New antitumor antibiotic

The present invention is a continuation of U.S. Patent Application No. 464,046 (CT-1970), filed January 12, 1990.

It is known that antibiotic agents can be prepared by fermentation of various types of microorganisms. In this way, some biologically active chromophores can be prepared by coupling them to a protein molecule to form a chromoprotein complex. The chromoproteins studied were described in [Neocarzinostatin chromophore, Napier, MA, et al., Biochem Biophys. Gap. Commun. 89: 635-642 (1979)]. Anti-tumor antibiotic C-1027 is a chromophore fraction derived from Streptomyces globisporus C-1027 microorganism. Its isolation, characterization and biological activity have been described by T. Otani et al., 1991, in the isolation and characterization of a non-protein chromophore and the degradation product of antibiotic C-1027, Journal of Antibiotics, 44, 564-568.

The present invention relates to a novel compound, BMY-46164, which can be produced by fermentation of the Actinomadura strain, to a process for the preparation thereof, to a process for the use and isolation of the active ingredient. The inventors include all pharmaceutically acceptable derivatives of the composition under the name of the compound.

The new fermentation product is based on high-resolution, fast atom bombardment mass spectroscopy (FABMS) data. ^c40^H43^N2°12^cIt is a colorless, amorphous solid which has the following characteristics.

The fermentation product of the present invention is a non-proteinaceous, non-covalently bound chromophore that binds to protein derived from Actinomadura strains during complexation.

The fermentation product has been shown to have antimicrobial activity against various gram positive microorganisms such as Enterococcus faecalis, Staphylococcus aureus, and Bacillus subtilis. It also has activity in the treatment of anti-tumor models, such as P388 leukemia.

The novel antibiotic can be prepared by fermentation of the BMY-46164 producing strain or a mutant thereof, which is carried out under immersed aerobic conditions in an aqueous medium until a significant amount of BMY-46164 is produced in said medium. The fermentation is followed by the recovery of BMY-46164 from the culture medium, which is essentially free of by-products.

The biologically pure culture of Actinomadura Q473-8 from which the compound of the present invention is derived was deposited at the American Type Culture Collection (AATCC) in Rockville, MD and the ATCC was assigned a permanent stock under code 53806.

Cultures of the above strain as lyophin were also maintained at the Bristol-Myers Squibb Pharmaceutical Research Institute Actinomycetes Culture Collection, Wallingford, Connecticut.

The ATCC deposit was made before the invention was filed, and

- 4 corresponds to the production of 35 USC 112 for all deposited microorganisms.

Figure 1 shows the UV spectrum of BMY-46164 in methanol; Figure 2 shows the IR spectrum of KBY-46164 (KBr); Figure 3 shows the proton NMR of BMY46164 in dg-DMSO. Figure 4 shows the NMR spectrum of carbon BMY46164 in dg-DMSO.

The fermentation product BMY-46164 of the present invention has an experimental formula of C4QH43N2O12Cl and a molecular weight of 778.

A preferred process for the preparation of BMY-46164 comprises the following steps:

1) fermentation of a suitable strain of actinomycet,

2) extraction of the protein-like fermentation product,

3) denaturing the product of step 2) and

4) Isolation of the product of formula C4QH43N2O2Cl having a molecular weight of 778.

The product is a colorless, amorphous solid, the chemical structure of which has not yet been demonstrated, but which has the following characteristics:

Mass Spectrum: Kratos MS 50 TC Mass Spectrometer.

FABMS: 778.2527.

Typical fragment ions are 294,1339 and 149,0603.

Ultraviolet spectrum: Hewlett Packard 8452A diode array spectrometer; Concentration: 1.0 mg / 100 ml methanol. The neutral solution has the following absorption maxima:

anax^nm(E^1%1 cm): 278 (635).

<td colspan="2">PerkinElmer</td><td> 1800</td><td>FTIR</td><td colspan="2">spectrometer</td><td>, KBr</td><td>cm 1:</td><td></td><td></td>

<td> 3424,</td><td> 3076,</td><td> 2934,</td><td> 2838,</td><td> 2170,</td><td> 1640,</td><td> 1578,</td><td> 1520,</td><td> 1490,</td><td> 1462,</td>

<td> 1424,</td><td> 1376,</td><td> 1292,</td><td> 1248,</td><td> 1184,</td><td> 1156,</td><td> 1112,</td><td> 1072,</td><td> 1046,</td><td> 952,</td>

<td> 880,</td><td colspan="3"> 832, 810, 754, 682</td><td> , 666,</td><td> 648,</td><td colspan="2"> 576, 525.</td><td></td><td></td>

900,

500 MHzÍH NMR:

Model AM-500 spectrometer.

Bruker mml. Solvent dg-DMSO.

Double carbon proton sample, 5

<td colspan="6">Chemical shifts (ppm): 9.18 (d 5, 1H),</td><td>8 09 (dd, 1H),</td>

<td> 7,40</td><td>(D,</td><td>1 H),</td><td> 7,15</td><td>(d, 1H), 6.97 (d, 1H)</td><td>H)</td><td>, 6.81 (d, 1H),</td>

<td> 6,58</td><td>(D,</td><td>1 H),</td><td> 6,37</td><td>(t, 1H), 5.41 (s, 1</td><td>H)</td><td>, 5.37 (dd, 1H),</td>

<td> 5,27</td><td>(M,</td><td>1 H),</td><td> 5,13</td><td>(s s, 1H), 4.91 (s</td><td> , 1</td><td>H), 4.89 (m, 1H),</td>

<td> 4,65</td><td>(D,</td><td>1 H),</td><td> 4,22</td><td colspan="2">(dd, 1H), 4.13 (ddd,</td><td>1H), 3.94 (m, 1H)</td>

<td> 3,80</td><td>(Dd,</td><td>1 H)</td><td> , 3,54</td><td>(s, 3H), 3.46 (dd,</td><td> 1</td><td>H), 3.21 (d, 1H),</td>

<td> 3,12</td><td>(S,</td><td>3 H),</td><td> 2,94</td><td>(ddd, 1H), 2.73 (m,</td><td> 1</td><td>H), 2.71 (dd, 1H),</td>

<td> 2,58</td><td>(Dd,</td><td>1 H)</td><td> , 2,31</td><td>(dd, 1H), 2.19 (s,</td><td> 3</td><td>H), 2.10 (s, 3H),</td>

<td> 1,26</td><td>(S,</td><td>3 H).</td><td></td><td></td><td></td><td></td>

125 MHz ¹³C NMR:

spectrometer.

Bruker Model AM-500

Proton decoupled spectrum: double carbon proton sample, mm.

Solvent: dg-DMSO. Chemical shifts (ppm):

131,3,

133,0

131,4,

168,4,

168.1, vol

153,3, 152,8, 139,3, 138,5,

<td> 131,0</td><td> , 130,8</td><td colspan="4"> ,126,8, 124,7, 124,3, 122,5,</td><td> 122,1,</td><td> 121,0,</td><td> , 109,</td>

<td> 97,5,</td><td> 95,7,</td><td> 94,3,</td><td> 94,2, 93,3,</td><td> 90,1,</td><td> 74,0,</td><td> 73,2,</td><td> 70,9,</td><td> 69,8,</td>

<td> 69,6,</td><td> 66,3,</td><td> 62,5,</td><td> 55,4, 54,4,</td><td> 52,3,</td><td> 43,2,</td><td> 37,5,</td><td> 33,6,</td><td> 22,7,</td>

16,1.

2,

19,5,

For all live hours described above, the following parameters were used.

«R ·

The solvents were not redistilled before use. The quality of methanol, ethyl acetate, chloroform, hexane, diethyl ether, dichloromethane and acetonitrile was in the grade of ÁCS reagent. The water used in HPLC HPLC was locally deionized using the Barnstead Nanopure II system. The HPLC was methanol and acetonitrile B&amp;J Brand HPLC grade solvents. The ammonium acetate used was Fisher HPLC grade. DEAE Cellulose Schleicher &amp; Schuell Anion Exchange Cellulose (Batches 2932 and 2893). The Tris buffer (tris (hydroxymethyl) aminomethane) enzyme was a high purity pure substance (Bethesda Research Laboratories) at a concentration of 0.05M, pH 7.4. Dicalite is a high speed filtration aid (Grefco Minerals).

Normal phase thin-layer chromatography (TLC) was performed on silica gel 60, F254 plates (EM Reagents, Cat. No. 5765, 5 x 10 cm, 0.25 mm thick). Reverse phase TLC analysis was performed on Whatman MKC1g plates (Cat. No. 4803-110, 0.2 mm thick). The plates were developed in Whatman cylindrical flasks with lids and 10 ml of eluent were used. The chromophores were visualized under UV using 254 nm ultraviolet light.

Preparative chromatography (TLC) was performed on silica gel 60, F254 plates with EM Reagents (Cat. No. 5766, 20x20 cm, 2 mm thick). The plates were developed in glass jars with lids, using 100 ml of eluent.

«· · · « • 4 · · ··· «««« « * · ·*··· ·· • · · · · «

- 7 The apparatus used for vacuum liquid chromatography consists of a Buchner funnel (Kontes, K-954100) and a sintered sintered glass plate (porosity M), a laterally fitted inlet for introduction of a vacuum and a receiving vessel with a 24/40 connection located at the bottom of the unit. The funnels were equilibrated by pouring the initial eluent onto a 5 cm thick absorbent bed under vacuum. Samples were pre-adsorbed onto the adsorbent, applied as a slurry to the funnel, and slurried with the initial eluent. The gradient elution steps are performed with a predetermined volume of increasingly polar eluent. After each volume of graduated eluent used, the adsorbent layer in the funnel is aspirated to dryness. The fractions were concentrated on a rotary evaporator and pooled based on the results of an in vitro biological assay as compared to HPLC-UV and TLC.

Dicalit cormatography refers to adsorption chromatography on silica. Samples were dissolved in chloroform / methanol (2: 1) and then adsorbed onto Dicalitra. The resulting powder is suspended in hexane and transferred to sintered glass filter vacuum chromatography funnels. Appropriate predetermined volumes of solvents were printed through Dicalin and the fractions were collected in a round bottom flask and evaporated on a rotary evaporator.

The particle size exclusion chromatography apparatus was as follows. Glenco column (2.5 in. X 100 cm), which • «• ·

- with 8 solvent-resistant Teflon ends and plates; fluid measuring apparatus (Inc. FMI Laboratory Pump (RP-G1501 Model); Glenco Glass Receptacle (500 mL); Isco 328 Model Fraction Collector. Columns are filled with 150 g Sephadex LH-20 (Pharmacia) pre-swollen ion exchange sludge in the eluent as eluent). Solvent is passed down the column at a rate determined by the laboratory pump.

HPLC purifications by HPLC were performed using the Beckman System Gold unit, which contains the following: 126 model solvent delivery units; 166 P Programmable Detector; solvent receiving kit; Altex injector; Dynamax 60 ° A metal prepared columns; normal phase silica gel (25 cm x 10 mm, 8 μ, Si-83-III-C) or (25 cm x 21.5 mm, 8 μ, Si-83-121-C) and reversed phase silica gel (25 cm x 10 mm) , 8 μ, C_Xg-83-211-C).

The antineoplastic antibiotic agent of the present invention may be prepared by fermentation of the actinomycet strain producing the appropriate BMY-46164 substance. A preferred microorganism is a strain obtained from a soil sample isolated in Athens (Greece) and bearing the symbol Q473-8. Biologically pure strain Q473-8 was deposited at ATCC as described above.

The taxonomic studies on strain Q473-8 are described in detail in U.S. Patent Application No. 464,046. Chemo-taxonomic data for strain Q473-8 · 9 · · · 9 · · · · · · · · · · · · · · · · · ·

9 and its morphological characteristics indicate that the micro-organism is a member of the genus Actinomadura; in more detail, morphology and charcoal utilization characteristics are similar to Actinomadura madurae. See, e.g., Williams et al., The Prokaryotes, Vol. II, 2103-2177 (1981), Starr, Stop, Truper, Balows, and Schlegel, Eds. Communication.

Further characterization, including menaquinone analysis, is required to determine whether the strain Q473-8 exhibits characteristics that are consistent with those of the newly deposited and proposed new genus, Nonomuria. See, e.g., Goodfellow et al., Biology of Actinomycetes (1988), 223-238 (1988), Okami, Beppu and Ogawara, eds. Communication.

It is to be understood that the present invention is not limited to the particularly preferred strain described above, or to microorganisms which are perfectly within the specification. In particular, any of the above-described microorganisms or mutants of the BMY-46164 material are contemplated to be within the scope of the invention, which may be prepared by conventional techniques such as X-rays, ultraviolet rays, nitrogen mustard, phage treatment or the like.

BMY-46164 can be prepared by culturing a strain of any Actinomadura species (producing BMY-46164), preferably strain Q473-8 or a mutant or variant thereof, under immersion, in aerobic aqueous medium.

k * ·

The microorganism is grown in a medium having an assimilable carbon source such as sucrose, lactose, glucose, rhamnose, fructose, mannose, melibose, glycerol or soluble starch. The medium further contains an assimilable source of nitrogen, such as peptone, fish nutrient, soybean meal, peanut nutrient, cotton seed nutrient, corn juice, yeast extract or ammonium salts. If desired, inorganic salts such as sodium chloride, potassium chloride, magnesium sulfate, calcium carbonate, phosphates, etc. may be added.

Trace elements such as copper, manganese, iron, zinc, etc. if desired, they may be added to the medium or may be added to the medium as impurities for other additives.

The production of BMY-46164 material can be carried out at any temperature that provides adequate growth of the producing microorganism and may be, for example, from about 16 ° C to about 41 ° C. The fermentation is preferably carried out at a temperature of about 25 ° C to about 35 ° C, more preferably about 27 ° C to about 37 ° C. The medium preferably has a neutral pH. The production of the antibiotic is usually carried out over a period of about 4 to about 5 days.

The fermentation may be carried out in flasks or in laboratory or industrial fermenters, which may have different capacities. When tank fermentation is required, it is preferable to form a vegetative inoculum in the medium by producing the vegetative inoculum by inoculation or by addition of a lyophilized culture in a small amount of medium.

Once this active inoculum has been prepared as described above, it can be aseptically transferred to the culture medium in the fermentation tank where large amounts of BMY-46164 are produced. The culture medium used to form the vegetative inoculum may be the same or different from the culture medium used in the container, within a range that ensures good growth of the microorganism. During the fermentation, mixing can be carried out using a mechanical propeller. During the fermentation, conventional antifoaming agents such as fatty oil or silicone oil may be used if desired.

Isolation and purification of the antibiotic BMY-46164 from the fermentation medium cannot be carried out by any conventional solvent extraction or chromatography. Instead, the isolation is carried out as described below.

The entire volume (90 L) was filtered through Dicalite filtration aid. To the filtrate was added 1 kg of DEAE cellulose with stirring and the mixture was cooled to 4 ° C. DEAE cellulose was recovered by filtration and rinsed with fresh Tris buffer cooled to 4 ° C.

The DEAE cellulose filtered cake was extracted for 1 hour

1 methanol-ethyl acetate (1: 1).

The extraction mixture was filtered and the resulting cellulose filter cake was rinsed with additional 3 L of ethyl acetate. To the combined organic extract was added 6 L of chilled water. The ethyl acetate layer was separated and concentrated in vacuo; 1 g of crude chromophore extract is thus obtained.

Alternatively, the chromophore extract is obtained by substituting dichloromethane for ethyl acetate in the partitioning step. In this case, 2 volumes of chilled water at 4 ° C and 1 volume of dichloromethane are added to a methanolic solution obtained by extraction of 1 volume of protein-bound DEAE cellulose filter cake. The organic phase is concentrated on a rotary evaporator to give the crude chromophore extract.

1.0 g of the chromophore extract was adsorbed onto 3.5 g of Universal silica gel (63-200 microns) and transferred to a 60 ml vacuum liquid chromatography funnel containing 25 g of Lichroprep Si 60 silica gel (EM Science, 9390, 25-40 micron). Chloroform-methanol step by step! gradient elution was performed using a 300 mL eluent volume. 3-5. fractions were determined by in vitro assays. The eluent used in the above fractions was chloroform containing 3% methanol, 5% methanol and 8% methanol. The combined fractions (202 mg) were further purified by preparative layer chromatography using chloroform-methanol (90:10) as eluent.

Recovered lane showing the main activity (R_f (0.37 mg) was subjected to further HPLC purification on a Dynamax semiprepared silica gel column (Si-83-III-C). The process used isocratic conditions (chloroform: methanol 94: 6) at a flow rate of 4 ml / min.

The peak was eluted after 13.3 minutes, which was detected by UV detection (254 nm). Collect the eluate corresponding to the peak and evaporate to dryness. The residue (28 mg) was applied to a second Merek preparative thin layer chromatography plate. The development was carried out using chloroform-acetonitrile (60:40) as eluent. The major recovered band (Rf = 0.15) is 16.5 mg.

The final purification was performed by reverse phase high performance liquid chromatography (HPLC) using a Dynamax semiprep column (C18-83-211-C).

The gradient elution was carried out with 20% A to 80% B to 40% A to 60% B eluent over 40 minutes, where A = acetonitrile, B = 0.1 m ammonium acetate-methanol (3: 1). Elution of the desired 11 mg of chromophore takes place in 30.9 minutes and the product is BMY-46164.

Other methods for purifying BMY-46164 material, starting from the crude chromophore extract, may be used as follows:

The crude extract (12.9 g) was dissolved in a 2: 1 mixture of chloroform-methanol and adsorbed onto 300 mg of dicalite adsorbent. The Dicalite filter cake was washed with 1 L of the following solvents, respectively: hexane, diethyl ether, ethyl acetate, dichloromethane and methanol.

BMY-46164 was found in the diethyl ether and ethyl acetate fractions, which were combined to give 0.6 g.

This residue was dissolved in 5 ml of chloroform-methanol (1: 1) and then pre-swollen with the above solvent mixture.

Loaded on a Sephadex LH-20 column. The elution flow rate was 1.5 ml / min. five fractions were collected. BMY-46164 is mainly obtained in fraction 3 at 0.7 brain volume. The fraction was evaporated to give 143 mg.

The final purification was carried out using normal phase high performance liquid chromatography (HPLC) using a 21.4 mm Dynamax preparative column (Si-83-121-0) and a flow rate of 10 ml / min. The gradient elution was carried out using chloroform-methanol (95: 5) -chloroform-methanol (85: 15) over 40 minutes. Detection was performed at 300 nm. 34 mg of BMY-46164 are obtained in 27.7 minutes.

Formulations containing BMY-46164 and / or an acid or base addition salt may also contain other appropriate ingredients employed in appropriate amounts. In general, from about 0.001% to 99.99% of one or more pharmaceutically acceptable carriers may be used in these compositions, which may be, for example, a filler, carrier, stabilizer, gelling agent, coloring agent, perfume and the like. The content of the active compound (s) in these compositions is generally from about 0.01% to about 10%, preferably from about 0.17% to about 5%.

The following examples illustrate the invention. The examples are not to be construed as limiting the scope of the invention.

First example

Fermentation of BMY-46164 in shake flasks

Strain Q473-8 is stored in and transferred to yeast extract-malt-extract agar-agar medium supplemented with calcium carbonate.

The medium contains 4.0 g of dextrose, 4.0 g of yeast extract, 10 g of malt extract, 1.5 g of calcium carbonate and 15 g of agar. The mixture of the above materials is made up to 1 liter with distilled water. For each transfer, agar-agar smear is incubated for 5 to 7 days at 28 ° C.

For the production phase, the inoculum is prepared by transferring a growing portion of the smear culture to a 500 ml Erlenmeyer flask containing 100 ml of vegetative medium. The vegetative medium contains 2% glucose,% fish nutrients and 0.5% calcium carbonate. This vegetative medium was incubated for 3 days at 28 ° C in a rotary shaker at 250 rpm.

ml of the above vegetative growth mixture is transferred to a 500 ml Erlenmeyer flask containing 100 ml of production medium containing 2% glucose,% peptone and 0.5% calcium carbonate. The production medium is incubated for 4 to 5 days at 28 ° C in a rotary shaker at 250 rpm. The culture produces maximal levels of BMY-46164 in about 4 days in the fermentation cycle.

• · * * · · · · • ·· »···* ··

Second example

Fermentation of BMY-46164 in a laboratory fermenter

1 A two-stage vegetative medium is used for fermentation in a nominal volume Biolafitte fermenter. Transfer 16 ml of the vegetative culture of Example 1 to a 2 L Erlenmeyer flask containing 400 ml of the second vegetative medium. The second vegetative medium 2^:Contains% glucose, 2% peptone and 0.5% calcium carbonate. The second vegetative culture was incubated for 3 days at 28 ° C in a rotary shaker at 250 rpm.

1200 ml of the above vegetative culture was transferred to a 4 L Vitro flask and inoculated into a 50 L nominal Biolafitte fermenter. The fermentor contains 30 L of production medium containing 2% glucose, 2% peptone and 0.5% calcium carbonate. The growth of the microorganism is assured under the following conditions: mixing: 250 rpm; temperature: 28 ° C; aeration: 30 rpm. An antifoam agent (polypropylene glycol 2000, Dow Chemical) is used to control foaming. Maximum production of BMY-46164 is reached on days 4-5 from the start of the fermentation cycle.

« * • ·· · ·« ·· • · · »«··· ··

Third example

Bacterial activity test

In accordance with the procedure described below, the efficacy of BMY-46164 was compared to that of ampicillin against various microorganisms. The results are shown in Table I below.

• 4

- 18 TABLE I

Anti-bacterial effect of BMY-46164

Primary MICs (/ µg / ml)

<td>MICRO-ORGANISM</td><td>BMY-46164</td><td>ampicillin</td>

<td>Enterococcus faecalis A2O688</td><td> 8</td><td> 0,25</td>

<td>E. faecalis A257O7</td><td> 4</td><td> 0,25</td>

<td>E. faecalis A257O8</td><td> 8</td><td> 0, 5</td>

<td>Staphylococcus aureus A9537</td><td> 4</td><td> 0,06</td>

<td>S. aureus / NCCLS strain</td><td> 16</td><td> 0,06</td>

<td>S. aureus</td><td> 16</td><td> 0,5</td>

<td>Escherichia coli A15119</td><td> &gt; 500</td><td> 1</td>

<td>E. coli / NCCLS strain</td><td> &gt; 500</td><td> 2</td>

<td>E. coli A9751</td><td> &gt;500</td><td> 0,25</td>

<td>Kiebsiella pneumoniae A2O468</td><td> &gt; 500</td><td> 16</td>

<td>K. pneumoniae A2O468</td><td> &gt; 500</td><td> 32</td>

<td>Proteus vulgaris A21559</td><td> &gt; 500</td><td> 32</td>

<td>Pseudomonas aeruginosa A9843</td><td> &gt; 500</td><td> &gt; 128</td>

<td>P. aeruginosa A2O235</td><td> &gt; 500</td><td> 32</td>

<td>P. aeruginosa / NCCLS strain</td><td> &gt; 500</td><td> &gt; 128</td>

<td>Bacillus subtilis A95O6-A</td><td> 64</td><td> 1</td>

The antimicrobial spectrum of BMY-46164 was determined by serial media dilution using medium (Difco).

. · ···: : :

• · » · · * ·· ····&lt; « ·* ·· ·

4th example

Test for antitumor activity

The anti-tumor activity of BMY-46164 P388 leukemia in mice was investigated. Figure 2 shows a

BMY-46164 and olivomycin A activity.

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<td></td><td>She</td><td></td><td>SHE</td><td></td>

<td></td><td>She</td><td></td><td>ex</td><td></td>

<td>stone</td><td></td><td></td><td></td><td></td>

<td></td><td>k.</td><td></td><td>ca.</td><td></td>

<td>SHE</td><td>Pd</td><td></td><td>PH</td><td></td>

<td>T-1</td><td>H</td><td></td><td>H</td><td></td>

<td>X</td><td> 00</td><td> &lt;&lt;</td><td>tendon</td><td>She</td>

<td></td><td> *.</td><td></td><td>CM</td><td>rh</td>

<td>r-d</td><td>She</td><td>She</td><td></td><td></td>

&gt; 1 r - 1

Φ • d tn 'Φ

Fri d

Ή

N cn • d ω 'Φ • P Φ!&gt; R — I

Φ PQ cn

<img file="HUT64397A_D0007.tif" />

Ö Ή

Ο • rl go&gt; • H r-1 o

CM rCM cn cn

I&gt; 1

Ski PQ

<td colspan="3">CM</td>

<td></td><td>OO o</td><td></td>

<td></td><td> &amp;</td><td>kd</td>

<td></td><td></td><td>r — d kO</td>

<td></td><td>r-</td><td></td>

<td></td><td>ί^-1</td><td> 1</td>

<td>tn</td><td>σ \</td><td></td>

<td>PQ</td><td>cn</td><td>Ski</td>

<td>E.G</td><td>SHE</td><td>PQ</td>

DMSO + PBS 5 15.5 155 * · »4 * I ·« · 4 · · · · · · · · · ··········································································································

II. The data described in Table II are obtained by the procedure described in Transplanted Animated Tumors, Bradner, WT, Cancer and Chemotherapy Vol. 1, 221-227 (1980), ST Crooke and AW Prestayko, Eds. literature.

The results illustrate the utility of the compound of the invention and its pharmaceutically acceptable derivatives in the treatment of bacterial infections and cancer in various host units.

Host refers to not only in vitro test cells and mice, but also to higher organisms such as mammals. Examples of preferred host units are human patients.

The compounds and compositions of the present invention may be administered to a suitable host unit by a variety of routes, for example, in patients with bacterial infections and / or cancer. The present invention includes oral, parenteral, topical, nasal, oral and ophthalmic compositions.

The present invention further encompasses modifications which are not substantially altered by those skilled in the art.