
room temperature or under cooling or heating. The cJeneral pro-
cedure for the amido bonding formation described in Rou-te A is
similarly applicable in the amido bonding formation of other
routes hereinafter described.
Route B
This route is effected by subjecting the glycylamide (IV)
and the amino acid (V) to the amido bonding forma-tioll. The
starting glycylamide being in a form of its acid addition salts
te.g. hydrobromide, hydrochloride) is also prepared by the
amido bonding formation of the amine (II) and glycine. The
amido bonding formation of this route is substantially carried
out as in Route A. For example, the glycylamide (IV) is treated
with phthalyl-glycyl chloride in a suitable solvent (.e.g.
dimethylformamide, hexamethylphosphoric triamide)to give the
phthalyl-glycyl-glycylamide (I), which is converted into the
Einal product (I) by hydrazinolysis.
Route C
This route is effected in -two steps by at Eirst reacting
the glycylamide (IV) preferably in a corm oE its acid addition
salt (e.g. hydrochloride, hydrobromide) with the halogenoacetyl
halid3(VI) to give the halogenoacetyl-g:lycylamicle (VII) and
secondly reacting the latter with the ammonia, alkali phthalimide
or amine (VIII). For elevating the reactivity of the intermediate
(VII), the halogen of the compound (VIII) may be substituted by
the other more active halogen before the second step, for
example, by treating with alkali halide (e.g. po-tassium iodide,

- ~9~6SZ
sodi~m iodidc, potassiu~ brom;d~). These reactions are effected
in an inert solvent (e.g. dimethylformamide, hexamethylphosphoric
triamide, tetrahydrofuran, acetone, chloroform, diglyme) at room
temperature or under cooling or heating in a conventional manner.
Route D
This route is effected in two steps by sub~ecting the
methylol compound (IX) and the glycine derivative (III) to the
amido bonding formation and then oxidizing the resulting peptide
(X). The starting methylol (IX) can be prepared by reducing the
corresponding carbonyl compound (II). The amido bonding formation
is carried out as in Route A, and the oxidation is carried out by
treating with such an oxidizing agent as Jone's reagent (chromic
acid/sulfuric acid/water), manganese dioxide, chromic anhydride
or the like in a conventional manner.
Route E
This route is effected in two steps by subjecting the 2~
aminomethylindole (XI~ and the amino acid (V) to the amido bonding
~ormation and then oxidizing the resulting amide (XII). The
amido bonding ~ormation is carried out as in Route A. The oxida-
tion is carried out by using such an oxidizing agent as oxygen,
ozono, hydrogen peroxide, chromic acid, peracid (e.~. peracetic
acid), potassium permanganese, manganese dioxide or sodium
periodate in a conventional manner for oxidizing a double bond
into carbonyl groups.
When the product (I) contains amino-protecting group, it
can be removed according to its necessity. Therefor a conventional
procedure for removing amino-protecting group from peptides can
be adopted as described in Route A.
The product ~I) can be converted into suitable acid addition
salts such as those of inorganic acid (e.g. hydrochloric acid,
... ~ ... ~ . . . , . . .: ... . .

1~91~5~
sulfurie aeid, n;tr;~ aeid, p~osrhor1e aeid, thioeyanic acid) or
those of organic acid (e.g. aeetic acid, suceinic acid, oxalic
acid, maleic aeid, malie aeid, phthalic acid, methanesul~onic
aeid) for the neeessity of preparation, crystalization, solubility
or improvement of stability.
Thus obtained dipeptide derivatives (I) and their aeid addi-
tion salts are useful as anxiolytics, sedatives, anticonvulsives,
hypnoties, musele relaxants, or their synthetie intermediates.
Pharmaeologieal aetivities of some dipeptide derivatives (I) are
shown in the following table in eomparison with ehlordiazepoxide
and diazepam.
1. Compound tested:
Compound No. Compound Name Note
1 2-benzoyl-4-chloro-N-methyl-
Naglyeyl-glyeinanilide
2 2-o-ehlorobenzoyl-4-ehloro-N-
methyl-Na-glycyl-glycinanilide
hydrate
3 2-o-fluorobenzoyl-4-ehloro-N-
methyl-N~-glyeyl-glyeinanilide
hydroehloride
4 Chlordiazepoxide Control
Diazepam Control
2. Test method:
1) Anti-pentylenetetrazol aetivity:
This test was measured on a group of 10 DS mal0 miee.
25 Within 15 minutes after subeutaneous injeetion of 125 mg/kg
o~ pentylenetetrazol, the mouse showed tonie eonvulsion which
eeased fatally. In this test, the test eompound was given
orally 60 minutes prior to the administration of pentylene
tetrazol. The observation was made for two hours after the
administration of pentylenetetrazol. The eriterion o~ anti-
, . . ~ ~ . , , , ^ .

109165Z
convulsant activity was determined as beina com~lete Protection
against mortality. The convulsions were disregarded. Results
were shown as ED50 [Goodman, et al.: J. Pharmacol., 108, 168
(1953)~.
2) Taming activity:
This test was measured on DS male mice.
When a 5 Hz square wave pulse (10 msc, 50 v) was delivered to a
pair of mice in a grid box, some pairs showed fighting posture 15
to 20 times for 3 minutes. These pairs of mice were selected in
the morning and used for drug test in the afternoon. The testcompound was administered to both mice of the pairs 60 minutes
prior to the experiment. Results were obtained as the mean per-
centage of inhibition of fighting responses in a group of 5 pairs
and shown by ED50 [Tedeschi, et al.: J. Pharmacol Exp. Thev.,
15 125, 28-34 (1959)~ .
3) Rotarod performance activity:
This test was measured on DS male mice.
The mouse was put on a scraped rod of wood, 3 cm in diameter,
turning at the rate of five rotations per minute. The mice that
could remain on the rod for three or more minutes in successive
trials were selected and placed in a group of 10 mice for each
dose. If the mouse fell down from the rod within less than 2
minutes, the test compound was considered to be effective.
Results were shown by ED50 [Dunham, et al.: J. Am. Pharm. Assoc.,
a5 ~6, 208 (1957)~.
4) Acute toxicity:
The test compound was orally administered to DS male mice in
different single doses. For each dose, 10 mice were used, their
weight ranging from 20 to 23 grams. The mice were observed for
72 hours after the administration of the compound. The mortality

~LOg~L65?
~as calculated by the Bliss method ~Bliss: Ann. Appl. Biol.,
22. 134-307 (1935): Qant. J. Pharmacol., 11, 192 (1938)~.
- 3. Result:
~able 1.
Compound Anti-pentylene- Taming Rotarod Acute
No. tetrazol acti- activity performance toxicity !
vity, ED50 ED50 ED50 LD50
(mg/kg) (m~/kg) (mg/kg)(mg/kg)
1 1.0 10.5 36.6 1309
2 0.58 3.0 27.7 1255
3 0.33 1.6 17.0 >1000
4 3.7 24.0 50.0 1090
1.19 6.0 9.03 1459
15 4. Conclusion: ~
Each of the five test compounds is very weak in the acute
toxicity, and any predominant difference i5 not observed between
them. Compounds on the subject o~ this invention (Compound Nos.
1-3) are about 2 to 4 times less potent in the disturbing effect
of motor coordination due to the rotarod performance than diaze-
pam (Compound No. 5). In the anti-pentylenetetrazol activity and
the taming activity, 2-benzoyl-4-chloro-N-methyl-N~-glycyl-g
cinanilide (Compound No. 1) is about 2 to 3 times moxe potent
than chlordiazepoxide (Compound No. 4), and 2-o-chlorobenzoyl-4-
chloro-N-methyl-NR-glycyl-glycinanilide hydrate (Compound No. 2)
and 2-o-fluorobenzoyl-4-chloro-N-methyl-N~-glycyl-glycinanilide
hydrochloride (Compound No. 3) are about 2 to 4 times more potent
than diazepam ~Compound No. 5).
The dipeptide derivatives (I) and their pharmaceutically
acceptable acid addition salts axe applied singly or in combination
; 9

~91t~52
w;th pharmaceutically suitable carriers such as wheat starch,
corn starch, potato starch, gelatin, etc. The choice of carriers
is determined by the preferred route of administration, the solu-
bility of the substance and standard pharmaceutical practice.
Examples of pharmaceutical preparations are tablets, eapsules,
pills, suspensions, syrups, powders, and solutions. These com~o-
sitions can be prepared in a conventional manner. A suitable
dosage of the dipeptide derivatives (I) or their pharmaceutically
acceptable acid addition salts for adults is in the order of about
1 mg to 30 mg per day.
Still, the dipeptide derivatives (I) and their acid addition
salts are useful as growth promotors of domestic cattles and fowls.
Presently preferred and practical embodiments of the present
invention are illustratively shown in the following examples.
Example 1
-
tl) To a solution of trityl-glycyl-glycine (5 g) in hexa-
methylphosphoric triamide ~24 ml), thionyl chloride (1.6 g) is
added dropwise at -8 to -2 C, and the resultant mixture is stirred
at -5 C for 20 minutes. The mixture is mixed with 2-amino-5
ehlorobenzophenone (3.08 g) and allowed to stand at room tempera-
ture overnight. The reaction mixture is neutralized with an
aqueous sodium bicarbonate solution and shaken with chloroform.
The organie layer is washed with water, dried and evaporated to
remove the solvent~ The residue is crystalliæed from ether to
give 2-benzoyl-4-chloro-N~-trityl-glycyl-glycinanilide (1.7 g).
The product is recrystallized from ethyl acetate to give needles
melting at 187 to 189 C. W:~ 237.5, 274 (sh.), 343 my (log
: 4.51, 4.03, 3.53).
(2) A suspension of 2-benzoyl-4-chloro-N-trityl-glycyl-
30 glycinanilide (1.7 g) in 50 % acetic acid (20 ml) is heated on a
c
.. , . . ~ , ., . . . ; ~ . . . .

gl652
water bath for 20 minutes. After coolin~, the preci~tat~ cry--
stals are filtered. The filtrate is neutralized with aqueous
sodium bicarbonate solution and shaken with chloroform. The
organic layer is washed with water, dried and evaporated to re-
move the solvent, whereby 2-benæoyl-4-chloro-N -glycyl-glycinani-
lide ~0.8 g) is obtained. The product is recrystallized from
ethyl acetate to give prisms melting at 135 to 136 C. W : A max
241, 275 (sh.), 340 m~ (log ~: 4.44, 4.03, 3.55).
Examples 2-5
Using the following starting materials (II) and (III) the
reaction is effected as in Example 1 to give the corresponding
products (Ia) and (Ib):
NHR 3 1
R R
CO ~ + R N-CHCON~ICH2COOH (III)
(Il)
R R 2
NCOCH2NHCOCHN 3
~ C~3 . ... .. _ ~
Cl (Ia)
R
NCOCH2NHCOCHNHR
~ C0\ ~
Cl (Ib)
wherein R represents amino-protecting group, and R, R and R
are as defined above.
11

9~L~;52
Table 2
. . ~
Example II III Ia Ib
No. - ---- - _ _ _ _ _
R Rl R2 R3 mp( C) mp(c)
_ ._ .. _ ~
2 ~I H H ~bz 163-164 135-1~6
3 Me H H Tri Amorph Amorph
4 H Me H Cbz 148-149 131-132
_ i-Bu H cbz 98-100 145-147
0 Note: The abbreviations in the table have the following signi-
ficance: H (Hydrogen), Me (Methyl group), su (Butyl group),
Cbz (Carbobenzoxy group), Tri (Trityl group), i- (iso-),
mp (Melting point), a) (L-form).
Example 6
(1) To a solution of carbobenzoxy-L-leucyl-glycine (4.05 g)
in dry meth~lene chloride (50 ml), triethylamine (1.75 ml) and
ethyl chlorocarbonate (1.2 ml) are added at -10 C, and the mixture
is stirred at the same temperature for 20 minutes. Still, a
solution of 2-amino-5-chlorobenzophenone (2.91 g) in dry methylene
chloride (50 ml) is added at 0 C gradually thereto, and the re-
sultant mixture is stirred under ice cooling for 15 minutes and
1 hour and at room temperature for 30 minutes and 1 hour, then
refluxed overnight. The reaction mixture is poured onto a mixture
of potassium carbonate and ice and shaken with methylene chloride.
The organic layer is washed with water, dried and evaporated to
remove the solvent. The residue is chromatographed on a column
of silica gel containing water (3 %), which is eluted with benzene
to recover the starting 2-amino-5-chlorobenzophenone (1.21 g) and
then eluted with benzene/ethyl acetate (9:1) to give a product.
The product is recrystallized from ether to give 2-benzoyl-4-chloro-
j 12

~as~s~
N -carbobenzoxy-L-leucyl-glycinanilide (3.13 g) as crystal~
melting at 98 to 10Q C. IR: 3425, 3315, 1700, 1640 cm (CHC13).
(2) In acetic acid solution (15 ml) containing hydrobromic
acid (24 %) 2-benzoyl-4-chloro-N~-carbobenzoxy-L-leucyl-glycin-
5 anilide (3.1 g) is dissolved under ice cooling, and the resultantsolution is stirred at room temperature for 1.5 hours. The solu-
tion is mixed with ether and allowed to stand for 30 min~tes.
The precipitate is filtered, dissolved in cold water and shaken
with methylene chloride/ether (1 2~. After removing the organic
layer, the aqueous layer is made alkaline with an a~ueous potas-
sium carbonate solution, saturated with sodium chloride and shaken
with chloroform. The chloroform layer i9 washed with water, dried
over anhydrous magnesium sulfate and evaporated to remove the
solvent. The residue is recrystallized from ether -to give 2-
benzoyl-4-chloro-N~-L-leucyl-glycinanilide (1.628 g) as crystals
melting at 145 to ]47 C. IR: 3325, 1685, 1639 cm (CHC13).
~D +50.7+0.9 (EtO~). Mass, m/e 401 (M~).
Examples 7 to 9.
Using the following starting materials (II) and (III), the
reaction is effected as in Example 6, whereby the corresponding
products ~Ia) and (Ib) are obtained:
NHR
~ CO ~ + 3/ N-OEICONHCH2COOH (III)
(II) Cl
R , /
NCOCH2NHCOCHN~ 3
CO ~ R _
Cl (Ia)
~0
13
` ;

65Z
R
NCOCH2NHCOCHNHR
- ~l,CO ~3
Cl (Ib)
wherein R, R , R and R are each as defined above.
Table 3
¦ Example II III Ia Ib
. _ -
No R Rl R2 R mp(C) m~t c) _ _
7 H H Me cb~ 131-133 143-145
8 H i-Pr H Cbz 158-168 119-12]
9 H Ph H Cbz 93-95 65-67
___
Note: The abbreviations in the table have the following si~ni~
ficance: Pr(Propyl group), Ph (Phenyl group), the others
are each as defined above.
Example 10.
To a solution of carbobenzoxy-glycine (1.05 g) in hexamethyl-
phosphoric triamide (8 ml), thionyl chloride (0.6 g) is added at
-4 to -6 C, and the mixture is stirred at -6 C for 10 mlnutes.
To the mixture 2-benzoyl-4~chloro-glycinanilide (1.44 g) is added,
and the resultant mixture is stirred at temperature below n C for
2 hours and allowed to stand overnight at room temperature. The
reaction mixture is made alkaline with aqueous sodium bicarbonate
solution and shaken with chloroform. The organic layer is washed
with water, dried and evaporated to remove the solvent. The
residue is recrystallized from ether/water to give 2-benzoyl-4-
chloro-N~-carbobenzoxy-glycyl-glycinanilide (1.9 g) as crystals
melting at 163 to 164C.
Example 11.
`i 14

~ 652
To a suspension of N,N-dimethylglycine hydrochLoride (0.97
g)in anhydrous pyridine (15 ml), triphenyl phosphite (2.15 g)
is added, and the resultant mixture is stripped at room tem~erature
overnight. A solution of 2-benzoyl-4-chloro-N-methyl.-N -glycyl-
anilide (2.0 g) in dry pyridine (10 ml) is added th~reto, and theresultant mixture is sitrred at room temperature for 103 hours. The
reaction mixture is evaporated under reduced pressure. The
residue i5 made alkaline with an aqueous potassium carbona-te
solution and shaken with me-thylene chloride/ether (1 : 2). The
organic layer is washed with water and evaporated. The residue
is made acidic with 3 N hydrochloric acid and shakerl with ether.
After removing -the ethereal layer, the aqueous layer is made
alkaline with an aqueous potassium carbonate solution and shaken
with ether. The ethereal layer is washed with a saturated saline
solution, dried over anhydrous sodium sulfate and evapora-ted to
remove the solvent. The residue (1.9 g) is dissolved in methanol
(2 ml), mixed with a solution of oxalic acid (0.6~ g) in water .
(2 ml) and evaporated under reduced pressure to dryness. The
obtained crystals are washed with e-ther Eour times to give 2-
benzoyl-4-chloro-N-methyl-N -dime-thylclycyl-ylycinanilide oxala-te
(1.75 g) as crystals melting at temperature above 90~C (decomp.).
IR: 3463, 1719, 1694 (sh.), 1668 (sh.), 1640 cm ~C[IC13),
Ex.ample 12
(1) To a solution oE N-carbobenzoxy-phenylalanine (3 g) in
hexamethylphosphoric triamide (16 ml), -thionyl chloride (1.2 g)
is added dropwise to -6 to -2C in S minutes, and the resultant
mixture is stirred at -6 to -8C for 10 minutes. To the mixture,
a suspension of l-methyl-2-amino-methyl-3-o-chlorophenyl-5
chloroindole hydrochloride (3~52 g) in ether (15 ml) previously
treated with triethylamine is added, and the resultan-t mixture is

- ~Wl~S;~
allQ~ed tn ~nd at room temperature overnight. The reaction mix-
ture is neutr211~ed ~ith an aqueous sodium bicarbonate solution
and shak~n with ether. The organic layer is dried and evaporated
to remove the ether. The residue is crystallized from ether ~o
give 1-methyl-2-(N~-carbobenzoxy-phenylalanylaminomethyl)-3-0-
chlorophenyl-5-chloroindole (3.15 g). This substance is recry-
stallized from ethyl acetate to give needles melting at 174 to
176 C. The yield is 54 %. W : ~E OH 232, 285 m~ (log ~=4.88,
3.2~).
(2) To a solution of l-methyl-2-(N~-carbobenzOxyphenylalanyl-
aminomethyl)-3-o-chlorophenyl-5-chloroindole (2.86 g) in acetic
acid (15 ml), a solution of chromic anhydride (1.5~ g) in water
(1.4 ml) is added dropwise at 13 to 21C for 5 minutes, and the
resultant mixture is stirred at room temperature for 4 hours. The
reaction mixture is mixed with icy water and shaken with chloro-
form. The organic layer is washed with water, dried and evapora-
ted to remove the solvent. The residue is chromatographed on a
column of silica gel, which is eluted with ether to give 2-o-
chlorobenzoy1-4-chloro-N-methyl-Na-carbobenzoxy-phenylalanyl-
glycinanilide (1.75 g) as a gelatinous substance. W : 1 max 256
(sh.), 298 (sh.) m~ (log f=4.01, 3.44).
(3) A solùtion o~ hydrobromic acid (21.8 %) in acetic acid
is added to 2-o-chlorobenzoyl-4-chloro-N-methyl-N -carbobenzoxy-
phenylalanyl-glycinanilide (1.65 g) and the resultant mixture is
stirred at room temperature for 1.5 hours. The reaction mixture
is mixed with dry ether, and the precipitated crystals are fil-
tered to give 2-o-chlorobenzoyl-4-chloro-N-methyl-Na-phenylalanyl-
glycinanilide hydrobromide hydrate (1.3 g) as crystals melting at
206 to 209 C (decomp.). W : ~ m 258 (sh.), 300 (sh.) m~ (log
~=3.97, 3.35).
16

- - ~3Ll)91652
Examr)l e 13
(1) usi7lg l~mf -hyl-2-aminomethyl-3-o-chlorophenyl-5-chloro-
indole hydrochloride and N-tritylglycine, the reaction is effected
as in Example 12 (1), whereby 1-methyl-2-(N-tri-tyl-glycyla~inO-
methylJ-3-o-chlorophenyl-5-chloroindole is obtained as crystals
melting at 198 to 200 C.
(2) To a suspension of l-methyl-2-(N-trityl-glycylamin-
methyl)-3-o-chlorophenyl-5-chloroindole (2.02 g) in acetic acid
(10 ml), a solution of chromic anhydride (0.81 g) in water
(0.6 ml) is added, and the resultant mixture is stirred at room
temperature for 22 hours. The reaction mixture is mixed with
water (22 ml), and the precipitated crystals are filtered. The
filtrate is mixed with 28 % aqueous ammonia solution (12 ml) and
shaken with chloroform. The organic layer is washed with water,
dried and evaporated to remove the solvent. The residue is dis-
solved in ethanol and mixed with a solution of oxalic acid in
ethanol. The precipitated crystals (0.5 g) are recrystallized
~rom ethanol to give 2-o-chlorObenæoyl-4-chloro-N-methyl-N-
glycyl-glycinanilide oxalate as crystals melting at temperature
below 167 C. W : ~ a H 253, 298 (sh.) m~ (log ~=3.98, 3.34).
Example 14
(1) Using l-methyl-2-aminomethyl-3-o-chlorophenyl-S-
chloroindole hydrochloride and N-carbobenzoxy-glycine, the reac-
tion is effected as in Example 12 (1), whereby 1-methyl-2-(N-
carbobenzoxy-glycylaminomethyl)-3-o-chlorophenyl-5-chloroindole
is obtained as crystals melting at 96 to 98 C.
(2) To a solution of l-methyl-2-(N-carbobenzoxyglycylamino-
methyl)-3-o-chlorophenyl-5-chloroindole (9.1 g) in acetic acid
(55 ml), a solution of chromic anhydride (5.5 g) in water (5.1 ml)
is added dropwise at temperature below 20 C, and the resultant
17
. .

~1~9~6SZ
~' soluti~n i~ allowe~ ~o stand at room ~mperature overnight. The
reaction mixture is mixed with icy water and shaken with ethyl
~ .
acetate. The organic layer is washed with water, dried and
~ evaporated to remove the solvent. The residue is chromatographed
i, 5 on a column of silica gel, which is eluted with ethyl acetate to
~,
give 2-o-chlorobenzoyl-4-chloro-N-methyl-N~-carbobenzoxy-glycyl-
glycinanilide (3.6 g) as a gelatinous substance. This substance
is mixed with a solution of hydrobromic acid (21.8 %) in acetic
acid (11. 5 ml) and stirred at room temperature for 1.5 hours.
~;.
'~ 10 The reaction mixture is mixed with ether to precipitate crystals.
1"f
~ The crystals are filtered, dissolved in water and neutralized
Jt'~ with an aqueous sodium bicarbonate solution. The precipitate is
. ..
filtered to give 2-o-chlorobenzoyl-4-chloro-N-methyl-N -glycyl-
,~ glycinanilide hydrate (1.8 g). This substance is recrystallized
. ~:
15 from aqueous alcohol to give prisms melting at 95-100 C.
Examples 15 to 21.
i
Using the ~ollowing starti~g compounds (XI) and ~V~, the
reactions are effected as in Example 1~, whereby the corresponding
- products (XII), (Ia) and (Ib) are obtained:
: ~
` ~ Rl
~: Cl ~ \ NCHCOOH (V)
R R R1 R2
I CH2NHCOCHN / 3 NcOcH2NHCOCHN < 3
ClJ~ ~ R Rl Cl
X NCOCH2NHCOCHNHR2
~ '' ? ~CO-~
~ 30 - ~ ~ Cl (Ib)
. , .
;' 18
.; .
~,...... .

1~91tiSZ
, ~
~able ~
Exam- X: ~ ¦ v ¦ XII ~ la I Ib
No. R ~ R ~3 mp( C) I mp(C) ¦ mp(C)(salt)
b) MeCl ~ Bz ~1 Cbz150-155 ~Syrup 116 ~(Oxalate~
16 MeCl ~ Bz H Cbz168-169 ,Syrup (HBr)
l _ . ~ .
17 Me Cl H H Cm158-159 1104-106 95-100 (H2O)
18 Me H i-Pr H I Cbz242-247 Syrup ~ 130(Hemi- I
I ... _ _, i
lga) Me H Bz H ~ Cbz255-257 Syrup 137-140 (HCl)
___ , . . ...
Me H H H ¦ Cbz150-153 Syrup -- 60
. I _ l .. .
21 ~ Me F H H j Cbz166-167 Syrup 80 ~ (HCl)
Note: The abbreviations in this table have the following signi.-
ficance: Cm (Carbomethoxy group), Cl (Chlorine), ~
(decomposition~, F (Fluorine), a) Levo, b) DextrO, and the
others are as defined above.
Exam~le_22.
(1) To a solution of crude l-methyl-2-aminomethyl-3-o-
20 chlorophenyl-5-chloroindole (9.97 g) in di.oxane (300 ml), potas-
sium carbonate (2.48 g) is added at room temperature with stirring,
and the mixture is mixed with N-phthalylglycyl chloride (8.036 g),
The resultant mixture is stirred at room temperature for 30
minutes. The reaction mixture is concentrated to a volume of
25 about 100 ml, which is mixed with n-hexane (100 ml). The preci-
pitated crystals are filtered, and dissolved in chloroform (2 L)/
methanol (100 ml) to give a solution, which is washed with water
dried over anhydrous sodium sulfate and evaporated to remove the
solvent. The residue is washed with ether to give l-methyl-2-
(N -phthalyl-glycylaminomethyl)-3-o-chlorophenyl-5-chloroindole
`; 19

~9~65~
t9.642 g). The qame ~r~dllc~ n m~ is o~tained from the diox-
ane/n-hexane mother liquor and the ethereal washings. The yield
is 62.8 %. This substance is recrystallized from methanol/
chloroform to give crystals melting at 253 to 254 C.
(2) To a solution of l-methyl-2-(N~-phthalylglycylamino-
methyl)-3-o-chlorophenyl-5-chloroindole (1.00 g) in acetic acid
(25 ml~, a solution of chromic anhydride (406 mg) in water (2 ml)
is added gradually with stirring. The resultant mixture is
stirred at 22 to 25C for 4 hours and concentrated under reduced
pressure to a~out half a volume. The residue is mixed with ice,
and the precipitate is ~iltered. The filtrate is shaken with
ethyl acetate, and the said precipitate is dissolved in the
ethyl acetate layer. The ethyl acetate layer is chromatographed
on a column of silica gel, and the eluate is evaporated to give a
precipitate, which is recrystallized from methylene chloride/
methanol to give 2-o-chlorobenzoyl-4-chloro-N-methyl-N -phthal~l-
glycyl-glycinanilide (580 mg) as crystals melting at 216 to 218 C~
The yield is 54.5 %.
(3) To a solution of 2-o-chlorobenzoyl-4-chloro-N-methyl-
N~-phthalyl-glycyl-glycinanilide (1.056 g) in dimethylformamide
(20 ml), a solution of hydrazine hydrate (180 mg) in dimethyl~or-
mamide (4 ml) is added at -8 to -6 C with stirring, and the
resultant mixture is stirred at -8 C to room temperature for 1
hour. A~ter cooling at 0C, the reaction mixture is mixed with
N-hydrochloric acid (4 ml) in 20 minutes and allowed to stand at
0 C ~or 17 hours. The reaction mixture is poured into a mixture
of icy water (200 ml) and ethyl acetate tlO0 ml) and made alkaline
to pH 8 with 28% aqueous ammonia solution. The ethyl acetate
layer is separated, washed with water, dried over anhydrous
sodium sulfate and evaporated under reduced pressure to give a
'; 20

~1652
residue (500 mg). The S3~ s~bstan~ ~410 ~ is ~bt~i.ned from
the aqueous layer and washings. Both are combined, dissolved in
ethanol (10 ml) and mixed with water (25 ml~ under cooling below
0 C. The precipitated crystals are filtered to give 2-o-chloro-
benzoyl-4-chloro-N-methyl-N~-glycyl-glycinanilide hvdrate (722
mg) as crystals melti.ng at 95 to 100C. The hemicitrate melt~ at
114 to 116 C. The vield is 87 ~/O.
ExamPles 23 to 25
. .
Using the follo~-ing com~ound (XI) an~ N-~hthalvl-glvcYl
chloride, the reactions are effected as in Exam~le 22, whereby.
the corresponding products (XII), (Ia) an~ (Ib) are obtàined:
C].J~-/~ Cl ,f3 ~0CH2 I!.T~3
(XI) (XII)
R ll
X NCOCH2NHCOCH2N ~
--;' ~co~ O
C1 (Ia)
X NcocH2NHcocH2Nll2
~ (Ib)
Cl
Table 5.
_ _ _ _ ~._
~ mple No. XI XII Ia Ih
_ .__ ___ _ ______._ ~___.__
_~ R _ _ _X _ _mp( C) mP(C~ --- m C)
23 MeH > 300 _ ~ 60 (AmorPh)
~ ____ ~_ ___ __ __._ ___._ .___ ____
. 24 2H > 300 232-234 (d~ trate~ _
_ _ . _
-CH2CN ~ Cl 288-289 184-186 139-143(d~
_ ! (d~ _ (Hemicitrate~
_ ___ _. _~__ ___ _ .______ .... _. __. _
; 21

65~
Note: The abbreviations are as defined above.
Example 26.
(1) To a solution of 2',5-dichloro-2-meth~laminobenzophen-
one (3.20 g) in benzene (80 ml), phthalyl-glycyl-glycyl chloride
(~.0 9) is adde~, and the resultant mixture is stirred at 70 to
80 C for 1 hour. The precipitated crystals are filtered, washed
with benzene and then ethanol and dried to give 2-o-chlorobenzovl-
4-chloro-N-methyl-N~-phthalyl-glycyl-glycinanilide (5~6 g), which
is recrystallized from ethanol to give crystals melting at 217 C.
(2) A suspension of 2-o-chlorobenzoyl-4-chloro-N-methyl-N
phthalyl-glycyl-glycinanilide (81.0 g) in ethanol (50 ml) is
mixed with hydrazine hydrate (20 ml), and the resultant mixture
is refluxed for about 30 minutes. After cooling, the reaction
mixture is filtered to remove the insoluble phthalhydrazide.
The filtrate is evaporated to remove the solvent, and the residue
is crystallized from dilute ethanol and washed with ether to give
2-o-chlorobenzoyl-4-chloro-N-methyl-N~-glycyl-glycinanilide
hydrate (55.3 g). This substance is recrystallized from dilute
ethanol to give crystals melting at 95 to 100 C.
Example 27.
(1) To a suspension of sodium borohydride (1.2 g) in tetra-
hydrofuran (10 ml), a solution of 2',5-dichloro-2-methylaminoben-
zophenone (3.12 g) in tetrahydro~uran (20 ml) is added dropwise.
The resultant mixture is mixed with water (5 ml) and stirred at
room temperature overnight. The reaction mixture is mixed with
a small amount o~ water and evaporated under reduced pressure to
remove the solvent. The residue is made to pH 8-9 with dilute
hydrochloric acid and shaken with chloroform. The organic layer
is dried and evaporated to give 2',5-dichloro-2-meth~laminobenz-
hydrol (3.05 g). This substance is recrystallized ~rom ether/n~
22

~9165~
hexane to give crystals melting at 105.5 to 106.5 C. The vieldis 97.1 %.
(2) To a solution of carbobenzoxy-glycyl-glycine (4.0 g)
in hexamethylphosphoric triamide (20 ml)/acetonitrile (lO ml),
thionyl chloride (1.77 g) is added dropwise at -18 C, and the
resultant mixture is stirred for 3 minutes at -18 C. A solution
of 2',5-dichloro-n-methylaminobenzhydrol (2.2 g) in hexamethyl-
phosphoric triamide (lO ml)/acetonitrile (5 ml) is added drop-
wise thereto at -18C, stirred at the same temperature for 8
hours and allowed to stand at -20C overnight. After the reac-
tion, the reaction mixture is mixed with water/ether, made
alkaline with an aqueous sodium bicarbonate solution and qhaken
with ether. The organic layer is dried and evaporated to remove
the solvent. The residue is chromatographed on a column of
silica gel, which is eluted with ethyl acetate to gi~e 2-o-chloro-
~-hydroxybenzyl-~-chloro-N-methyl-~ -carbobenzoxy-glycyl-
glycinanilide (3.31 g) as crystals melting at 57 to 60 C.
(3) To a solution of 2-o-chloro-~-hydroxybenzyl-N-methyl-
N -carbobenzoxy-glycyl-glycinanilide (21.8 g) in acetone (300 ml),
~ones reagent (a solution of chromic acid and sulfuric acid in
water) is added dropwise until the reaction mixture keeps red.
The reaction mixture is filtered to remove the precipitate. The
red ~iltrate is mixed with isopropanol until the red solution
becomes green. The mixture is filtered, and the filtrate is
neutralized with an aqueous sodium bicarbonate solution and eva-
porated. The residue is mixed with water and shaken with chloro-
form. The chloroform layer is purified with active c~rbon to
give 2-o-chlorobenzoyl-4-chloro-N-methyl-~ -carbobenzoxy-glycyl-
glycinanilide (21.3 g). The yield is 9~.5 %.
Examples 28 to 33.
23
.. . , .. ~ . . . . . . . .

lC~91652
Using the following compound (IX), the reaCtiOnS are effec-
ted as in Example 27 but when phthalyl group is adopted for
amino-protection, it is removed by hydrazinolysis, whereby the
f~llowing compounds (X), tIa) and (Ib) are obtained:
R R 3~1 R2
X ¦ ~ ~COCH2NHCOCHN ~ 3
CH~ ~ (X)
R I ~
0 ' NCOCH2NHCOCHN ~ 3
~ (~CO~/ \>
(Ia)
Cl
R
R
,X NCOCH2NHCOCHNH2
~ ~ CG- ~ (Ib)
:~
Cl
~ 20 Table 6.
.: _
Example IX X Ia ¦ Ib
No. R X R R R ~ mp~ C) mp( C) I mp( C)
28 Me H H H Cbz ! 75-78 45-50 ~60 ~Amorph)
_ __
. 29a) MeCl Bz H Cbz 70 Amorph 110 ~ (Amorph) _
MeCl H Ft 200-201 223-226 95-100 (H2O)
.~ .
31 Me F H Ft 193-194 1 213-214 80~ (HCl)
_ (d) _
32 De F H Ft 166-168 186-187 Amorph
_. _ _ ___ .
33 ¦ Me ¦ Cl i- r H Cbz 172-173 _ 100 (HCl)
Note: The abbreviations have the following significance: Ft
'; 24

~L09165~
,~
(Phthalyl group), De (Diethylaminoethyl group), a) Leve,
and the others are as defined above.
Examp le 3 4 .
(1) To a solution of 2-o-chlorobenzoyl-4-chlOro-N-methyl-
glycinanilide hydrobromide (1.8 g) in hexamethylphosp~oric tria-
mide (10 ml), chloroacetyl chloride (0.73 ~) is added under ice
cooling. The resultant mixture is stirred under ice cooling for
2 hours and at room temperature for 3 hours. The reaction mi~-
ture is shaken with ether and the organic layer is made alkaline
with an aqueous ammonia solution, washed with water and evaporated
to give 2-o-chlorobenzoyl-4-chloro-N-methyl-N-chloroacetyl-
glycinanilide (1.6 g). This substance is recrystallized from
ethyl acetate to give colorless n~eedles melting at 134 to 136 C.
(2) A mixture of 2-o-chlorobenzoyl-4-chloro-N-methyl-N -
chloroacetyl-glycinanilide (6.2 g), potassium iodide (2.74 g~
and acetone t60 ml) is refluxed for 1 hour. The reaction mixture
is evaporated to remove the acetone, and the residue is dissolved
in chloroform. The organic layer is washed with water, dried and
evaporated. The residue is washed with ether to give 2-o-chloro-
benzoyl-4-chloro-N-methyl-Na-iodoacetyl-glycinanilide (6.9 g).
This substance is recrystallized from ethyl acetate to give
colorless needles m~lting at 168.5 to 169.5 C.
~3) Into a suspension of 2-o-chlorobenzoyl-4-chloro-N-
methyl-N~-iodoacetyl-glycinanilide tl.l g) in tetrahydrofuran
(20 ml) ammonia gas is introduced for 30 minutes, and the mixture
is stirred at room temperature for 5 hours. The reaction mixture
is evaporated to remove the tetrahydrofuran. The residue is
dissolved in chloroform. The organic layer is washed with an
aqueous sodium bicarbonate so]ution and then water, dried and
evaporated to remove the chloroform. The residue is chromato-
;- 25

L6S2
~J.a~hed on a column of silica gel, which is eluted with methanol
to give 2-o-chlorobenzoyl-4-chloro-N-methyl-N~-glycyl-glycinani-
lide.
Example 35.
(1) The reaction is effected as in Example 34 (1) by using
bromoacetyl bromide in lieu o~ chloroacetyl chloride, whereby
2-o-chlorobenzoyl-4-chloro-N-methyl-N -bromoacetyl-glycinanilide
is obtained as colorless needles melting at 153 to 155C. The
yield is 69 %.
(2) To a solution of 2-o-chlorobenzoyl-4-chlorO-N-methyl-
N -bromoacetyl-glycinanilide (1.01 g) in dimethylformamide
(10 ml), potassium phthalimide (0.34 g) is added, and the resul-
tant mixture is stirred at room temperature for 3 hours and
allowed to stand at room temperature overnight. The reaction
mixture is mixed with water (100 ml), and the precipitate is
filtered and washed with water to give 2-o-chlorobenzoyl-4-
chloro-N-methyl-N -phthalyl-glycyl-glycinanilide (1.0 g).
Example 36.
(1) ~o a solution of 2-o-chlorobenzoyl-4-chlorO-N-methyl-
20 glycinanilide hydrobromide (0.628 g) in dimethylformamide (7 ml),
phthalyl-glycyl chloride (0.437 g) is added, and the res~lltant
mixture is stirred for 3 hours. The reaction mixture is evaporated
to remove the solvent. The residue is shaken with chloroform,
and the chloroform layer is washed with water, dried and evapora-
ted. The residue is washed with ether to give 2-o-chlorobenzoyl-
4-chloro-N-methyl-N -phthalyl-glycyl-glycinanilide (0.71 g~. The
yield is 93.5 %.
(2) The above product is treated with hydrazine hydrate to
give 2-o-chlorobenzoyl-4-chloro-N-methyl-N -glycyl-glycinanilide
hydrate.
, 26

~'.xam~ s 37 to ~G. ~:IL09165Z
g thc ~ol]owinc3 amilles ~ ie~ f allllnonia t~-e rea(:tions
are effected as in ~xample 34 (3), wl1ere~)y ~h~ coLresponding
pr o d u c t s ( I ) a r ~ oll t a
Me
Cl 2 2
j- CO--~ ~> R NTlR (VIII)
(VXI) Cl Me R2
ICl Ncocl~2~ (;ocll2~ 3
].n <~co~;
cl ( I)
Tcll)] ~ 7 .
E:~a~n~)le VIII
No. ~ __..... _ . . ..... ...... _..... ..
15 ~_._ ___ ___ ____ __ ____.____ _ rnp ( C ) ( sal t !
37 Pipcrid;.nc ~l-B:3
_ _.. _.. _ ~ .,__ . _.. ........ ...... . ... ....
3~ D.i.eth~, lam;.ne ll.3-l]S
~ . _.. __ . . .. . . .. _ . .. . . _ .. . . . .. .. .... . . . I
39 4-hyd3. o~ ( p-c11 loropllell~ 1) 1 4 4-1~16
p .iper :i d in e
. _._ __. ._____ . _ ---- ----- -- l-- -- -- -- - -- --
~û Dlmel:llyJ~t~ e I 132-1:33
~. .___ __ _ ______ _ _.... ...... ...... _ ..... ......... .
~11 Mor pl~o 1 ine :l 20- 12~ 1
_ __ _________._.___ ~ .. _ .. ........ .. ..... . .. _ .. _ _.. ...
~12 Methy].clllli.ne I ~. on- L02
~3 4-~ cn~ ;i.pc3 a7Aine ¦ 155-164 (d) (2l~t,'1) i
_~ _ ___._ _. ._ __.. _ _.. .. _.. . . .. I
4~1 4-met11ylpiperazin~ ¦ 226-.!28 (cl) (2~C1)
__ t ~ --------~ ~ ~ ~ -
2 5~15 I so prol?~ l c~ .in e I 19 7 - 2 (~(~ ( d ) ( ~C l )
~__ _ ___ ___._______. _, _ .. ._ .... ....... ... .... . . ....
_ _ Pllell~tllylalllille ! L/6--.1. 1~ (()xal a.t~)
__ _._ .. _ . _ _ __ _ _ .. _.. _ .. _ ___ ._ _ ._.. ___ _ .. _ _ .. __ _~ .. _ . _ . _ .___ .. _ . .. . _ .. ~ . .
Notc: Th~ ab~)reviations arc as d~E.inc-3 al~c~ve
Exam~)lcs 47 to 5 0.
Thc reactions ~re ~f~fccted cas in E:~alllPle J., w~lereby tl
30 followi.nc~ pro~ucts ( I) are obtai.nc~l:
27
.
. .
.. . .

16~iZ
Example Compound Name mp(C)
47 2~ picolinoyl)-4-chloro-N~-glycyl-glycin- 192-194(d)
anilide dihydrobromide
48 3-benzoyl~5-ethyl-2-(N~-glycyl-glycyl)amino-
thiophene hydrochloride 190-192
49 2-o-chlorobenzoyl-4-chloro-N-methyl-N -di-
glycyl-glycinanilide 100
2-o-chlorobenzoyl-4-chloro-N-methyl-N -tri-
glycyl-glycinanilide Amorph
" 28