Note: Descriptions are shown in the official language in which they were submitted.
5 ¦ Background of the Invention
Oleandomycin, its productlon in fermentation broths and its use as an
antibacterial agent were first described in U.S. Patent 2,757,123. The natu-
rally occurring co~pound is known to have tha following structure:
)2
OC~3
The conventionally accepted numbering scheme and stereochemical representation
for oleandomycin and similar compounds is shown at a varlety of positions.
Several synthetic modifications of this compound are known, particu-
larly those in which from one to three of the free hydroxyl groups found at the
2',4" and ll-positions are esterified as acetyl esters. In addition, there
are described in U.S. Patent 3,022,219 simiiar modifications in which the acety
in the above-mentioned esters is replaced with another, preferably unbsanched
lower alkanoyl of three to six carbon atoms.
~ I
7 ~ 1
¦ Summary of the Invention
¦ The semi-synthetic oleandomycin antibacterial agents of this invention
¦are represented by the formulae:
~ 'R'b'~
~ f~
2 3 ~ ~ NH2
OCH3 OCH3
IV V
; RO,~
VI OCH3
11's.5';;~3
and the pharmaceutically acceptable acid addition salts thereof, wherein R and
Rl are each hydrogen or alkanoyl having two to three carbon atoms; R2 is hydro-
gen or methyl; and R3 is hydrogen or alkyl having from one to six carbon atoms
provided that when R2 is met~yl, R3 is methyl.
A preferred group of compounds within this class of chemotherapeutic
agents are those qf Formula ~V. Especially preferred within this group are
those compounds wherein R2 and R3 are each hydrogen and R is acetyl. Also
preferred are compounds of Formulae V and VI wherein R is acetyl.
A second class of compounds of the present invention, useful as in-
termediates leading to the antibacterial compounds of For~ulae IV, V and VI,
¦ are of the formulae:
~3~2 ~ ~ )
~- OCU3
3~2
I~I ~ 3
11~5'7l8
¦wherein R and Rl are each hydrogen or alkanoyl having fr~m two to three carbon
¦atoms; and X is 0, N-OH, N-OCH3 or N-O-CCH3.
Preferred within this group of intermediates are those of Formula I
l wherein X is O, N-OH or N-o-8C~3. Also preferred are those intermediates of
5¦ Formula II wherein X is O, N-OH or N-o-8CH3. Finally, preferred within these
intermediates are those compounds of Formula III wherein X is O, N-OH or
N-O-CCH3.
Also within the scope of the present invention is a process for pre-
paring a compound selected from the formulae:
RO---.~ ROn~ 2
~ ~0 '~0
~ ~C~
RO~
o
OCH3
. III
. 1.
-4-
i '.
. I ,
11'`:~i'7 ~ ~
wherein Ac is alkanoyl having two to three carbon atoms and R is hydrogen or
alkanoyl having two to three carbon atoms, which comprises reacting, respec-
tively, a compound selected from those of the formulae:
~ ~ ~ 3)2 _ Ac ~ ~H3)2
RO-" ~ " ~ ~""~ ~ "` ~
OH ~ ~ "'OH
OCH3 OCH3
IA IIA
~0,;~
¦ IIIA
fl
-5-
~1
11'~5'74Y
¦with one mole each of ~-chlorosuccinimide and dimethylsulfide in a reaction-
¦inert-solvent at about O to -25C. followed by contacting the reaction mixture
¦with one mole of triethylamine.
¦ A preferred feature of the clai~ed process is the oxidation of com-
¦pounds of Formula IA wherein the solvent is toluene.
¦ The compounds II, III, V and VI, although all derived from the natural
occurring oleandomycin, differ in the structure at the 8-position. In the na-
¦tural material, I and IV, this structure is an epoxide ring depicted as follows:
Compounds related to II and V contain a methyl group at the 8-positio~
¦ with the indicated sterochemistry and are depicted as follows:
The nomenclature used ~o name the modified oleandomycins of Formulae
II and V is 8,8a-deoxy-8,8a-dihydro-oleandomycin.
Those compounds of Formulae III and VI which contain a cyclopropyl
~ ring at the 8-position are named as 8,8a-deoxy-8,8a-methylene-oleandomycin, and
15 ar depicted aB foll =s:
-6- ,
13 ,~.5'7 }8
¦ Detailed Description of the Invention
¦ In accordance with the process employed for synthesizing the 4"-deoxy
¦ 4"-amIno-oleandomycin derived antibacterial agents of the present invention the
¦ following scheme, starting ~ith a 11,2'-dialkanoyl- or 2'-alkanoyloleandomycin,
5 ¦ is illustrative:
~ ~ 3)Z ~ 3~Z
~ ~ "0~ X'
~3 OCH3
IA
(R = hydrogen or alkano~l of (X = O)
two to three carbon atoms; Ac =
alkanoyl of two to three carbon
atoms)
(X = N-OH, N-OC~3 or ~-O-CCH3)
l ~Continued. . .)
(~ N-OR, N-OCR3 or 3-OCCR3) R ~ 2
The above described scheme is equally applicable to the conversion of
compounds IIA and IIIA to the products V and VI, respectively5 said compounds
being of the formulae:
RO~." ~ __~AA_~ RO~
"OR
IIA V
--8--
I ~ L~ I
¦and
~3~2 ~ 2
IIIA VI
wherein R and Ac are as previously defined.
The initial reaction in these sequences is the selective o~idation of
l the 4"-hydroxy group and is the claimed process of the present invention. Said
5 ¦ process comprises reacting the compounds IA, IIA or IIIA with ~-chlorosuccinimid a
and dimethylsulfide, followed by the addltion of a tertiary amine, such as
triethylamine.
In practice, the N-chlorosuccinimide and dimethylsulfide are first
combined together in a reaction-inert-solvent at about 0C. After ten to
twenty minutes the temperature of the resulting mixture is adjusted to about
O to -25C., and substrate IA, IIA or IIIA is added while maintaining the afore-
mentioned temperature. After two to four hours of reaction time the tertiary
amine is added and the cooling bath removed.
Regarding the quantities of reactants, for each mole of alcohol sub-
strate employed one mole each of N-chlorosuccinimide and dimethylsulfide are
required. Experimentally, it is advantageous to employ a 1-20 fold excess of
the succinimide and sulfide reactants in order to hasten the co~pletion of the
reaction. The tertiary amine employed should correspond to the molar amount of~
succinimide used.
_g_
, i '.
~ .5~
¦ The reaction-inert-solvent utilized in the claimed process should be
¦ one which appreciably solubilizes the reactants and does not react to any appre-
¦ ciable extent with either the reactants or the products formed. Since the
¦ reaction is conducted at about 0 to -25C. it is preferred that, in addition
5 ¦ to haying the above characteristics, it should possess a freezing point below
the reaction te~perature. Such solvents or mixtures thereof which meet these
l criteria are toluene, ethyl acetate, chloroform, methylene chloride or tetra-
¦ hydrofuran. Solvents which meet the above requirements but which have a freez-
I ing point above the reaction temperature can also be employed in minor amounts
10 ¦ in combination with one or more of the preferred solvents. The especially
preferred solvent for the claimed process is toluene containing benzene.
The claimed process is viewed as unique since the oxidation takes
place at the 4"-position leaving the ll-position virtually uneffected when R is
hydrogen.
Removal of the alkanoyl group at the 2'-position is carried out throu h
a solvolysis reaction wherein the 2'-alkanoyl-4"-deoxy-4"-oxo-oleandomycin re-
lated compound is allowed to stir with an excess of methanol overnight at room
temperature. Removal of the methanol and subsequent purification of the resi-
dual provides for compounds of Formula I, II or III wherein Rl is hydrogen and
g ~s 0.
The hydroxy groups at positions 11 (R=H) and 2' (~l=H) of the ketones
(~-0) I, II or III can be acylated by treating said compounds with two moles
of pyridine and an excess of the alkanoic anhydride at ice bath temperatures.
In practice, the hydroxy containing compound is added to cooled alkanoic anhy-
25 ¦ dride followed by the addition of the pyridine. When the additions are com-
plete the ice bath is removed and the mixture allowed to stir overnight at room¦
temperature. The product is obtained by hydrolysis of the reaction mixture wit
water and subsequent extraction of the product with ethyl acetate. Alternately~
the excess alkanoic anhydride solvent can be removed under vacuum and the resi-
3o ! dual material purified by conventional means.
-10-
ll
. ~ .
~1~5'~
As previously indicated, the compounds I, II and III wherein X - O
and ~ and Rl are as previously defined are useful intermediates leading to the
4"-amino antibacterial agents of the present invention. Preferred as intermed-
l iates within this group are 11,2'-diacetyl-4"-deoxy-4"-oxo-oleandomycin, 11-
¦ acetyl-4"-deoxy-4"-oxo-oleandomycin, 4"-deoxy-4"-oxo-oleandomycin, 2'-acetyl-
4"-deoxy-4"-oxo-oleandomycin, 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-
4"-oxo-oleandomycin, 11-acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleando-
mycin, 8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin, 2'-acetyl-8,8a-
I deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin, 11,2'-diacetyl-8,8a-deoxy-8,8a- .
1 methylene-4"-deoxy-4"-oxo-oleandomycin, 11-acetyl-~,8a-deoxy-8,8a-methylene-
4"-deoxy-4"-oxo-oleandomycin, 8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-olean-
domycin and 2'-acetyl-8,~a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin.
Several synethtic pathways can be employed in the preparation of the
1 4"-deoxy~4"-amino-oleandomycin derived compounds. The first compr~seY initial
¦ conversion of the 4 -deoxy-4 -oxo compounds to an oxime or oxime derivative,
i.e., X ~ ~-OH, N-0C~3 or N-O~CH3, followed by reduction of the oxime or deri-
vative thereof to the amine of Formula IV (R2, R3 = ~), V or VI.
The oximes of the ketones (X=03 are prepared by reacting said ketones
with hydroxylamine hydrochloride in a solution of methanol-water at room tem-
perature. In practice, it is preferred that an excess of hydroxylamine beemployed, and as much as a three fold excess provides the desired intermediate
in good yields. Employing ambient temperatures and an~excess of the hydroxyl-
amine allows for the preparation of the desired oxime derivative in a reaction
period of one to two hours. The product is isolated by addition of the reac-
tion mixture to water followed by basification to pH 9.5 and extraction with ater-~m=iaclble solvene such aa etbylacetate.
-11-
l ll
¦ When O-methylhydroxylamine hydrochloride is employed in place of
¦hydroxylamine hydrochloride, the reaction provides the O-methyloxime derivative
¦When using O-methylhydroxylamine, it is preferred to extend the reaction time tc
¦ six to twelve hours. Isolation of the product is carried out in the same manne
¦ as previously described for the oxime derivative.
Preparation of the O-acetyloxime compounds (X = N-O~CH3) is effected
by acetylation of the corresponding oxime. Experimentally, one mole of the
oxime is reacted with one mole of acetic anhydride in the presence of one mole
l of pyridine. The use of an excess of the anhydride and pyridine aid in the
10 ¦ completion of thereaction and an excess of two to three fold is preferred. The
reaction is best conducted in an aprotic hydrocarbon solvent such as benzene or
toluene at room temperature overnight. On completion of the reaction, water is
added and the product is separated in the hydrocarbon layer. Alternatively,
O-acetyl derivatives can be prepared by treating the requisite ketone with O-
acetylhydroxylamine hydrochloride under reaction conditions operable in the
preparation of the oxime derivatives.
The preferred oxime and oxime derivatives which are useful interme-
diates leading to the 4"-deoxy-4"-amino-oleandomycin derived antibacterial
agents include 11,2'-diacetyl-4"-deoxy-4"-oxo-oleandomycin oxime, 11-acetyl-4"-
20 deoxy-4"-oxo-oleandomycin oxime, 11,2'-diacetyl-4"-deoxy-4"-oxo-oleandomycin
O-acetyloxime, ll-acetyl-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime, 11,2'-
diacetyl-8,ôa-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin oxime, ll-acetyl-
8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin oxime, 11,2'-diacetyl-
8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime, ll-acetyl-
25 8,8a-deoxy-8~8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime, 11,2'-
diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin oxime, 11-
acetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin oxime, 11,2'-di-
acetyl-8,8a-deoxy-8,8a-methylene-4'l-deoxy-4"-oxo-oleandomycin O-acetyloxime
and ll-acetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin O-acetyl-
30 ¦ o e. -12-
1 1~1.;,5'7 ~8
l Reduction of the ketone derivatives (X=N-OH, N-OCH3 or N-OCCH3) is
¦ carried out by a catalytic hydrogenation wherein a solution of the oxime or
¦ derivative thereof in a lower alkanol, such as isopropanol, and a Raney nickel,
¦ 10%-palladium-on-charcoal or platinum oxide catalyst is shaken in a hydrogen
¦ atmosphere at an initial pressure of 50 p.s.i. at room temperature overnight.
Filtration of the spent catalyst followed by removal of the solvent from the
l filtrate provides for the isolation of the desired 4"-deoxy-4"-amino substitute
¦ antibacterial agent of the FormulaeIV, V or VI. If methanol is employed as the
l reduction solvent, solvolysis of the 2'-alkanoyl group will occur. To avoid
¦ removal of this moiety, isopropanol is the preferred solvent.
The second, and preferred, route from the ketones (X=O) of Formulae
I, II and III to the primary amines of Formulae IV, V and VI comprises the con-
densation of said ketones with the ammonium salt of a lower alkanoic acid and
the subsequent reduction of the in situ generated imine. In addition to am-
monium salts of lower alkanoic acid being operable, other ammonium salts suchas those of inorganic acids can also be employed.
In practice, a solution of the ketone I, II or III (X=O) in a lower
alkanol such as methanol is treated with an ammonium salt of an alkanoic acid
such as acetic acid and the cooled reaction mixture treated with the reducir.g
agent sodium cyanoborohydride. The reaction is allowed to proceed at room tem-
perature for several hours, and is subsequently hydrolyzed and the product iso-
lated.
Although one mole of ammonium alkanoate is needed per mole of ketone,
it is advantageous to add an excess in order to ensure a rapid formation of the
imine. As large as a ten fold excess can be employed without effecting the
quality of the final product.
Regarding the amount of the reducing agent to be employed per mole ofl
ketone, it is preferred that about two moles of sodium cyanoborohydride per mole
of ketone be used.
The reaction time for the reduction varies from two to three hours at~
ambient temperatures. -13-
i ~ 4~ 7~
As previously mentioned, the preferred solvent is methanol while the
preferred ammonium alkanoate is ammonium acetate. Isopropanol can also be used
as a solvent, and is especially desirable when solvolysis of the 2'-alkanoyl
l group is to be avoided.
S ¦ In isolating the desired 4"-deoxy-4"-amino-oleandomycin derivatives
from any non-basic by-products or starting material, advantage is taken of the
basic nature of the final product. Accordingly, an aqueous solution of the
product is extracted over a range of gradual increasing pH such that neutral
or non-basic materials are extracted at lower pH's and the product at a pH of
about 9. The extracting solvents, either ethyl acetate or diethyl ether, are
backwashed with brine and water, dried over sodium sulfate and obtained by re-
oval of the solvent.
Additional purification, if necessary, can be effected by column
chromatography on silica gel according to known procedures.
The aforementioned reductive amination can be carried out with other
educing conditions besides the use of sodium cyanoborohydride. Certain noble
metal catalysts, such as palladium-on-~harcoal, can be employed with hydrogen
nd an ammonium alkanoate to effectively provide for the conversion of compounds
f Formulae I, II and II~ ~X=~) to those of Formulae IV, V and VI, respectively.
2~ Experimentally, a solution of the appropriate ketone in a lower al-
anol, such as methanol or isopropanol, is treated with an ammonium alkanoate,
uch as ammonium acetate, and 10% palladium-on-charcoal, and the resulting
uspension shaken in a hydrogen atmosphere at temperatures of about 25~50~C.
ntil the theoretical amount of hydrogen has been absorbed.
Regarding the ratio of reactants, it is preferred that a ten fold
xcess of the ammonium alkanoate be employed to ensure complete reaction in a
easonable time period. The amount of the catalyst can vary from 10% to 50%,
n a weight basis, of the starting ketone. The initial pressure of the hydro-
en is not critical, and a Dressure from one atmosphere to 500 D.S.i. iS nre-
erred to shorten the reaction time. Employing the aforementioned parameters,
he reaction time will vary between two to six hours.
11'~'5'7 ~ 1
At the conclusion of the reductive amination reaction, the spent
catalyst is filtered and the filtrate concentrated to dryness. Purification
of the product is carried out by the aforementioned procedure wherein sodium
l cyanoborohydride is used as the reducing agent.
5 ¦ Synthesis of antibacterial compounds of Formula IV wherein R2 is
hydrogen and R3 i9 alkyl of one to six carbon atoms is conveniently achieved
from the ke~one I (X=0) and the appropriate amine, R3NH2, using sodium cyano-
borohydride as the reducing a8ent. In order to maintain the pH between about
l 6 and 7, a molar amount of an alkanoic acid, such as acetic acid, equal to that
10 ¦ of the amine is employed. Alternately, a corresponding amount of hydrogen
chloride gas can also be employed in place of the alkanoic acid.
The raeio of reactants, reaction temperature and time and work-up of
the reductive amination reaction are the same as the corresponding parameters
for the reaction leading ~o those compounds wherein R2 and R3 are each hydrogen
and sodium cyanoborohydride is employed as the reducing agent.
The antibacterial compounds IV wherein R2 and R3 are each methyl are
prepared by the reductive alkylation of the 4"-deoxy-4"-amino-oleandomycin IV
wherein R2 and R3 are each hydrogen using formaldehyde, hydrogen and 10%
palladium-on-charcoal.
The ratio of reactant~, reaction temperature, solvent, pressure,
time and work-up are the same as those parameters for the reaction leading to
I rq whereln R2 and R3 are each hydrogen and hydrogen gas and 10% palladium are
! employed as the reducing agent.
l As previously mentioned, solvolysis of the 2'-alkanoyl moiety can be
25 ¦ effected by allowing said derivative of the amine related to IV, V or VI to
stir in a =ethanol soluti~n ~verni~ht at ambient te=peratures.
I -15-
;
"
.~
Preferred among these compounds because of their antibacterial utilit~
are 4"-deoxy-4"-amino-oleandomycin, 11-acetyl-4"-deoxy-4"-amino-oleandomycin,
11,2'-diacetyl-4"-deoxy-4"-amino-oleandomycin, 11-acetyl-8,8a-deoxy-8,8a-di-
hydro-4"-deoxy-4"-amino-oleandomycin, 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-
4"-deoxy-4"-amino-oleandomycin,ll-acetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-
4"-amino-oleandomycin and 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-
4"-amino-oleandomycin.
In the utilization of the chemotherapeutic activity of those compoundb
of the present invention which form salts, it is preferred, of course, to use
pharmaceutically acceptable salts. Although water-in~olubility, high toxicity,~
or lack of cryqtalline nature may make some particular salt species unsuitable ¦or less desirable for use as such in a given pharmaceutical application, the
water insoluble or toxic salts can be converted to the corresponding pharmaceu-
tically acceptable bases by decomposition of the salt as described above, or
alternately they can be converted to any desired pharmaceutically acceptable
acid addition salt.
Examples of acids which pro~ide pharmaceutically acceptable anions
are hydrochloric, hydrobromic, hydroiodic, nitric, sulfuric, or sulfurous,
phosphoric, acetic, lactic, citric, tartaric, succinic, maleir, gluconic and
aspartic acids.
The stereochemistry of the starting materials leading to the anti-
bacterial agents of the present invention is that of the natural material. The
oxidation of the 4"-hydroxyl group to a ketone and the subsequent conversion
of said ketone to the 4"-amines presents an opportunity for ehe stereochemistry
of the 4"-substituent to change from that of the natural product. Accoridngly,
when the compounds I, II and III (X=0) are converted to amines by one of the
hereinbefore described procedures, it is possible that two epimeric amines are
formed. Experimentally, it is ob~erved that both epimeric amines are present
in the final product in varying ratios depending on the choice of synthetic
7 ~
¦method. If the isolated product consists predominantly of one of the epimers,
¦said epimer can be purified by repeated recrystallization from a suitable sol-
¦vent to a constant melting point. The other epimer, the one present in smaller
l amounts in the originally isolated solid material, is the predominant product
¦ in the mother liquor. I~ can be recovered therefrom by methods known to those
skilled in the art, as for example, the evaporation of the mother liquor and
repeated recrystallization of the residue to a product of constant melting
point or by chromatography.
l Although said mixture of epimers can be separated by methods known
¦ to those skilled in the art, for practical reasons it is advantageous to use
said mixture as it is isolated from the reaction. However, it is frequently
advantageous to purify the mixture of epimers by at least one recrystallization
from an appropriate solvent, subjecting it to column chromatography, solvent
partitioning or by trituration in an appropriate solvent. Said purification,
I .~
while not necessarily separating the epimers, removes ~uch extraneous materials
as starting materials and undesirable by-products.
The absolute stereochemical assignment for the epimers has not been
completed. Both epimers of a given compound, however, exhibit the same type
of activity, e.g., as antibacterial agents.
The novel 4"-deoxy-4"-amino-oleandomycin derivatives described herein
exhibit in vitro activity against a variety of Gram-positive microorganisms
such as Staphylococcus aureus and Streptococcus pyogenes and against certain
Gram-negative microoganisms such as those of spherical or ellipsoidal shape
Ccocc~). Their activity is readily demonstrated by in vitro tests against
various microoganisms in a brain-heart infusion medium by the usual two-~old
serial dilution technique. Their in vitro activity renders them useful for
topical application in the form of ointments, creams and the like; for sterili-
zation purposes, e.g., sick-room utensils; and as industrial antimicrobials,
for example, in water treatment, slime control, paint and wood preservation.
For in vitro use, e.g., for topical application, it will often be
convenient to compound the selected product with a pharmaceutically-acceptable
carrier such as vegetable or mineral oil or an emollient cream. Similarly,
they may be dissolved or dispersed in liquid carriers or solvents, such as
water, alcohol, glycols or mixtures thereof or other pharmaceutically-acceptabl~
inert media; that is, media which have no harmful effect on the active ingre-
dient. For such purposes, it will generally be acceptable to employ concen-
trations of active ingredients of from about 0.01 percent to about 10 percent
by weight based on total composition.
~5 Additionally, many compounds of this invention are active versus Gram-
positive and certain Gram negative microorganisms in vivo such as Pasteurella
multocida and Neisseria sicca via the oral and/or parenteral routes of adminis-
tration in animals, including man. Their in vivo activity is more limited as
regards susceptible organisms and is determined by the usual procedure which
comprises mice of substantially uniform
1~'45'7 ~
weight with the test organism and subsequently treating them orally or subcu-
taneously with the test compound. In practice, the mice, e.g. 10, are given
an intraperitoneal inoculation of suitably diluted cultures containing approxi-
mately 1 to 10 times the LD~oo (the lowest concentration of organisms required
to produce 100% deaths). Control tests are simultaneously run in which mice
receive inoculum of lower dilutions as a check on possible variation in viru-
lence of the test organism. The test compound is administered 0.5 hour post-
inoculation, and is repeated 4, 24 and 48 hours later. Surviving mice are
held for four days after the last treatment and the number of survivors is
noted.
When used in vivo, these novel compounds can be administered orally
or parenterally, e.g., by subcutaneous or intramuscular injection, at a dosage
of from about l mg./kg. to about 200 mg./kg. of body weight per day. The
favored dosage range is from about 5 mg./kg. to about lO0 mg./kg. of body weighl
per day and the preferred range from about 5 mg./kg. to about 50 mg./kg. of
body weight per day. Vehicles suitable for parenteral injection may be either
aqueous such as water, isotonic saline, isotonic dextrose, Ringers' solution,
or non-aqueous such as fatty oils of vegetable origin (cotton seed, peanut oil,
corn, sesame~, dimethylsulfoxide and other non-aqueous vehicles which will not
interfere with therapeutic efficiency of the preparation and are non-toxic in
the volume or proportion used (glycerol, propylene glycol, sorbitol). Addi-
tionally, compositions suitable for extemporaneous preparation of solutions
prior to administration may advantageously be made. Such compositions may in-
clude liquid diluents, for example, propylene glycol, diethyl carbonate, gly-
2 cerol, sorbitol, etc.; buffering agents, hyaluronidase, local anesthetics andinorganic salts to afford desirable pharmacological properties. These compound<
may also be combined with various pharmaceutically-acceptable inert carriers in- .
cluding
li.'~57-~
¦ olld dlluents, aqueous vehicle:, non-toxic organlc solvents in the for= of
¦capsules, tablets, lozenges, troches, dry mixes, suspensions, solutions, elixir~
and parenteral solutions or suspensions. In general, the compounds are used
l in various dosage forms at concentration levels ranging from about O.S percent
5 ¦ to about 90 percent by weight of the total composition.
The following examples are provided solely for the purpose of illus-
tration and are not to be construed as limitations of this invention, many
variations of which are posslble without departing from the spirit or scope
~ _19_
l `.
, I ~
ll.Z5'i'~8
EXAMPLE 1
2'-Acetyl-4"-deoxy-4"-oxo-oleandomycin
¦ Dimethylsulfide (0.337 ml.) is added to a turbid solution of 467 mg.
¦ of N-chlorosuccinimide in 20 ml. of toluene and 6 ml. of benzene cooled to -5C
¦ and maintained under a nitrogen atmosphere. After stirring at 0C. for 20 min.
¦ the mixture i~ cooled to -25C. and 1.46 g. of 2'-acetyloleandomycin and 15 ml.
of toluene are added. S~irring is continued for 2 hrs. at -20C. followed by
the addition of 0.46 ml. of triethylamine. The reaction mixture is maintained
l at -20C. for an additional 5 min. and then allowed to warm to 0C. The mix-
10 ¦ ture i8 poured, with stirring, into 50 ml. of water and 50 ml. of ethyl aceta~e
The pH of the aqueous mixture is adjusted to 9.5 by the addition of aqueous
sodium hydroxide ~olution. The organic layer is subsequently separated, dr~ed
over sodium sulfate and concentrated in vacuo to a white foam (1.5 g.). Tri-
I turation with diethyl ether gives 864 mg. of crude product, which on recrystal-
¦ lization twice from methylene chloride-diethyl ether gives 212 mg. of the pure
product, m.p. 183-185.5C.
37 61 13 C, 61.1, H, 8.5; N, 1.9.
Found: C, 60.9; ~, 8.4; N, 1.9.
lNMR (~, CDC13)o 5.60 (lH)m, 3.50 (3H)s, 2.73 (2H)m, 2.23 (6H)s and
2~ ¦ 2.03 ~3H)s.
Employing the above proc~dure and starting with 2'-propionyloleandomy _
cin, there is obtained 2'-propionyl-4tl-deoxy-4'l-oxo-oleandomycin.
li!~.`r~'^9' ~ 1
¦ EXANPLE 2
4"-Deo~2~4"-oxo-oleandomycin
l A solution of 1.0 g. of 2t-acetyl-4ll-deoxy - 4"-oxo-oleandomycin in
¦ 20 ml. of methanol is allowed to stir at room temperature overnight. The solu-
5 ¦ tion is concentrated in vacuo to give the desired product as a white foam
937 mg.
NMR t~, CDC13): 5.60 (lH)m, 3.50 (3H)s, 2.85 (2H)m and 2.26 (6H)s.
EXAMPLE 3
11 2'-Diacetyl-4"-d~ 4"-oxo-oleandomycin
10 ¦ A. Via Acety~ation
To 4.0 ml. of acetic anhydride under a nitrogen atmosphere and coole
to 0C. in an ice bath is added 727 mg. of 2'-acetyl-4"deoxy-4"-oxo-oleandomyci 1.
After 5 min .158 ml. of pyridine is added and the hazy suspension allowed to
l stir overnight at room temperature. The resulting ~olution is quenched in
15 ¦ water layered over with ethyl acetate, and the pH adjusted to 7.2 by ~he ad-
dition of solid sodium bicarbonate and then to 9 5 using a lN sodium hydroxide
solution. The organic layer is separated, washed successively with water and
a saturated brine solution and dried over sodium sulfate. Removal of the sol- ¦
l vent under reduced pre~sure gives 588 mg. of the desired product.
2U ¦ NMR ~, CDC13~: 3.48 (3H)s, 2.63 (2H)m, 2.26 (6H)s and 2.06 (6H)s.
Repeating this procedure and employing the requisite 2'-alkanoyl-4"-¦
deoxy-4"-oxo-oleandomycin and aoylating reagent, the following co~pounds are
synthesized: ll-acetyl-2'-propionyl-4"-deoxy-4"-oxo-oleandomycin9 ll-propionyl-
2'-acetyl-4'~-deoxy-4"-oxo-oleandomycin and 11,2'-dipropionyl-4"-deoxy-4"-cxo-
¦ o andQmyci=.
-21-
11. ~ 5 1 i8
l 8. Via Oxidation
l _ !
To 4.5 g. of N-chlorosuccinimide, 50 ml. of benzene and 150 ml. of
toluene in a dry flask fitted with a magnetic stirrer and nitrogen inlet and
l cooled to -5C. is added 3.36 ml. of dimethylsulfide. After stirring at 0C.
for 20 min., the contents are cooled to -25C. and treated with 5.0 g. of
11,2'-diacetyl-oleandomycin in 100 ml. of toluene. Cooling and stirring are
continued for 2 hrs. followed by the addition of 4.73 ml. of triethylamine.
The reaction mixture is allowed to stir at 0C. for 15 min., and is subse-
quently poured into 500 ml. of water. The pH is adjusted to 9.5 with lN
aqueous sodium hydroxide and the organic layer separated, washed with water
and a brine solution and dried over sodium sulfate. Removal of the solvent
in vacuo gives 4.9 g. of the desired product, identical to that prepared in
Example 3A, as a foam.
NMR (~, CDC13): 3.48 (3H)s, 2.61 (2H)m, 2.23 (6H)s and 2.03 (6H)s.
EXAMPLE 4
ll-Acetyl-4''-deoxy-4ll--o~o-oleandomycin
3'
A solution of 4.0 g. of 1132'-diacetyl-4"-deoxy -4"-oxo-oleandomycin
in 75 ml. of methanol is allowed to stir at room temperature overnight. The
reaction mixture is concentrated under reduced pressure to give the product as
a foam. A diethyl ether solution of the residue, on treatment with hexane,
gives 2.6 g. of the product as a white solid, m.p. 112-117C.
NMR (~, CDC13): 3.43 (3H)s, 2.60 (2H)m, 2.23 (6~)s and 2.01 (3H)s.
Si~ilarly, by employing 11,2'-dipropionyl-4"-deoxy-4"-oxo-oleandomy-
cin or ll-propionyl-2'-acetyl-4"-deoxy-4"-oxo-oleandomycin in the above proced-
ure, 11-propionyl-4"-deoxy-4"-oxo-oleandomycin is prepared.
il~5'7~1
¦ EXAMPLE 5
¦11,2'-Diacetyl-4"-deoxy-4"-oxo-oleandomycin
I
¦ A reaction mixture comprising 1.0 g. of 11,2'-diacetyl-oleandomycin,
¦ 7.09 ml. of dimethylsulfoxide and 9.44 ml. of acetic anhydride is allowed to
5 ¦ stir at room temperature for 4 days. The resulting yellow colution i~ added
to water layered over with benzene. The pH is subsequently adjusted with
aqueous lN sodium hydroxlde to 9.5 and the organic layer separated, dried over
sodium sulfate and concentrated in vacuo to dryness. The yellow oily foam
l ~1.14 8-) is chromatographed over 20 8. of silica gel using chloroform-acetone~
10 ~ 9:1 as the eluate. Removal of the solvent from the fractions gives 800 mg. of
the desired product and 110 mg. of a by-product.
In a ~imilar manner, when ll-acetyl-2'-propionyloleandomycin, 11,2'-
dipropionyloleandomycin or ll-propionyl-2'-acetyloleandomycin is employed a~ ¦
l the starting material in the aoove oYidation procedure, ll-acetyl-2'-propionyl
15 ¦ 4"-deoxy-4"-oxo-oleandomycin, 11,2'-dipropionyl-4"-deoxy-4"-oxo-oleandomycin
l and ll-propionyl-2'-acetyl-4"-deoxy-4"-oxo-oleandomycin are obtained, respec- ¦
I t~vely.
-23-
I
.1,
5~
¦ EXAMPLE 6
¦ 2'-Acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
¦ To a dry flaRk fitted with a magnetic stirrer and nitrogen inlet con-
¦ taining 11.6 g. of N-chlorosuccinimide, 750 ml. of toluene and 25u ml. of
¦ benzene and cooled to -5C. is added 6.0 ml. of dimethylsulfide, and the re-
I sulting solution allowed to stir for 20 min. The tempera~ure is further lowere 1
¦ to -20C. and 25 g. of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-oleandomycin in 500 ml .
¦ of toluene is added. After stirring for 2 hrs. at -20C., 11.4 ml. of triethyl _
¦ amine i~ added and the reaction mixture gradually allowed to warm to O~C. It
¦ is then poured into 1500 ml. of water and the pH adjusted to 9.5 with lN sodium
¦ hydrox$de solut~on. The organic layer is separated, washed succes3iYely with
water (3x) and a saturated brine solution (lx) and dried over ~odium sulfate.
Removal of the solvent under reduced pressure gives a foam, which on recry-
l stallizat$on from diethyl ether provides 13 g. of the pure product, m.p. 197-
15 1 199C.
l NMR (~, CDC13): 5.11 (lH~m, 3.51 (3H)s, 2.25 (6H)s and 2.03 (3H)s.
~ .5'~ ~
¦ EXAMPLE 7
¦ 8,8a-Deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
¦ A suspension of 2.0 g. of 2'-acetyl-8,8a-deoxy-8,8a-dihydro-"-deoxy-
¦ 4"-oxo-oleandomycin in 100 ml. of methanol is allowed to stir at room tempera-
¦ ture for 20 hrs. The reaction mixture is concentrated under reduced pressureto provide 1.8 g. of the desired product as a white foam.
NMR (~, CDC13): 5.30 (lH)m, 3.51 (3H)s and 2.26 (6H)s.
EXAMPLE 8
l 11,2'-Diacet~1-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
10 To a suspension of 13.0 g. of 2'-acetyl-8,8a-deo~y-8,8a-dihydro-4"-
deo.Yy-4"-oxo-oleandomycin in 65.0 ml. of acetic anhydride cooled in an ice bath
is added 2.8 ml. of pyridine; The bath is removed and the reaction mixture
allowed to stir at room temperature overnight. The resulting solution is sub-
sequen~ly added to 500 ml. of water and 300 ml. of ethyl acetate. The pH is
15 adjusted to 7.0 with solid sodium bicarbonate and then to 9.5 with 4N aqueous
sodium hydroxide. The organic layer is separated,washed with water (2x) and
brine solution ~lx), and dried over sodium sulfate. Removal of the solvent
in vacuo gives tne crude product as a foam.
Since chromatography results on the crude sample suggest incomplete-
ness of reaction, the crude foam ls recombined wi~h 28 ml. of pyridine and 79ml. of acetic anhydride and allowed to stir at room temperature for 72 hrs.
The reaction mixture is worked up as above to give 12.4 g. of the desired prod-
~uct.
~MR ~ DC13): 3.51 (3H~s, 2.26 (6H)s and 2.10 (6H)s.
-:~5-
EXAMPLE 9
ll-Propionyl-2'-acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
The procedure of Example 8 i9 repeated starting with 6.5 g. of 2'-
acetyl-8,8a-deoxy~8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin, 40 ml. of pro-
pionic anhydride and 14 ml. of pyridine. After a reaction time of 72 hrs.,the mixture is worked up as indicated in Example 8 to provide the desired
product.
~gAMPLE 10
ll-Acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
A solution of 11.5 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4'l-
deoxy-4"-oxo-oleandomycin in 100 ml. of methanol is allowed to stir overnight
at room temperature. Concentration of the reaction mixture to dryness under
reduced pressure gives 10.6 g. of the crude product as a foam. The crude
material ic dissolved in chloroform and placed on a silica gel column. After
3 1. o~ chloroform has passed through the column, the product i~ eluted with
chloroform/methanol (19:1). Eight hundred drop fractions are taken on an
automatic fraction collector. Fractions 50-56, 57-62, 63-69 and 70-80 are
combined and concentrated in vacuo to dryness to give 2.9 g. of the pure prod-
uct.
NMR (&, CDC13): 3.55 (3H)s, 2.31 (6H)s and 2.05 (3H)s.
llr~57~
EXAMPLE 11
ll-Propionyl-8,8a-deoxy~8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
Starting with ll-propionyl-2'-acetyl-8,8a-deoxy-8,8a-dihydro-4"-
I deoxy-4"-oxo-oleandomycin of Example 9 and following the procedure of Example
¦ 10, the desired compound is prepared.
EXAMPLE 12
2 ~Acetyl-8 8a-deoxy-8 8a-dihydro-oleandomycln
To a solution of 5.0 8. of 8,8a-deoxy-8,8a-dihydro-oleandomycin in
l 15 ml. of benzene is added .73 ml. of acetic anhydride and the resulting reac-
¦ tion mixture allowed to stir at ambient temperature for 1.5 hrs. The solutionis added to 100 ml. of water and the pH adjusted to 7.5 with solid sodiu~ bi-
I carbonate and then 9.5 with 1~ aqueous sodium hydro~ide. After 10 min. stir-
¦ ring the organic layer is separated, washed successively with water (2x) and
l a satura~ed brine solution (lx) and then dried over sodium sulfate. Removal
¦ of the solvent under reduced pres~ure gives 4.9 g. of the desired product9
.p. 202-204C.
NMR (~, CDC13): 5.05 (l~)m, 3.40 (3H)s, 2.25 (6E)s and 2.05 (3H)s.
Similarly, by replacing the acetic anhydride ~ith an equivale~t
l amount of propionic anhydride, 2'-propiQnyl-8,8a-deoxy-8,8a-dihydro-oleandomy-
¦ is prepared.
-Z7-
~ ~ J~
EXAMPLE 13
2'-Propionyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
To a solution of 375 ml. of toluene and lZ5 ml. of benzene is added
5.8 8. of N-chlorosuccinimide and the mixture allowed to stir at room tempera-
ture for 15 min. The reaction mixture is cooled to -5C. and 3.0 ml. of di-
methyl ~ulfide is added, and stirring co~tinued for an additional 20 min. The
temperature is lowered to -20C. followed by the addition of 12.8 g. of 2'-
propionyl-8,8a-deoxy-8,8a-dihydro-oleandomycin in 250 ml. of toluene. After 2
hrs., 5.7 m}. of diethylamine is added and the cooling bath removed. When
~he reaction temperature reaches 0C., the mixture ~s quenched in 750 ml. of
water. The pH is adjusted to 9.5 with lN sodium hydroxide solution and the
organic layer separated. Ater washing wi~h water (3x) and a saturated brine
solution, the organic layer is dried and concentrated under reduced pressure
to give the desired product.
-28-
5'~ ~
EXAMPLE 14
11~2'-Diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin
To a turbid solution of 434 mg. of N-chlorosuccinimide in 15 ml. of
toluene and 5 ml. of benzene cooled to -5C. is added .327 ~1. of dimethyl-
S sulfide. After stirring for 20 min. at 0C., the reaction mixture is cooledto -25C. and 500 mg. of 11,2'-diacetyl-8,8a-deoxy-8,8a-me~hylene-~leandomycin~
and 10 ml. of toluene are added. Stirring is continued for 2 hrs. a~ -20C.
followed by the addition of .46 ml. triethylamine and 1 ml. of toluene. The
cooling bath is removed and the reaction mixture allowed to warm to 0C. It -
is then poured into 50 ml. of water and 50 ml. of ethyl acetate. The pH is ¦carefully adjusted to 9.5 and the organic layer separated, dried and concen-
trated to dryness. In this manner 520 mg. of the slightly wet desired productis obtained as a white foam.
NMR (~, CD~13): 3.50 (3H)s, 2.30(6H)s, 2.06 (6H)s and 0.58 (4H)m.
-29-
I
EXAMPLE 15
¦ Starting with the appropriate 8,8a-deoxy-8,8a-methylene-oleandomycin¦ and employing the procedure of Example 14, the following 8,8a-deoxy-8,8a-
thylene-4"-deo~y-4"-oxo-oleand~ycln derlvaelves are prepared:
3)~
OCU3
ca3c- . c~3ca2c-
. CH3C~2C- C~3c- O
CH3CH2C- CH30,H2C-
H- CH3C- O
lt H- CH3CH2C-
-3~-
1~
EXAMPLE 16
ll-Acetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin
A solution of 400 mg. of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-
4"-deoxy-4"-oxo-oleandomycin in 10 ml. of methanol, after stirring at room
temperature overnight, is concentrated under vacuum to glve 270 mg. of the
desired product.
NMR (~, CDC13): 3.46 (3H)s, 2.26 (6~s, 2.03 (3H)s and 0.56 (4H)m.
EXAMPLE 17
The procedure of Example 16 is repeated starting wi~h the appropriate
2'-alkanoyl-8,8a-deoxy-8,8a-methylPne-4"-deoxy-4"-oxo-oleandomycin to provide
the following compounds:
"~'~0
oc~3
~1 11;5~
¦ Starting Material Product
¦ R Rl R R
I O O O
l CH3C- C~3CH2C- CH3C- H-
I O O O
I ,. .. ..
l CH3CH2C- CH3C- 3 2
I O O O
5¦ CH3CH2C-CH3CH2C- CH3CH2C
l l O
l l ~- CH3C- H- H-
l l O
H- 3 2C H H-
¦ EXAMPLE 18
l 11,2l-Diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin
10 ¦ In a dry flask equipped with a nitrogen inlet is introduced 18 ml. of
methylene dichloride and 1.97 ml. of dimethylsulfoxide, and the resulting solu-
tion cooled to -60C. Trifluoroacetic anhydride (3.9 ml.) is gradually added
and stirring continued in the cold for 10 min. The reaction mixture is further
l cooled to 70C. and 5.34 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-
¦ oleandomycin in 27 ~1. of methylene dichloride is added dropwise at such a rate
ehat the temperature does not rise above -50C. After the reaction mixture is
ooled back to -70C., 9.69 ml. of triethylamine is added and cooling continued
for lQ min. The reaction is allowed to warm to -10C., and is then poured into
1 75 ml. of water. The p~ i9 adjusted to 9.5 with lN sodium hydroxide solution
~ and the methylene dichloride separated. The organic layer is subsequently
I
"~J~5~71~
washed with water (2 x 30) and a saturated brine solution (1 x 20) and dried
over sodium sulfate. Removal of the solvent in vacuo give~ 6.2 g. of product
which, except for a trace amount of impurities, is identical with the product
of Example 14.
E~AMPLE l9
The procedure of Example 18 is repeated, starting with the requisite
2'-alkanoyl-8,8a-deo~y-8,8a-methylene-oleandomycin, to give the following 4"-
deoxy-4"-oxo derivatives:
~ 3)2
"`~0
OCH3
R R
CH3CH2C- CH3C-
E- CH3CH2C-
CH3C- CH3CH2C_
CH3CH2C- CH3C~2
-33-
,,
11..~5'7 ~
EXAMPLE 20
11~2~-Diacetyl-4''-deoxy--4~-o~o-oleandomycin oxime
To a ~olution of 18.1 g. of hydroxylamine hydrochloride in 300 ml.
1 of water and 200 ml of methanol is added 50 g. of 11,2'-diacetyl-4"-dPoxy-4"-
5 ¦ oxo-oleandomycin and the reaction mix~ure allowed to stir at room temperature
for one hour. The resulting ~olution is added to water and the pH subsequently
adjusted to 7.5 with solid sodium bicarbonate and then 9.5 with lN sodium hy-
droxide. The product is extracted into e~hyl acetate and the dried extracts
concentrated to about 170 ml. Hexane is added to the heated ethyl acetate
qolution to the cloud point and the hazy solution cooled. The precipitated
product i~ filtered and dried, 29.8 g., m.p. 223.5-225C.
NMR (~, CDC132: 3.30 ~3H)~, 2.65 (2H~m, 2.35 (6H)s and Z.10 (3H)s.
In a si-~ilar manner, by starting wi~h 0-acetylhydroxylamine hydro-
chloride and the requisite ketone and following the above procedure there is
obtained 2'-acetyl-4"-deoxy-4"-oxo-oleandomycin 0-acetyloxime and 2'-propionyl-
-~4-
¦ EXAMPLE 21
¦ The procedure of Example 20 is repeated, starting with the appropria
4"-deoxy-4"-oxo-oleandomycin, to give the following oximes:
RO,
OCH3
,0,
H- CH3C- ,0,
oH~ CH3CH~C-
C~3C- CH3CH2C-
O O
CH3CH2o,- CH3C-
o CH3CH2C-
103 H-
O
CH3CH2C- H-
-35-
1~ 5~
EXAMPLE 22
ll-Acetyl-4"-deox~-4"-oxo-oleandomycin oxime
A solution of S00 mg. of 11,2'-diacetyl-4"-deoxy-4"-oxo-oleandomycin
oxime in 100 ml. of methanol, after stirring 72 hrs. at room temperature, is
concentrated ~o drynes~ under reduced pressure. The resulting foam is re-
crystallized from ethyl acetate-hexane, 372 mg., m.p. 184-186C.
NMR (~, CDCl3): 3.30 C3H2s, 2.66 (2H)m, 2.36 (6H)s and 2.10 (3H)s.
EXAMPLE 23
Starting with ~he requisite oxime and employing the procedure of
Example 22, the following compounds are prepared:
1~0~
OC~3
~ I
-36-
St rtin~ Material Product
R Bl R R
l O
H- CH3C- 0 H- H-
H- CH3CH2C- H- H-
O O O
.. 1~ ..
3 C~3CH2C- CH3C- H-
O O O
3 2 CH3C 3 2 H
O O O
.. .. ..
CH3CH2C- CH3CH2C- CH3C~2
EXAMPL~ 24
11 r 2l-Diacetyl-4''-deoxy--4l'-oxo-oleandomycin O~acetyloxime
10 To a stirring turbid solution of 20 g. of 11,2'-diacetyl-4"-deoxy-4"-
oxo-oleandomycin oxime in 250 ml. of benzene is added 8.21 ml. of pyridine fol-
lowed by 9.62 ml. of acetic anhydride, and the resulting reaction ~ixture al-
lowed to stir ae room temperature overn~ght. The solution is poured into water
and the organic layer, after washing successively with water and a saturated
brine solution and drying, is concentrated to dryness. Recrystallization of
the residual foam from ethyl acetate-hexane gave 13.4 g. of the pure product,
m.p. 198-202C.
NMR (~, CDC13): 3.38 (3H)s, 2.66 (2H)m, 2.33 (6H)s, 2.26 (3H)s and
2.10 (6H)s.
J~7~
I
EXAMPLE 25
The procedure of Example 24 is again repeated, starting ~ith the
appropriate oxime, to give the following products:
3)Z
N-O-CCH3
OCH3
CH3C- O CH3CH2C-
C~3C,H2C- CH3C- ~,
CH3C- ,O, H- i
E~3CH2C EI
5',~
EX~MPLE 26
1 11,2'-Diacetyl-4"-deoxy-4"-oxo-oleandomycin 0-methyloxime
I
¦ To 50 ml. of water and 50 ml. of methanol is added 1.25 g. of methoxy
¦ amine hydrochloride and 2.5 g. of 11,2'-diacetyl-4"-deoxy-4"-oxo-oleandomycin
5 ¦ and the resulting reaction mixture allowed to stir at room temperature over-
¦ night. The solution is added to water and the pH adjusted to 7.5 with solid
¦ sodium b~carbonate and then 9.5 with lN aqueous sodium hydroxide solution. The
¦ product is extracted into ethyl acetate and the extract dried over sodium sul-
¦ fate. Removal of the solvent in vacuo gives 2.4 g. of the desired product as
10 ¦ a white foam.
NMR ~, CDC13): 3.88 (3H)s, 3.26 (3H)s, 2.56 (2H)m, 2.30 (6H)s and
l 2.06 (6H)s.
¦ E~AMPLE 27
l Employing the procedure of Example 26 and starting with methoxyamine
15 ¦ hydrochloride and the requisite 4"-deoxy-4"-oxo-oleandomycin, the following
-¦ compounds are prepared: 2'-acetyl-4"-deoxy-4"-oxo-oleandomycin 0-methyloxime,
2'-propionyl-4"-deoxy-4"-oxo-oleandomycin 0-methyloxime, 11-acetyl-2'-propionyl
4"-deoxy-4"-oxo-oleandomycin 0-methyloxime, 11-propionyl-2'-acetyl-4"-oxo-
l oleandomycin 0-methyloxime, 11-acetyl-4"-deoxy-4"-oxo-oleandomcyin 0-methyloxi~ e
20 ¦ and 11-propionyl-4"-deoxy-4"-oxo-oleandomycin 0-methyloxime.
ll'~. S'i' }~ ~
¦ EXAMPLE 28
~ 2'-Diacetyl-8~8a-deoxy-8~8a-dihydro-4"-deoxY-4"-oxo-oleandomycin oxime
¦ A solution of 49.0 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-4"-
¦ deoxy-4"-nxo-oleandomycin and 18.1 g. of hydroxylamlne hydrochloride in 300 ml.
¦ of water and 300 ml. of methanol is allowed to stir at room temperature for
1.5 hrs. The resulting solution is added to 250 ml. of water and the pH ad-
justed to 7.5 and 9.5 with solid sodium bicarbonate and lN sodium hydroxide
solution, respectively. The product is extracted with ethyl acetate, which is
l then dried over sodium sulfate and concentrated to dryness Recrystallization
10 ¦ of the residue from ethyl acetate-hexane gives the desired product.
E~AMPLE 29
Employing the procedure of Example 28 and starting with the appro-
priate 4~-deoxy-4~-oxo-oleandomycin and requisite hydroxylamine derivatives
the following compounds are prepared:
-b,O_
.
i~;5';'~ 1
H- CH3C- N-OH
CH3CH2CCH3C N-OH
l 3 E- ~-OH
5 ¦ CH3CH2C- o
O
CH3CH2C-C~3C- ~-OCH3
~E3C-C~30- ` N-OCH3
l H- CH3C- ~ OCH3
10 ¦ CH3C- H- N-OCH3
CH3CH2C-H- N-OCH3
l l
. I
I
11 ;~'7~8
EXAMPL~ 30
11,2'-Diacetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin
O-acetYloxime
l To a ~olution of 9.9 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-dihydro-
¦ 4"-deoxy-4"-oxo-oleandomycin oxime and 4.1 ml. of pyridine in 125 ml. of ben-
zene is added 4.81 ml. of acetic anhydride and the resulting reaction mixture
allowed to stir at room temperature overnight. The reaction i~ poured into
~ater and the pH adjusted to 7.5 and 9.5 with solid sodium bicarboante and lN
l sodium hydroxide, respectively. The benzene layer is separated, dried over
10 ¦ ~odium sulfate and concentrated to give the deslred product as a white foam.
I
EXAMoeLE 31
The procedure of Example 30 is repeated, starting with the appro-
priate oxime9 to give the following compounds: 2'-acetyl-8,8a-deoxy-8,8a-di-
l hydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime, ll-propionyl-2'-acetyl-
15 ¦ 8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetylcxime, ll-propiony 1-
8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime, 2'-propiony 1-
8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime and 11-
acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-oxo-oleandomycin O-acetyloxime.
-42-
EXAMPLE 32
ll-Propionyl-8~8a-deoxy-8~8a-dihydro-4~-deoxy-4~-oxo-oleandomycin oxime
A solution of 500 mg. of 11-propionyl-2'-acetyl-8,8a-deoxy-8,8a-
dihydro-4"-deoxy-4"-oxo-oleandomycin oxime in 100 ml. of methanol is allowed
to stir at room temperature overnight. The solution is concentrated to dryness
and the residual foam purified by recrystallization from ethyl acetate-hexane.
In a similar manner is prepared 8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-
oxo-oleandomycinoxime from 2'-acetyl-8,8a-deoxy-8,8a-dlhydro-4"-deoxy-4"-oxo-
oleandomycin oxime.
EXAMPLE 33
11,2'-Diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin
0-methvlo~ime
.._ - _
A solution of 90 mg. of 11,2'-diacetyl-8,8a-deoxy-8,8a-met~ylene-4"-
deoxy-4"-oxo-oleandomycin and 45 mg. of methoxyamine hydrochloride in 2 ml. of
water and 2 ml. of methanol is allowed to stir at room temperature overnight.
The solution is poured into water and the pH adjusted to 7.5 and 9.5 with solid
sodium bicarbonate and lN sodium hydroxide, respectively. The product is ex-
tracted with ethyl acetate and the organic layer subsequently dried and con-
centrated to give 89.2 mg. of the desired product.
NMR (~, CDC13): 5.56 (3H)s, 3.33 (1.5H)s, 3.26 ~1.5H)s, 2.28 (6a)
2.06 (6H)s and 0.56 (4H)m.
5~ 7~
EXAMPLE 34
¦ 11,2'-Diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin oxime
The procedure of Example 33 is repeated employing 10.0 g. of 11,2'-
l diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin, 3.4 g. of
S ¦ hydroxylamine hydrochloride, 50 ml. of methanol and 50 ml. of water to give
¦ on work-up 9.2 g. of the desired product which can be further purified by re-
crystallization from ethyl acetate , m.p. 177-180C.
NMR (~, CDC13): 3.35 (1.5H)s, 3.25 (1.5~)s, 2.33 (6H)s, 2.06 (6~)s
l and 0.53 (4H)m.
10 ¦ EgAMPLE 35
Employing the procedure of Example 33 and starting with the re-
quisite hydroxylamine hydrochloride and appropriate 8,8a-deoxy-8,8a-methylene-
4~-deoxy-4!~-oxo-oleandomycin the following co~pounds are synthesized:
il,,.57i~ 1
I
I
3t2
OCH3
~ ~1, X
CH3C- CH3CH2C- N-OH
CH3CH2C- CH3C- ~-OH
o -I
H- CH3C- N-OH
EI- CH3CH2C- N-Oa
C~3C- ,ol H- N-OH
3 2 C~3C N OCH3
3 H2C CH3CH2C- N-OCH3
o CH3C- N-OCH3
3 H- N-OCH3
1 1~25;L~ !
EXAMPLE 36
¦ 11,2'-Diacetyl-8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-oxo-oleandomycin
0-acet~loxime
l To a suspension of 1.0 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-methylene-
¦ 4"-deoxy-4"-oxo-oleandomycin oxime in 10 ml. of benzene is added .18 ml. of
pyridine followed by .24 ml. of acetic anhydride. After 2 hrs. of stirring at
room temperature an additional .09 ml. of pyridine and .12 ml. of acetic an-
hydride are added and the stirring continued overnight. The reac~ion mixture
is poured into water and the pH adjusted to 7.5 then 9.5 by the addition of
solid sodium bicarbonate and lN sodium hydroxide, respectively. The benzene
layer is separated, dried over sodium sulfate and concentrated under vacuum
to dryness to give 890 mg. of the desired product.
NMR ~, CDC13): 3.31 (1.5H)s, 3.25 (1.5H)s, 2.25 (6H)s, 2.16 (3H)s,
2.01 (6~)s and 0.55 (4H)m.
Similarly the oximes o Example 35 are converted to their 0-acetyl
derivatives.
d 5 f ~
EXAMPLE 37
ll-Acetyl-4"-deoxy-4"-amino-oleandomycin
To a suRpension of 10 g. of 10% palladium-on-charcoal in 100 ml. of
methanol is added 21.2 g. of a~monium acetate and the resulting ~lurry is
treated with a solution of 20 g. of 11-acetyl-4"-deoxy-4"-oxo-oleandomycin in
100 ml. of the same solvent. The suspension is shaken at room temperature in
a hydrogen atmosphere at an initial pressure of 50 p.s.i. After 1.5 hrs., the
catalyst is filtered and the filtrate is added with stirring to a mixture of
1200 ml. of water and 500 ml. of chloroform. The pH is adjusted from 6.4 to
4.5 and the organic layer is separated. The aqueous layer, after a further
extraction with 500 ml. of chloroforml is treated with 500 ml. of ethyl acetate
and the pH adiusted to 9.5 with 1~ sodium hydroxide. The ethyl acetatP lay~r
is separated and the aqueous layer extracted again with ethyl acetate. The
ethyl acetate extracts are combined, dried over sodium sulfate and concentrated
to a yellow foam (18.6 g.), which on crystallization from diisopropyl ether,
provides 6.85 g. of the purified product, m.p. 157.5-160C.
NMR (~, CDC13): 3.41 (3~)s, 2.70 (2H)m, 2.36 (6~)s and 2.10 (3H)s.
The other epimer, whlch exists in the crude foam to the extent of
20-25%, is obtained by gradual concentration and filtration of the mother
20 ~ 1 ~or~.
~,
l~ _47_ '
11~5~
¦ EXAMPLE 38
¦ Employing the procedure of Example 37 and starting with the appro-
¦ priate 4"-deoxy-4"-oxo-oleandomycin, the following amines are prepared:
E;O~q~
R Rl ~MR (~, CDC13)
l _ _ .
5¦ C~3C- CH3C- 3.43 (3H)s, 2.70 (2H)m, 2.30 ~6H)s
and 2.10 (6H)s.
H- H- 5.60 (lH)m, 3.36 (3H)s, 2~83 (2H)m
and 2.30 (6H)s.
l H- ~13C- 5.80 (lH)m9 3.43 (3~)s, 2.80 (2H)m,
lQ ¦ 2.30 (6H)s and 2.10 (3E)s.
~ ~ 7~
EXAMPLE 39
The procedure of Example 37 i5 again repeated starting with the re-
quisite 4"-deoxy-4"-oxo-oleandomycin and using isopropanol as the solvent to
give the following products: 2'-propionyl-4"-deoxy-4"-amino-oleandomycin, 11-
acetyl-2'-propionyl-4"-deoxy-4"-amino-oleandomycin, 11-propionyl-2'-acetyl-4"-
deoxy-4"-amino-oleandomycin and 11,2'-dipropionyl-4"-deoxy-4"-oxo-oleandomycin
EXAMPLE 40
ll-Acetyl-4"-deoxy-4"-amino-oleandomycin
To a stirred suspension of 50 g. of 11-acetyl-4"-deoxy-4"-oxo-olean-
domycin and 53 g. of ammoniu~ ace~ate in 500 ml. of methanol cooled to -10C.
is added dropwise over a one hour period a solution of 3.7 g. of 85% sodium
borocyanohydride in 200 ml. of methanol. After stirring for 2 hrs. in the
cold, the reaction is poured into 2.5 1. of water and 1 1. of chloroform. The
pH is adjusted from 7.2 to 9.5 by the addition of lN sodium hydroxide and ~he
organic layer separated. The aqueous layer is washed once with chloroform and
the organic layer combined. The chloroform solution of the produc~c ls treated
with 1.5 1. of water at pH 2.5, and the water layer separated. The pH of the
aqueous layer is adjusted from 2.5 to 7.5 and then to 8.25 and is followed by
an ethyl acetate extraction. These extracts are discarded ~nd the pH is finallY
raised to 9.9. The aqueous layer is extracted (2 x 325 ml.) with ethyl acetat~
and the extracts combined and dried over sodium sulfate. Removal of ~he sol- !vent under reduced pressure gives 23.9 g. of the product as a foam.
NMR ~C, CDC13~: 3.41 (3H)s, 2.70 (2H)m, 2.36 (6H)s and
2.10 (3~)s.
_49_
ll
il~5'7~
EXAMPLE 41
4~-Deoxy-4 ~amino-oleandomYcin
A solution of 20 g. of 2'-acetyL-4"-deoxy-4"-oxo-oleandomycin in
l 125 ml. of methanol, after stirring at room temperature overnight, is treated
with 21.2 g. of ammonium acetate. The resulting solution is cooled in an ice
bath and treated with 1.26 g. sf sodium cyanoborohydride. The cooling bath is
then removed and the reaction mixture allowed to stir at room temperature for
2 hrs. The reaction i~ poured into 600 ml. of water and 600 ml. of diethyl
ether and the pH adjusted from 8.3 to 7.5. The ether layer is separated and th~
aqueous extracted with ethyl acetate. The extracts are set aside and the pH
of the aqueous adjusted to 8.25. The die~hyl ether and ethyl acetate extracts
made at this pH are also set aside, and the pH raised to 9.9. The diethyl ethe
and ethyl acetate extracts at this pH are combined, ~ashed successively with
water (lx) and a saturated brine solution and dried over sodium sulfate. The
latter extracts, taken at pH 9.9, are concentrated to a foam and chromatographe
on 160 g. of silica gel, using chloro~orm as the loading solvent and initial
eluate. Af~ar eleven fraction, which amounts to 12 ml. per fr~ction, are ~aken
the eluate is changed to 5% methanol- 95% chloroform. At fraction 370 the
eluate is changed to 10% methanol -90% chloroform and at fraction 440, 15%
20 methanol -85% chloroform is u~ed. Fractions 85-260 are combined and concen-
trated in vacuo to dryness to provide 2.44 g. of the desired product.
~2 (~, CD~13): 5.56 (ld)=, 3.36 (3d)s, 2.3 (2d)= a~d 2.26 (6d)s,
-50-
~1~57~8
¦ EXAMPLE 42
Employing the procedure of Example 40 and starting with the appro-
priate 4"-deoxy-4"-oxo-oleandomycin and using isopropanol as the solvent, the
l following compounds are prepared: 11,2'-diacetyl-4"-deoxy-4"-amino-oleandomyci] I,
5 ¦ 2'-propionyl-4"-deoxy-4"-amino-oleandomycin, 2'-acetyl-4"-deoxy-4"-amino-
oleandomycin, ll-acetyl-2'-propionyl-4"-deoxy-4"-amino-oleandomycin, ll-pro-
pionyl-2'-acetyl-4"-deoxy-4"-amino-oleandomycin and 11,2'-dipropionyl-4"-deoxy-
4"-amino-oleandomycin.
EXAMPLE 43
10 4"-Deoxy-4"-amino-oleandomycin
A solution of 300 mg. of 2'-acetyl-4"-deoxy-4"-amino-oleandomcyin
in 25 ml. of methanol is allowed to stir under a nitrogen atmosphere overnight
at room temperature. The reaction mixture is concentrated in vacuo to give
286 mg. of the desired product as a white foam.
NMR (~, CDC13): 5.56 (lH)m, 3.36 ~3H)s, 2.90 (2H)m and 2.26 (6H)s.
~1'.,5'i~
I EXAMPLE 44
I
¦ The procedure of Example 43 is repeated, starting with the requisite
2 -alkanoyl-4 -deoxy-4 -amino-oleandomycin to give the following compounds:
~o,~ 2
l Starting Material Product
5 1 R Rl R
l O O O
.. ,- .,
C~3C- CH3C- CH3C- H-
l O O O
l - .. "
~3C- C~3CH2C- CH3C- H-
l O O O
l . " .,
CH3CH2C- CH3C- C 3CH2C H
l O O O
I
l CH3CH2C- CH3CH2C-CH3CH2C- H-
l O
10 1 H- 3 2 ~ H-
~ -52-
~,
I
ili~5~
EXAMPLE 45
Acetyl~8,8a-deoxy-8,8a-dihydro-4"-deoxy-4"-amino-oleandomycin
A solution of 2.15 g. of 11-acetyl-8,8a-deoxy-8,8a-dihydro-4"-deoxy-
1 4"-oxo-oleandomycin and 2.31 g. of ammonium acetate in 15 ml. of methanol cooled
5 ¦ to 20C is treated with 136 mg. of sodium cyanoborohydride. After stirring
for 45 min. at room temperature the reaction mixture is poured into 60 ml. of
water and 60 ml. of diethyl ether~ and the pH adjusted from 8.1 to 7.5. The
ether layer is separated and discarded~ and the pH of the aqueDus raised to 8.0
l Fresh ether is added, shaken with the aqueous layer and di carded. The pH i~
10 ¦ adjusted to 8.5 and the process repeated. Finally, the p~ is adjusted to 10.0
¦ and 60 ml. of ethyl acetate is added. The aqueous layer is discarded and the
ethyl acetate treated with 60 ml. of fresh water. The pH of the water layer is
adjusted to 6.0 ~ith lN hydrochloric acid and the ethyl acetate layer di~carded
The aqueous layer is successively exeracted at pH 6.5, 7.0, 7.5, 8.0 and 8.5
with ethyl acetate (60 ml.) and the organic extracts set aside. The pH i~
finally raised to 10.0 and the aqueous extracted with ethyl acetate. The ex-
tracts taken at pH 8.0, 8.5 and 10.0 are combined and concentrated under vacuum
to give 585 mg. of a white foam, which consists of a pair of 4"-epimers.
NMR (~, CDC13): 3.38 and 3.35 (3H) 2 Yinglets, 2.31 and 2.28 (6H)
2 singlets and 2.03 (3H).
11~57~13
EXA~PLE 46
8,8a-Deoxy-8,8a-dihydro-4"-deoxy-4''-am no-oleandom~cin
l Sodium cyanoborohydride (126 mg.) is added to a solution of 1.86 g. o~
¦ 8,8a-deoxy-8,8a-dihdyro-4"-deoxy-4"-oxo-oleandomycin and 2.1 g. of ammonium
5 ¦ acetate in 10 ml. of methanol at room temperature. After one hour the reaction
mixture is cooled to 0C. and allowed to stir for 2.5 hrs. The reaction mix-
¦ ture is poured into 60 ml. of water and 60 ml. of diethyl ether and the pH ad- ¦
justed to 7.5. The ether layer ic discarded and the aqueous layer adjusted
l successively to 8.0 and 8.5 being extracted with ether following each ad~ust-
10¦ ment. The aqueous is finally ad~usted to pH 10.0 and is extracted with ethyl
acetste. Fresh water is added to the ethyl acetate extracted and the pH ad-
justed to 6Ø The ethyl acetate layer is discarded and the aqueous layer
adjusted successively to pH 6.5, 7.0, 8.0, 8.5 and 10.0, the aqueous layer being
l extracted after each pH adjustment with ethyl acetate. The ethyl acetate ex-
15¦ tracts at pH's 7.5, 8.0 and 10.0 are combined and concentrated to a foam which !
is reconstituted in ethyl acetate and extracted with fresh water at p~ 5.5.
The acid aqueous layer is successively adjusted, as before, to pH's 6.0, 6.5,
7 0~ 7 5~ 8.0 and 10.0 being extracted after each adjustment with diethyl
l ethe~. The ether extracts at pH 7.5, 8.0 and 10.0 are combined and concentrated
201 to dryness in vacuo to give 166 mg. of the desired product.
¦ ~MR ~, CDC13): 5.48 (lH)m, 3.40 (3H)s and 2.30 (6H)~.
I
l 1.
11~'5~
EXAMPLE 47
l The procedure of Example 45 is repeated, starting with the requisite
¦8,8a-deoxy-8,8a-dihydro-4'~-deo~y-4"-oxo-oleandomycin and using isopropanol as
the solvent to ~ive the following compounds:
- R ~ (CH3)2
¦ RO~", ~ ~
~ ' ~ ~NH2
; I OCH3
H- ~o~ C~3c~-
- CH3C-
CH3CH2C- C~3C-
~H3~N~C- ~-
11~
EgAMPLE 48
ll-Acet 1-8,8a-deQxy-8,8a-methylene-4"-deoxy-4"-amino-oleandomycin
_~
To a methanol C30 ml.) solution of S.0 g. of 11-acetyl-8,8a-deoxy-8,8 _
methylene-4"-deoxy-4"-oxo-oleandomycin and 5.2 g. of ammonium acetate cooled to
20C. is added 300 mg. of sodium cyanoborohydride. The reaction mixture is
allowed to stir at room temperature for one hour, and is then poured into 120
ml. of water and 120 ml. of diethyl ether. The aqueous layer is adjusted to
pH 7.5, 8.0, 8.5 and lO.0, successively, being extracted after each pH adjust-
ment with ethyl acetate. The final organic extract made at pH 10.0 is treated
with water and ehe pH adjusted to pH 6. The aqueous layer is treated again as
above and the pH adjusted to 7.0, 7~5, 8.0, 8.5 and 10.0, being extracted with
ethyl acetate subsequent to the p~ change. The ethyl acetate extracts follow-
ing p~ changes at 8.0, 8.5 and 10.0 are combined and concentrated in vacuo to
gi~e 1.5 g. of the desired product.
15 ¦ RMR ~, CDC13): 3.38 (35)o, 2.30 (65)o, 2.05 (3~)s nd 0.65 (4H)m.
l ~
¦ E~aMple 49
l Starting with the approprlate 8,8a-deoxy-8,8a-methylene-4"-deoxy-4"-¦ oxo-oleandomycln and isopropanol as the solvent and employing the procedure of¦Example 48, the ~ollowing compounds are synthesl~ed:
R0", ~ C~3)2
~N2
5 ¦ R Rl
30- ~H3C~ Q
CH3C- 0 CH3CH2C-
CE3CH2C,- CH3C-
l CH3Ca2C- CH3C.H2C-
10 ¦ H- CH3C- ,0,
3~ CU3CU2C-
_57_
~ 7'~
¦ EXAMPLE 50
11~2~-Diacetyl-4''-deoxy-4~-amino-oleandomycin
A suspension of 1 g. of Raney nickel, washed with isopropanol, in 25
l ml. of isopropanol containing 250 mg. of 11,2'-diacetyl-4"-deoxy-4"-oxo-olean- ¦
¦ domycin O-acetyloxime is shaken in a hydrogen atmosphere at an initial pressure
of 50lp.s.i. at roo~ temperature overnight. The reaction mixture is filtered
and the filtrate concentrated under reduced pressure to give 201 mg. of the
desired product.
The entire 201 mg. in methanol (10 ml.) is refluxed for one hour givin
11-acetyl-4"-deoxy-4"-amino-oleandomycin identical with that prepared in Example¦
37.
EXAMPLE 51
Starting with the indicated 41'-deoxy-4"-oxo-oleandomycin derivative
and employing the procedure of Example 50 with the designated catalyst, the fol-
lowing 4"-deoxy-4"-amino-oleandomycins are prepared:
~ C~3)2
'"~'¢~ I
OCH3
-i I 1~'~5~
l R Rl Derivative Catalyst
I o o
C~30,- CH3C- oxime Ni
CH30,- CH3CH2C- oxime Pd/C
CH3C- H- oxime PtO2
3 2C H oxime PtO2
H- H- oxime Pd/C
CH3C- CH3C- 0-acetylox~me Ni
CH3C- CH3Ca2C- 0-acetyloxime Ni
H- CH3C- 0-acetyloxime Pd/C
" CH3C- 0-methyloxime pto2
CH3C- CH3CH2C- 0-methyloxime PdtC
3CH2C ~ 0-methyloxime Pd/C
E~AMPLE 52
Employing the procedure of Example 50, 8,8a-deoxy-8,8a-dihydro-4"-
deoxy-4'l-amino-oleandomycins are prepared which correspond to the reduction
products of the following 4"-oxo derivatiYes using the indicated catalyst:
11257'~
Ro~CEI3) 2
"`~'¢6x
OCH3
R Rl X Catalys t
H- CH3C- N-OH Pd/C
.. ~-
3 2 CH3C N-OH Ni
CH3C- H- N-OH Ni
3CH2C H N-OH pto2
H- CH3CE~2C- N-OH Ni
CH3CH2C- CH C- N-OCa3 pto2
. ~~ Ca3C- N-OCH3 Pd/C
C113C- H- N-OCa3 Pd /C
3 2 ~ N-OCH3 PtO2
O O O
CH3C- CH3C- N-O-CCH3- pto2
(Continued. . . ~
-60- !
~ 1~;5i^~
!
(Continued)
R Rl X Catalyst
CH3CH2C- CH3C- N-O-CCa3- Ni
" N-O-CCH3- Pd/C
l CH3C- H- N-O-CCH3- Ni
5 1 E~AMPLE 53
The procedure of Example 50 is again repeated, starting with the in-
dicated 8~8a-deoxy-8 8a-methylene-4~-deoxy-4~r-oxo-oleandomycin derivative and
catalyst to give the following compounds:
OC~3
-61-
I
l ~1~5 7 ~?~
Sta~ting
R Rl Derivative Catalyst
CH30- CH3C- oxime Ni
CH3C- CH3CH2C- oxime PtO2
l CR3CH2C- CH3CH2C- oxime PtO2
5 I H- CH3C- oxime Pd/C
0 0 oxime PtO2
CH3CH2C- CH3C- 0-methyloxime Pd/C
¦ CH30,- H- lol 0-methyloxime Ni
¦ . " CH3C- 0-acetyloxime Ni
CH3C- H- 0-acetyloxime Ni
C~3~- CH3C- 0-acetyloxime Pd/C
-62-
Z5~
¦ EXAMPLE 54
¦ 4"-Deoxy-4"-ethylamino-oleandomycin
l To 25 ml. of methanOl containing 4.59 g. of 4l~-deoxy-4ll-oxo-oleando-
¦ mycin, 6.6 ml. of a 5 M solution of ethylamine in ethanol and 1.89 ml. of acetic
5 ¦ acid is added 365 mg. of sodium cyanoborohydride in 50-60 mg. portions. After
stirring at room temperature for one hour, the reaction mixture is poured into
110 ~1. of water and 120 ml. of ethyl acetate.
The aqueous layer is adjusted to pH 7.5, 8.0, 8.5 and 10.0, success-
ively, being extracted after each pH adjustment with ethyl acetate. The final
lQ organic extract made at pH 10.0 is treated with water and the pH adjusted to
pH 6. The aqueous layer is treated again as above and the pH adjusted to 7.0,
7.5, 8.0, 8.5 and 10.0, being extracted with ethyl acetate subsequent to the
pH change. The ethyl acetate extracts following pH changes at 8.0, 8.5 and
10.0 are combined and concentrated in vacuo to give a foam. The product is
lS further purified by chromatographing on 75 g. of silica gel using acetone
eluates. Fractions 62-104, each fraction comprising 4 ml., are combined and
ur~ - 9-~ r~
-63-
11257.~ 1
EXAMPLE 55
ll-Acetyl-4"-deoxy-4"-ethylamino-oleandomycin
I
In a manner similar to Example 54 366 mg. of sodium cyanoborohydride
l is added portionwise to a solution of 5.82 g. 11-acetyl-4"-deoxy-4"-oxo-olean-
5 ¦ domycin and 16 ml. of a 5.0 solution of ethylamine in ethanol in 27.4 ml. of a2.92 M solution of hydrogen chloride in ethanol. After stirring at room tem-
perature for 1.5 hrs., the reaction mixture is poured into 120 ml. of water and
12Q ml. of ethyl acetate and worked up as in Example 54 to give 1.2 g. of the
desired product.
EXAMPLE 56
ll-Acetyl-4"-deoxy-4"-n-hexylamino-oleandomycin
The procedure of Example 54 is repeated, starting with 4.8 g. of 11-
acetyl-4"-deoxy-4"-oxo-oleandomycin, 6.7 g. of n-hexylamine, 3.78 ml. of acetic
acid, 302 mg. of sodium cyanoborohydride and 25 ml. of metnanol, to give, after
chromatographing an 80 g. of silica gel using chloroform as the eluate, 1.3 g.
of the desired product.
~5~
¦ EXAMPLE 57
¦ Starting with the appropriate 4"-deoxy-4"-oxo-oleandomycin, isopropano L
as the solvent and amine, and employing the procedure of Example 54, the follow-¦ing compounds are prepared:
RO,~
R Rl R3
CH3C,- H- CH3-
CH3C,- H- n~C3H7~
CH30,- H- i-C3H7-
CH3C,- H- - 5 11
CH3C- CH3C,- C2H5-
CH3C,- CH3C,- l-C3H7-
cH3c,- CH3C- 0 t-C4Hg-
CH3C- CH3CH2C- CH3-
CH3C- CH3CH~C- - C6 13
(Continued. . . )
~ 1~7~ 1
(Example 57 Continued)
R R1 R3
CH3C- O CH3CH2C- i~C5Hl1-
" n 3 7
CH3CH2 n C4 9
CH3CH2 H- n-C6H13
" " CH3-
~13CH2C- CH3C,- i-C3H7-
CH3CH2C- CH3Co- n C5 11
,. ,.
CX3CH2C- CH3C- O 1_C5H11_
CH3cH2c- CH3CH2 C2H5-
CH3CH2C CH3~H2 n C6 13
CH3cH2c- CH3cH2c- i C4 9
H- H- CH3-
H- H- C2H5-
H- H- t C4 9
H- H- 1_C5~11
H- H- n C6 13
~ 7'~1~
EXAMPLE 58
ll-Acetyl-4"-deoxy-4"-dimethylamino-oleandomycin
Two grams of ll-acetyl-4"-deoxy-4"-amino-oleandomycin, 1 g. of 10%
palladium-on-charcoal and 2.06 ml. of formalin solution are combined in 40 ml.
of methanol and shaken in a hydrogen atmosphere at an initial pressure of 50
p.s.i. overnight. The spent catalyst i5 filtered and the filtrate concentrated
to dryness under reduced pressure. The residual product (1.97 g.) is chroma-
tographed on 40 g. of silica gel using chloroform as the initial eluate. After
fraction #25, which comprised 650 drops per fraction, the eluate is changed to
3% methanol in chloroform. Fractions 36-150 are rombined and concentrated in
vacuo to give 704 mg. of the desired product as a white foam.
~ MR (~, CDC13): 3.33 (3H)s, 2.63 (2H)m, 2.30 (12H)s and 2.10 (3H)s.
EXAMPLE 59
Employing the procedure of Example 53, with the exception that iso-
propanol is used as the solvent, and starting with the requisite 4"-deoxy-4"-
amino-oleandomycin, the following compounds are synthesized:
11'i,5~
~ Cll~
Cll C53cR~l o
CH3C- CH30,H2C-
C53C~12C- CR3C
CH3CH2o,- H-
CH3CH2C- CH3CH2C- 1¦
R- 5-
-68-
i~,~'57'~
EXAMPLE 60
ll-Acetyl-4"-deoxy-4"-amino-oleandomycin dihydrochloride
To 7.28 g. of 11-acetyl-4"-deoxy-4"-amino-oleandomycin in S0 ml. of
dry ethyl acetate is added 20 ml. of a lN ethyl acetate solution of hydrogen
chloride, and the resulting solution concentrated to dryness under reduced pres
sure. The residual material is triturated with ether and filtered to give the
de~ired ~alt.
By a similar procedure the amine compounds of the present invention
are converted to their di-acid addition salts.
EXAMPLE 61
11,2'-Diacetyl-4"-deoxy-4"-amino-oleandomycin hydrochloride
The procedure of Example 60 is repeated with the exception that 10 ml
of a 1~ ethyl acetate solution of hydrogen chloride is added. The solution is ¦
concentrated to dryness in vacuo and the residual mono-hydrochloride salt is
tri~urated with ether and filtered.
By a cimilar procedure the amine compound~ of the present invention t
~re converted to their mono-acid addition salts.
,...
.
I
-69- ~
S~
EXAMPLE 62
ll-Acetyl-4"-deoxy-4"-amino-oleandomycin aspartate
To 960 mg. of 11-acetyl-4"-deo~y-4"-amino-oleandomycin in 6 ml. of
acetone at 40C. is added 18 ml. of water followed by 175 mg. of aspartic acid.
The mixture is heated to reflux until a hazy solution is affected. The mixture
is filterPd hot and the clear filtrate i8 concentrated to remove the acetone.
The remaining solution is then freeze-dried to gi~e the product as ~he residual
material.
_70~
~1
:~Z~74~ 1'
PREPARATION A
2'-Acetyl-8,8a-deoxy-8 8a-dihyd~o-oleandomycin
la. 2'-Acetyl-8,8a-deoxy-oleandomycin
A 250 ml , 3-necked round bottom flask is charged with zlnc dust
Clo g.~ and mercuric chloride Cl g.). After the solids are mixed well, lN HCl
~25 ~1.) is added and the mixture is stirred vigorously for 15 min. The aqueouc
supernate is removed and fresh lN HCl (25 ml.~ added and the flask placed under
a carbon dioxide atmosphere. A filtered solution of chromium trichloride
(50 g. in 65 ml. of lN ~Cl~ is added rapidly to the zinc amalgam. The mixture
0 i9 stirred under a carbon dioxide atmosphere for 1 hr. during which time a
light blue color develops indicating the presence of chromous chloride (CrC12)~
Stirring is discontinued after 1 hr. and the zinc amalgam allowed to settle to
the bottom of the flask.
A solution of 2'-acetyl-oleandomycin (29.2 g.) in acetone (200 ml.)
and water (100 ml.) is placed in a dropping funnel attached to a 600 ml., 3-
necked round bottom flask equipped with an overhead mechanical stirrer. To
this flask is added, under a carbon dioxide atmosphere and with stirring, the
solution of 2'-acetyl-oleandomycin and the previously prepared solution of
chromous chloride. The solutions are added simultaneously at quch a rate that
2Q both finished at the same time. The addition takes about 12 min. After 35 min
of stirring at room temperature, water (100 ml.) and ethyl acetate (100 ~1.)
are added to the reaction and stirring continued for 15 min. The ethyl acetate
layer is separated and washed with water (80 ml.~. The ethyl acetate is se-
parated and the aqueous extracts combined and washed with fresh ethyl acetate
25 ~ ( ml.). The ethyl acetate layer is sep~rated and washed with water (100
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5~
The organlc phaae is separated and the aqueous washe~ co bined and treated with !
¦sodium chloride (75 g.2. The additional ethyl acetate which separates is sypho d
¦off and com~ined ~ith the ~ther ethyl acetate extracts. Water ic added to the
¦combined ethyl acetate extracts and adjusted to pH 8.5 with sodium bicarbonate.
5 ¦The organic layer is separated, washed with water, saturated sodium chloride anc
¦dried over anhydrous sodium sulfate. Filtration and evaporation of the solvent
¦under reduced pressure affords a white solid which is crystallized from ethyl
¦acetate/heptane to give the desired compound, 8.4 g., m.p. 183.5-185C.
¦ i 37 63 13 C, 62.2; H, 9.0; N, 2Ø
lQ ¦ Found: C~ 62.0; H, 8.9; N, 2Ø
NMR C~, CDC13): 5.63 (lH~s, 5.58 (lH)s, 3.43 (3~)s, 2.36 (6H)s and
2.08 (3~)s.
lb. 2 t -Acetyl-8,8a-deoxy-8,8a-dihydro-oleandomycin
l Aluminum foil (4.0 g.) cut $~to 1/4" pieces and covered with 290 ml.
15¦ of an aqueous mercur~c chlorlde solution is stirred for 30-45 sec. The solu-
tion is decanted and the ~malgamated aluminum washed successively with water
~2x) isopropanol ~lx2 and tetrahydrofuran (lx~ The pieces are layered over
with 45 ml. of tetrahydrofuran, 45 ml. of isopropanol and 10 ml. of water and
l subsequently cooled to 0C. in an ice bath. A solution of 2.0 g. of 8,8a-deoxy
2'-acetyl-oleandomycin in tetrahydrofuran, isopropanol and water is added drop-
wise to the amalgamated aluminum at such a rate that the temperature remains at
0C. When the addition is complete the bath is removed and the reaction mixtur
allowed to stir at room temperature overnight. The solids are filtered and the
filtrate concentrated in vacuo to dryness. The residue is treated ~ith ethyl
a tate - tater ard the pd adjasted to 9.0 ~Ith a ~aturated sodi~- carbonate
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l ~57 ~
! ~lotion. The organic phase is separaced, wa~hed with water and a Jatura~ed
brine sulution and dried over sodium sulfate. Removal of the solvent gives
2.27 g. of the desired product.
PREPARATION B
I . _
5 ¦ 2a. 11,2'-Diacety~-8,8a-deoxy-8,8a-methylene-oleandomycin
In a flame drièd 20Q ml three-nPcked flask equipped with a dropping
funnel, magnetic stlrrer and a positi~e-pressure nitrogen inlet is combined
16.4 g. of trlmethylsulfoxonlum iodide and 3.4 g. of a 50% oil dispersion of
sodlum hydride. The solids are mixed well and 43.2 ml. of dimethylsulfoxide
0 i5 added via the dropping funnel. After one hour, when the evolution of hydro-
gen has stopped, the su~pension is cooled to 5-10C. and a solution of 22.6 g.
of 11,2'-diacetyl-8,8a-deoxy-oleandomycin in 32 ml. of tetrahydrofuran and 16 m~ .
of diemthylsulfoxide is added over a 10 min. period. The suspension is stirred
at room temperature for 90 min. and poured in 300 ml. of water and extracted
with two 3ao ml. portions of ethyl aceta~e. The organic extracts are washed
with water, saturated solution of sodium chloride, dried over anhydrous sodium
sulfate, filtered, and evaporated to dryness under reduced pressure. The resi-
due is crystallized from ether to give 8.9 g. of 11,2'-diacetyl-8,8a-deoxy-8,8a-
methylene-oleandomycin.
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