CA1140923A - Antitumor agents - Google Patents

Abstract

ABSTRACT OF THE DISCLOSURE
This disclosure describes symmetrical 1,4-bis-(substituted-amino)-5,8-dihydroxyanthraquinones useful as chelating agents and for inhibiting the growth of transplanted mouse tumors.

Description

114~923 This invention relates to new organic compounds and, more particularly, is concerned with novel symmetri-cal l,4-bis(substituted-amino)-5,8-dihydroxyanthraquinones which may be represented by the following general formula:

OH NH -Q-N ~ 1 OH NH-Q-N

wherein Q is a divalent moiety selected from the group consisting of those of the formulae:

lCH3 IH3 1CH3lH3 -(CH2)n~ ~ -CH-CH2- , -CH2-CH- , -CH-CH- , IC2Hs IC2H5 lCH3 -CH-CH2- ,-CH -CH- -CH-CH2-CH2-ClH3 ICH3 -cH2-cH-cH2- and wherein n is an integer from 2 to 4, inclusive; Rl and R2 are each individually selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms, mono-~, -1-1~4~9Z3 hydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, dihydroxyalkyl having from 3 to 6 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, formyl, alkanoyl having from 2 to 4 carbon atoms, trifluoroacetyl and moieties of the formula:

- (CH2) n-CN , - tCH2) n~O~R and (CH2) n N ~R

wherein n is an integer from 2 to 4, inclusive, R is alkyl having from 1 to 4 carbon atoms, and R3 and R4 are each individually selected from the group consisting of hydro-gen, alkyl having from 1 to 4 carbon atoms, and monohy-droxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hy-droxy group, and R3 and R4 taken together with their associated N(itrogen) is morpholino, thiomorpholino, pi-perazino, 4-methyl-1-piperazino or a moiety of the formula:
~
-N (CH2)m J , wherein m is an integer from 2 to 6, inclusive; with the first proviso that the ratio of the total number of car-bon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in the side chains at the l-position and the 4-position may not exceed 4; provided that when Q is -(CH2)n- then:

~'

-2-a) Rl, and R2 may not simultaneously be hydrogen;
b) when n is 2 then Rl/R2 may not be CH3/CH3, C4Hg/C4Hg, or -(CH2)s-;n is 3 then Rl/R2 may not be H/H, CH3/CH3, C3H7/C3H7, or -(CH2)2-0- (CH2)2-; n is 4 then Rl/R2 may not be C2H5/C2H5, C4Hg/C4Hg, or -(CH2)4-; and c) only one of Rl and R2 may be alkanoyl.
The compounds of the above general formula I
wherein A-B is CH2-CH2 are generally stable compounds and are known as the leuco forms of the corresponding anthra-quinones. These leuco froms are known to exist in their respective tautomeric forms, and all such forms are equivalent for the purposes of the invention. The leuco forms (herein, leuco bases), and the tautomers thereof may be represented by the following general formula:

OH C) I~IH - Q- N ~ 1 C (II, leuco bases) OH (~ NH-Q-N~

~ ~ ~ -N~ 1 ~ ~ ,Rl (III, tautomeric form) OH o N-Q-N

-3-1 ~ 4~ ~ Z3 - The invention also relates to processes for preparing the novel compounds represented by the above formulae I, II and III.
Thus, one process contemplated by the instant invention comprises reacting a compound represented by the following general formula:

~5 ~ [~ Q~s - X

~A
~ N~ -Q~-S - -X

with a compound of the formula: Y[ Q]T Z
wherein Rs is hydroxy, chloro, or alkanoyloxy one of X and Y is NRlR2 or NHCOCF3 and the other is selected from the group consisting of ORlo, Cl, Br, I, N(R10)2~ NO2, SO3Rlo~ S2R10~ SR10~ SR10~ N3, ONO, and tetrazolyl, wherein Rlo is selected from the group consisting of hydrogen, alkyl of from 1 to 6 carbon atoms, phenyl, para-tolyl and benzyl; S and T are different and are chosen from 0 to 1;
Z is selected from the group consisting of hydrogen, alkyl of from 1 to 4 carbon atoms, and NRlR2 or a group convertible thereto; provided:
a) that only X may be NHCOCF3 and only when S
is O and Z is not hydrogen or alkyl;
b) except as in a), when S is O, then Rl/R2 is H/H and Z is not hydrogen or alkyl of from 1 to 4 carbon atoms;

.':

114~Z3 c) when T is O and Y is NRlR2 then Z is hydrogen;
d) when T is O and X is NRlR2 at least one of Rl/R2 is hydrogen and Z is alkyl of from 1 to

4 carbon atoms;
e) only one of Rl and R2 may be alkanoyl; and when Rs = chloro or alkanoyloxy converting them to hydroxy, and when X is NHCOCF3, re-moving the COCF3 group, and when Z is a group convertible to NRlR2 effecting its conversion thereto; and, if desired: converting A-B when it is CH2-CH2 into CH=CH; and contacting the product with a pharmacologically acceptable salt-forming reagent under salt-forming con-ditions.
The invention also relates to processes for preparing the novel compounds of the following general formula:

NH-Q-N ~1 N~l-Q-N ~

and the pharmacologically acceptable acid-addition salts ~hereof; wherein Rl, R2, and Q are as hereinabove defined.
The invention also relates to the acid-addition salts of the novel compounds of formulae I, II and III.
The invention also relates to pharmaceutical therapeutic preparations and compositions comprising, ,~
. _5_ ll~V923 compounds of formula I, II and III and their pharmaceu-tically acceptable acid-addition salts, as well as to mixtures thereof as the active ingredients.
The invention also relates to methods for using the compounds and pharmaceutical preparations and compo-sitions comprising the compounds disclosed herein.
The novel compounds of the instant invention are represented by the following general formula:

~H ~ N1~-Q~

~N~

the tautomeric forms and the pharmacologically accepta~le acid-addition salts thereof, wherein A-B is selected from the group consisting of CH=CH and CH2-CH2;
Q is a divalent moiety selected from the group consisting of those of the formulae:

~113 fll3 ~113CI~3 (C1]2 )n ~ ~ C~32- , -C112-CII- , , -Cll-C~
~2~15 C2}15 C,,3 25- 11-C112- ; -C112-~ 11-C1l2-c~l2 ' f~l3 - C~3 -CH2-CI1-C1~2- and 2 2 114(3~Z3 wherein n is an integer from 2 to 4, inclusive; Rl and R2 are each individually selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms; mono-hydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, dihydroxyalkyl having from 3 to 6 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, formyl, alkanoyl having from 2 to 4 carbon atoms, trifluoroacetyl and moieties of the formulae:

-~CH2)n-CN , -(CH2)n-O-R and ~(CH2)n- ~ 3 wherein n is an integer from 2 to 4, inclusive, R is alkyl having from 1 to 4 carbon atoms, and R3 and R4 are each individually selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms and monohydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group; and Rl and R2 taken together with their associated N(itrogen) atom is morpholino, thiomorpholino, piperazino, 4-methyl-1-piperazino or a moiety of the formula:

-N ~ 2)m wherein m is an integer from 2 to 6, inclusive, provided that the ratio of the total number of carbon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in the side chains at the - 114~9Z3 l-position and the 4-position may not exceed 4;
provided that when Q is -(CH2)n- then:
a) Rl and R2 may not simultaneously be hydrogen;
b) when n is 2 then Rl/R2 may not be CH3/CH3, C4Hg/C4H9, or -(CH2)5; n is 3 then Rl/R2 may not be H/H, CH3/CH3, C3H7/C3H7, or -(CH2)2-;n is 4 then Rl/R2 may not be C2Hs/C2Hs, C4Hg/C4Hg, or -(CH2)4-;
and c) only one of Rl and R2 may be alkanoyl.
These compounds are generally obtainable as reddish brown to blue black crystalline materials having characteristic melting points and absorption spectra and which may be purified by leaching with lower alkanols since the free bases are for the most part insoluble in water and some of them are insoluble in most organic sol-vents. The organic bases of this invention (I, II and III), form non-toxic addition salts with a variety of pharmacologically acceptable organic and inorganic salt--forming reagents. Thus, acid-addition salts, formed by admixture of the organic free base with 1,2 or up to eight equivalents of an acid, suitably in a neutral sol-vent, are formed with such acids as sulfuric, phosphoric, hydrochloric, hydrobromic, sulfamic, citric, lactic, malic, succinic, tartaric, acetic, benzoic, gluconic, ascorbic, and the like. The preferred acids are hydro-chloric and acetic. For purposes of this invention the free bases are equivalent to their non-toxic acid-addition salts. The acid-addition salts of the organic bases of the present invention are, in general, crystalline solids, relatively soluble in water, methanol and ethanol but ~r~

1~4~23 relatively insoluble in non-polar organic solvents such as diethyl ether, benzene, toluene, and the like.
It will be understood by those skilled in the art that all such salts may be reconverted to their re-spective free forms represented by formulae I, II and III
by methods known to those skilled in the art, and all such salts and free forms are therefore equivalent for purposes of the invention.
A preferred embodiment of the novel compound aspect of the present invention may be represented by the following general formula:

ON O NH-Q-N

and the pharmacologically acceptable acid-addition salts thereof, wherein A-B and Q are as hereinbefore defined;
Rl is hydrogen, alkyl having from 1 to 4 carbon atoms or monohydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atoms alpha to the nitrogen from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, dihydroxy-alkyl having from 3 to 6 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group or a moiety of the formula:
,_,R3 ~ (C1~2 ) n~

_g_ 114q~923 wherein n, R3 and R4 are as hereinbefore defined; with the proviso that the ratio of the total number of carbon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in each of the side chains at the l-position and the 4-position may not exceed four.
A second preferred embodiment of the novel com-pound aspect of the present invention may be represented by the following general formula:

01~ ~ Nll-Q- l_ (C112) n 011 ~\~ '1 }~ o Nl~-Q-~l- (Cl12) -011 and the pharmacologically acceptable acid-addition salts thereof, wherein R is hydrogen or alkyl having from 1 to 4 carbon atoms and A-B, n, and Q are as hereinabove defined with the proviso that the ratio of the total of number of carbon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in each of the side chains at the l-position and the 4-posi-tion may not exceed 4.
A third preferred embodiment of the novel com-pound aspect of the present invention relates to compounds of the following formula:

~ ,j ,~ -10-114~23 ~~ 1~ C l2 O NH-(C~ )-N
2 n \R

wherein n, Rl, R2 and A-B are as defined above for the first preferred embodiment and the pharmacologically acceptable acid-addition salts thereof.
A fourth preferred embodiment of the novel compound aspect of the present invention relates to com-pounds of the following formula:

~ 2CIl2~cl~2cl12O

NllC112C~l2I C112C~21 Rl wherein Rl is hydrogen or -CH2CH2OH, and the pharmacologi-cally acceptable acid-addition salts thereof. The pre-ferred salts are the hydrochloride and the acetate.
The novel compounds of the present invention represented by formulae I, II and III may be readily prepared by the following general process.
A compound of the formula:

~

114~Z3 OH O OH
~A

OH OH

is reacted with a compound of the formula H N-Q-N

wherein A-B, Q, Rl and R2 are as defined above and if desired converting the product into the tautomer thereof;
and if desired converting A-B when it is CH2-CH2 into CH-CH and contacting the product with a pharmacologically acceptable salt-forming reagent under salt-forming condi-tions.
The reaction is preferably carried out by heating the reactants at a temperature of from about 40 to 110C. for a period of from about 2 to about 10 hours in a reaction inert solvent such as water, a lower alkanol, for example, methanol, ethanol, propanol, i_ -propanol, any of the isomeric butanols and the like, or in an amide solvent such as formamide, dimethyl formamide and the like, or in a solvent such as N,N,N',N',-tetra-methyl ethylenediamine or mixtures thereof. The above temperatures, times and solvents and combinations thereof will most normally suffice for carr~ing out the instant process. ~ther solvents and reaction parameters may at times be desirable or required and the choice of such 114~9Z3 additional solvents and reaction parameters is within the skill of the art and are all considered equivalents for the purposes of the instant invention.
In most instances, the product will be a solid and will crystallize spontaneously or upon seeding or scratching, from the reaction solvent upon cooling and may be collected by filtration or decantation. In other instances the reaction mixture may be concentrated, for example at elevated temperature under vacuum, and upon cooling the product will crystallize and may be collected by filtration or decantation as above. In certain other instances it may be necessary to evaporate the solvent to dryness in order to collect the product or alterna-tively to mix or dilute the reaction mixture with another miscible solvent such as water and proceed to collect the product by for example filtration or extraction. Those skilled in the art will be able to choose which procedure to follow and all such procedures are considered equiva-lent for the purposes of this invention. The product, once collected, may be purified by for example, crystal-lization, chromatography (thin layer or column) or pre-ferably by leaching with a lower alkanol. Other proce-dures such as maceration or crushing in a solvent, for example, an organic solvent such as ethanol may be utilized and all purification procedures are to be con-sidered as equivalents for purposes of the invention.
Those skilled in the art will recognize that when a product in the leuco form is desired, the tricyclic starting material should be in the leuco form, and care should be taken to protect such materials, especially 1~4~23 when at elevated temperatures (i.e. above room tempera-ture), from oxidizing agents such as, for example oxygen.
Thus, when a leuco product is desired the reaction is normally and e~ficiently carried out in an atmosphere to the exclusion of air. Thus, the reaction S may be carried out in an atmosphere of nitrogen or argon, for example, and this precaution should also be taken during purification procedures especially those requiring elevated temperatures. When an aromatic product is desired and an aromatic starting material is utilized no sùch precautions are normally necessary.
It is well known in the art that the leuco form may be readily converted to the aromatic form when desired by a variety of methods. Thus, air oxidation is one such method, other methods are treatment with for example, hot nitrobenzene, chloranil, hydrogen peroxide, or sodium perborate. All such methods, and other methods for conversion of the leuco form to the aromatic form, are to be considered equivalents for purposes of the invention.
Those skilled in the art will also understand that when the substituent Z is a group convertible to - ~ that the said conversion will be able to be 2S carried out by obvious procedures, well known to those skilled in the art and within the skill of the art. Con-version of the group Z to N ~ Rl by all of these , ~14~23 procedures is therefore contemplated by the instant inven-tion and all such procedures are to be considered equiva-lent for purposes of the instant invention. The preferred procedures contemplated by the instant invention for converting the group Z into ~ ~1 are exemplified below. It is to be understood that these are merely examples of the said conversion, and are not intended to delimit the scope of the instant invention. Other con-version procedures will be known to those skilled in the art and are to be considered full equivalents for the purposes of the instant invention. Thus, when Z is an aldehyde or ketone group, i.e. -~-R wherein R is chosen from hydrogen or alkyl, treatment with an amine of the formula / Rl wherein Rl and R2 are as herein IIN
\ ~2 defined under reducing conditions such as hydrogen with Raney nicket catalyst or sodium borohydride will accom-plish the desired conversion.
Also contemplated for Z are moieties of the formula ,~
-N O
(~C1~2) ) wherein m is 2 or 3. Treatment of this group with acid, for example aqeuous-ethanolic hydrogen chloride at 40--80C. for from 1 to 6 hours will convert it to -NH-(CH2)m-OH which is a moiety contemplated by the group N,-Rl 1~4V~Z3 Also contemplated for Z are efficient leaving groups, such as Cl, Br, I, para-tosyl, OSO2CH3 and the like. When Z is an above disclosed leaving group or an equivalent, treatment with an amine of the formula Rl in for example a lower alkanol solvent ~ R2 will effect the desired conversion.
In addition, when Z is a primary or secondary amine, i.e. -NH2 or -NHRl, then further alkylation will convert it to the desired group ~ Rl . Well known 1~2 alkylating agents are known which will readily effect this conversion. Such agents as alkyl halides, alkyl sulfates, substituted and unsubstituted acrylonitriles, and the like are contemplated. Aldehydes and ketones condensed under reducing conditions may also be utilized.
All such conversions and other conversion procedures are considered equivalents for the purposes of the invention.
A preferred embodiment of the primary process aspect of the present invention may be represented by the following reaction scheme:

o fH

S/~ + H N ,- R

R5 ~H (V ) \
lIV) ~

~ / ~ Q-N

(II) ~~~

R5 O NH-Q-N~

~I) wherein Rl, R2, Rs, and Q are as hereinabove defined.
In accordance with this reaction scheme, leuco 1,4-di-hydroxyanthraquinone (IV) is condensed with an appropri-ate alkylene diamine (V) in a solvent such as N,N,N',N'--tetramethylethylenediamine, ethanol, water, dimethyl-formamide, or mixtures thereof at from about 40C. to about 60C. under an atmosphere of nitrogen for several hours to produce the corresponding leuco bases (II).
7,- ~ 30 114~23 The leuco bases (II) may be readily oxidized to the fully aromatic derivatives (I) by a variety of methods such as air oxidation or treatment with hot nitrobenzene, or treatment with chloranil, hydrogen peroxide, or sodium perborate. The product may then be contacted with a pharmacologically acceptable quaternizing or salt-forming reagent under quaternizing or salt-forming conditions.
A preferred subgeneric embodiment of the primary process aspect of the invention may be repre-sented by the following reaction scheme A:
OH O OH

H r 2 Q ~R ~ ,R
OH O NH-Q-N
' ~ ~ ~r ~ ~`

I OH O NH-Q-N

OH O NH-Q-N~ 1 ~, ' r~ `~

OH O NH-Q-N
~R2 Scheme A
wherein Rl, R2 and Q are as hereinabove defined.
A more preferred subgeneric embodiment of the primary process aspect of the invention is represented ~30 4~9Z3 by the above reaction scheme A wherein Q is as herein-before defined; Rl hydrogen or alkyl having from l to 4 carbon atoms; R2 is monohydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atoms alpha to the nitrogen atom may not bear an hydroxy group, dihydroxy-alkyl having from 3 to 6 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group or a moiety of the formula:-(cll2)n-N 3 ~R4 wherein n, R3 and R4 are as hereinbefore defined; with the proviso that the ratio of the total number of carbon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in each of the side chains at the l-position and the 4-position may not exceed four, and contacting the product with a pharma-cologically acceptable acid, preferably acetic or hydro-chloric, to prepare the pharmacologically acceptable acid addition salts thereof.
A second more preferred subgeneric embodiment of the primary process aspect of the invention is represented by the above reaction scheme A wherein R
is hydrogen or alkyl having from 1 to 4 carbon atoms and R2 is -(CH2)nOH and n and Q are as hereinabove defined with the proviso that the ratio of the total number of carbon atoms to the sum of total number of oxygen atoms plus the total number of nitrogen atoms in each of the side chains at the l-position and the 4-position may not exceed four, and contacting the product with a pharmacologically acceptable acid, pre-ferably acetic or hydrochloric, to prepare the pharma-114t3~23 cologically acceptable acid addition salts thereof.
The most preferred subgeneric embodiment of the primary process aspect of the invention is represented by the above reaction scheme A wherein Q is -(CH2)n- wherein n is an integer of from 2 to 4, preferably 2, and Rl is hydrogen or -CH2CH2OH and R2 is -CH2CH2OH, and contacting the product with a pharmacologically acceptable acid, preferably acetic or hydrochloric to prepare the pharma-cologically acceptable acid addition salts thereof.
The invention also contemplates conversion of the products into acid-addition salts. Many methods for salt formation are known to those skilled in the art and are to be considered equivalent for purposes of the in-vention. Thus, for example, the product may be dissolved or suspended in a solvent such as a lower alkanol (e.g. methanl, ethanol, i-propanol) and treated with the salt forming reagent, itself in solution in the same or in a different solvent. Thus, a suspension or solution of the product in ethanol for example may be treated with dilute or concentrated acetic acid, hydrochloric acid or the like, or ethanolic-HCl and the corresponding salt may be collected by for example filtration. The salt forming reagent may also be added in pure form. Thus, the solution or suspension of the product may be treated with glacial acetic acid or gaseous HCl and the corres-ponding acid addition salt collected.
The instant invention also contemplates the use of the compounds of formulae I, II and III and the pharmacologically acceptable acid-addition salts thereof, as well as mixtures thereof, as the active ingredients . _ _ 1~4~9Z3 in pharmaceutical therapeutic preparations and composi-tions.
The therapeutic compositions of this invention inhibit transplanted mouse tumor growth and induce regression and/or palliation of leukemia and related cancers in mammals when administered in amounts ranging from about 5 mg. to about 200 mg. per kilogram of body weight per day. A preferred dosage regimen for optimum results would be from about 5 mg. to about 50 mg. per kilogram of body weight per day, and such dosage units are ranging from about 5 mg. to about 200 mg. per kilo-gram of body weight per day. A preferred dosage regimen for optimum results would be from about 5 mg. to about 50 mg. per kilogram of body weight per day, and such dosage units are employed that a total of from about 350 mg. to about 3.5 grams of the active compound for a subject of abount 70 kg. of body weight are administered in a 24-hour period. This dosage regimen may be adjusted to provide the optimum therapeutic response. For example, several divided doses may be administered daily or the dose may be proportionally reduced as indicated by the exigencies of the therapeutic situation. A
decided practical advantage is that the active compound may be administered in any convenient manner such as by the oral, intravenous, intramuscular, or subcutaneous routes.
The active compounds may be orally admin-istered, for example, with an inert diluent or with an assimilable edible carrier, or they may be enclosed in hard or soft shell gelatin capsules, or they may be ~l~V923 compressed into tablets, or they may be incorporated directly with the food of the diet. For oral therapeutic administration, the active compounds may be incorporated with excipients and used in the form of ingestible tab-lets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compo-sitions and preparations should contain at least 0.1%
of active compound. The percentage of the compositions and preparations may, of course, be varied and may con-veniently be between about 2 to about 60% of the weight of the unit. The amount of active compound in such therapuetically useful compositions is such that a suitable dosage will be obtained. Preferred compositions or preparations according to the present invention are prepared so that an oral dosage unit form contains be-tween about 5 and 200 milligrams of active compound.
The tablets, troches, pills, capsules and the like may also contain the following: A binder such as gum tragacanth, acacia, corn starch or gelatin; excip-ients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate;
and a sweetening agent such as sucrose, lactose or sac-charin may be added or a flavoring agent such as peppermlnt, oil of wintergreen, or cherry flavoring.
When the dosage unit form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier Various other materials may be present as coatings or to otherwise modify the physical form of the dosage unit. For instance, tablets, pills, or capsules may be coated with shellec, sugar or both. A syrup or 1141~)923 elixir may contain the active compound, sucrose as a sweetening agent, methyl and propylparabens as preserva-tives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any dosage unit form should be pharmaceutically pure and substantially non-toxic in the amounts employed. In addition, the active compounds may be incorporated into sustained-release preparations and formulations.
The active compounds may also be administered parenterally or intraperitoneally. Solutions of the active compound as a free base or pharmacologically acceptable salt can be prepared in water suitably mixed with a surfactant such as hydroxypropylcellulose. Dis-persions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof and in oils.
Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms.
The pharmaceutical forms suitable for inject-able use include sterile aqueous solutions or dispersions and sterile powders for the extemporanous preparation of sterile injectable solutions or dispersions. In all cases the form must be sterile and must be fluid to the extent that easy syringability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of micro-organisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils.
'?~ '?L
~J -23-114{)~23 The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars or sodium chloride. Prolonged absorption of the injectable compositons can be brought about by the use in the compo-sitions of agents delaying absorption, for example, aluminum monostearate and gelatin.
Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the various sterilized active ingredient into a sterile vehicle which contains the ba~ic dispersion medium and the required other ingredi-dents from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze-drying technique which yield 2S a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.
As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic ~ 2 3 and absorption delaying agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except insofar as any conven-tional media or agent is incompatable with the active ingredient, its use in the therapeutic compositions is contemplated. Supplementary active ingredients can also be incorporated into the compositions.
It is especially advantageous to formulate parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the mammalian subjects to be treated; each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the novel dosage unit forms of the invention are dictated by and directly dependent on (a) the unique characteristics of the active material and the particular therapeutic effect to be achieved, and (b) the limitations inherent in the art of compoundng such an active material for the treat-meot of disease in living subjects having a diseased condition in which bodily health is impaired as herein disclosed in detail.
Regression and palliation of cancers are attained, for example, using intraperitoneal administra-tion. A single intravenous dosage or repeated daily dosages can be administered. Daily dosages up to about

5 or 10 days are often sufficient. It is also possible to dispense one daily dosage or one dose on alternate or ';~~~
~ -25-114~9Z3 less frequent days. As can be seen from the dosage regimens, the amount of principal active ingredient administered is a sufficient amount to aid regression and palliation of the leukemia or the like, in the absence of excessive deleterious side effects of a cyto-toxic nature to the hosts harboring the cancer. As used herein, cancer disease means blood malignancies such as leukemia, as well as other solid and non-solid malig-nancies such as the melanocarcinomas, lung carcinomas, and mammary tumors. By regression and palliation is meant arresting or retarding the growth of the tumor or other manifestation of the disease compared to the course of the disease in the absence of treatment.
The novel compounds of the present invention may be readily prepared in accordance with the following reaction scheme:

il409Z3 OH O OH
~ J H 2 N Q N 1 ~
OH OH (V) (IV) OH O NH-Q~

(II) ~R2 OH O NH_Q_N~ l NH-Q-~
(I) _ _ wherein Rl, R2 and Q are as hereinabove defined. In accordance with this reaction scheme, leuco 1,4,5,8--totrahydroxyanthraquinone (IY) is condensed with an appropriate alkylene diamine (V) in a solvent such as N,N,N',N'-tetramethylethylenediamine, methanol, ethanol, water, dimethylformamide, or mixtures thereof at from about 40C. to about 60C. under an atmosphere of nitro-gen for several hours to produce the corresponding leuco bases (II). The leuco bases (II) may be readily oxidized to the fully aromatic derivatives (I) by a variety of methods such as air oxidation or treatment with hot nitrobenzene, or treatment with chloranil, hydrogen peroxide, or sodium perborate.

~14V923 The novel compounds described herein are useful as chelating, complexing or sequestering agents. The complexes formed with polyvalent metal ions are partic-ular~y stable and usually soluble in various organic solvents. These properties, or course, render them useful for a variety of purposes wherein metal ion con-tamination presents a problem; e.g., as stabilizers in various organic systems such as saturated and unsaturated lubricating oils and hydrocarbons, fatty acids and waxes, wherein transition metal ion contamination accelerates oxidative deterioration and color formation. They are further useful in analyses of polyvalent metal ions which may be complexed or extracted by these materials and as metal carriers. Other uses common to sequestering agents are also apparent for these compounds. In addi-tion, the leuco bases (II) are useful as intermediates in the preparation of the fully aromatic derivatives (I).
The novel compounds of the present invention also possess the property of inhibiting the growth of transplanted mouse tumors as established by the following tests.
Lymphocytic leukemia P388 test The ar.imals used are DBA/2 mice all of one sex, weighing a minimum of 17 g. and all within a 3 gram weight range. There are 5 or 6 animals per test group.
The tumor transplant is by intraperitoneal injection of 0.1 ml. of dilute ascitic fluid containing 106 cells of lymphocytic leukemia P388. The test compounds are ad-ministered intraperitoneally on days one, 5 and 9 (relative to tumor inoculation) at various doses. The aminals are weighed and survivors are recorded on a ~ 2 3 regular basis for 30 days. The median survival time and the ratio of survival time for treated (T)/control (C) animals are calculated. The positive control com-pound is 5-fluorouracil given as a 60-mg./kg. injection.
The results of this test with representative compounds of the present invention appear in Table I. The criterion for efficacy is T/C x 100 > 125%.

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The criterion for efficacy is T/C x 100 ~ 125%.

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A one-gram portion of melanotic melanoma B16 tumor is homogenized in 10 ml. of cold balanced salt solution and and 0~.5 ml. aliquot of the homogenate is implanted intraperitoneally into each of the test mice. The test ; compounds are administered intraperitoneally on days one through 9 (relative to tumor inoculation) at various doses.
The animals are weighed and survivors are recorded on a regular basis for 60 days. The median survival time and the ratio of survival time for treated (T)/control (C) animals are calculated. The positive control compound is 5-fluorouracil given as a 20~mg./kg. injection. The results of this test with representative compounds of the present invention appear in Table III. The criterion for efficacy is T/C x 100 ~ 125%

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1~4~23 Ridgway Osteogenic Sarcoma The animals used are AKD2Fl/J mice, all of the same sex, weighing a minimum of 17 g, and all within a three-gram weight range. There are normally 8 animals per test group. The tumor is administered subcutaneously by trocar as five 2 mm. fragments per mouse. The test compounds are administered intraperitoneally every 4 days for a total of 6 inoculations beginning on day 15 (relative to tumor inoculation) at various doses. The animals are weighed and survivors are recorded on a regular ~asis for 90 days. ~The regression of tumors is recorded in all test animals. Table IV gives the result o~ thi& test with a representative compound of this invent;on in terms of the percentage of animals showing tumor regression.

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This invention will be described in greater detail in conjunction with the following specific examples.
Example 1 Leuco-1,4-bis~(2-dimethylaminoethyl)animo~-5,8--dihydroxy-anthraquinone A reaction mixture comprising 10.58 g. of N,N--dimethylethylenediamine, 60 ml. of N,N,N',N'-tetramethyl-ethylenediamine and 10.96 g. of leuco-1,4,5,8-tetra-hydroxyanthraquinone is flushed with nitrogen and stirred under nitrogen for 2 hours while heating with an oil bath kept at 49-51C. The mixture is allowed to cool under nitrogen. The solid is collected and washed with ethanol giving 14.78 g. of the desired product as a dark red-brown solid.
Example 2 1~4-Bis~(2-dimethylaminoethylLamino]~5-r8 dihydroxy-anthraquinone A 12.00-g. portion of leuco-1,4-bis[(2-dimethyl-aminethyl)amino-5,8-dihydroxy-anthraquinone in 100 ml. of nitrobenzene is heated under reflux for 15 minutes and then filtered while hot. The filtrate is reheated to boiling, allowed to cool, and the solid is collected and washed with ethanol giving 8.44 g. of the desired product as blue-black crystals, mp. 236-238C.
Example 3 Leuco-1,4-bis(2-morpholinoethylamino)-5,8-dihydroxy-anthraquinone A solution of 15.62 g. of N-(2-aminoethyl)-morpholine in 40 ml. of N,N,NI,N'-tetramethylethylenedi-amine is de-aerated by bubbling nitrogen through it '" ? 3 - 54 -11~0923 for 15 minutes. A lQ,~7~g. portion of leuco-1,4,5,8--tetrahydroxyanthraquinone is added slowly and the suspension is treated as described in Example 1, giving 18.07 g. of the desired product as an olive solid, mp, 223 227C.
Example 4 1,4-Bis(2-morpholinoethylamino~-5,8-dihydroxy-anthraquinone A 13.90-g. portion of leuco-,4-bis(2-morpholino-ethylamino)~5,8-dihydroxy-anthraquinone in 100 ml. of nitrobenzene is oxidized as described in Example 2 giving lQ.3Q g. of the desired product as black rods, mp. 241-243C.
Example 5 Leuco-1,4-bic~(2-diethylaminoethyllamino]-5,8--dih.ydroxyanthraquinone The procedure of Example 3 is repeated using 13.qS g. of N,N-diethylethylenediamine in place of the N-(2-a~inoethyl)-morpholine, giving 13.97 g. of the 2~ desired product as a red-brown solid, mp. 182~185C.
Exam~le 6 1!4~Bis L (2-diethylaminoethyl)amino]-5,8-dihydroxy-anthraquinone A 10.90 g. portion of leuco-1,4-bisl(2-diethyl-aminoethyl)amino]-5,8-dihydroxyanthraquinone is oxidized as described in Example 2 giving 6.35 g. of the desired product as blue-black needles, mp. 202-204C.
Example 7 Leuco-1,4-bis[2-(1-pyrrolidinyl)ethylamino~--5,8-dihydroxyanthraquinone 114~Z3 The procedure of Example 3 is repeated using 12.Q5 g, of N-2-pyrrolidinoethylamine, in place of the N~(2-aminoethyll-morpholine, and 90 ml. of the N,N,N',N'--tetramethylethylenediamine, giving 13.24 g. of the desired product as a red-brown solid, mp. 180-185C.
Example 8 1!4 Bisf ?- ~l-pyrrolidinyl)ethylamino]-5,8--dihydroxyanthraquinone An 8.61 g. portion of leuco-1,4-bis~2-1(1-lQ -pyrrolidinyllethyl]amino]-5,8-dihydroxyanthraquinone is oxidized as described in Example 2. The reaction mixture is eYaporated to dryness and the residue recrystallized from toluene, giving 5.12 g. of the desired product as ~lue-hlack crystals, mp. 193-196C.
Example 9 Leuco-1,4-bisl2-(methylamino ethylamino~-5j8--dihydroxyanthraquinone The procedure of Example 7 is repeated using 8,QQ g. of N-methylethylenediamine in place of the N-2-2Q ~pyrrolidinoet~ylamine, giving 13.73 g. of the desired product a~ a dark green solid, mp. 157-160C.
Example 10 Leuco-1l4-bis[(3-dimethylaminopropyl)amino]-5~8 -dihydroxyan hraquinone Nitrogen is bubbled through an 80-ml. portion of dimethylaminopropylamine for 15 minutes. A 10.97 g.
portion of leuco-1,4,5,8-tetrahydroanthraquinone is added slowly with stirring. The mixture is heated under nitrogen at 50-52C. for 2 hours and then allowed to cool. The solid is collected and washed with cold Ei ethanol giYing 5,59 g. of dark~ orange-red crystals, 1~4U~Z3 mp. 115-118C.
Example 11 1~4-BisL(~3-dimethylaminopropyl~amino3-5 ! 8 -dihydrox~-anthraquinone A suspension of 6ØQ g. of leuco~l,4-bis~3~di methylaminopropyl)amino~-5,8-dihydroxyanthraquinone in 60 ml. of N, N',N'-_etramethylethylenediamine is heated on a steam bath.under reflux while air is bubbled in for 12 hours. The solution is cooled, producing a solid which is collected and washed twice with.heptane and once with petroleum ether. This solid is recrystallized by extrac-tlng wi.th 35Q ml. of hot heptane, filtering and concentra-ting to 3Q0. ml. Crystallization and washing with petro-le~m ether gives 3.72 g. of the desire product as black lS needle~ mp. 154`157C, Exam~le 12 Leuco-1,4-bis~2-aminoethylamino)-5~8~dih~droxy~
anthraquinone A reaction mixture comprising 10..97 g. of 2Q leuco-1,4,5,8-tetrahydroxyanthraquinone in 80 ml. of de-aerated ~,N,N'N'-tetramethylethylenediamine containing 7.22 g. of ethylenediamine is heated and stirred under nitrogen at 8Q-SQC. for one hour. The mixture is allowed to stand under a slow flow of nitrogen, pro-ducing a solid which is collected and washed with ethyl acetate, acetonitrile and petroleum ether giving 13.8 g.
of t~e desired product as a red-black solid.
Example 13 Leuco-1~4~bis(3-aminopropylamino~5f8-dihydroxy 30. anthraquinone ,. J

il4~923 A suspension of 10.97 g. of leuco l,4,5,8--tetrahydroxyanthraquinone in a de-aerated solution of 8.9 g. of 1,3-diaminopropane in 80 ml. of N,N,N',N'--tetramethylethylenediamine is stirred and heated at 49C.
for one hour under nitrogen, then allowed to cool. The resulting solid is collected and washed with cold ethanol giving 14.21 g. of the desired product as a black solid.
Example 14 Leuco-1,4-bis~ ~ rox~ethyla no)ethylamino~
lQ -5~8-dihydroxyanthraquinone A suspenion of 12.5 g. of 2-(2-aminoethyl-animo)ethanol in 40 ml. of N,N,N',N'-tetramethylethylene-diamine is stirred and de-aerated by bubbling nitrogen in for 15 minutes. A 10.97-g. protion of leuco-1,4,5,8-tetrahydroxyanthraquinone is gradually added with stir-ring. The suspension is heated and stirred under nitro-gen in an oil bath at 5Q-52C. for 5 hours. The mixture is allo~ed to stand and cool under nitrogen for 12 hours.
The solid is collected by decantation, macerated in etha-2Q nol, collected and wash with ethanol giving 15.06 g. of the de~ired product as a green-gray solid, mp. 129-131C.
~xam~le 15 Leuco~1,4-his~2-~di(~hydr no]-5~8-dihydroxyanthraquinone A solution of 17.8 g. of N,N-di~hydroxyethyl)-ethylenediamine in 100 ml. of methanol is cooled with an ice ~ath, stirred, and de-aerated by bubbling in nitrogen for 15 minutes. A 10.97-gram portion of leuco-1,4,5,8--tetrahydro~yanthraquinone is gradually added with stir-ring and continued cooling. The suspension is heated ~t~ ~

and stirred under nitrogen in an oil bath at 50-52C.
for one hour and the mixture is then allowed to stand and cool under nitrogen overnight. The solid is collected and washed with ethanol giving 14.8 g. of a red-brown solid, m.p. 165-168C.
Example 16 1,4-Bis[2-methylamino)ethylamino]-5,8-dihydroxy-anthraquinone_dihydrochloride To a suspension of 11.60 g. (0.03 mole) of lQ leuco-1,4-bisL2-(methylamino)ethamino]-5,8-dihydroxy-anthraquinone in 200 ml. of 2-methoxyethanol was added gradually with stirring 15 ml. of 8N ethanolic hydrogen chloride, The system was chilled with an ice bath and gtirred as 7.50 g. (0.0305 mole) of chloranil powder was ~radually added. The mixture was stirred overnight at room temperature and diluted with 600 ml. of ether.
The solid was collected and washed with tetrahydrofuran.
The product (14.16 g.) was recrystallized by dissolving it in 13Q ml. of water and adding 650 ml. of acetone to 2Q giYe 13.15 g. of a blue-black solid.
Example 17 1,4-Bis~2-(2-aminoethylamino)ethylamino]-5 ! 8 dihydroxyanthraq-uinone Following the general procedure of Example 3, a mixture of 10.97 g. of leuco-1,4,5,8-tetrahydroxyanth-raquinone, 80 ml. of N,N,N',N'-tetramethylethylene-diamine and 21.84 g. (0.24 mole) of diethylenetriamine soon gave a thick, congealed mass which prevented effec-tive stirring so the reaction time was extended to 24 hours. The mixture was allowed to cool and the superna-tent liquid was decanted and discarded. A solution of E~q - 59 -1~4~923 ~he congealed mass in 100 ml. of methanol was filtered, then allowed to oxidize in the air for four days in a partially covered flask. The gelantinous mass which had separated became solid when the oxidation mixture was agitated with 20n ml. of acetonitrile and then allowed to stand for one hour. After the solid was collected and ~ashed first with acetonitrile, then with ether, it amounted to 10.88 g. of blue-black powder.
Example 18 Leuco-1,4-bis(4-aminobutylamino)-5,8-dihydroxy _nthraquinone Following the general procedure of Example 3 hut u8ing 45 ml. of 1,4~diaminobutane as the primary amine component, there was obtained 12.20 g. of product a~ a dull grey-green solid.
Example 19 Leuco~lr4~bis~2-dimethylaminopropylamino]-5~8-di~
h.ydroxyanthraquinone The reaction of 12.26 g. of 2-dimethylamino-2a propylamine ~ith 10.97 g. of leuco-1~4,5,8-tetrahydroxy-anthraquinone is 100 ml. of ethanol for one hour by the procedure of Example 1 gives 7.29 g. of red-brown crys-tal~.
Example 20-Leuco-1,4-bis[2-(2-methylaminoethylamino)ethyl-amino-5,8-dihydroxyanthraquinone To a solution of 14,10 g. of l-methyl diethy~
lenetrimaine in 50 ml. of ethanol and 40 mol. of N,N,N',N'-tetramethylethylenediamine is added 10.97 g.
3Q of leuco-,4,5,8-tetrahydroxyanthraquinone as in Example 1. The mixture is heated at 50 and stirred under nitro-gen for one hour, chilled with an ice bath, the solid 114~5~3Z3 collected and wash with.cold ethanol to give 7.23 g.
of green-black crystals, m.p. 108-111C.
Example 21 Leuco-1~4-his~2-(2-dimethylaminoethylamino)ethyl-amino,l-5 ! 8-dihydroxyanthraquinone The reaction of N-(dimethylaminoethyl)ethy-lenedia~ine with.leuco-1,4,5,8-tetrahydroxyanthraquinone by the procedure o Example 20 gives the title comp,ound.
Bxample 22 Leuco-lL4~bis~?~ piperazinyl)-ethylamino]~5r8 -dihydroxyanthraquinone-The procedure of Example 20 applied to 15.50 g. of N~ aminoethyl).piperazine gives 3.92 g. of a hlack.powder which does not melt by 35QC. and is dis-ca~ded. The mother liquor and ethanol washes, on stan-ding and partly evaporating during two weeks in an un-atoppered flask, deposit a solid which is collected and was.hed with.ethanol to giYe 6.19 g. of the title cQmpound as a black soli.d, m.p. 200-203C.
2Q Bxample 23 1,4~is.(2-aminoethylamino?,-5r8-dihydroxyanthraqui-none dihydrochloride Oxidation with chloranil of 28.25 g. of the product of. Bxample 12 hy the procedure of Bxample 16 giYes 29.66. g. of a crude, hlue.-black solid which is then extracted hy stirring for 14 hours with 800 ml. of water.
Soli.d& are remoYed by centrifugation and the supernatent ~oluti.on freeze.dried, leaYing 16.38 g. of a blue-black &oli,d which is unmelted by 350C.

11~4~923 Example 24 lt4~is~2~-~2~hydrQxyeth~aminoLethylamino~5~8;dihydroxy anthraqui e Di~ydrochloride Chloranil oxidation of 17.86 g. of the product of Example 14 by the procedure of Example 16 gives ~with-out recrystallizationl 21.34 g. of blue-black solid, m.p.
2Q3-205C.
Example 25 lr 4~B~s~2~(2 meth~laminoethylamino)~ethylamino~-5~8-dihyd 10roxyanthraquinone Tetrahydrochloride The product of Example 20 (11.70 g.) is oxi-dized with chloranil by the procedure of Example 16, gi-ving 18.03 g. of hlue-black solid, m.p. l9Q-203C.
Example 26 15lt4~Bi~2~ hydroxyeth~lamino)-ethylamino~-5~8 dihydroxyanthraquinone In a modification of the synthesis of Example 14 the solYent used is lQQ ml. of ethanol. The mother liquor from the leuco product is allowed to stand for two 2Q week~ in an unstoppered flask, whereupon the oxidized product separates. It is collected and washed with etha-nol, giving ~lue-black crystals, m.p. 175-177C.
Example 27 Leuco-1,4-bis~3-(?-hydroxyethylamino)-l-propyl-a-ml-no]
255 ! 8 ~ dihydroxyanthraquinone The procedure of Example 15 is used with a solution of 14.18 g. of 2-(3-aminopropylamino)ethanol in lQQ ml. of ethanol. The resulting solution is filtered and the filtrate diluted with 300 ml. of ether, preci-3Qpitating the product as a goo. After decantation of the 62 _ ~' 114~923 supernatent solution the goo is caused to crystallize by agita,ting it with lQQ ml. of tetrahydrofuran. Wash-in~g ~ith,ethanol giYe~ 12.56 g. of green hlack solid, m.p. lQ1C~lQ4C.
Example 28 1,4~is~3 ~2~h~droxyethylaminoL~ ropylamino]
5,,8 dihydrQ~yanthraqu~none dih;y rochloride Oxidation of q.~S g. of leuco-1,4-bis~3-~('2-hydro~yethylamino)propylamino~ 5,8 dihydroxyanthra-quinone with chloranil as in Example 16 gives 11.70 g.
o,f a blue solid which does not melt by 350C.
Example 23 Le~co 1~4 bis~2-(,3~hydroxy-l-propylamino ?- ethylamino~-5,8~dihydroxyanthraquinone lS The procedure of Example lS is paralleled with 14.18 g, of N-(3-hydroxypropyl)ethylenediamine in 100 ml.
of ethanol to give 14.63 g. ~f red-brown crystals, m.p.
58-60,C.
Example 30 2 Q 1 ! 4-Bis~2-(3-hydroxy-l-propylamino)ethylamino]
5r8-dih,ydroxy'anthraquinone dihydrochloride Choranil oxidation of 10.77 g. of the product of Example 2~. by the procedure of Example 16 yielded 11.64 g. of a dark hlue solid, m.p. 210-216C.
Example 31 Leuco-1,4-bis[2-(,2-hydroxy-l-propylaminolethylamino]
5 ! 8-dih~droxyanthraquinone With 14.18 g. of 1-(2-aminoethylamino)-2-propanol in lQ0 ml. of ethanol the procedue of Example 17 3Q yields 17.61 g. of green-black crystals, m.p. 50-60C.

~ .

Exa~ple 32 1!4-~ia~2-(2~h~droxy-1-propylamino)ethylamino~
5~8-dihydroxyanthraquinone dihydrochloride A filtered solution of 14,44 g. of leuco-,4-~isl2-(2-hydroxy-1-propylaminto),ethylamino~-4-dihydroxy-anthraquinone in 215 ml. of 2-methoxyethanol is oxidized with,7.65 g. of chloranil by the procedure of Example 16, affording 16.75 g. of purple solid, m.p. 177-185C.
Example 33 Leuco-1!4-his~2-(,2-h~droxyethylamino)ethylamino]
eth~lamino~ ~ 8-dihydroxyanthraquinone The procedure of Example 15 used with a solu tion of 17,67 ~. of 2-~2 (2-aminoethylaminolethylamino]~
e,thanol in lOQ ml. of methanol gives a solution which is filtered, then diluted with 300 ml. of ether, precipi-tating a goo which,hardens on standing overnight. Har-dening is completed by thorough maceration of the solid in the solYent. The &olid is collected and washed with ether! yielding 16.82 g. of a green-black solid. This 20, solid remains, granular if stored at -25C., but coales-cea into a solid cake if stored at 25C.
Example 34 1,4-Bis~2-(,2-hydroxyethylami~olethylamin ethylamlno] 5t8-dihydr-o-xxanthraquinone tetrahydrochloride Chloranil oxidation of 12.lQ g. of the product of Example 33 hy the method of Example 16, including three additional washings of the solid with methanol, ~ives 12.46 g. of dark, bluet solid product.

~ 64 -- `` 1141rJ9Z3 Exam~le 35 1!4~Bis~2 (2/3~dlhydroxy~ropylamino?ethy~amlno]~
5,.8-di.h~droxyanthraquinone dih~drochloride By the procedure of Example 15 a solution of 16..... 10. g. of 3 (2-aminoethylaminol~1,2-propanediol ~A. R. ~urrey, C. M. Sutter and J, S. Buck, J. Am. Chem.
Soc.! 74~ 41Q2 U9,52).~ in lQ0. ml. of methanol gives a goo which.is separated from solvent b.y chilling with an ice bath.~ then decanting. The goo is washed four times by lQ. stirring 1,5 hours at 25 with.lQQ-ml. portions of me.thanol, chilling with.an ice bath., then decanting. A
fi.ltered s.olution of the goo in 280. ml. of 2-methoxyetha-nol is oxidized with.10..01 g, of chloranil by the method of Example 16. The product is additionally washed with e,thanol! giving 15.25 g. of blue-hlack solid, m.p. 191--19.3C.
Example 36 Leuco-1!4~bis~2 (l~aziridino?.ethylamino~-5,8-dih ~ oxyanthraqu_~one 2Q ~ith.lQ,33 g. of N-(,2-aminoethyl).aziridine in 8Q ml. of N!N,NIrN'-tetramethylethylenediamine the procedure.of Example 15 gives. a stiff gum. The next day the, aupernatent solution is discarded, 100 ml. of ether is added and the,gum periodically macerated therein for another day, when the gum is mostly hardened. Hardening is completed by maceration during three washings of the solid with.ether, giYing 17.66 g. of blue-black, granu-lar powder.

Example 37 1,4-Bis~2~ aziridino)ethylamino~-5,8-dihydroxy-anthraquinone To a suspension of 4.10 g. of the product of Example 36 in 40 ml. of chloroform is added a solution of 1.74 g. of diethyl azddicarboxylate in 25 ml, of chloro-form, The mixture is stirred for 20 minutes, the resul-ting dark blue solution is filtered, and the filtrate is eYaporated at ~ 30. A solution of the residue in 40 ml.
of chloroform is stirred five minutes with 2 g. of lQ decolorizing carbon, filtered and washed through with another 25 ml. of chloroform. Addition of 100 ml. of ether to the filtratefi precipitates a gum which is eliminated by decantation-filtration. The filtrates depoait crystals which are washed sparingly with acetone.
The chloroform-ether mother liquor, chilled at -60C., deposits a second crop of crystals which is washed with ethe~r and with methanol. A solution of both crops of crystals in 2Q ml. of chloroform is stirred with decolo-rizing carbon, filtered, evaporated at ~ 25C. to a 2Q Yolume of 5 ml., diluted with 20 ml. of ether, then chilled at -60C. The resulting blue-black crystals, washed ~ith ether, amount to 0.64 g., m.p. 168-170C.
In thin-layer chromatography on silica gel the product is mo~ed as a blue spot by chloroform-triethylamine-5 methanol, 27/3/1 (xatios by volumel.Example 38 1!4-B.iS[2-~2-(l-morph41ino).ethylamino]ethylamino]-5!8 dihydroxyanthraquinone tetrahydrochloride A solution of 20.80 g. of N-(morpholinoethyl)-Q eth~lene diamine in lOQ ml. of ethanol is used in the 114~923 procedure of Example lS to give a solution which isfiltered and diluted with 90.0 ml. of ether, precipating a goo. The supernatent solution is dec~ted, the goo dis.solved in 175 ml. of 2-methoxyethanol and oxidized ~ith 5.2~ g. of chloranil by the method of Example 16, giYing 17.7 g. of dark blue solid.
Example 39 Leuco-1,4-Bis~2-(acetamido).ethylamino.]-5,8-dihydroxyanthraquinone A solution of 12.26 g. of N~acetylethylene di.a~ine in lOQ ml. of ethanol in the procedure of Example 15 giYes 15.27 g, of dark, red-brown solid, m,p. 125C.
Example 40.

1,4-~isi2-(acetamido)ethylamino.]~
5~8-dih.ydrQxyanthraquinone A suspension of ll.9S g. of leuco-1,4-bis[2-~acetami.do)-ethylamino~-5,8-dihydroxyanthraquinone is oxidized with.6.76 g. of chloranil during 61 hours by 2~ the.~ethod of Example 61, giving a very acidic hydro-chloride. salt which.is converted to the free base by four ~ashings with water. Crystallization from 110 ml.
of dimethyl sulfoxide (~oiling only 2 minutes and not attempting a hot filtration), then washing with dimethyl sulfQxide.and with.ethanol gives 7.76 g. of blue-black solid, m.p. 273-274C.
Example 41 1, 4-~is f 2-~N-(2-hydroxyethyl triflouracetamido]-ethylamino]-5~8-dihydroxyanthraquinone 3Q A suspension of 1.50 g. of 1,4-bis[2-(2-114~923 -hydrQxyethylamino)ethylamino]-5,8 dihydroxyanthra-quinone in 75 ml. of ethyl triflouroacetate and 75 ml.
of methanol is stirred for 10 minutes. Evaportation of the resulting solution ln vacuo at 30C. leaves a residue which is washed and macerated with methylene chloride, giYing 2.11 g. of blue-black solid, m.p.
162C.
Example 42 1!4-Bis~2-amino-2-carboxyethylamino]-5r8-dihydr anthraquinone . 3/4 HCl To a solution of 6.23 g. of dl-a,~-diaminopro-pionic acid in 3Q ml. of warm water is added 1.078 g. of lithium hydroxide and 60 ml. of dimethyl sulfoxide. The system is flushed with nitrogen and 4.12 g. of leuco-1,4,-lS 8-tetrahydroxyanthraquinone is added gradually with stir-ring, The mixture is stirred and heated with an oil bath at 5Q, first for 15 hours under nitrogen, then for 21 hours as the initial product is oxidizéd by bubbling in a stream of air. Thin-layer chromatography on silica gel with methanol~water-concentrated ammonia (25/5/11 by volume) shows all the product spots to be blue when the oxidation is complete. After the mixture is cool the solids are removed by filtration and washed once with dimethyl sulfoxide -water (2/1). Addition of 400 ml.
of methanol to the filtrates precipitates a solid which is collected and washed with methanol. Further washing with a total of 13. ml. of 0.01 N aqueous acetic acid dissolYes virtually all of the solid. Addition of 3 ml.
of concentrated h~drochloric acid to the acetic acid filtrates precipitates a blue-black solid which is washed with acetone to giye 0.24 g. of the product.
Example 43 Leuco-1!4-bisI2-(2-methoxyethylamino ethylamino]-5,8-dihydroxyanthraquinone An ethanol solution of N-(2-methoxyethyl)-ethylenediamine (U.S. Pat. 3,454,640) reacts in the procedure of Example 51 to give the title compound.
Example 44 1,4-Bis[2 (1,3-oxazolidin-1-yl)ethylamino]-5,8-dihydroxy-anthraquinone A solution of 1.62 g. of 37% aqueous formal-dehyde solution in 50 ml. of water is stirred overnight with 4~44 g. of 1,4-bis~2-(2-hydroxyethylamino)ethylaminoI--5,8-dihydroxyanthraquinone. The resulting solid is washed with water to give the product.
Example 45 1,4-Bis[2-(tetrahydro-1,3-oxazin-1-yl)ethylamino]-5,8-dihydroxyanthraquinone A solution of 1.62 ml. of 37~ aqueous formal-dehyde in 50 ml. of 0.4N aqueous sodium hydroxide is stirred overnight with 5.45 g. of 1,4-bis[2-(30hydroxy-1-~propylaminolethylamino]-5,8-dihydroxyanthraquinone di-hydrochloride. The product is obtained by washing the resulting solid with water Example 46 1,4-Bis[2-(1,3-oxazolidin-2-one-1-yl)ethylamino]-5,8-dihydroxyanthraquinone A solution of 0.020 g. of sodium in 25 ml. of methanol is stirred and heated under reflux overnight with 75 ml. of diethyl carbonate and 4.44 g. of 1,4-bis-114~9Z3 ~2~(2~h~droxyethylamino)~ethylamino]-5,8edihydrQxyanth-raquinone. The mixture is allowed to cool. It is stirred with Q.l ml. of acetic acid, the solid is collected by filtration and washed with methanol to give the product.
Example 47 1!4-Bis~2-(1,3-oxazin-2-one-1-yl?,ethylamino~-5,8-dihydrQxyanth-raquinone A solution o o.48 g. of sodium in 25 ml. of me*hanol is stirred and heated overnight with 75 ml. of diethyl carbonate and 5.45 g. of 1,4-bis~2 (3-hydroxy-1-~propylamino)ethylamino~-5~8-dihydroxyanthraquinone di-hydrochloride. After the mixture cools it is stirred with Q.l ml. of acetic acid. The solid product is collected by filtration and washed with ~ethanol and then with water.
Example 48 1!4-Bls[2-.[di (,~-h~droxyethyl)amino~ethylamino]
5 ! 8rdihydroxyanthraquinone dihydrochloride Chloranil oxidation of 10.77 g. of the product 2Q o~ Example 15 by the method of Example 16 gives 11.64 g.
of a dark blue solid, m.p. 216C.
Example 49 Preparation of 50 mg. Tablets Per Tablet Per 10 ! Tablets 250,Q50 gm. 1~4-bis(~-aminopropyalamino)-5,8-dihydroxyanthraquinone ................ 500 gm.
0,.0,80 gm. Lactose ... ,....................... 800 gm.
Q,010 gm. Corn Starch (for mix)........................ 100 gm.
0.008 gm. Corn Starch ~for paste) ....... ,............. .75 gm.
Q`.~4'8 gm 1475 gm.
Q.Q02 gm Magnesium Stearate (1%) ................. ..... .15 gm 0`.150'gm 1490 gm ~' The 1,4-bis(3~aminopropylamino)~5 ! 8-dihydroxy-anthraquinone ! lactose and corn starch (for mix2 are ~lended together. The corn starch (for paste~ is sus-pended in 6~0 ml. of water and heared with stirring to form a paste. This paste is then used to granulate the mixed powders. Additional water is used if necessary.
The wet ~ranules are passed through a No. 8 hand screen and dried at 120F. The dry granules are then passed through a No. 16 screen. The mixture is lubricated with lQ 1% magnesium stearate and compressed into tablets in a suitable tableting machine.
Example 5Q
Preparation of Oral Sus~ension Ingredient Amount Leuco-1~4-bis~3-aminopropylaminol~5,8-dihydroxy-anthraquinone ....,................................ 500 mg.
Sorbitol solution (70~ N.F.) ....................... 40 ml.
Sodium benzoate ..,................................ 150 mg.
Saccharin ........................................ , 10 mg.
Red dye ............................................ 50 mg.
Cherry flavor ....,................................. 50 ml.
Distilled water... qs...ad.......................... 100 ml.

The sorbitol solution is added ta 40 ml. of distilled water and the leuco-1,4-bis(3-aminopropylamino)--5~8-dihydroxyanthraquinone is suspended therein. The saccharin, sodium benzoate, flavor and dye are added and dissolved. The volume is adjusted to 100 ml. with dis-tilled water. Each ml. of syrup contains 5 mg. of leuco--1,4-b (3-aminopropylamino)-5,8-dihydroxyanthraquinone.
Example 51 Preparation of Parenteral solution In a solution of 700 ml. of propylene glycol 3Q and 200 ml. of water for injection is suspended 20.0 ~ grams of 1,4~bis~3-(dime.thylamino propylamino.l -5,8~di-: hydroxyanthraqui.none.dihydrochloride.with.stirring.
After suspension is complete, th.e pH. is adjusted to 5.5 with hydrochloric acid and the volume is made up to 1000 ml with.water for injecti.on. The formulation is steri.
li.zed, filled into 5.Q ml. ampoules each.containing 2 0 ml CrePresenting 40 mg. of drug). and sealed under nitrogen Exam~le 52 10. 1!4-Bis~2 (2-hydroxyethylamino)ethylamino~-5l8 -dihydroxyanthraquinone disuccinate salt .

A mixture of 222 mg. of 1,4 bis~2-(.2 hydroxy-ethylaminoLethylamino.]-5,8.dihydroxyanthraquinone, 118 mg. of s.uccinic acid, and 5Q ml. of ethanol is heated under reflux for 3Q minutes to give the title compound.
Example 53 .

1,4-Bis~2~q3~hydroxypropylamino)ethylamino~5~8 -dihydr~xyanthaquinone dimalate.salt A mixture of 228 mg. of 1,4-bis~2;(.3-hydroxy-20. propylamino)ethylamino.~5,8-dihydroxyanthraquinone, 134 mg, of DL-~alic acid~ and 50. ml. of ethanol is heated under re.flux for 3Q minutes to give the title compound.
Example 5~
1!4-Bis~2-(.2-hydroxypropylamino)ethylamino~-5,8-dihydroxyanthraquinone dilactate salt A mixture of 228 mg. of 1,4-bis[2-(2-hydroxy-propylaminolethylamino)-5,8-dihydroxyanthraquinone, 120 mg. of 80% DL-lactic acid, and L0 ml. of ethanol is heated on a steam bath for lO.minutes, cooled, treated with.50.ml. of acetone and cooled to obtain the title compound.

Claims (8)

27,449 THE EMBODIMENTS OF THE INVENTION IN WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. A process for preparing chemical compounds of the formula:

the tautomers and the pharmacologically acceptable acid-addition salts thereof:
wherein A-B is CH=CH or CH2-CH2;
Q is a divalent moiety selected from the group consisting of:
-(CH2)n- , , , , , , , and wherein n is an integer of 2 to 4 inclusive;
R1 and R2 are each individually selected from the group consisting of hydrogen, alkyl having from 1 to 4 carbon atoms, mono-hydroxyalkyl having from 2 to 4 carbon atoms, and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, dihydroxyalkyl having from 3 to 6 carbon atoms, and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group, formyl, alkanoyl having from 2 to 4 carbon atoms, trifluoroacetyl and moieties of the formulae:

-(CH2)n-CN, -(-CH2)n-O-R, and wherein n is an integer from 2 to 4, inclusive, R is alkyl having from 1 to 4 carbon atoms, and R3 and R4 are each individually selected from the group consisting of hydrogen, alkyl having 1 to 4 carbon atoms and mono-hydroxyalkyl having from 2 to 4 carbon atoms and wherein the carbon atom alpha to the nitrogen atom may not bear an hydroxy group; and R1 and R2 taken together with the associated N(itrogen) atom and R3 and R4 taken together with the associated N(itrogen) atom is morpholino, thio-morpholino, piperzino, 4-methyl-1-piperazino or a moeity of the formula:

wherein m is an integer from 2 to 6, inclusive; with the proviso that the ratio of the total number of carbon atoms to the sum of the total number of oxygen atoms plus the total number of nitrogen atoms in the side chains at the l-position and the 4-position may not exceed 4;
provided that when Q is -(CH2)-n- then:
a) R1 and R2 may not simultaneously be hydrogen;
b) when n is 2, then R1/R2 may not be CH3/CH3, C4H9/C4H9, or -(CH2)5-;
n is 3 then R1/R2 may not be H/H, CH3/CH3, C3H7/C3H7, or -(CH2)2-; n is 4 then R1/R2 may not be C2H5/C2H5, C4H9/C4H9, or -(CH2)4-; and c) only one of R1 and R2 may be alkanoyl.
which comprises reacting a compound of the formula;

with a compound of the formula wherein A-B, Q, R1 and R2 are as defined above;
and if desired, converting A-B when it is CH2--CH2 into CH=CH; and if desired, converting the product into the tautomer thereof; and if desired, contacting the product with a pharmacologically acceptable salt--forming reagent under salt-forming conditions.

2. A compound of the formula:

wherein Q, R1 and R2 are as defined in Claim 1 whenever prepared according to the process of Claim 1 or by an obvious chemical equivalent thereof.

3. A process for the preparation of leuco-1,4-bi[(2-dimethylaminoethyl)amino]-5,8-dihydroxy--anthraquinone which comprises reacting N,N-dimethyl-ethylenediamine with leuco-1,4,5,8-tetrahydroxy-anthra-quinone.

4. The compound of leuco-1,4-bis[(2-dimethyl-aminoethyl)amino]-5,8-dihydroxy-anthraquinone whenever prepared by the process of Claim 3 or by an obvious chemical equivalent thereof.

5. A process for the preparation of 1,4-bis-[2-(hydroxyethylamino)ethylamino]-5,8-dihydroxyanthra-quinone which comprises reacting 2-(2-aminoethylamino)-ethanol with leuco-1,4,5,8-tetrahydroxyanthraquinone followed by chloranil oxidation to obtain the desired product.

6. The compound 1,4-bis[2-(2-hydroxyethylamino)-ethylamino2-5,8-dihydroxyanthraquinone whenever prepared by the process of Claim 5 or by an obvious chemical equivalent thereof.

7. A process for the preparation leuco-1,4-bis(2-aminoethylamino)-5,8-dihydroxyanthraquinone which comprises reacting leuco-1,4,5,8-tetrahydroxy-anthraquinone with ethylenediamine.

8. The compound leuco-1,4-bis(2-aminoethyl-amino)-5,8-dihydroxyanthraquinone whenever prepared by the process of Claim 7 or by an obvious chemical equivalent thereof.