This invention relates to new organic compounds and, more particularly, is concerned with novel arithracene- -9,10-bis-carbonyl-hydrazones which may be represented by. the following general formula:

wherein Z is a moiety of the formulae:

R

(CH ₂ ) _n —c= or -CH- -CIJ=

wherein n is 0, 1, 2 or 3 and R is hydrogen, alkyl having up to 4 carbon atoms, cycloalkyl having from 3 to 6 carbon atoms, phenyl or benzyl; R, is hydrogen or alkyl having up to 4 carbon atoms; R^ is & moiety of the formulae:

wherein p is 1, 2 or 3, q is 0, 1 or 2, R' is hydrogen o ^* alkyl having up to 4 carbon atoms, R" is alkyl having up to 4 carbon atoms, R _? is a moiety of the formula:

wherein m is 2, 3, 4 or 5, R-. R' and.R" are as ±ierein— before defined and - he moiety -NR'R" may be pyrrolidino, piperidino, morpholino or N-imethylpiperazino, X ^' is oxo (O=), thioxo (S=) or imino (R ^** -N= wherein R' is as herei before defined); and R„, R., R _g and R _g are each individually selected from the group consisting of hydrogen, halogen (F, Cl, Br _r I), hydroxy, nitro, ainj.no, sulfonamido, alkyl having up to 3 carbon atoms and alkox having up to 3 carbon atoms. The hydrazono substituents pendant from the anthracene—9,10 ^" —bis—carbonyl nuclei may be the same or different and may be in the s ^' yn or ^{' "} an ^' i forms. Additio ally, where the hydrogen atoms or .other substituents at • positions 1, 4, 5 and 8 of the anthracene nucleus cause restricted rotation of the bonds extending from C ^' and C _Q of the anthracene nucleus, then the entire units

cis (both extending out from the same face of the ■ anthracene nucleus) or trans (extending out from the opposite faces of the anthracene nucleus)'.

__0Λ

The novel compounds of the present invention are obtainable as yellow crystalline materials having character¬ istic melting points and absorption spectra and which may be purified by recrystallization from common organic sol¬ vents such as lower alkanols, dimethylformamide, tetrahydro- furan, methyl isobutyl ketone, and the like.

The organic bases of this invention form non-toxic acid-addition and quaternary ammonium salts with a variety of pharmacologically acceptable organic and inorganic salt- -forming reagents. .Thus, acid-addition salts, formed by admixture of theOrganic free base with one or more equiva¬ lents of an acid, suitably in a neutral solvent, are formed with such acids as sulfuric, phosphoric, hydrochloric, hydrobromic, sulfamic, citric, lactic, malic, succinic, tartaric, acetic, benzoic, gluconic, ascorbic, and the like. Quaternary ammonium salts may be formed -by reaction of the free bases with one or more equivalents of a variety of organic esters of sulfuric, hydrόhalic and aromatic sulfonic acids. The organic reagents employed for quaternary ^' ammonium salt formation are preferably lower alkyl halides. However, other organic reagents are suitable for quaternary ammonium salt formation, and may be selected from among a diverse class of compounds including benzyl chloride, phenethyl chloride, naphthylmethyl chloride, dimethyl sul- fate, methyl benzenesulfonate ethyl toluenesulfonate, allyl chloride, methallyl bromide and crόtyl bromide. For pur- ' poses of this invention, the free bases are equivalent to their non-toxic acid-addition and quaternary ammonium salts. The acid-addition and quaternary ammonium salts of the organic bases of the present invention are, in general, crystalline solids, relatively soluble in water, methanol and ethanol but relatively insoluble in non-polar organic solvents such as diethyl ether, benzene, toluene, and the like.

The novel compounds of the present invention possess the property of inhibiting the growth of transpl mouse tumors as established by the following tests.

Lymphocytic leukemi P388 ^: ;test The animals used are mice all of one sex, weig a minimum of 17 g. and all within a 3 gram weight range. There are 5 or 6 animals per test group. The tumor tran plant is by intraperitoneal injection of 0.1 ml. or 0.5 of dilute ascitic fluid containing 10 cells of lymphocy leukemia p388. The test compounds are administered intr peritoneally on days one, 5 and 9 (relative to tumor ino lation) at various doses. The animals are weighed and s vivors are reocrded on a regular basis for 30 days. The median survival time and the ratio of survival time for treated (T)/control (C) animals are calculated. The pos tive control compound is 5-fluorouracil given as a 60 mg. injection. . The results of this test with representative compounds of the present.invention appear in Table I below. The criterion for efficacy is T/C x 100^125%

^• TABLE I

Lymphocytic Leukemia P388 Test

TABLB I. , cont'd. Lymphocytic Leukemia P'388 Test

Median

Dose Survival T/C x 100

Compound (mg./kg.) (Days) (Percent)

9,10-Anthracenedicarboxaldehyde, bis (4,4-dimethyl-2- AJ-. ■ 22.0 220 -imidazolin-2-ylhydrazone) dihydrochloride 6 . 25 21.0 210

3. 12 20.0 200

1. , 56 19.0 190

0. .78 17.5 175

Control 0 10.0 5-Fluorouracil 60 21.0 210

9,10-Anthracenedicarboxaldehyde, bis[(1,4,5,6-tetra- 200 28.0 255 hydro-5,5~dimethyl~2~ρyrimidinyl)hydrazone] dihydro¬ 100 33.5 305 chloride 50 31.0 282

25 23.5 214

12.5 22.0 200

Control 0 11.0 5-Fluorouracil 60 18.0 164

The novel compounds of formula (I) and their pharmacologically acceptable acid-addition and quaternary ammonium salts would be expected to show activity against a broad range of cancer diseases, and especially ^' blood cancer diseases such as leukemia, in standard test animals at doses substantially below toxic levels. The modes con¬ templated for administration are essentially parenteral . and intraperitoneal.

The pharmaceutical compositions can be in forms suitable for injectable use, which forms include sterile aqueous- solutions or dispersions and sterile powders for the extemporanous preparation of sterile injectable solu¬ tions 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 manufac¬ ture and storage and must be preserved against the contamina¬ ting action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, .ethanol,' polyol (for example, glycerol, . propylene glyco ^' l, and liquid polyethylene glycol, and the

- 8 - like), suitable mixtures thereof, and vegetable oils. 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 micro¬ organisms can be brought about by various antibacterial and antifungal gaents, 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 absorp¬ tion of the injectable compositions can be brought about by the use in the compositions of agents delaying absorptio for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared ±>y in- corporating the active ingredient or ingredients 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 basic dispersion medium and the required other ingredients 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 a powder of the active ingredient plus ^' any additional desired ingredient from a previously sterile-filtered solution thereof.

As used herein, "pharmaceutically acceptable carr¬ ier" includes any and all solvents, dispersion media, coat- ings, antibacterial and antifungal agents, isotonic 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 conventional media or agent is incompatable with the active ingredient, its use in the present compositions is contemplated. Sup¬ plementary active ingredients can also be incorporated into the inventive compositions.

It is especially advantageous to formulate compo¬ sitions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used in the speci¬ fication and claims herein refers to physically discrete units suited as unitary dosages for the animal subjects to be treated, each unit containing a predetermined quantity of active material calculated to produce the desired thera¬ peutic effect in association with the required pharmaceutical carrier. The specification for the. ovel dosage unit forms of the invention are dictated by the 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 compounding such an active material for the treatment of disease in living subjects having a diseased condition in which bodily health is impaired as disclosed in detail in this specification.

The dosage of the principal active ingredient for the treatment of the indicated conditions depends upon the ^• age, weight and condition of the subject being treated; ^' the particular condition and its severity; the particular form ^* of the active ingredient and the route of administration. A daily dose of from about one to about 100 mg./kg. of body weight given singly or in divided doses of up to 5 times .a day embraces the effective range for the treatment of most _. conditions for which the novel compounds are effective and substantially non-toxic. For a 75-kg. subject, this trans¬ lates into between about 75 and about 7500 mg./day. If the dosage is divided, for example, into 3 individual dosages, these will range from about 25 to about 2500 g. of the active ingredient. The preferred range is from 2 to about 50 mg./kg. of body weight/day with about 2 to about 30 g./ ■ kg./day being more preferred.

The principal active ingredient is compounded for . convenient and effective administration in effective amounts with a suitable pharmaceutically-acceptable carrier in dosage unit form as hereinbefore disclosed. A unit dosage form can, for example, contain the principal active ingre- UREi- /

- 10 - dient in amounts ranging from about 0.1 to about 400 mg., with from about one to about 30 mg. being preferred. Ex¬ pressed in proportions, the active ingredient is generally present in from about 0.1 to about 400 mg./ml. of carrier. In the case of compositions containing supplementary active ingredients, the dosages are determined by reference to the usual dose and manner of administration of the said ingre¬ dients. ^{' *} .

Regression and palliation of cancers are attained, for example, using intraperitoneal administration. 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 dail dosage or one dose on alternate or less frequent days. .As can be seen from the dosage regimens, the amount of princi¬ pal active ingredient administrated 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 cytotoxic nature to the hosts harboring the cancer.. As used herein, cancer means blood malignancies such as leukemia, as well as other solid and non-solid malignancies 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.

Most of the novel compounds of the present in¬ vention may be readily prepared as set forth in the follow¬ ing reaction scheme:

wherein Z, R-, , R _? , R-,, R4.,' R5 _c and R6,- are as hereinabove de- fined. In accordance with the above reaction scheme, an appropriately substituted 9,10-anthracenedialdehyde or di- ketone (III) is reacted with a hydrazine derivative of the formula: H ₂ N- R,R ₂ to ^' provide the 9,10-anthracene-bis-anthra- cene-bis-hydrazones (I) . The reaction may be carried out in a lower alkanol in the presence of an acid such as hydro¬ chloric, hydriodic or acetic (or glacial acetic acid may be used as the sole solvent) usually at the reflux tempera¬ ture of the reaction mixture.

The starting 9,10-anthracenedialdehydes and ketones may be obtained commercially or prepared in accordance with ny one of the reaction schemes denominated (A) nd (B) below wherein R ₃ , R., R ₅ and R_- are as hereinabove defined.

(VI)

In accordance with the above reaction scheme, the anthracene derivative (IV) suspended in dioxane and concen¬ trated hydrochloric acid and saturated with. HCl, is treated with paraformaldehyde at the reflux temperature for 2-6 hours to obtain the 9,10-bis-(chloromethyl) anthracene deri¬ vative (V). This 9,10-bis-(chloromethyl)anthracene, sus¬ pended in dry dimethyl sulfoxide under nitrogen at room temperature, is treated with sodium in ethanol and worked up as described in the examples to obtain the desired 9,10-anthracene dicarboxaldehyde (VI).

O

(VI )

In accordance with the above reaction scheme _r the anthracene- derivative (IV) is heated with excess vinylene carbonate under reflux for about 20 hours to give the cyclic carbonate (XI). Hydrolysis of. the cyclic carbonate (XI) with aqueous-ethanolic potassium hydroxide at 75°C. for about 2 hours produces the diol (XII) which in turn is treated with lead tetraacetate in acetic acid at 35°C. for about 2 hours to give the 9,10-anthracenedicarboxaldehyde <VI) .

Certain anthracene bis-hydrazones are more readily prepared by methods other than the general method previously outlined. One of these is outlined as reaction scheme (C) below wherein Rg, R ₄ , R ₅ and R _Q are as .hereinabove defined.

In accordance with the above rea fcXon scheme, a mixture ^" of 9\,10-anthracenediacetonitrile derivative (XVI) and a substituted hydrazine hydrochloride, sodium acetate and Raney—nickel in ethanol is reduced with hydrogen unti 2 molar equivalents of hydrogen have been absorbed to giv the desired anthracene ^' bis-hydrazone (XVII). The novel monohydroxya thracene—9,lO'—b s— hydrazones and the dihydrox anthracene—9 _τ 10—bis-hy ^' drazones of the present invention may be prepared as set forth in the reaction scheme (B) after having converted the hydrox . roups to tri methylsilyloxy derivatives, as by treatment with trimethylsilyl chloride in dry tetrahydrofuran in the presence of triethylamine.

The invention is illustrated by the following specific examples.

Example" 1. 1,1'- f9,10-Anjhrylenebis (methylidynenitrilo) jbis- [3- ^' (2-dimethylaminoeth l)guanidine] tetrahydrochloride _.

A mixture of 3.5 g. of 9,10-anthracenεdicarboxalde- hyde, 8.3 g. of l-amino-3-(dimethylaminoethyl) guanidine tri- hydrochloride, 150 ml. of ethanol and 8 ml. of water is heated under reflux for 2.5 hours, treated with charcoal and filtered. This solution is evaporated to dryness and reevaporated with ethanol. The residue is slurri ^' ed with .hot isopropanol, cooled _. and filtered to give 9.0 g. of orange solid. .

Example 2 ^' 9 ^* ,10-Anthracenedicarboxaldehyde, bis [4-(3-dimethyl- aminopropyl)-3-thiosemicarbazonej dihydrochloride

A mixture of 2.9 g. of 9,10-anthracenedicarboxalde- hyde, 5.0 g. of 4-(3-dimethylaminopropyl) -3-thiosemicarba- zide, 100 ml. of ethanol* and 12 ml. of 8M ethanolic hydrogen chloride is heated under reflux for 3.5 hours, filtered hot and cooled to give 5.7 g. of orange solid. This is recry- •stallized from 85 ml. of dimethylformamide to give 4.5 g. of product. - After drying at 75°C. in a pistol it has a .p. of 221-223°C.

Example ^* * ^■ - ^• 9 , 0-Anthracenedicarboxaldehyde, bis [4-(3-dimethyl- aminoethyl) -3-thiosemicarbazone] dihydrobromide

A mixture of 3.5 g. of 9,10-anthracenedicarboxalde- hyde, 5.4 g. of 4—(dimethylaminoethyl) hiosemicarbazide, 250 ml. of ethanol and.2 ml. of 48% hydrogen bromide is heated under reflux for 4 hours and filtered hot to give 6.4 g. of solid. This is resuspended in 175 ml. of boiling ethanol and treated with 20 ml. of 48% hydrogen bromide to obtain complete solution. This is cooled, treated with 100 ml. of ether and cooled further to give 3.1 g. of the product in the form of orange-red crystals, m.p. 289-290°C.

" Example 4 ^' 9,10-Anthracenedicarboxaldehyde bis (4 4-dimethy -2- -imidazolin-2-ylhydrazone) dihydrochloride

A mixture of 2.3.4 g. of 9,10-anthracenedicarboxalde hyde, 4.4 g. of 4,4-dimethylimidazolin-2-ylhydrazine hydro- bromide and 50 ml. of n-propanol is boiled for 2 hours. The yellow-orange solid is collected, washed with n-propanol and dried. This hydrobromide salt is dissolved in water ^* and basified with sodium bicarbonate ^' to give a solid which is collected, redissolved in 60 ml. of hot n-propanol and treated with 0.6 ml. of 7N hydrochloric acid in isopropanol and cooled to give 1.2 g. of the title compound, m.p. 310- -315°C.

Example 5 9, 10-Anthracenedicarboxaldehyde, bis (1,4,5, 6-tetra- hydro-5,5-dimethy -2-pyrimidinyl)hydrazone, dihydro¬ chloride / A mixture of 0.94 g. of 9,10-anthracenedicarboxalde hyde, 1.7 g. of 5,5-dimethy1-1,4,5,6-tetrahydropyrimidin-2- -ylhydrazine and 50 ml. of n-propanol is.heated to boiling for 0.5 hours, then cooled to give 1.7 g. of .a yellow cry¬ stalline product, m.p. 340°C. (dec).

Example ^* 6

'

^• ^3-thiosemicarbazide ^" ] dihydrochToride

A mixture of 3.5 g of l-(2-morpholinoethyl)thio semicarbazide, 1.75 g of 9 ,10- ^* -an hracenedicarboxaldehyde, 250 ml ethanol and 2 ml of concentrated hydrochloric acid is .heated under reflux for 5.5 hours and filtered hot to give 5.5 g of product, m.p. 229-231°C. This is recrystallized from a methanol->-water-ether solution to give 5.0 g of orange product: m.p. 235-237°C.