Glyoxylic acid hydrocarbylsulfonylhydrazones and therapeutic compositions

BACKGROUND OF INVENTION
The invention described herein was made in the course of or under a contract with the U.S. Department of Health, Education, and Welfare.
The nearest known prior art is the article of Herbert O. House and C. John Blankley, "Preparation and Decomposition of Unsaturated Esters of Diazoacetic Acid" published in The Journal of Organic Chemistry, Vol. 33, No. 1, January, 1968, pages 53-60. This article discloses the compound p-toluenesulfonylhydrazone. No therapeutic utility is ascribed to this compound or to the others which are disclosed.
SUMMARY OF INVENTION
The present invention rests in part on the discovery that glyoxylic acid hydrocarbylsulfonylhydrazone compounds have utility as anticancer agents when administered to cancerous warm-blooded animals. In one embodiment, this aspect of the invention relates to a process for treating leukemia which comprises administering to a warm-blooded animal having an abnormal proportion of leukocytes a therapeutic amount of a glyoxylic acid hydrocarbylsulfonylhydrazone compound. The latter compounds can be administered per se, or in association with a pharmaceutically acceptable diluent or carrier. The invention accordingly also provides a pharmaceutical composition in dosage unit form comprising from about 0.1 to 500 mg of the active compound, per dosage unit, together with a pharmaceutically acceptable nontoxic, inert carrier or diluent therefor. Lastly, the present invention discloses particular novel compounds coming within the scope of the broad class of glyoxylic acid hydrocarbylsulfonylhydrazone compounds which have utility as anticancer agents .
The novel compounds of the invention are those wherein the hydrocarbyl group is an aryl, alkyl or alkenyl group, whether substituted or unsubstituted, as set forth below in Table 1. In said table, the old compound of the prior art is given the number (1), with the novel compounds being numbered from (2) through (17).
TABLE 1______________________________________Hydrocarbyl-SO.sub.2 NHN.dbd.CH--CO.sub.2 HCompound No. Hydrocarbyl Group______________________________________1 p-tolyl2 phenyl3 p-chlorophenyl4 p-methoxyphenyl5 m-nitrophenyl6 2,5-dichlorophenyl7 .beta.-naphthyl8 m-aminophenyl9 p-acetamidophenyl10 2,4,6-trimethylphenyl11 phenyl,1,3-bis12 2-thienyl13 styryl14 benzyl15 methyl16 n-butyl17 n-hexadecyl______________________________________
In general, the novel compounds of the present invention can readily be prepared by reacting the known hydrocarbylsulphonylhydrazide with glyoxylic acid, usually present in excess. The acid is normally employed in monohydrate form. Typical reaction temperatures range from about 5.degree. C. to room temperature. The reaction is conducted for one or more hours, usually in the presence of an organic solvent, though in some cases aqueous solutions are employed. The product can be recovered by evaporation of the solvent. When aqueous media are employed, the product precipitates out as formed, it being water insoluble except when present in salt form (compound 8). The products are all white crystalline solids which melt with decomposition. They are soluble in many organic solvents such as acetonitrile, DMF, DMSO, dioxane, ethanol and methanol.

The examples which follow detail the preparation of each of the compounds shown in Table 1. The example numbers correspond to those by which each such compound is identified in said table.
EXAMPLE 1
GLYOXYLIC ACID p-TOLUENESULFONYLHYDRAZONE
A mixure of p-toluenesulfonylhydrazide (1.86 g, 10 mmol), glyoxylic acid monohydrate (1.20 g, 13 mmol), concentrated hydrochloric acid (1 cc, 12 mmol), and acetonitrile (40 cc) was stirred overnight at room temperature. The solvent was removed in vacuo and the residue triturated with water. The resulting white crystalline solid was collected by filtration and air dried to give 2.2 g of compound 1, m.p. 150.degree. (dec.). Here, as in the other examples, the product compound is identified by elemental analysis as well as by IR and NMR methods.
EXAMPLE 2
GLYOXYLIC ACID BENZENESULFONYLHYDRAZONE
To a stirred, room temperature solution of 1.48 g (0.02 M) glyoxylic acid monohydrate in 20 cc of water was added, in one portion, a room temperature solution of 3.44 g (0.02 M) of benzenesulfonylhydrazide in 10 cc of 2.5 N HCl. The mixture turned milky and was followed by a white precipitate within a minute. After stirring for 2 hrs, the mixture was filtered and the precipitate was collected, washed with water and (sparingly) with methanol in which the compound is very soluble. After drying the desired product was obtained (3.54 g) as a white crystalline powder, m.p. 130.degree.-3.degree. (dec.).
EXAMPLE 3
GLYOXYLIC ACID p-CLOROBENZENESULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 6.2 g (30 mmol) of p-chlorobenzenesulfonylhydrazide was reacted with 3.6 g (39 mmol) of glyoxylic acid monohydrate in the presence of 3 cc conc. HCl and 120 cc acetylnitrile. The product obtained (7.3 g) was a white crystalline material having a melting point of 163.degree. (dec.).
EXAMPLE 4
GLYOXYLIC ACID p-METHOXYBENZENESULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 6.06 g (30 mmol) of p-methoxybenzenesulfonylhydrazide was reacted with 3.6 g (39 mmol) of glyoxylic acid monohydrate in the presence of 3 cc conc. HCl and 120 cc acetonitrile. There was recovered 5.5 g of a white crystalline product having a melting point of 163.degree. (dec.).
EXAMPLE 5
GLYOXYLIC ACID m-NITROBENZENESULFONYLHYDRAZONE
The subject compound was prepared using the general method of Example 1 except that 2.17 g (10 mmol) of n-nitrobenzenesulfonylhydrazide was reacted with 1.2 g (13 mmol) glyoxylic acid monohydrate in the presence of 1.0 c conc. HCl and 40 cc acetonitrile. There was recovered 2.4 g of the desired product, m.p. 164.degree. (dec.).
EXAMPLE 6
GLYOXYLIC ACID 2,5-DICHLOROBENZENESULFONYLHYDRAZONE
The subject compound was prepared using the general method of Example 1 except that a mixture of 2,5-dichlorobenzenesulfonylhydrazide (2.41 g, 10 mmol) and glyoxylic acid monohydrate (1.20 g, 13 mmol) was stirred overnight at room temperature in the presence of conc. HCl (1 cc, 12 mmol) and acetonitrile (40 cc). The solvent was removed in vacuo and the residue was triturated with water. The resulting white, crystalline solid was collected by filtration and air dried to give 2.8 g of the desired product, m.p. 144.degree. (dec.).
EXAMPLE 7
GLYOXYLIC ACID .beta.-NAPHTHYLSULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 4.4 g (20 mmol) of .beta.-naphthylsulfonylhydrazide was reacted with 2.4 g (26 mmol) of glyoxylic acid monohydrate in the presence of 1 cc conc. HCl and 100 cc acetonitrile. The desired white crystalline product was obtained in a yield of 5.1 g, m.p. 149.degree. (dec.).
EXAMPLE 8
GLYOXYLIC ACID 3-AMINOBENZENESULFONYLHYDRAZONE HYDROCHLORIDE
Glyoxylic acid monohydrate (966 mg, 10.5 mmol) was added all at once to a stirred mixture of 3-aminobenzenesulfonylhydrazide (1.8 g, 10 mmol), conc. HCl (1.9 cc, 22 mmol), and dioxane (50 cc). The reaction mixture was stirred for 1 hr, cooled, and the precipitated white crystalline solid collected by filtration. The product was washed with dioxane, then with ether and air dried to give 2.4 g of the desired product, m.p. 123.degree. (dec.).
EXAMPLE 9
GLYOXYLIC ACID p-ACETAMIDOBENZENESULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 4.58 g (20 mmol) of glyoxylic acid p-acetamidobenzenesulfonylhydrazide was reacted with 2.4 g (26 mmol) of glyoxylic acid monohydrate in the presence of 1/2 cc conc. HCl and 200 cc acetonitrile. The desired product was obtained in a yield of 5.4 g, m.p. 136.degree. (dec.).
EXAMPLE 10
GLYOXYLIC ACID 2,4,6-TRIMETHYLBENZENESULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 6.43 g (30 mmol) of 2,4,6-trimethylbenzenesulfonylhydrazide is reacted with 3.5 g (39 mmol) diglyoxylic acid monohydrate in the presence of 15 drops conc. HCl and 150 cc acetonitrile. The desired white crystalline product was obtained in a yield of 7.7 g, m.p. 153.degree. (dec.).
EXAMPLE 11
DIGLYOXYLIC ACID 1,3-BENZENESULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that no conc. HCl was used. In the reaction, 4.8 g (52 mmol) of glyoxylic acid monohydrate were reacted with 5.3 g (20 mmol) of benzenesulfonylhydrazide in the presence of 200 cc acetonitrile. The method gave the product compound in the amount of 6.2 g as a white crystalline solid m.p. 178.degree. (dec.).
EXAMPLE 12
GLYOXYLIC ACID 2-THIENYLSULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that no conc. HCl is employed. In this operation, 7.13 g (40 mmol) of 2-thienylsulfonylhydrazide is reacted with 4.42 g (48 mmol) of glyoxylic acid monohydrate in the presence of 160 cc acetonitrile. The desired product is recovered as a white crystalline material, m.p. 103.degree. (dec.).
EXAMPLE 13
GLYOXYLIC ACID .beta.-STYRENESULFONYLHYDRAZONE
In this operation a suspension of 1-formyl-2-(.beta.-styrenesulfonyl)hydrazine (4.98 g, 22 mmol) in 6 N HCl (44 cc) was stirred for 4.5 hr at 50.degree.-55.degree. C. After cooling a small amount of insoluble material was removed by filtration. The filtrate was added all at once to a stirred solution of glyoxylic acid monohydrate (2.4 g, 26 mmol) in water (180 cc), and stirring was continued for 3 hr. The white crystalline solid which had precipitated was collected by filtration and air dried to give 3.0 g of the product compound, m.p. 141.degree. (dec.).
EXAMPLE 14
GLYOXYLIC ACID BENZYLSULFONYLHYDRAZONE
The subject compound is prepared using the general method of Example 1 except that 3.72 g (20 mmol) of benzylsulfonylhydrazide is reacted with 2.39 g (26 mmol) glyoxylic acid monohydrate in the presence of 0.5 cc conc. HCl and 75 cc of acetonitrile. There is recovered 4.6 g of the desired white crystalline product compound, m.p. 140.degree. (dec.).
EXAMPLE 15
GLYOXYLIC ACID METHANESULFONYLHYDRAZONE
In this operation methanesulfonylhydrazide (2.20 g, 20 mmol) was added all at once to a vigorously stirred suspension of glyoxylic acid monohydrate (2.02 g, 22 mmol) in dioxane (2 cc) and ether (8 cc). After stirring for 0.5 hr the reaction mixture was diluted with additional ether (10 cc). The upper solvent layer was removed by decantation and the residue was triturated with cold ether. The resulting white crystalline solid was collected by filtration and air dried to give 1.8 g of the desired white crystalline product, m.p. 154.degree. C. (dec.).
EXAMPLE 16
GLYOXYLIC ACID 1-BUTANESULFONYLHYDRAZONE
A mixture of 1-butanesulfonylhydrazide (6.0 g, 40 mmol), glyoxylic acid monohydrate (4.4 g. 48 mmol), and ethyl acetate (90 cc) was stirred for 1 hr and then diluted with additional ethyl acetate (90 cc). The organic solution was twice washed with water using 20 cc portions. The organic solution was dried (MgSO.sub.4) and the solvent was removed in vacuo to give a solid. After trituration with benzene, the white crystalline solid was collected by filtration and air dried to give 5.1 g, m.p. 142.degree. C. (dec.).
EXAMPLE 17
GLYOXYLIC ACID n-HEXADECANESULFONYLHYDRAZONE
The subject compound was prepared using the general method of Example 1 except that no conc. HCl is employed. The preparation is carried out by reacting 3.2 g (10 mmol) of n-hexanedecanesulfonylhydrazide with 1.3 g (14 mmol) glyoxylic acid monohydrate in the presence of 200 cc acetonitrile. There is recovered 3.4 g of the desired product compound in the form of a white crystalline material, m.p. 144.degree. (dec.).
The compound of example 8 was prepared in the form of a water-soluble, acid addition salt. While HCl was employed in this example as the salt forming acid, other pharmaceutically acceptable, non-toxic addition salts with acids such as nitric, sulfuric, phosphoric, glycolic could be used.
The compounds used in a practice of this invention, including any salts thereof, can be administered to the animal by any available route, including oral and parenteral (intravenous, intraperitoneal, subcutaneous, and intramuscular) administration. The amount administered is sufficient to ameliorate the leukemia or other type of cancer against which the compounds hereof may prove to be effective, and will depend upon the type of cancer, the species of animal, and the weight of the animal. A dosage of a compound of the present invention within the range from about 0.1 mg to about 500 mg per day per kg of body weight is therapeutic to ameliorate leukemia in the treatment of warm-blooded animals having an abnormal proportion of leukocytes. The upper dosage limit is that imposed by toxic side effects.
To facilitate administration, the compounds employed in a practice of this invention, including the salts thereof, can be provided in composition form, and preferably in dosage unit form. While any compound selected can be administered per se, it is normally administered in conjunction with a pharmaceutically acceptable carrier therefor, which dilutes the compound and facilitates handling. The term "pharmaceutically acceptable" means that the carrier (as well as the resulting composition) is sterile and nontoxic.
The carrier or diluent can be solid, semisolid, or liquid, and can serve as a vehicle, excipient, or medium for the anti-cancer agent. Exemplary diluents and carriers include lactose, dextrose, sucrose, sorbitol, mannitol, starches, gum acacia, calcium phosphate, mineral oil, cocoa butter, oil of theobroma, alginates, tragacanth, gelatin, syrup, methyl cellulose, polyoxyethylene sorbitan, monolaurate, methyl- and propyl-hydroxybenzoate, talc or magnesium stearate.
For convenience in handling, the compounds employed in a practice of this invention and the carrier or diluent can be enclosed or encapsulated in a capsule, sachet, cachet, gelatin, paper or other container, especially when intended for use in dosage units. The dosage units can for example take the form of tablets, capsules, suppositories or cachets.
The following examples illustrate various forms of dosage units in which the compound of Example 2 can be prepared, said compound being typical of the other compounds which can be employed in a practice of this invention.
EXAMPLE 18
______________________________________Tablet formulation Mg/tablet______________________________________Compound (2) 15Lactose 86Cornstarch (dried) 45.5Gelatin 2.5Magnesium stearate 1.0______________________________________
Compound 2 is powdered and passed through a mesh sieve and well mixed with the lactose and 30 mg of the cornstarch, both passed through a sieve.
The mixed powders are massed with a warm gelatin solution, prepared by stirring the gelatin in water and heating to form a 10% w/w solution. The mass is granulated by passing through a sieve and the moist granules dried at 40.degree. C.
The dried granules are regranulated by passing through a sieve and the balance of the starch and the magnesium stearate is added and thoroughly mixed.
The granules are compressed to produce tablets each weighing 150 mg.
EXAMPLE 19
______________________________________Tablet formulation Mg/tablet______________________________________Compound (2) 100Lactose 39Cornstarch (dried) 80Gelatin 4.0Magnesium stearate 2.0______________________________________
The method of preparation is identical with that of Example 2 except that 60 mg of starch is used in the granulation process and 20 mg during tableting.
EXAMPLE 20
______________________________________Capsule formulation Mg/tablet______________________________________Compound 2 250Lactose 150______________________________________
Compound 2 and lactose are passed through a sieve and the powders well mixed together before filling into hard gelatin capsules of suitable size, so that each capsule contains 400 mg of mixed powders.
EXAMPLE 21
______________________________________Suppositories Mg/suppository______________________________________Compound 2 50Oil of Theobroma 950______________________________________
Compound 2 is powdered and passed through a sieve and triturated with molten oil of theobroma at 45.degree. C. to form a smooth suspension. The mixture is well stirred and poured into molds, each of nominal 1 g capacity, to produce suppositories.
EXAMPLE 22
______________________________________Cachets Mg/cachet______________________________________Compound 2 100Lactose 400______________________________________
Compound 2 is passed through a mesh sieve, mixed with lactose previously sieved and fitted into cachets of suitable size so that each contains 500 mg.
EXAMPLE 23
______________________________________Intramuscular injection(sterile suspension inaqueous vehicle) Mg______________________________________Compound (2) 10Sodium citrate 5.7Sodium carboxymethyl-cellulose (low viscositygrade) 2.0Methyl para-hydroxybenzoate 1.5Propyl para-hydroxybenzoate 0.2Water for injection to 1.0 ml______________________________________
EXAMPLE 24
______________________________________Intraperitoneal, intravenousor subcutaneous injection(sterile solution in aqueouscarrier system) Mg______________________________________Compound (8) hydrochloricacid addition salt 15Sodium citrate 5.7Sodium carboxymethyl-cellulose (low vis-cosity grade) 2.0Methyl para-hydroxybenzoate 1.5Propyl para-hydroxybenzoate 0.2Water for injection to 1.0 ml______________________________________
The other compounds useful in a practice of this invention can be prepared in dosage unit form in the same general fashion as that described above for compound 2.
BIOLOGICAL TESTS
Biological testing data for the compounds of this invention are presented in the table given below. Said data were obtained when these compounds were tested against lymphocytic leukemia P388 implanted in mice under the auspices of the National Cancer Institute (NCI) and according to protocols which use the increased survival time of treated animals compared to controls as the measure of antitumor efficiency. In carrying out these tests, the various doses were administered ip in the form of aqueous dispersions or emulsions, the latter being formed in many cases with the aid of hydroxypropylcellulose.
Table 2__________________________________________________________________________BIOASSAY DATA OF SUBJECT COMPOUNDSHydrocarbyl-SO.sub. 2 NHN.dbd.CH--CO.sub.2 H Activity vs. Leukemia P388 in Mice.sup.bCpd. qd 1-9, % T/C for Doses (mg/kg)No. NSC No..sup.a Hydrocarbyl Group 400 200 100 50 25 12.5 6.25__________________________________________________________________________1 176331 p-tolyl 165.sup.c2 245422 phenyl tox 99 138 131 113 163.sup.d 153 137 1283 267212 p-chlorophenyl 1/6.sup.e 2/6 0/6 134 134 122 99 159 142 126 1014 267213 p-methoxyphenyl 0/6 2/6 tox 158 149 141 tox 159 162 128 1165 268244 m-nitrophenyl 3/6 160 141 134 122 138 3/6 tox 137 117 119 4/6 tox 130 131 1186 268245 2,5-dichlorophenyl tox tox 130 132 119 114 tox 137 155 122 122 102 tox tox tox 121 119 1097 274883 .beta. -Naphthyl 4/6 153 196 164 146 135 0/6 tox 90 161 144 1368 274884 m-aminophenyl 1/6 tox 117 119 116 1119 276745 p-acetamidophenyl 4/6 4/6 tox tox 135 135 3/6 tox 146 137 118 11110 278173 2,4,6-trimethylphenyl 1/6 3/6 tox 137 122 112 tox 154 137 126 11711 279507 phenyl, 1,3-bis 0/6 3/6 tox 128 130 119 tox tox 143 133 12712 279506 2-thienyl 1/6 2/6 149 137 127 119 0/6 4/6 136 130 13013 285698 styryl 0/6 tox 186 155 140 12914 273425 benzyl tox 166 185 142 129 127 tox 154 147 146 128 13915 276744 methyl 0/6 0/6 tox 125 119 114 tox 159 126 127 118 0/6 3/6 157 160 13416 276743 n-butyl 0/6 3/6 tox 132 126 114 tox 172 162 135 124 119 0/6 tox tox 136 13617 283459 n-hexadecyl 0/6 2/6 4/6 145 125 116 0/6 tox 140 131 129 .sup.a NSC accession number of the National Cancer Institute. .sup.b Ip P388 murine leukemia treated ip on QD19 schedule according to standard NCI protocols. Assay described in R. I. Geran, N. H. Greenberg, M. M. MacDonald, A. M. Schumacher and B. J. Abbott, Cancer Chemother. Rep., Part 3, 3 (No. 2), 9 (1972), Protocol 1,200. T/C = ratio of survivial time of treated mice to that of untreated controls times 100. Untreated controls survive about 9 days. .sup.c Average of 28 assays. .sup.d Dosages were 75, 50, 32 and 20 mg/kg for this test. .sup.e The indicia such as 0/6, 1/6, 2/6, etc. show no. survivors/no. tes animals as toxicity day survivors (4 days after day of first injection).

Glyoxylic acid hydrocarbylsulfonylhydrazones and therapeutic compositions

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Non-Patent Literature Citations (2)

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House "Chemical Abstracts", vol. 68, (1968), p. 39184s.
Blankley "Chemical Abstracts", vol. 72, (1970), p. 120977g.