SARCOMA 180 SCREENING DATA1

C. Chester Stock, Donald A. Clarke, Frederick S. Philipsand Ralph K. Barclay

Division of Experimental ChemotherapySloan-Kettering Institute andSloan-Kettering Division of

Cornell University Medical College

Although the report on Sarcoma 180 screening data in Supplement 2

(1955) of Cancer Research, page 179, presented a full account of the tech

nique, it will be repeated for the convenience of having it accompany the new

data reported at this time and to include minor modifications or additions to

the procedure. The following is the method we have employed for ability to

retard or inhibit the growth of Sarcoma 180 in mice:

1. Mice. Swiss females, 18-22 gms., from the Blue Spruce, Carworth,

Harpaul, Millerton Research, Rockland and Taconic Farms, and from Joseph

E. Stocker have been used. The mice have been given Purina Laboratory

Chow supplemented with rolled oats and sunflower seeds and water ad lib.

The mice are weighed prior to tumor implantation and at the end of therapy.

2. Tumor transplantation. Tumor fragments, approximately 1.5 mm.

in any dimension, are cut from non-necrotic portions of the donor tumor.

1. The screening program for anti-tumor activity has been supported since1947 by institutional grants from the American Cancer Society. Therealso has been support in part by a grant-in-aid (CH-22) from the AmericanCancer Society upon recommendation of the Committee on Growth of theNational Research Council. Additional support has been given by a researchgrant C-2329 from The National Cancer Institute of the National Institutesof Health, U. S. Public Health Service.

49

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SO Cancer Research Stock et al.

Such fragments are implanted subcutaneously by trocar into the axillary

region. Samples of each tumor are cultured for detection of any possible

bacterial contamination.

3. Therapy. Twenty-four hours after tumor implantation therapy is

initiated by the intraperitoneal route. Maximum tolerated doses of compounds

are injected on a repeated basis. In nearly all cases the toxicities of the

compounds were determined by either of two methods. The maximum

tolerated doses by intraperitoneal injections repeated daily for five days have

been found. Although exceptions have been encountered, in general, one-fourth

to one-third the single dose LD50 has been found to be a dose satisfactorily

approximating the maximum that is tolerated on repeated injection. In later

studies an arbitrarily selected dose of 125 mg/K/day has proven adequately

satisfactory as an initial dose in the anti-tumor test without prior toxicity

data. In the tables the doses are reported in mg/K except for many of the

liquids which have been measured by volume and are listed in cmm/K. It

will be obvious that some of the data reported have been obtained at dose

levels which have caused some toxic manifestations. This toxicity is not

being overlooked if it is also noted in the Remarks column that the compound

is toxic at a higher level. Injections are usually given twice daily for seven

1. The toxicities of most of the compounds included in this report andmany other compounds were obtained by support in part by a researchgrant (C-415) from The National Cancer Institute of the National Institutesof Health, U. S. Public Health Service. The data are on file at theChemical - Biological Coordination Center, National Research Council,Washington 25, D. C.

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Stock et al. Cancer Chemotherapy Screening Data 51

successive days, but in some instances single injections have been given

daily. These include the materials prepared fresh daily and, in some

instances, those prepared in organic solvents. In these data the number of

injections will be listed in the tables as seven or fewer.

4. Repeat testing. The test is repeated at the same or a different

dose level depending upon the result of the first trial. If the first dose has

been too toxic, a lower dose is tried: if the first dose appears to be well

tolerated, a higher one may be tried. For each compound reported herein

as negative there are usually additional supporting data at lower or higher

dose levels or both; however, most of the compounds have not been tested

at dose levels higher than 500 mg/K/day. In order to make more data

available at this time, the standards followed in our reports in the first two

supplements have been relaxed somewhat for some of the present data. In

certain instances negative results obtained in a single test at 500 mg/K

have been reported, and in some cases data have been included without the

usual confirmatory data obtained for most of the compounds included in the

present report.

Results of testing materials have been evaluated as follows:

No effect, when the average diameter was 3/4 ormore of the average diameter of the control tumors.

+ Slight inhibition, when the average diameter was 1/4to 3/4 of the average diameter of the control tumors.

+ Marked inhibition, when the diameter was 1/4 orless of that of the control tumors; actually, thevolume of such a tumor would be 1/64 or less ofthe control if tumors may be considered spherical.

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52 Cancer Research Stock et al.

Over 16,000 compounds and an equal or greater number of materials of

natural origin have been screened for ability to inhibit Sarcoma 180. Fourteen

compounds have met the requirements for the grading of +. They are:

2,4,6-tris-ethyleneimino-s-triazine

4-aminopteroylglutamic acid

4-amino-N -methylpteroylglutamic acid

4-aminopteroylaspartic acid (DL)

4-amino-9-methylpteroylglutamlc acid

4-aminopteroylthreonine

4-amino-9,10-dimethylpteroylglutamic acid

4-aminopteroyltriglutamic acid

5-bis-(ß-chloroethyl)aminoethyl-3-hydroxy-4-methoxym ethyl -2- me thy Ipyr idi ne dihydr ochiori de

N, N', N"-triethylenephosphoramide

N, N-diethyl-N1, N"-diethylenephosphoramide

N-pentamethylene-N', N"-diethylenephosphoramide

O-diazoacetyl-L-serine

6-diazo-5-oxo-L-norleucine

Most of them have already been reported: Annals of the New York

Academy of Sciences 52, 1360-78 (1950); Cancer Research U_, 432-36 (1951);

Proc. Soc. Exp. Biol. Med. 78, 299-305 (1951); Cancer j>, 144-52 (1952);

Nature 173, 71 (1954); Proc. Am. Assoc. for Cancer Research 2, 100 (1956).

More than 200 compounds are in the + category. Included are 6-mer-

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Stock et al. Cancer Chemotherapy Screening Data 53

captopurine, 6-chloropurine, thioguanine, purine, actidione, N-methyl-forma-

mide, urethane, 3,3-dimethyl-l-phenyl triazene, 2-ethylamino-l, 3,4-thia-

diazole, HN2 and certain other nitrogen mustards, some 2,4-diamino-pyrimi-

dines and some anti-folic acids not as effective as those listed above. The

rest of the _+ compounds are being studied as possible leads to more ef

fective, related compounds. All of the other compounds we have tested against

Sarcoma 180 have thus far been negative, but many require retesting at

higher dose levels. The present report lists only negative results.

In the accompanying tabulated data the solvents have been coded as follows:

Solvent Coding

Code No. Solvent

1 saline, physiological

2 carboxymethylcellulose (Cellulose gum) high viscosity,Type 120, Hercules Powder Co., 0.5% in saline

3 gum acacia (gum arable) 1% or 10% in saline

4 propylene glycol

5 butyl Buccinate

6 peanut oil

7 mineral oil

It isa pleasure to acknowledge the many sources of the compounds we

have used. All of the cooperating laboratories have been most helpful in

attempting to provide adequate supplies of compounds, many of which would

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54 CancerResearch Stock et al.

not otherwise have been available to us. The sources of the compounds have

been designated in the tables as listed below.

C Dr. John R. Dice, Parke, Davis and Company

C, Dr. John Ehrlich, Parke, Davis and Company

D Dr. Stanton A. Harris, Merck and Company

DI Dr. Karl Folkers, Merck and Company

Ü2 Dr. Karl Pfister, Merck and Company

Dj Dr. Lewis H. Sarett, Merck and Company

E Eastman Kodak Company

F Dr. F. E. Ray, Cancer Research Laboratory, University of Florida

G Southern Research Institute

H Sterling Winthrop Research Institute

Hj Winthrop Laboratories

Ij Dr. Thomas D. Fontaine, Biologically-Active Compounds DivisionU. S. Department of Agriculture

Ia Dr. H. L. Haller, Insecticide Investigations, U. S.Department of Agriculture, Beltsville, Maryland

Ic Eastern Regional Research LaboratoryU. S. Department of Agriculture

L Monsanto Chemical Company

M Bound Brook Division of American Cyanamid Company

M£ Dr. R. O. Roblin, American Cyanamid Company

N Lilly Research Laboratories

Q E. I. duPont de Nemours and Company

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Stock et al. Camer Chemotherapy Screening Data 55

S S. B. Penlck and Company

T Prof. Alexander Haddow, Chester Beatty Research Institute

W Abbott Research Laboratories

Z William S. Merrell Company

AA B. F. Goodrich Company

AE National Research Council, Chemical-Biological

Coordination Center

AG Lederle Laboratories Division of American Cyanamid Company

AJ Hoffman-LaRoche, Inc.

AM The Squibb Institute for Medical Research

AN National Aniline Division of Allied Chemical and Dye Corporation

AP Esso Research and Engineering Company

AQ Endo Products, Inc.

AR Commercial Solvents Corporation

AW Sloan-Kettering Institute personnel or related sources:

AWj Dr. Liebe Cavalieri

AW2 Dr. Kenneth Savard

AW3 Dr. Aaron Bendich

AW5 Dr. Ralph K. Barclay

AW6 Dr. John Davoll

AW? Dr. M. Earl Balis

AW10 Dr. C. Chester Stock

AW. Dr. Jack J. Fox

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56 Cancer Research Stock et al.

AWj3 Dr. Le onida Santamaría

AW16 Dr. Paul Roll

AW.- Dr. William Regelson (now at Roswell Park Memorial Institute)

AW19 Dr. Milton Gordon

AW24 Dr. Alexander Hampton

AW25 Mr. Norman Haber

AZ Dr. Kenneth M. Campbell (now at Mead Johnson and Company)

BA Dr. C. K. Cain (now at McNeil Laboratories)

BB Chas. Pfizer and Company, Inc.

BD Dr. Karl Dittmer, Department of ChemistryFlorida State University

BE Dr. George Hitchings, Wellcome Research Laboratories

BI Dr. J. Philip Mason, Boston University

BL Remington Rand, Inc.

BM Dr. Robert C. Elderfield, University of Michigan

BO Ayerst, McKenna and Harrison

BP Sharpies Chemicals, Inc.

BV Union Carbide and Carbon Corporation (Mellon Institute Fellowship)

BX Bristol Laboratories

BY The Upjohn Company

CA Dr. N. H. Cromwell, Department of Chemistry,University of Nebraska

CB Dr. Henry A. Rutter

CC G. D. Searle and Company

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Stock et al. Cancer ChemotherapyScreening Data 57

CE, Dr. Louis L. Fieser, Harvard University

CF General Mills, Inc.

CG Dow Chemical Company

CI General Aniline and Film Corporation

C L Boots Pure Drug Company

CM, Dr. Leslie Hel 1er man, Johns Hopkins University

CO Dr. Henry Richter, University of Colorado

CQ Sharp and Dohme, Inc.

CT National Drug Company

DB Virginia - Carolina Chemical Corporation

DC Wallace Laboratories, Inc.

DF Ciba Pharmaceutical Products, Inc.

DJ Campbell Pharmaceutical Company

DK Dr. Louis H. Goodson, Midwest Research Institute

DN Dr. W. T. Sumerford, Mead Johnson and Company

DP Warner - Chilcott Laboratories

DQ Dr. Irving A. Kaye, Brooklyn College

DW American Smelting and Refining Company

EC Rohm and Haas Company

EF Dr. L. H. Briggs, Auckland University

EG Dr. Jerry Milionis, Purdue University

EH N. V. Organen

El Dr. Wilson M. Whaley, Pabst Laboratories

EJ Dr. Marvin D. Armstrong, University of Utah

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58 CancerResearch Stock et al.

EL Dr. Karl W. Eishold, Farbwerke Hoechst

EN Dr. Selman Waksman, Rutgers University

EU Dr. Adrien Albert, The Australian National University

EW Dr. D. S. Tarbell, University of Rochester

EX Wallace and Tiernan Products

EY Dr. P. E. Fanta, Illinois Institute of Technology

EZ Dr. Richard Wiley, University of Louisville

FB Alfred I. duPont Institute of The Nemours Foundation

FC Dr. E. R. Weidlein, Jr., Mellon Institute of Industrial Research—¿�v

FCj Dr. Marcus Morgan

FC2 Dr. Horace A. DeWald

FÜ3 Dr. Robert Tipson

FC4 Dr. Alice RenfrewParke, Davis and Company, Fellowshipsin Medicinal Chemistry at Mellon Institute

Dr. Alexander Moorewf

FC6 Miss Arveta MeKim

FD Standard Brands, Inc.

FE Prof. Dr. F. Bustinza, University of Madrid

FF Dr. Sidney Farber, Children's Cancer Research Foundation, Boston

FG Dr. Ernst R. Kirch, University of Illinois

FH Dr. Charles F. Geschickter

FK Dr. Patrick J. Hannan, Fungus Control Section Materials Branch,Engineer Research and Development Laboratories,Fort Belvoir, Virginia

FQ Prof. Dr. H. Oettel, Badische Anilin and Soda-Fabrik AG.

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Stock et al. Cancer ChemotherapyScreening Data 59

FR Falrfield Chemical Division of Food Machinery and Chemical Corp.

FU Dr. T. Lloyd Fletcher, University of Washington

FW Dr. Ng. Ph. Buu-Hoi, Radium Institute of Paris

GA Dr. Donald Visser, University of Southern California

GC Dr. Glenn S. Skinner, University of Delaware

GD Dr. Ralph L. Shriner, University of Iowa

GJ Ethyl Corporation Research Laboratories

GM Dr. Henry Gilman, Iowa State College

GP Dr. H. H. Burchalter, University of Kansas

GS Heifetz & Co.

GV Dr. O. Ferno, Halsingborg, Sweden

HA Carbide and Carbon Chemicals Company (Boyce Thompson Institute)

HC Dr. Roland K. Robins, New Mexico Highlands University

HE Nepera Chemical Company

HF Dr. Clifton E. Miller, North Dakota Agricultural College

HG Dr. Alfred Burger, University of Virginia

HH Verona Chemical Company

HI Dr. Morris Levine, Universal Detergents, Inc.

HK Ethicon Suture Laboratories

HM Colgate-Palmolive Company

HN Allen & Hanburys Ltd.

HQ Hooker Electrochemical Company

HT Dr. Walter H. Härtung, University of North Carolina

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60 CancerResearch Stock et al.

HU Barrett Division of Allied Chemical and Dye Corporation

HV Dr. Samuel P. Massie, Fisk University

HW van Ameringen-Haebler, Inc.

JB Dr. A. Jonsson, Royal Institute of Technology, Sweden

JG Dow Corning Corporation

JH Dr. LeRoy H. Klemm, University of Oregon

JK Dr. D. M. Carney, University of Detroit

JP Phillips Petroleum Company

JQ Geigy Pharmaceuticals Division of Geigy Chemical Corporation

JR Dr. C. S. Rondestvedt, Jr., University of Michigan

JT Air Reduction Company

JU Dr. Erwin Sheppard, New York Hospital

JV Lever Bros., Research and Development Division

JZ Dr. G. Shaw, New South Wales University of Technology

KG E. L. Clark

KE Dr. E. B. Christens en, Oregon State College

KH Cyclo Chemical Corporation

KL Pennsylvania Salt Manufacturing Company

KT Dr. R. P. Mariella, Loyola University, Chicago

KZ Philips Export Company

LA The Richardson Company

LB Norda Essential Oil and Chemical Company

LC Nippon Seibutsu Kagaku, Kenkyusho, Ltd.

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Stock et al. Cancer ChemotherapyScreening Data 61

LD J. T. Baker Company

LE Jefferson Chemical Company

LF Department of Neoplastia DiseasesJefferson Hospital, Philadelphia

LG Dr. Hans Lettre, Heidelberg, Germany

LH Wyeth Institute of Applied Biochemistry

LI Dr. L. F. Audrieth, University of Illinois

LJ Specific Pharmaceuticals, Inc.

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62 CancerResearch Stock et al.

The Sarcoma ISO studies since 1947 have been made possible through

the effective help of many technicians, who faithfully carried out the routine

work even on week-ends. In addition to those mentioned in the second report

in this series (Supplement No. 2 Cancer Research 1955, page 181), the

following have also participated in studies on the compounds included in the

present report :

Mary Ann Albrycht Doris KwokErika Di Cyan Minnie MikellElinor Evans Alice O'NeillCynthia Folkers Ann OpenheimerJoan Friedman Barbara RapaportMarian Gold Ana RothGeorgetta Harrar Nancy SimmsAnne Haywood Mary Frances SoriaAnn Infangar Inese SulcsWinifred Kine Barbara WeissPhyllis Korn Evelyn Wilbur

Others who have helped in the assembly of the data, in proof-reading,

and in various administrative details are:

Amir Askari Julia KeaneNancy Chllson Rosalie MenegasDietrich Fischer Sophronia MyronWilliam Elton Ann ParkerEsther Garfinkel Mildred PhillippsMargaret Hasson Edward PodvollMargaret Jeter

We are particularly indebted to Miss Martha Nelson, Miss Elisabeth

Platt, and Mrs. Mariam Anthony for careful typing of the data for the photo

offset reproduction.

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Stock et al. CancerChemotherapyScreening Data 63

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1958;18:49-224. Cancer Res   C. Chester Stock, Donald A. Clarke, Frederick S. Philips, et al.   Sarcoma 180 Screening Data

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