Note: Descriptions are shown in the official language in which they were submitted.
08~
This invention relates to ~ew chemical compounds,
to processes for their preparation and to the use o~
these compounds as sterilising agents and preservatives,
especially in the cosmetics industry, and as antiseptics
~or e~mple in dermatology. -~
.- .
SU~RY OF TI~ INVENTION
-- . .
The compounds according to the present i~vention
haYe the follol~ing ~ormula:
R - X - CH2 - CH(OH) - CH2NH2 (I)
in which
R represents a saturated or ~thylenically or acetylenically
unsaturated straight-chain C~-C18 aliphatic radical and
X represents -O-, -S-, -NH- or CH2-9 preferably an
o~ygen or sulphur atom.
The present inventio~ alsa relates to the salts and -
esters o~ these compounds, more espeoially their
hydrochlorides, hydrobromides arld acetates, and also
to their alkylated quaternar~!r a~monium derivatives~
The nonyl, decyl and dodecyl radicals are
.~
mentioned in particular as e~amples o~ the saturated
aliphatic radical which m~y be represented by the radical
Rn
More particularly, the present inYention relates -
to compounds corresponding to the formula~
Rl - Xl - CH2 - CII(O~) - CH2NH2 (Ia) ~;
in which
Rl is an unbranched C5 to C14 al~yl chain and ~;
Xl is -O- or -S-,
~ ~mt
and even more particularly to the compound o~ formula Ia
in whieh Rl is the decyl radical and Xl is oxygen, iOe. -~3-decylo~y-2-hydroxy-1-aminopropane in the form of its
hydrochloride, hydrobromide or acetate.
The compounds according to the in~en~ion and in
particular the compounds corresponding to formula Ia may
be prepared by processes similar to those used ~or
preparing the lower homologues (cf. in particular:
Chem. Abs. 1953, 47, lOI~ 195l~, 48, 108i; 1954~ 48,
7549b; 1955, 49, 10850g; 1956, 50, 10700f; 1965, 62,
16783e; and 1971, 757 44187u).
~owe~er, the compounds of ~ormula (I), in which
X is oxygen or sulphur, are pre~erably prepared by the
followin~ prooess: `
(l)n~Xl~l 2-c~{-c~2~l2~ xl-c~2-~Fi
(III) (II)
(2)1~-Xl-C~2-C -C~2~3 ~ ~H2-N~
(II) 0
~ C, NH
R~ CH2--1~--CH2N~2 ~ 1 IT ..
H ~ ~ CI NH
(I) 1 ,
; 15 In these ~ormulae, ~1 represents -0- or -S-, R
has the same meaning as in ~ormula (I) and R3 and R2
together represent a~ epo~y radical or R3 represents
a hydro~y radical and R2 a halogen atom~ pre~erably
chlorine or bromine.
~.
~ ~mt
'
' ~
~3~ 33
The reaction (1~ i9 carried out in the presence of
a compound, such as potassium or sodium carbonate, which
is capable of making the phthalimide react in the ~orm o~
its sodium or potassium derivative.
The hydrolysis reaction (2) is carried out in the
presence of hydrochloric acid.
The compound of formula (III) may be prepared for
example by reactin~ an epihalohydrin:
C~2-CH-CH2Hal (IY)
0 '
with an nlcohol in the presence o~ a catalyst, suoh as
stanic chloric, zinc chloride, ferric chloride, the boron
fluoethe~ate complex or tosylic acid. The product obtained
is a hal~igenated alcohol.
The epo~ide of formula (III) may be prepared by
cyclizing the above halogenated alcohol or b~ other ~ ~;
knol~ methods. -~
The compounds (I~ accordin~ to the invention may
also be prepared by reacting ammonia or an amine with a
halogenated alcohol of formula (III) or with an epoxide
o~ formula (III~. However, these reactions generally
result in the formation of a mixture of amines. The
bromine derivatives are more suitable than the other
halogen derivatives ~or obtaining primary amines, but
unfortunately they are more expensive. Derivatives
of the alkanolamine type are preferably obtained by
reducing the corresponding cyanhydrin with a compou~d ~-~
o~ the lithium aluminium hydride type. The compounds
according to the invention are then separated from the
reaction mi~ture and puri~ied by known methods, such
as liquid-liquid extraction or ~ractional crystallisation.
- 4 - kmt
., . , . ~. , . ~,
1~39~3~
I)I~S(IILI~rlON ()1~ 5'1~E1l~ 1) F~11301~1~11N'I'S
The preparation of the compounds accordin~ to the
invention is illustrated by, but by no means limited to,
the following Ex~mplesO
EXA~iPLE 1
Pr~aration of 3-decyl~Y~-2-hydro~y=~ inopropa e
300 cc of decanol, 90 cc (106 g) of freshly
distilled epichlorhydrin and 1.5 g of anhydrous ferric
chloride are introduced into a 1 litre ~lask.
After the reaction mixture has been heated at 1~5C
for 10 hours, the 2-decyloxy-1-chloromethyl ethanol is
distilled, first in a water jet vacuum and then in a
vane-pump vacuum.
- B P. of the chlorine derivative at 0.06mm:
128-130C
- yield: 65~ -
20 g of the chlorine derivative and a ~inely -
powdered mixture of 14.8 g o~ phthalimide and 8.4 g o~
potassium carbonate are introduced into a reactor equipped
with a mechanical stirrer.
This mixture is heated under reflux for 5 hours at ~
190C in an oil bath. A~ter removal o~ the heat, 100 cc ~;
of hot ethanol are introd~ced and the mixture filtered to
eliminate the potassium chloride which has formed.
The ethanolic solution obtained is treated while
stirring with a magnetic stirrer, with 5 cc of 98~
hydrate hydrazine, for 30 minutes at ambient temperature
and then for 2 hours under refluxO An insol~ble precipitate
is formed. This mi~ture is then acidified with hydrochloric
acid and rerluxed for 15 minute~. A~ter cooling and
filtration, the solution obtai~ed is evaporated in vacuo.
The evaporation residue is taken up in 50 cc o~
5 kmt
': '`
33
water and, after standing in a re~rigera-tor for 3 hours,
the solution is filtered.
The filtrate is treated with a 10~ soda solution
until a basic pH is obtained. The 3-decyloxy 2-hydro~y-1-
aminopropane is then e~tracted with ether, the ether
phase is washed and dried over anhydrous sodium sulphate
and filtered.
A stream of hydrochloric acid is introduced into the
ethereal solution and, after standing in a re~rigerator,
the product is centrifuged to obtain crystals of 3-decylo~y-
2-hydro~y-1-aminopropane.
The oompound obtained 1s soluble in water and-begins
to melt at a temperature of i~ooc.
EXAMPLES 2~
The procedure is as in Example 1 except that the
decanol is replaced by the alcohol or thiol indicated~
The following compounds are obtained~
myristic alcohol gives 3-tetradecyloxy-2-hydro~yamino~
l-propane hydrochloride (the corresponding chlorinated
; 20 alcohol distils at 160-180C/0.5 mm Hg~ which has a
melting point of 65C (softening), is soluble in water,
and possesses the following elemental analy~is: ;
calculated: H 11.74; C 63.06; N 4.32; Cl lO.g7
observed: H 11.88; C - ; N 4.54; Cl 10.86
n-octadecyl alcohol gives 3-octadecyloxy-2-hydroxy~
ami~opropane hydrochloride, which is soluble in water;
ethanol gives ~-ethoxy-2~hydroxy-1-aminopropane
hydrochloride
decane thiol gives 3-decanethio-2 hydro~y-1-amino-
propane hydrochloride which has a melting point o~ 270~
is soluble in alcohol, and has the following elemental analysis ;
~j _ kmt ~
~, :
.. .
~ 3
calculated: C 55002; H 10.58; N 4.94; Cl 12052; S11.29
observed: C 5~.47; H 10.10; N 5.3 ; Cl 15.71, S10.05.
EX~`IPLE 6
PrepAration o~ ~ode~o~y~2-h~ aminopropane
hydrochloride
By replacing the 3~decyloxy-2-hydroxy-1-chloropropane
(compound III~ used in Example 1 by 3-dodecyloxy-1,2~epoxy
propane, 3-dedecyloxy-2-hydroxy-1-aminopropane is obtained
in the form o~ its hydrochloride: melting point: 60C
~softening).
EXAMPLE 7
Preparation of 3-dec~lo ~-2-hydroxy-N,N,N-trimethyl-l-
ammonium chloride
A m:ixture of 15 g o~ 3-decyloxy-1-chloro-2-propanol
prepared L~ accordanoe with Example 1, 20 cc oi trimethylamine
and 20 cc o~ benzene is heated ~or 6 hours at 120C in an
autocla~e. ~;
After cooling and treatment with ether, the mi~ture
preoipitates a product which is centrifuged. 8.4 g of the
2~ title compound are obtained by recrystallisation in acetone~
It has a melting point of 88~C, is soluble in water, and has
an elemental analysis as ~ollows:
calculated: C 62~03; H 11.63; N 4.52; Cl 11.47
observed: C ~3.20~ Hi11.68; N 4.49; Cl 11.32
EXA~IPLE,,8
Preparation o~ 3-dec~lo~y~2-h~droxy-1-aminopropane hydro-
bromide
A methanolic solution of 28.7 g o~ 3-decyloxy-2-hydro~y-
l-bromopropane (prepared in accordance with Example 1 ~rom
epibromhydrin) and e~cess ammonia is heated for 2~ hours ~ ,~
120C in an autoclave.
~ ~ -
- '~ - kmt
After the reaction produce has been evaporated to
dryness and taken up in ether, the title product is obtained
in a yield of 22.3 g: melting point: 120~C (softening at
60C). The product is soluble in water, and has an
analysis of:-
M.theoretical: 312.3
argentimetry: 306.5
nitrogen: 310.9 ~.
EXA~IPLE 9
Preparation of 3-octyloxy-2-hydro~y-1-amino2ropane
hydrochloride
The procedure is as in Example 8 using as starting -
product 3-octyloxy-2-hydroxy-1-bromopropane (prepared in
accordance with E~ample 1 from epibromhydrin a~d octanol).
The hydrobromide obtained is pa~sed through a column `
of ion e.~change resin (CG-400-Cl) so as to obtain the
corresponding hydrochloride. The title compound is
obtained after recrystallisatioII from acetone and has
~; the follolring properties:
melting point: 55C (so~tening),
soluble in water, ~.
elemental analysis:
calculated: Cl 14.80; N 5~64
observed: Cl 14.88; N 5.95
EXAMPLE 10 :~
Preparation of 3-dodecylo~y-2-hydroxy-1-ami~ro~ane
hydrochloride :~
,
A methanolic solution containing 10 g of 3-dodecyloxy-
1,2-epo~ propane and an excess of ammonia is heated for
4 hours at 100C in an autoclave~ After cooling, evaporation, ~`
treatment with ethe:r, filtration and treatment with gaseous
_ ~ _ kmt
hydrochloric acid, a product is obtained which, after
recrystallisation from methyl ethyl ketone, gives 4.65 g
o~ the title compound.
EX~LES ll ~nd l2
The reduction in ether in the presence of lithium
aluminium hydride of the cyanhydrin of decanal, followed
by recrystallisation from a mixtl7re of isopropyl ether
and methanol, gives 2-hydroYy-l-aminodecane.
- 2-hydroxy-l-amino tridecane is obtained from laurie
aldeh~de by the same process.
The present invention also relates to the use of
eompounds of formula ~I) and more particularly of
~ormula (Ia) as !;terilising agents and/or preservati~es.
The compounds according to this invention have a
~r de ran~e of aetivity against microorganism~, i.e. not
only bac1eria, ~ungi and yeasts, but also baeterial spores.
Their wide range of activity, their very low to~ieity
and the faet that they do not attach the skin, or mueosa
nor are corrosive to metals~ makes them suitable for use as
sterilising agents and preservatives, espeeially in
eosmetology and surgery.
However, the eompounds aeeording to the invention are
antiseptie medicaments intended in particular ~or the
treatment and prevention of disorders eaused by gram-negative
baeteria, in human or veterinary medieine,
The eompol~ds may be used either on their own or in
admixture, in partieular with other baeterieides, in order
to widen the range of activity of the compositions obtained.
- 9 - kmt -
3~i9~
By virtue of their outstandin6 solubility in water,
the compounds according to the invention are pre~erably
used in the form of aqueous solutions, although they may
also be pac~aged in other forms, such as pastes (soaps for
example ), creams, soluble granules, powders, alcoholic
solutions, according to the particular application for
~hich they are inten~ed. For example, the liquid
formuiations will be particularly suitable for the
sterilisation of instruments or apparatus.
The studies reported below were carried out in
particular with 3-decylo~y-2-hydroxy-1-aminopropane
hydrochloride which will be referred to hereinafter as
"compound 1".
Toxicit~
The acute to~icity o~ compound 1 was d~termined by
intragastric administration to male rats of the Wister
~ AF-EOPS strain weighing from 120 to 130 g, and to male
mice o~ the N~I-Han strain weighing between 22 and 25 g
- and aged about 6 weeks.
The LD50 was calculated by the graphic method of
J.T. Lichtfield and F~ lYilcoxon (J. Pharm. Exp. Ther. 194
96 : 99-133).
The LD50 i~ comparable in both animals, and amounts to
1.30 g/kg both after 4~ hours and a~ter 14 days.
Acute to~icity was again determined in male mice by
intravenous administration.
The LD50 of the compound, as determined by intraYenous
administration, both after 3 minutes and after 14 days is
54.5 mg/kg with safety limits of P = 0.05 of 44.2 and 67.3
mg/kg.
- 10 - kmt ;~
3~
Deter~ination of ocular irritation
The ~ethod used is the method described in the
Official Journal o~ 21st April, 1971, page 3863) and
comprises applying to the right eye of each animal 0.1 mm
of a 0.5~ solution. The general total of the results is
O which confirms the high tolerance of the product men~ioned
aboveO
Determinntion of ~ressiveness to the s~in by repeated
aPpli~cation
The method used is the method described in the
Official Journal of 28th April, 1971 for the analysis
of cosmetics and beauty products, and comprises applying
0.5 cc of 2~ solution over a period of 1 month to the
t~o 20nes treated, one normal, the other raw, a control
zone bein~ reserved on the hindquarters of the animal.
Good local tolerance was observed in all the
animals, and histological examination o~ pieces o~ skin
taken from the animals did not r~veal any significant
modi~ication. ~aematological exa~ination carried out
on blood taken from the ear ~ein on completion of the
test showed a normal blood f~rmula.
Stud~ of bacteriostatic activit~
Compound 1, dissolved in distilled water9 is
introduced into a conventional gelose medium, pH 7 to
~5 7.2, optionally containing 20% of horse serum.
The medium containing the solution to be tested is
inoculated in ~he form of three single str~ae with a 105
times dilution of a bacterial culture aged 16 to 18 hours
grown in salted peptone broth at a temperature of 37C.
After inoculation, the media are stored at 37C
for 48 hours, a~ter which the bacterial growth is inspected.
~ kmt
3~
The ~IC ~minimum inhibiting concentrations) are expressed
in micrograms of pure product per cubic centimetre o~
gelose medium required for inhibiting the culture (~g/cc).
The control substance used is ben2alconium chloride. The
results obtained are shown in Table I below.
It is pointed out that the bacteriostatic activity
o~ the product is particularl~ significant i~ the case of
gram-negative bacteria, especially Escherichia coli and
Pseudomonas aeru~inosa.
,
TABLE I
, , , ,, i. .
BACTERIA WITHOUT SEnUM WITH 20~ ~ SE~UM r
Compound 1 Control Compound 1 Control
~g/cc~ ~g/cc) ~pg/ccJ ~/ccJ
Staphylococcus aureus 60 1.5 120 12
9acillus subtilis 30 2.5 50 705
~Escherichia eoli 20 125 75 300
Pseudomonas aerugi~osa ~0 600 150 850
Pneumobacille 35 35 75 75
Serratia 20 10 75 3o
Proteus vulgaris 40 35 150 75
Enterobaeter eloaeae 30 20 80 75
Mor æella glueidolytioa 75 75 150
Determination o~ ~aeterioidal Aetivity
Baeterieidal activity is determined by the following
methods:
A titrate suspension of bacteria is brought into
contact over a period o~ ~ hours at the temperature of
the laboratory with inereasing dilutions o~ the test
product. The number of bacteria still alive are counted
after passage throug~ a ~iltering membra~e. The results
~ 12 - kmt
i3~33
obtained are as follows:
T~BLE II
, . , , _ ,
BACTERIA ~SIC
(}1~7~C )
, _ .
Escherichia coli 50
Pseudomonas aeruginosa 50
Bacillus subtilis 50
M pyogenes aureui 100
An MIC study in a solid medium was carried out on
35 strains of Bacillus pyoc~aneus, and ~howed that, for
the ma~ority o~ these strains, the minimum inhibiting
concentrution was 62,5 ~g/cc.
Determination of th~ sporostatic activi
a) 1 cc of a suspension of spores o~ B.subtilis var.
Niger containing approximately 200 spores is placed i~ `
one Petri dish, whilst 1 oc o~ a ~olution containing ~ g/
100 co o~ the product (mother solution) or o$ the same
mother solution diluted to l/lOth, l/lOOth, etc. up to 1/
lOO,OOOth, are placed in other Petri dishes. Following
the addition o~ a culture medium, the dishes are agitated9 -
oooled and then placed in an o~en. The n~ber of colonies ;~
obtained are then determined. The results are set out in
the ~ollo~ing Table:
~BLF. III
_ _ __ _ _ . ::-
NI~E~ OF DILUTIONS TO NU~ER OF COLONIES
1/10 OF CO~POUND 1 (ave)age of 2 tests No
= _ . __ _ _ ~ _ r
. 18
3 ~ 200
4 ~200
~200
__ :,:
- 13 - kmt ~
, ~ :
. ;
q~ 3
,
b) A porous membrane 0.45 ~ thick is plaeed
aseptically in a sterile filter support, eovered with
10 cc o~ sterile distilled l~ater eontaining 1 cc o~
mother solution of the product, followed by the addition
o~ 1 cc of a suspension of spores o~ B. subtilis var.
Ni~er eontaining appro~imately 40 spores. These
liquids are drawn through the membrane, after which
the membrane is transferred aseptically to a culture
medium and placed in an oven. The operation is
recommeneed using up to 5 rinses o~ the membrane per
10 ce of sterile distilled water. The number of colonies
developing is then determined. The results obtained
are set out in the following Table:
TABLE IV
. . .. ... ... - .
NUMBER OF RINSES OF ~IE NUMBER OF COLONIES
M~n~ No = 37
3 `
1 4
2 35
3 38
4 48
5 _ - 55
Taking into aeeount the dilution by the gelose eulture
medium, it ean be seen, espeeially from Table III, that ;
20 ~g/ee of compound 1 still show sporostatie activity.
The sporocidal aetivity of eompound 1 was also tested
and ~ound to be positive, especially at temperatuxes above
35C and at a pH around 7.
Tests were also carried out to determine the bacterio-
static aetivity o~ other compounds aecording to the
invention by the method deseribed above and in comparison
with known substances " n partieular with benzaleonium chloride
tCompound T in the Tables~,
- 14 - kmt `
1~J~ 'r3~
.
TABI.E V
Compound R-x-cH2-cH(oH)-cH2NH2~A
.
~ . " . . . . ~
. COMPOUND~ R X A CONSTANTS .
, ~:
1 C10H21 O HCl
2 C12H25 O HCl
3 C10H21 O HBr M.p. 120C ?`
4 C6H13 CH2 HCl ~ CH
C10H21 O \CH3 M.p. 88C ~;
6 C8H17 O HCl (sof-t)
7 C7H15 CH2 .. M.p. 195C :~.
8 9 19 CH2 ,l M.p. 200C
9 C14H29 O l M.p. 65C
C18H37 O ,l indeterminate :~
11 CH3 O ll M.p. 92C :~
12 C2H5 O . M.p. 60C --
13 C10H21 O AcOH M.p. 63C
14 C10~21 5 _ M~p. Z70C
.:
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It is pointed out that the variations in the
bacteriostatic activity o~ compounds 1 and T correspond
to variations in the e~perimental conditions and also
in the strains tested, and it is for this reason that
the control compounds were retested at the same time
as the test compound.
In vitro tests on the activity of -the compound o~
E~ample 1 were also carried out so as to determine the
activity of that compound on Bacillus pyoc~aneus. The
results of determining the minimum inhibiting concentration
on 100 different strains are as follows:
25 ~g/cc ~or 20 strains
50 p~/cc for 70 strains
100 ,ug/cc ior 10 strains ~`
On the other hand, tests on 6 different strains
showed that the bactericidal concentration was very ~ -
similar to the MIC.
; The activity of the compound of E~ample 1 was also
tested in vivo, in particular the compound was tested for
its effect on Bacillus p~ocyaneus in the following manner: `
Rats are intra~enously injected with 1 ml o~ a 108
solution of Ps. aeru~inosa. A 20 cm2 cutaneous e~cision
was then made in the animals.
~,0 minutes after the injection, the bacteria injected
were detected in the vicinity of the wound by the replica
method on a solid nutriti~e medium.
One group of rats was sprayed with the compound of
E~ample 1 taqueous solution)^in the vicinity o~ the wounds
(1 to 3 times daily).
A group of control rats were sprayed with sterile
distilled water.
- 19 - kmt
".
. . . . . . . . .
In the treated rats, it was observed that 75
of the replica cultures were totally negative, and
the positive cultures showed only a very small number
of colonies in relation to the control ratsO
Local tolerance was good, and the healing
process was normal.
Accordingly, the compound of E~ample 1 would
appear to be effective to a certain extent in controlling
the infection of excised wounds in rats infected with
Ps. aeru~inosa.
Accordingly, the above tests clearly demonstrate
the e~cellent activity of th~s compound according to
the present invention, in particular with respect to
gram-negative baoteria.
. ' "'~. ~'~
'~.'
,' , "; .
`~
. '~
- 20 - kmt
~ ' ~
