Vindesine) in Rodents and Mitotic Accumulation Studies in Culture'

[CANCER RESEARCH 38, 2886-2891 , September 1978] 0008-5472/78/0038-0000$02.00 Antitumor Activity of Deacetyl Vinblastine Amide Sulfate (Vindesine) in Rodents and Mitotic Accumulation Studies in Culture'

Martin J. Sweeney, George B. Boder, George J. Cullinan, Hlllman W. CuIp, William D. Daniels, Richard W. Dyke, Koert Gerzon,2 Robert E. McMahon, Robert L. Nelson, Gerald A. Poore, and Glen C. Todd

The Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, Indiana 46206 tM J. S., G. B. B., G. J. C., H. W. C., W. D. D., K. G., R. E. M., G. A. P.J; The Greenfield Laboratories, Eli Lilly and Company, Greenfleld, Indiana 46140 (G. C. TI; and Lilly Laboratories for Clinical Research and the Department of Medicine, Indiana University Schoolof Medicine, Indianapolis, Indiana 46202(R. W. D., R. L. N.J

ABSTRACT Since then VLB has demonstrated effectiveness against other lymphomas, neuroblastomas, and histiocytosis X; Vlndesine (VDS; deacetyl vinblastine amide sulfate) VCR has been used in acute lymphocytic leukemia and has giveni.p. dailycompletelyinhibitedthe growthof boththe been incorporated into several combination chemotherapy Ridgewayosteogenicsarcomaat 0.4 mg/kgandthe Gard regimens for the treatment of beukemias and solid tumors. ncr IymphosarcÃ³maat0.25 mg/kg. In contrast,the parent VLB and VCR were also found to differ in their side alkaloid, vinbbastine(VLB), at 0.5 mg/kg was inactive effects; the dose-limiting effect of VLB is leukopenia, and againstthesetwotumors.VDScausedinhibitionofgrowth for VCR it is neurotoxicity. of the Mecca lymphosarcomathatwas comparableto the The differences between VLB and VCR in clinical use and inhibitionby vincristine(VCR),(60 versus68%)while VLB side effects would not have been predicted either from the caused 41% inhibition. The Ca115 Shlonogi mammary differences in their structural formulas (they differ in only 1 carcinomadid not respondto VDS,but bothVLBand VCR functional group) or from their biological activity in preclin causedinhibitionsof62 and67%, respectively.TheCa755 icalstudies(12). mammaryadenocarcinomashoweda moderateresponse The dimeric Vinca alkaloid molecule presents several to VDS (46%) and VCR (39%), and no responseto VLB. sites for chemical modifications that could further alter the The Sarcoma 180 ascites tumor (solid, s.c.), the X5563 spectrum of tumor response, change the therapeutic index, myeloma, and the C3H mammary carcinoma were not or reduce toxicity. A study on a series of 4-acyb and a- responsiveto VDS,VLB,or VCR. aminoacetyl analogs of VLB has been reported by Johnson VDScauseda 113%prolongationoflife in micewiththe eta!. (13). B16 melanoma(i.p.), and there were three of ten 45-day This report summarizes the experimental antitumor activ survivorsin one study,and ten of ten 45-daysurvivorsin ity and mitotic studies with VDS (Chart 1). anotherstudy.VLBhadtwo of ten andeightof ten 45-day survivorsin the same studies. Animals inoculatedwith the P388leukemiaandthe Walker256 and Ehrlichascites MATERIALS AND METHODS tumors had greater than 100% prolongationof life with VDS was prepared from VLB by either the preferential manyindefinitesurvivorswhentreated with VDS, VLB,or ammonolysis of the C-23 carbomethoxy group or the pref VCR. The L1210, AKR, L5178Y, and C1498 leukemias erentiab hydrazinobysis and subsequent Raney nickel hydro showsdno responseto VDSgivendaily at I .0 mg/kg. genolysis. These modifying procedures also effect the loss In tissue culturesof Chinese hamsterovary cells, the of the 4-O-acetyl group. Prior to its use in bioassays, VDS minimumeffectiveconcentrationofVDSthat caused10to was converted to the sulfate salt by treatment with dilute 15%accumulationofcells in mitosiswas 2.3 x 1O@N,for sulfuric acid solution at pH 4.2 in alcoholic solution. Prelim VLB it was 2.2 x 1O@M,and for VCR ft was 7.3 x 1O@M. mary details of these preparations have been reported (6), The concentrationof VDS needed for 40 to 50% accumu and additional chemical aspects have been published (3). lation of cells in mitosiswas 2.8 x 1O@M;for VLB it was VLB, the parent alkaloid, VCR, and deacetyl-VLB (11) were 8.3 x 1O-@M;and for VCR ft was 2.4 x 1O@N. included for comparisons and were given as the sulfate salts. INTRODUCTION TumorTransplantationandMeasurements.Solidtumor fragments, 1 to 2 mm square, were implanted s.c. by trocar When VLB3 and VCA were introduced into clinical prac in the axillary region of mice or rats. Treatment with an tice in the early 1960's, both of these Vinca alkaloids experimental compound was delayed for 8 days in animals showed promising activity against Hodgkin's disease (1, 2). implanted with ROS; in all other animals treatment began 24 hr after implantation. The alkaloids were administered I Presented in part at the 69th Annual Meeting of the American Associa tion of Cancer Research, March 27 to 29, 1974, Houston, Texas (17). i_p. once daily, except for the study on the routes of 2 Present address: Department of Pharmacology, Indiana University administration, to all animals for the number of days mdi Schcol of Medicine, Indianapolis, Ind. 46202. cated; control animals received 0.85% sodium chloride 3 The abbreviations used are: VLB, vmnblastmne; VCR, vincristine, VDS, vindesine; ROS, Ridgeway osteogenic sarcoma; GLS, Gardner lymphosar solution at times corresponding to drug treatment. One day coma; D,,,,,, the dose of each alkaloid that caused 10 to 15% mitotic accu after the final dose, the inhibition of tumor growth was mulation; D@,,, the dose of each alkaloid that caused a 40 to 50% mitotic accumulation. determined by comparing the mean diameter of the tumors Received August 8, 1975; accepted June 5, 1978. in the treated group of 5 animals to the mean diameter of

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Table 1 Inhibition of the ROSÂ°byVOScompared to other Vincaalkaloids % inhibition of growth of tumor at the following daily dose, mg/kg i.p. for 8 days

pound0.050.100.150.200.250.300.400.51.0VDS322366696786100100TxbVLB19000000VCR187290100100100TxDeacetyl

IHI 07734588384VLB CH3 t@ONH2 a The host was the AKR mouse. Chart 1. Structure of VDS (deacetyl-VLB amide). b Tx toxicity; 60% of the mice died before the ninth day. the tumors of 5 animals in the control group. The difference is expressed as the percentage of inhibition of tumor Table2 growth. A dose that caused 3 or more deaths in a group of Inhibitionofalkaloids%the GLSaby VDScompared to other Vinca 5 animals was considered toxic. dailyCom inhibition of growth of tumor at the following Ascites tumors and beukemias were initiated by an i.p. days0.05 mg/kg i.p. for 8 inoculation of 1 x 10@cells. Treatment was begun 24 hr 1.0VDS29pounddose, 0.100.15 0.20 0.25 0.30 0.40 0.50 after inoculation. The response to therapy was determined by comparing the mean life span of the 5 treated animals to TxVLB10 22 70 100 100 100 Txb 7VCR11 0 5 27 47 that of the 5 control animals; prolongation of life in the TxDeacetyl 69 90 100 Tx treated animals beyond that of the controls is expressed as TxVLB 0 0 0 28 84 91 100 a percentage. MitoticStudies.Chinesehamsterovarycellswere main a Host was the C3H mouse. tamed in Medium F-12 supplemented with 10% fetal calf b Tx toxicity; 60% deaths before the ninth day. serum in plastic tissue culture vessels. Twenty-four hr prior to testing, the cells were transferred to Leighton tubes at a density of 2 x 10@cells/mI (2.0 mI/tube). noma did not respond to VDS (12%), but both VCR and VLB The alkaloids were dissolved in complete growth me inhibited the growth of this tumor (67 and 62%, respectively) dium, serially diluted to the appropriate concentrations, at 0.2 mg/kg. Another mammary tumor, the Ca755 mam and added in duplicate to the cells, which were then mary adenocarcinoma, showed a moderate response to incubated in the presence of the alkaloid for 5 hr. The both VDS (46%) and VCR (39%) with no response to VLB at medium was then removed by gentle aspiration, and the 0.2 mg/kg. The solid s.c. form of the Sarcoma 180 ascites cells were washed very carefully with Hanks' balanced salt tumor, the X5563 myeloma, and the C3H mammary carci solution, fixed with Carnoy's solution, and stained with noma did not show adequate responses to VDS, VCR, or hematoxylin-eosin. The mitotic index (percentage of mitotic VLB (Table 3). cells/1000 cells) was determined by microscopic observa VDS and VLB had comparable activity against the i.p. tion of coded slides. form of the B16 melanoma. When given daily for 9 days, VDS caused up to 119% prolongation of life and three of RESULTS ten 45-day survivors in 1 study and as many as ten of ten 45- day survivors in another study. VLB caused as high as 85% Antitumor Activity. The responsesof various murine prolongation of life with 5 of 10 indefinite survivors and up tumors to VDS are compared to those with VLB, VCR, and to 8 of 10 survivors in the same 2 studies. VCR was less deacetyb-VLB. The lowest dose of VDS that completely effective then was either VDS or VLB, causing 60% probon inhibited the ROS was between 0.3 and 0.4 mg/kg i.p., gation of life and only three of ten 45-day survivors. When compared to 0.15 to 0.20 mg/kg for VCR. VLB did not given at large doses on Days 1, 5 and 9, VDS caused up to inhibit the growth of the ROS at 0.4 mg/kg, and deacetyl 82% prolongation of life at 1 .0 mg/kg, and as many as 10 of VLB showed 84% inhibition at 0.4 mg/kg (Table 1). 10 indefinite survivors at 1.67 mg/kg. VLB and VCR had no VDS also showed complete inhibition of the GLS between more than 4 indefinite survivors or 72% prolongation of life 0.20 and 0.25 mg/kg, which was similar to the 0.15- to 0.20- (Table 4). mg/kg dose of VCR required for complete inhibition. VLB The percentage of prolongation of life was used as the showeda minimal47% inhibitionat 0.3 mg/kg, anddeace measure of effectiveness of each of the Vinca alkaloids tyl-VLB at 0.4 to 0.5 mg/kg completely inhibited the GLS against the solid s.c. form of the P1534-J leukemia. VDS (Table 2). showed an erratic response (0 to 70%); VLB showed 93% Several other murine tumors showed response to VDS, and 2 of 4 indefinite survivors at 0.4 mg/kg; VCR showed VCR, and VLB, but none were as responsive as the ROS the best activity with 4 of 4 indefinite survivors at 0.25 mgI and GLS. The Mecca lymphosarcoma showed comparable kg (Table 5). responses to VDS (60%) and VCR (68%), and VLB had a AntibeukemiaandAscites Activity.The P388 leukemia lesser effect (41%) than did VDS. All 3 Vinca alkaloids were showed responses to VDS, VCR, and VLB. VDS at 0.6 mgI given at 0.2 mg/kg. The CallS Shionogi mammary carci kg caused a 213% prolongation of life of the C57BL x DBA/

SEPTEMBER1978 2887

2 F, (hereafter called BD2FJ) mice with 4 of 10 mice against the GLS. No response was seen when VDS was surviving 45 days. VCR at 0.5 mg/kg caused a 263% probon given p.o. The i.v. route was also slightly better than the i.p. gation of life with 5 of 10 survivors, and VLB caused a 263% route, and no p.o. activity was seen against the ROS in prolongation of life with 5 of 10 survivors. The L1210, AKR, mice with VDS (Table 7). Neither VLB nor VCR has shown L5178Y, and C1498 leukemias did not respond to VDS, any oncolytic activity in previous studies in mice when VCR, or VLB. The Walker 256, Ehrlich, and Sarcoma 180 givenp.o. ascites tumors were inhibited by VDS, VLB, and VCR, Delayed Therapy. The GLS was implanted in 8 groups of which resultedinmostly45-daysurvivors,withtheexcep mice. VDS at 0.3 mg/kg daily was started in 1 group 24 hr tion of the lack of response by the Sarcoma 180 ascites to after the GLS implantation with an untreated control group. VCR in this study (Table 6). in other groups, no VDS was given until 3, 5, or 8 days after Routesof Administration.VDSis a water-solublesulfate the GLS was implanted, thereby allowing the GLS to estab salt that can be given i.v. or p.o. to mice. When given i.v., lish itself. This was an attempt to mimic the clinical situation VDS showed a slightly better effect than when given i.p. where well-advanced tumors are presented; GLS tumors that had grown on 5 and 8 days without treatment repre Table 3 sented an advanced stage of this tumor. When VDS was Comparativeactivity VDSand related Vincaalkaloids aofdditional murine tumorsagainst%inhi given starting 1 or 3 days after implantation, 100% inhibition of growth was seen at Days 14 to 16. When VDS was bitionDaily delayed for 5 days, there was a decrease in the mean tumor doseof tu diameter from 5.5 mm at Day 5 to 3.2 (â€”42%)at Day 16, (mg/kgmorTumorapoundCom- compared to an increase in the control tumors from 10.5 days)growthMeccaVDS Host x mm on Day 6 to 26 mm on Day 16 (+248%). In the 8-day 760lymphosarcomaVCR AKR 0.25 x delay study, there was a large difference on Day 8 between 0.25 x 7 the mean tumor diameter of the control tumors, 4.7 mm, 41Ca115 VLB 0.25 x 768 compared to 11.1 mm forthe tumors in mice to be treated. mammaryVDS dds 0.2 x 10 Treatment with VDS, 0.3 mg/kg, was started on Day 8. On VCR 0.2 x 10 67 Day 10 the control tumors had grown to 20 mm, and the 62Ca755 VLB 0.2 x 1012 tumors in the treated mice had grown to 17 mm. By the 16th day or after 8 days of treatment with VDS, the tumors mammaryVDS C57BL/6 0.2 x 8 VCR 0.2x8 39 in the mice given VDS had regressed to 13.9 mm (â€”19%) 0Sarcoma VLB 0.2x846 while the control tumors were 21.6 mm (+177%; Chart 2).

180 (solid)VDS Swiss 0.2 x 9 Table5 VCR 0.2x9 2 Prolongation of life in mice with the P1534(solid) by VOS 20X5563 VLB 0.2x931 compared to other Vinca alkaloids

myelomaVDS C3H 0.2 x 9 daily0.150.20prolongation of life of BD2FI mice at the fo VCR 0.2x9 12 dose, mg/kg i.p. for 8 daysIlowing 9C3H VLB 0.2x92 Com pound%0.40.50.6VDS2116 0.25 0.30 mammaryVDS C3H 0.2 x 9 0TxTxVLB 70(1) 5 VCR 0.2x9 9 38 36 93(2) VLB 0.2x919 0 VCR32 1 (1)@'1278 (3) 0 (4) Txb TxTx Tx0 Tx

Il The mean tumor diameters of the control groups were: Ca755 a Numbers in parentheses,45-day indefinite survivors out of 4 mammary adenocarcinoma, 17.0 mm; X5563, 20.9 mm; C3H, 19.0 mice. mm; Sarcoma 180, 13.1 mm; Ca115, 26.7 mm; MLS, 24.0 mm. b Tx toxicity; 6O% deaths before the ninth day of the test.

Table 4 Prolongation of life of the miceaVDS% with the B16melanomaby lifemg/kg prolongation of 9Drug0.216 i.p. daily for 9 doses mg/kg i.p. on Days1, 5, and 1.67VDS9fJ(l)b 0.36 0.60 1.0 0.6 1.0 1.25 TxVCR56 119 (3) 25(3) 67(1) 82(2) @ 0VLB85(5) 74 Txâ€• 33 69 (2) Tx 72(1) 57 79 VDS 26(6)0(10)Tx(1)71(8)67(9)0(10)34(1)0(2)Tx3536(2)18(3)39 VCR VLB (7)0(8)Tx 53(4)59 (1)53(3) @ host was the C57BL/6 mouse. b Numbers in parentheses, number of 45-day survivors of the 10 mice beginning the study.

C Tx, toxicity; 60% deaths before the ninth day.

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Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1978 American Association for Cancer Research. VDS and Murine Tumors Table 6 ascitesLeukemia/ascitesHostDailyComparativeactivity of VDS and related Vincaalkaloids againstexperimental leukemiasand IifeaVDSprolongation of dose (mg/kg VLBP388leukemiat'BD2F10.6,0.5,0.6k'213 for 9 days)% VCR (5/10)L1210 (4/lo)d 263 (5/10) 263 7AKR leukemiaDBA/20.2, 0.2, 0.20 ND@ 18L5178YleukemiaAKR0.25, 0.25,0.250 ND 5Cl498leukemiaC57BL/60.20,0.20,0.206leukemiaDBA/20.40, 0.40, 0.406 0 20 27

WalkerascitesSD0.15,0.15,0.15330(4/5)Ehrlich 200 (2/5) 346(4/5)0(5/5)0 0.250(5/5)SarcomaascitesSwiss0.25, 0.25, 0 (5/5) 180ascitesSwiss0.25, 0.20, 0.2541(3/5) 24 (1/5)

a Average life span in days for the controls: P388, 11.0; L1210, 14.8; AKR, 11.7; L5178, 10.7; C1498, 14.5;(4/4)Walker ascites, 9.2; Ehrlich ascites, 21 .2; Sarcoma 180, 26.3. b Data presented for the P388 are abstracted from data supplied by Dr. R. Montgomery of the Southern Research Institute, Birmingham, Ala. Indefinite survivors are counted at 40 days. C Dose of VDS, VCR, and VLB, respectively. d Numbers in parentheses, number of 45-day survivors over the number of animals in each group. e ND, not done.

Table 7 Activity of VDSby variousroutes of administration against the GLSand ROS

mg/0.050.10.150.20inhibition of growtha at the followi daily dose, kg for 8 daysng

- TumorRoute%0.300.40.51.0GLSi.p. 0.25 100 100 i.v. 13 79 92 100 100 100 100 100 9ROSi.p. p.o.4 3229 122 170 0 9 28100 7Txb22Tx 67 86 i.v. 60 51 100 100 P.O.322366 069 0 0 0100100Tx a The measurements were made on Day 9 for the GLS and Day 11 for the ROS. The mean tumor diameters for the GLS control tumors were: i.p., 26.0 mm; i.v., 26.5 mm; p.o., 23.8 mm. For the ROScontrol tumors the mean tumor diameters were: i.p., 11.0 mm; i.v., 9.2 mm; p.o., 9.6 mm. b Tx toxicity; 60% deaths before the ninth day.

A B Table 8 28 28 _I 5OAY MitoticcultureCom accumulation in Chinesehamsterovary cells in 1 CONTROL 241 3OAY ofpoundacidSubstitution @ CONTROL 8 DAY (M)(N)VDS functionD1@,..1, @2O @2O CONTROL x 10-. x 10-s @ 16 CONTROL 16 VLBb COOCH@ 2.2 x 10@ 8.3 x 10@ @@ 8DAY VCRbCONH,[email protected] 7.3 x 10-'2.8 2.4 x 10-s @ 12 12 TREATED a Control cells had 3 to 7% of cells in mitosis. b VLB and VCR have the 4'-acetyl group.

@ 4 lsnd3DAY 5DAY effect, and D@. The D,@,..,5forVDS was 2.3 x 10@ M, which ./ â€œ..s TREATED TREATED was 10-fold lower than the 2.2 x 10@ N D1,@15forVLB and 3 @@@@ 0 i'o1'214 16 18 0 times lower than the D,@..15forVCR, 7.3 x 10@ M. The D4@ DAYS for VDS was comparable to the D@ for VCR, 2.8 x 10@ M Chart 2. Delayed therapy with VDS against the GLS. The GLS was im versus 2.4 x 10-s M, respectively, and 3 times lower than planted s.c. in several random groups of AKR mice. VDS treatment was starled 1@3,5, or 8 days after the GLS was implanted. Each treatment group D4@ for VLB, 8.3 x 10@ (Table 8). had Its own control group. At D1@,5VDS-treated cells had few postmetaphase cells compared to VLB- or VCR-treated cells, and the nuclei were MetaphaseArrest.Becauseallof theVincaalkaloidsthat swollen with dispersed chromosomes unlike the condensed have been evaluated in the clinic against cancer cause cells chromosomes and sunken nuclei seen with VCR (13). These to accumulate in mitosis, VDS was compared with VLB and â€œball-typemetaphasesâ€•havebeen seen with VCR and other VCR for mitotic inhibition of Chinese hamster ovary cells in antitububin compounds in animal cell populations and culture. Two effects were measured: D1@,..15,aminimum mammalian cultured cells treated with these agents (9). In

SEPTEMBER1978 2889

Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1978 American Association for Cancer Research. M. J. Sweeney et a!. most cases the pyknotic cells resulting from VCR treatment given nonlethal leukopenic doses of the alkaloids. They rapidly degenerate, and time-lapse micrographic evaluation compared VDS with VCR in the White Rock chicken and the indicates that these cells are incapable of reentering the rhesus monkey, and signs of neurotoxicity were produced cell cycle. in the animals given VCR. VDS given i.v. twice weekly at 0.8 mg/kg for 1 week and DISCUSSION then at 1.6, 2.4, and 2.4 mg/kg for the next 3 weeks did not produce neurotoxic signs in the chicken. However, VCR at ExperimentalAntitumorActivity.Advancesin chemical 0.4, 0.4, 0.4, and 0.8 mg/kg for the 4 weeks produced signs techniques, especially improved separation by high-pres of ataxia, wing drop, leg weakness, and an inability to stand sureliquidchromatography,havefacilitatedmoreextensive erect or to hold the head erect. modification of VLB and VCR. The recent chemical altera VDS given i.v. at final doses of 0.3 mg/kg over 9 months tions attempted to provide clinical agents that would have a did not produce any signs of neurotoxicity in the rhesus wider tumor spectrum, more favorable therapeutic indices, monkey. However, VCR at 0.25 mg/kg produced signs of and minimal side effects. The search for an agent with a neurotoxicity at 5 to 7 months consisting of marked incoor spectrum of oncolytic activity resembling or surpassing that dination,reluctancetomove, inabilitytousethehindlegs, of VCR but with minimal or no neurotoxicity was a major followed by loss of use of the hands with contraction of the aim of the modification program. flexor muscle, and formation of a permanent fist. VDS was selected for both biological and chemical con Metabolism.The studiesof CuIp et a!. (7) showedthat siderations as a clinical candidate (3). First, the ROS tumor when [3H]VDS was given i.v. at 0.5 mg/kg, it was readily has been described by the National Cancer Institute Drug distributed to all tissues except those of the central nervous Development Program as a solid tumor system with a system. Tritium levels fell rapidly over the first hr, and a satisfactory degree of clinical predictiveness. Therefore, second phase of drug disappearance from the blood the activity of VDS against the ROS and the inactivity by showed a half-life of about 10 hr. The tritium bevel in the VLB showed a qualitative change in the spectrum of activity spleen was higher than in any other tissue, but this was also from the parent VLB. Second, VDS caused inhibition of the true for VLB (4) and VCR (5). The organs involved in the B16 melanoma, another tumor that shows reasonable clini excretory function, such as the liver and kidney, also had cal predictiveness of experimental drugs. Third, there was high levels of tritium as well as did the lung. The presence a potential for less neurotoxicity with VDS than with VCR. of unmetabolized [3H]VDS was shown in bile, but no other The in vitro studies by Ochs and Worth (16) showed that metabolites were found. When [3H]VDS was given p.o., VDS had less effect on axoplasmic transport in the sciatic there were no detectable levels of [3H]VDS in the serum of nerve than did VCR. Third, various aspects of chemical the rats. stability, feasibility of preparation, and pharmaceutical ac Toxicology.The studiesby Toddet a!. (18) showedthat ceptability favored VDS over other VLB modification prod the dose lethal to 50% of the rats for VDS given i.v. is 6.3 ucts(3). mg/kg which is between the doses lethal to 50% of the rats Mitotic Accumulation.The differencesin mitotic accu of VLB, 10.0 mg/kg, and of VCR, 2.0 mg/kg. When rats mulationmay indicateapossibledifferenceinstrengthof were given i.v. doses of VDS totaling 0.15 mg/kg/week for binding to tubulin, assuming that binding to tubulin is the 3 months, they developed no remarkable signs of toxicity. mechanism of metaphase arrest. At minimal effective doses Doses of 0.3 mg/kg or greater per week produced anorexia, all 3 alkaloids differ from each other in potency and cytobog depressed blood cell counts, atrophic intestinal mucosa, ical effects. in addition to binding strength to tubulin, 2 and inhibition of spermatogenesis. Groups that received factors may influence the results seen with these 3 alkaloids 0.1 or 0.16 mg/kg had leukopenia, slight erythropenia, on mitotic accumulation, the rate of entry of cells into inhibition of spermatogenesis, and focal skeletal muscle mitosis and also the rate at which cells can proceed through degeneration. There was no evidence of functional or mitosis during the 5-hr exposures. At the higher concentra structural changes in neural tissues. The previous effects tions, an effect of antitubulins on progress of cells through are common to animals given VCR at bower doses and for other portions of the cell cycle, especially S and G@,arewell shorter periods of time. Todd et a!. concluded that VDS was known (8, 15). This effect would result in a lowered mitotic less toxic than VCR in animals on a mg/kg basis. index if cells were not permitted to progress through S and/ Preliminary Clinical Studies. Dyke and Nelson (10) have or G@into mitosis. A lowered mitotic index may also result published on the Phase 1 studies on VDS and have pre from mitotic delay followed by recovery at low doses. Time sented the following summary on other Phase 1 and 2 lapse micrographic observations of the effect of antitubu studies. The Phase 1 clinical trial initiated at the Lilly lineson Chinese hamster ovaryproliferationinthe pres Laboratories for Clinical Research and continued elsewhere ence of high and low doses of antitubulins indicate that established that adult patients tolerate with minimal toxicity both factors are involved in a low or high mitotic index. a dose of 3 to 4 mg/sq m body surface area given once Neurotoxicity Studies in VIvo. Todd et a!. (19)@have weekly by rapid i.v. injection. This schedule produces mild reported on an extensive study in which several species leukopenia in the average patient without previous marrow were studied for the development of neurotoxicity when damage with a nadir of 3 to 5 days and recovery of the pretreatment WBC by the seventh day (10). Nonprogressive 4 G. C. Todd, W. J. Griffing, W. R. Gibson, and D. M. Morton. Animal neuropathy appears with this dose but does not in itself Models for the Comparative Assessment of Neurotoxicityfollowing Repeated Administration of Vinca Alkaloids. Submitted for publication to Cancer Treat decrease performance status. Our Phase 1 to 2 trial has ment Reports. produced a complete remission (16 months) in a patient

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Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1978 American Association for Cancer Research. VOS and Murine Tumors with metastatic (skin and lymph nodes) malignant mela 4. Beer, C. T., and Richards, J. F. The Metabolism of Vinca Alkaloids. II. The Fate ofTritiated Vinblastine in Rats. Lloydia, 27: 352-360, 1964. noma and partial remission in 2 patients with non-Hodgkin's 5. Castle, M. C., Margileth, D. A., and Oliverlo, V. T. Distribution and lymphoma. Two adults (ages 70 and 73) with acute myelo Excretion of [3H]Vincristine in the Rat and Dog. Cancer Res., 36: 3684- monocytic leukemia have been maintained in complete 3689, 1976. 6. Cullinan, G., Gerzon, K., Poore, G. A., and Sweeney, M. J. Inhibition of remission induced by other chemotherapy for significant Experimental Tumor Systems by Desacetyl Vinblastine Amide and Con periods of time. Elsewhere, several children with acute genera. In: Proceedings of the Eleventh International Cancer Congress, leukemia unresponsive to chemotherapy protocols includ Florence, Italy, October 23 to 26, 1974. Milan: Casa Editrice Ambrosiana, 1974. ing Oncovin have undergone complete remission when 7. CuIp, H. W., Danlels, W. D., and McMahon, R. E. Disposition and Tissue treated with VDS (14). Phase 3 trials are in progress. Levels of (3HjVindesine in Rats. Cancer Res., 37: 3053-3056, 1977. 8. DahI, W. N., Oftebro, R., Pettersen, E. 0., and Brustad, T. Inhibitory and Cytotoxic Effects of Oncovin (Vincristine Sulfate) on Cells of Human ACKNOWLEDGMENTS Line NHIK 3025. Cancer Res., 36: 3101-3105, 1976. 9. De Brabander, M. J., Van de Veire, R. M. L., Aerts, F. E. M., Borgers, We express our appreciation to V. Bothwell, S. Miller, E. Priller, and J. M., and Janssen, P. A. J. The Effects of Methyl [5-(2-Thienylcarbonyl)- Richardson for their technical assistance In the studles on the antitumor 1H-benzimidazol-2-yl]carbamate R 17934; NSC 238159), a New Synthetic activity and to R. W. Johnson for this technical assistance in the studies on Antitumoral Drug Interfering with Microtubules, on Mammalian Cells mitotic inhibition and accumulation. Cultured In vitro. Cancer Res., 36: 905-916, 1976. 10. Dyke, A. W., and Nelson, R. L. Phase I Anti-Cancer Agents. Vindesine Addendum (Desacetyl Vinblastine Amide Sulfate). Cancer Treat. Rev., 4: 135-142, 1977. Since the acceptance of this paper, Eldesine was designated as the 11. Hargrove, W. W. Preparation and Activities of Chemically Modified registered trademark for VDS. Also, the following clinical report was recently Dimeric Catharanthus Alkaloids. Llyodia, 27: 340-345, 1964. published: Mathe, G., Misset, J. L., DeVassel, F., Gouveia, J., Hayat, M., 12. Johnson, I. S., Armstrong, J. G., Gorman, M., and Burnett, J. P., Jr. The Machover, D., Belpomme, D., Pico, J. L., Schwarzen, L., Ribaud, P., ymcaAlkaloids:ANewClassof OncolyticAgents.CancerRes.,23: Musset, M., Jasmin, Cl., and De Luca, L. Phase II Clinical Trial with Vindesine 1390-1427,1963. for Remission Induction in Acute Leukemia, Blast Crisis of Chronic Myeloid 13. Johnson, I. S., Hargrove, W. W., Harris, P. N., Wright, H. F.. and Boder, Leukemia, Lymphosarcoma, and Hodgkin's Disease: Absence of Cross G. B. Preclinical Studles with Vinglycinate, One of a Serles of Chemically Resistance with Vincristine. Cancer Treat. Rept. 62: 805-810, 1978. Derived Analogs of Vinblastlne. Cancer Res., 26: 2431-2436, 1966. 14. Lilly, Eli & Co., data on file. REFERENCES 15. Madoc-Jones, W., and Moura, F. Interphase Action of Vinblastine and Vincristine: Differences in Their Lethal Action through the Mitotic Cycle I . Armstrong, J. G., Dyke, R. W., and Fouts, P. J. Initial Clinical Experience of Cultured Mammalian Cells. J. Cellular Physiol., 72: 185â€”195,1968. with Leurocristine, A New Alkaloid from Vinca rosea Linn. Proc. Am. 16. Ochs, S., and Worth, R. Comparison of the Block of Fast Axoplasmic Assoc. Cancer Res., 3: 301, 1962. Transport in Mammalian Nerve by Vincristine, Vinblastine and Desacetyl 2. Armstrong, J. G., Dyke, R. W., Fouts, P. J., and Gammer, J. E. Hodgkin's Vinblastine Amide Sulfate (DVA). Proc. Am. Assoc. Cancer Res., 16: 70, Disease,Carcinoma of the Breast, and Other Tissue Treated with 1975. Vinblastine Sulfate. Cancer Chemotherapy Rept., 18: 49-71 , 1962. 17. Sweeney, M. J., Cullinan, G. J., Poore, G. A., and Gerzon, K. Experimen 3. Bamett, C. J., Cullinan, G. J., Gerzon, K., Hoying, A. C.. Jones, W. E., tal Antitumor Activity of Vinblastine Amides. Proc. Am. Assoc. Cancer Newlon, W. N., Pcore, G. A., Robison, R. L. Sweeney, M. J. and Todd, Res., 15: 37, 1974. G. C. Structure-Activity Relationships of Dimeric Catharanthus Alka 18. Todd, G. C., Gibson, W. R., and Morton, D. M. Toxicology of Vindesine bids. 1. Desacetylvinblastine Amide (Vindesine) Sulfate. J. Med. Chem., (Desacetyl Vinblastine Amide) In Mice, Rats and Dogs. J. Toxicol. 21: 88-96, 1978. Environ. Health, 1: 843â€”849,1976.

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Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1978 American Association for Cancer Research. Antitumor Activity of Deacetyl Vinblastine Amide Sulfate (Vindesine) in Rodents and Mitotic Accumulation Studies in Culture

Martin J. Sweeney, George B. Boder, George J. Cullinan, et al.

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